{"paper_id":"d7297f9d-cee9-4858-ac3a-9ba2fbe74f3e","body_text":"828\n© Aluna Publishing\nWiadomości Lekarskie Medical Advances, VOLUME LXXVIII, ISSUE 4, APRIL 2025    \nINTRODUCTION\nEndometriosis is a chronic inflammatory condition \ncharacterized by the growth of tissue similar to the \nlining of the uterus outside the uterus, usually in areas \nsuch as the peritoneum, ovaries, and cervix. Clinical \nsymptoms often include progressive dysmenorrhea, \nchronic pelvic pain, profound dyspareunia, and infertility, \nwhich significantly affect the patient’s quality of life [1]. \nIt is estimated that approximately 10% of women of \nreproductive age suffer from endometriosis [2].\nAlthough the exact cause and development of \nendometriosis remain unclear, the theory of retrograde \nmenstruation proposed by Sampson in 1921 is \nwidely accepted. Other hypotheses, such as coelomic \nmetaplasia and vascular/lymphatic metastasis [3], have \nalso been proposed, but cannot fully explain all forms of \nthe condition. In addition, factors such as the immune \nsystem, hormones, genetics, and the environment are \nbelieved to play an important role in the pathogenesis \nof endometriosis [4].\nGiven the involvement of natural killer (NK) cells in \nendometriosis due to reduced toxicity, one potential \ntreatment approach is to activate these cells. In animal \nmodels, intraperitoneal injections of Lactobacillus \ngasserii OLL2809, a probiotic that stimulates IL-12 \nproduction, resulted in NK cell activation and reduction \nof ectopic endometrioid lesions. A randomized, \ndouble-blind, placebo-controlled trial also showed \nthat this probiotic can alleviate pain associated \nwith endometriosis [5]. Adhesion of endometrial \nfragments to other tissues is considered to initiate a \nlocal inflammatory response that over time develops \ninto chronic inflammation. The inflammatory response \nin the pelvic cavity largely involves the activation of \nmacrophages, which produce a number of growth-\nregulating substances. There is evidence that some \nof the inflammatory factors also stimulate the growth \nof ectopic endometrial cells in the early stages \nof endometriosis [6]. These compounds can also \naffect fertility, as well as nociceptors, thus causing \nCorrelation between MRNA IL1Β and type, duration of \ninfertility in women with endometriosis on the stage preparing \nto assisted reproductive technologies using probiotics\nOksana V. Bakun, Kristina V. Dyak, Oksana V. Kolesnik, Tetiana Protsak,Yevheniia A. Dudka \nBUKOVINIAN STATE MEDICAL UNIVERSITY , CHERNIVTSI, UKRAINE\nABSTRACT\nAim: To investigate the correlations between the level of IL1β mRNA gene expression and the type and duration of infertility and to study the levels of IL1β \nmRNA gene expression in women with endometriosis associated with infertility. \nMaterials and Methods: For mRNA gene expression analysis IL1β and determination of relative normalized mRNA expression \nIL1β used the real-time reverse transcription polymerase chain reaction method (RT-PCR).. Examined group consists of 30 infertile \nwomen undergoing assisted reproductive technologies. The main group consisted of 20 women diagnosed with endometriosis undergoing assisted reproductive \ntechnologies. The control group consisted of 10 healthy women. \nResults: In the main group, the level of IL1β mRNA gene expression before preparing was 26.7877±0.01, which was significantly higher than the level after \npreparing (0.1610±0.01* ). \nAnalized results, it’s found out that Mean level of mRNA IL1β in women with endometriosis associated infertility 1-st degree is 8.53 c.u., at the same time Mean \nlevel of mRNA IL1β in women with endometriosis associated infertility 2-nd degree is 1.0 c.u. \nConclusions: The inclusion of probiotics in a comprehensive regimen of preparation for assisted reproductive technologies leads to a noticeable improvement \nin the patient’s well-being. \n  KEY WORDS: endometriosis, probiotics, assisted reproductive technologies, infertility, IL1β\nWiad Lek. 2025;78(4):828-837. doi: 10.36740/WLek/203893 DOI\nORIGINAL ARTICLE CONTENTS\n\n829\nCorrelation between MRNA IL1Β and type, duration of infertility in women with endometriosis on the stage preparing...\ninfertility and pain. Cytokines are regulatory peptides \nor glycoproteins that can be produced by virtually \nevery type of nucleated cell in the body and have \npleiotropic regulatory effects on many cell types. Unlike \nhormones, cytokines usually act as paracrine and/\nor autocrine signals, only occasionally entering the \ncirculation, where they can act as endocrine mediators \n[7]. Macrophages are among the major producers of \ncytokines, especially interleukins-1 and 6 (IL-1, IL-6) and \ntumor necrosis factor-α (TNFα); this is probably not the \ncase under normal conditions, but after stimulation \nby various substances [8]. Interleukins are considered \nmodulators of cell proliferation and as inducers of other \ncytokines, as a cascade in acute inflammation [9].\nCytokines produced in the uterine environment \nare involved in the regulation of endometrial growth \nthrough steroid-cell and cell-cell interactions [10]. \nCytokines may also contribute to the pathophysiology \nof endometriosis in at least two ways, namely by \nenhancing the establishment and proliferation of \nectopic endometrial implants and by influencing \ncytokine secretion by macrophages, which can lead \nto adverse changes. The cytokines IL-1β, IL-6 and TNFα \nare of great interest because they are partly hormonally \nregulated and play important roles as mediators of \ninflammation. IL-1 is involved in the regulation of the \nimmune response and inflammation. There are two \ndifferent forms of IL-1, α and β, with similar biological \nactivities [7]. IL-1α is present in the endometrium, in \nboth epithelial and stromal cells, at least in the late \nsecretory phase. IL-1β has a similar distribution, usually \nappearing in lower amounts. IL-1β mRNA is expressed \nin the endometrium in the late secretory phase and cor-\nresponds to serum IL-1β levels, which vary throughout \nthe cycle with maximum values during the secretory \nphase [11].\nAIM\nTo study mRNA gene expression level IL1β and \nestimated correlation between IL1 mRNA β and type, \nduration of infertility in women with endometriosis \non the stage of preparing for assisted reproductive \ntechnologies using probiotics\nMATERIALS AND METHODS\nFor gene expression analysis IL1β mRNA and \ndetermination of relative normalized mRNA expression \nIL1β used the real-time reverse transcription \npolymerase chain reaction method(RT-PCR).The \nobject for molecular genetic studies using RT-PCR was \na fraction of mononuclear cells isolated from whole \nblood of patients with endometriosis. In this study, we \nconducted a retrospective analysis of case histories of \n30 infertile women undergoing assisted reproductive \ntechnologies. The main group consisted of 20 women \ndiagnosed with endometriosis who underwent assisted \nreproductive technologies. In addition to standard \npreparation for assisted reproductive technologies, \nwomen in the main group received a probiotic \ncontaining Lactobacillus 10 10 manufactured by Unic \nBiotech Ltd, India. They took one tablet twice a day \nfor one month as part of the general treatment before \nundergoing assisted reproductive technologies. We \ndetermined the level of IL1β expression before and after \nthis stage of preparation. The control group consisted \nof 10 women who had tubal infertility due to a previous \ninflammatory disease, but according to the results of \na comprehensive clinical and laboratory examination \nthey were equated to healthy women. These women \naged 21 to 42 years with a mean age of 29.75 years did \nnot undergo our proposed preparation for ART with the \ninclusion of a probiotic. This study was conducted at the \nBukovinian State Medical University and the “Medical \nCenter of Infertility Treatment” clinic.\nRESUL TS\nThe average age of women in the control group (who \ndid not take the probiotic – 28.78±5.09 years) and the \nmain group (who took the probiotic) 29.54±2.04 (p \n>0.05). Women in the main and control groups were \nexamined and expression levels were determined IL1β \nmRNA genes. Expression level IL1β mRNA genes in \nwhole blood in women before preparation for assisted \nreproductive technologies are given in Table 1\nExamining the data presented in Table 1, we can \ndistinguish two clear subgroups: the main group, \nconsisting of women with endometriosis who received \nour proposed training for assisted reproductive \ntechnologies, including probiotics, before and after \ntraining, respectively. In the main group, the level \nof IL1β mRNA gene expression before training was \n26.7877±0.01, which was significantly higher than the \nlevel after training (0.1610±0.01).\nWe performed analysis of level expression mRNA IL1β \nbefore treatment conditioning on group.\nAccording to the presented table 2, when comparing \nof IL1β before treatment, statistically significant differ-\nences were revealed depending on group (p < 0.001) \n(applied method: Mann-Whitney U-test).\nAnalized results according Fig.1, Mean level of mRNA \nIL1β in women with endometriosis is 10.35 c.u., at the \nsame time Mean level of mRNA IL1β in women control \ngroup is 1.0 c.u.\n\nOksana V. Bakun et al. \n830\nWe also performed analysis of IL1β after treatment \nconditioning on group.\nAccording to the presented table 3, when comparing of \nlevel expression mRNA IL1β after treatment, statistically \nsignificant differences were revealed depending on group \n(p < 0.001) (applied method: Mann-Whitney U-test).\nAnalized results according Fig.2., Mean level of mRNA \nIL1β in women with endometriosis is 0.14 c.u., at the \nsame time Mean level of mRNA IL1β in women control \ngroup is 1.0 c.u. \nWe also performed analysis of level expression mRNA \nIL1β before treatment conditioning on group.\nIn accordance with the presented table 4, when com-\nparing of IL1β level before treatment, statistically sig-\nnificant differences were revealed depending on group \n(p < 0.001) (applied method: Pearson’s chi-square test).\nTable 1. Expression level IL1β mRNA genes in whole blood in women before preparation for assisted reproductive technologies  (M±m)\nGroup\nExpression level IL1β mRNA genes in whole blood \nP\nBefore preparing (treatment) After preparing (treatment)\nMain 26,7877±0,01 0,1610±0,01 <0,001\nTable 2. Analysis of level expression mRNA IL1β before treatment conditioning on group\nLevel expression mRNA IL1β before treatment\nVariable Categories Me Q₁ – Q₃ n p\nGroup\nControl 1.00 1.00 – 1.00 10 < 0.001*\nEndometriosis 10.35 2.70 – 21.62 20\n* – differences are statistically significant (p < 0.05).\n \nFig. 1. Analysis of level expression mRNA IL1β before treatment conditioning on group. \n \n \nFig. 2. Analysis of level expression mRNA IL1β after treatment conditioning on group. \n \nFig. 1. Analysis of  \nlevel expression \nmRNA IL1β \nbefore treatment \nconditioning on \ngroup.\nFig. 2. Analysis of \nlevel expression \nmRNA IL1β \nafter treatment \nconditioning on \ngroup.\n\n831\nCorrelation between MRNA IL1Β and type, duration of infertility in women with endometriosis on the stage preparing...\ncally significant differences were revealed depending \non infertility degree (p = 0.003) (applied method: \nMann-Whitney U-test).\nAnalized results according Fig.6., Mean level of mRNA \nIL1β after treatment in women with endometriosis asso-\nciated infertility 1-st degree is 0.14c.u., at the same time \nMean level of mRNA IL1β in women with endometriosis \nassociated infertility 2-nd degree is 1.0 c.u.\nAnalysis of level expression mRNA IL1β before treat-\nment was performed conditioning on infertility degree.\nWhen comparing of level expression mRNA IL1β \nbefore treatment depending on infertility degree no \nstatistically significant differences were revealed (p \n= 0.075) (applied method: Pearson’s chi-square test) \n(Table 8).\nAnalized results according Fig.7., normal level of \nmRNA IL1β before treatment in women with endome-\ntriosis associated infertility 1-st degree is in 33,3%, high \nlevel – in 61,1%, low level in 5,6%, at the same time \nnormal level of mRNA IL1β in women with endome -\ntriosis associated infertility 2-nd degree is in 75% and \nhigh level is in 25%. \nWe performed analysis of level expression IL1β level \nafter treatment conditioning on infertility degree.\nAccording to the presented table 9, when comparing \nof level expression IL1β after treatment, statistically \nsignificant differences were revealed depending on \ninfertility degree (p = 0.008) (applied method: Fisher’s \nexact test).\nOdds of low were 13.000 times less in 2nd degree \ngroup than in 1st degree group, the relative difference \nin odds was statistically significant (OR = 0.077; 95% CI: \n0.012 – 0.482).\nAnalized results according Fig.3., normal level of \nmRNA IL1β in women with endometriosis is in 25% \npatients, high level is observed in 70% patients , low \nlevel is in only 5% patients., at the same time Mean \nlevel of mRNA IL1β in women control group is normal \nin 100 % patients.\n Analysis of level expression mRNA IL1β level after \ntreatment was performed conditioning on group.\nIn accordance with the presented table 5, when com-\nparing of level expression mRNA IL1β after treatment, \nstatistically significant differences were revealed de -\npending on group (p < 0.001) (applied method: Fisher’s \nexact test).\nOdds of low were 59.182 times greater in women \nwith endometriosis comparing with control group, the \nrelative difference in odds was statistically significant \n(95% CI: 2.949 – 1187.719).\nAnalysis of level expression mRNA IL1β before treatment \nwas performed conditioning on infertility degree (Fig. 4).\nAccording to the data obtained when comparing of \nlevel expression mRNA IL1β before treatment statisti-\ncally significant differences were revealed depending \non infertility degree (p = 0.050) (applied method: \nMann-Whitney U-test) (Table 6).\nAnalized results according Fig.5., Mean level of mRNA \nIL1β in women with endometriosis associated infertility \n1-st degree is 8.53 c.u., at the same time Mean level of \nmRNA IL1β in women with endometriosis associated \ninfertility 2-nd degree is 1.0 c.u. \nAnalysis of level expression mRNA IL1β after treat -\nment was performed conditioning on infertility degree.\nAccording to the presented table 7, when comparing \nof level expression mRNA IL1β after treatment, statisti-\nTable 3. Analysis of level expression mRNA IL1β after treatment conditioning on group\n Level expression mRNA IL1β after treatment\nVariable Categories Me Q₁ – Q₃ n p\nGroup\nControl 1.00 1.00 – 1.00 10 < 0.001*\nEndometriosis 0.14 0.09 – 0.24 20\n* – differences are statistically significant (p < 0.05).\nTable 4. Analysis of level expression mRNA IL1β  before treatment conditioning on group\n        Variable       Categories\nGroup\np\nControl endometriosis\nIL1β  level before treatment\nNormal 10 (100.0) 5 (25.0) < 0.001*\nHigh 0 (0.0) 14 (70.0)\nLow 0 (0.0) 1 (5.0)\n* – differences are statistically significant (p < 0.05).\n\nOksana V. Bakun et al. \n832\n \nFig. 3. Analysis of level expression mRNA IL1β level before treatment conditioning on group. \n \nFig. 3. Analysis of \nlevel expression \nmRNA IL1β level \nbefore treatment \nconditioning on \ngroup.\nFig. 4. Analysis of \nlevel expression \nmRNA IL1β  \nafter treatment \nconditioning on \ngroup. \nFig. 4. Analysis of level expression mRNA IL1β  after treatment conditioning on group. \n \nFig. 5. Analysis of \nlevel expression \nmRNA IL1β \nbefore treatment \nconditioning on \ninfertility degree. \nFig. 5. Analysis of level expression mRNA IL1β before treatment conditioning on infertility \ndegree. \n \n\n\n833\nCorrelation between MRNA IL1Β and type, duration of infertility in women with endometriosis on the stage preparing...\nmodel, 5.9% of the observed variance of IL1β before \ntreatment were explained.\nWe performed a correlation analysis of the association \nbetween infertility, duration and IL1β after treatment. \nA weak correlation positive association between level \nexpression mRNA IL1β after treatment and infertility \nduration was estimated (Fig. 8).\nObserved dependence of level expression mRNA IL1β \nafter treatment from infertility duration is described by \na linear regression equation:\nYIL1b after treatment = 0.053 × Xinfertility, duration + 0.164\nDuring our research we found out that there was no \nassociation between level expression mRNA IL1β before \ntreatment and infertility duration.\nObserved dependence of level expression mRNA IL1β \nbefore treatment from infertility, duration is described \nby a linear regression equation:\nYIL1b before treatment = 4.283 × Xinfertility, duration - 4.223\nWith an 1 increase of infertility, duration 4.283 change \nof IL1β before treatment should be expected. According \nto the coefficient of determination R² of the resulting \nTable 5. Analysis of level expression mRNA IL1β  after treatment conditioning on group\nVariable       Categories\nGroup\np\nControl Endometriosis\nIL1β level expression mRNA \nafter treatment\nNormal 10 (100.0) 5 (25.0)\n< 0.001*\nLow 0 (0.0) 15 (75.0)\n* – differences are statistically significant (p < 0.05).\nTable 6. Analysis of level expression mRNA IL1β before treatment conditioning on infertility degree\nVariable Categories\nLevel expression mRNA IL1β before treatment\nn p\nMe Q₁ – Q₃\nInfertility degree\n1st degree 8.53 2.16 – 18.98 18 0.050*\n2nd degree 1.00 1.00 – 3.36 12\n* – differences are statistically significant (p < 0.05).\nTable 7. Analysis of level expression mRNA IL1β after treatment conditioning on infertility degree\nVariable Categories Level expression mRNA IL1β after treatment n p\nMe Q₁ – Q₃\n18\nInfertility degree\n1st degree 0.14 0.10 – 0.24 0.003*\n2nd degree 1.00 0.30 – 1.00 12\n* – differences are statistically significant (p < 0.05).\nTable 8. Analysis of level expression mRNA IL1β before treatment conditioning on infertility degree\nVariable Categories Infertility degree p\n1st degree 2nd degree\nLevel expression mRNA IL1β  \nbefore treatment\nNormal 6 (33.3) 9 (75.0) 0.075\nHigh 11 (61.1) 3 (25.0)\nLow 1 (5.6) 0 (0.0)\nTable 9. Analysis of level expressionIL1β level after treatment conditioning on infertility degree\nVariable Categories Infertility degree            P\nLevel expression  mRNA IL1β level \nafter treatment\nNormal\n       1st degree          2nd degree\n0.008*\n5 (27.8) 10 (83.3)\n  Low 13 (72.2) 2 (16.7)\n* – differences are statistically significant (p < 0.05).\n\nOksana V. Bakun et al. \n834\nDISCUSSION\nIn this study, our main aim was to examine gene \nexpression levelsIL1β mRNAin whole blood in patients \nwith endometriosis associated with infertility and to \nestablish correlations between the type of infertility, \nits duration at the stage of preparation for assisted \nreproductive technologies, using a probiotic.\nEndometriosis is a common benign gynecological \ndisease characterized by the presence of ectopic \nendometrium, which causes dysmenorrhea, chronic \nWith an 1 increase of infertility, duration 0.053 change \nof level expression IL1β after treatment should be ex -\npected. According to the coefficient of determination \nR² of the resulting model, 6.3% of the observed variance \nof level expression mRNA IL1β after treatment were \nexplained( Fig. 9).\nTherefore, our proposed preparation for assisted \nreproductive technologies with the inclusion of a \nprobiotic is quite effective, as the levels of IL1β mRNA \ngene expression decreased sharply (Fig. 10).\n \nFig. 6. Analysis of level expression mRNA  IL1β after treatment conditioning on infertility degree. \n \n \nFig. 8. Analysis of level expression mRNA IL1β  after treatment conditioning on infertility degree. \n \nFig. 7. Analysis of \nlevel expression \nmRNA IL1β \nbefore treatment \nconditioning on \ninfertility degree.\nFig. 6. Analysis of \nlevel expression \nmRNA  IL1β \nafter treatment \nconditioning on \ninfertility degree.\n\n835\nCorrelation between MRNA IL1Β and type, duration of infertility in women with endometriosis on the stage preparing...\nTherefore, our study aimed to investigate this \nrelationship and its potential implications for \nclinicopathological features. Our results show that \nIL-1β mRNA gene expression levels have a negative \ncorrelation with the duration of infertility, significantly \nincreased IL-1β mRNA gene expression levels in women \nwith primary infertility than with secondary infertility, \nand may serve as non-invasive markers in women with \nendometriosis associated with infertility [9].\nCONCLUSIONS \nThe extremely increased expression of IL1β mRNA \ngenes indicates a close relationship between the \npelvic pain, and infertility, and is associated with \ninflammation and immune disorders, as well as changes \nin ovarian steroid hormone production. The growth and \nmaintenance of the endometrium and endometrioid \ntissue is regulated by several cytokines and growth \nfactors, such as interleukin (IL) 6, 8, tumor necrosis factor \n(TNF)α, and vascular endothelial growth factor (VEGF). \nRetrograde menstruation into the abdominal cavity \nthrough the fallopian tubes plays an important role in \nthe pathogenesis of endometriosis. Menstrual fluid is \ncomposed of blood cells, endometrial tissue, and waste \nproducts that are sources of endometrial cells. However, \nthe profile of bioactive molecules in menstrual blood \nis unclear [8].\n \nFig. 10. Regression line characterizing the dependence of level expression mRNA IL1β after \ntreatment from infertility  duration. \n \n \nFig.7. Analysis of level expression mRNA IL1β before treatment conditioning on infertility \ndegree. \n \nFig. 9. Regression \nline characterizing the \ndependence of level \nexpression mRNA IL1β \nbefore treatment from \ninfertility duration.\nFig. 8. Analysis of \nlevel expression \nmRNA IL1β  \nafter treatment \nconditioning on \ninfertility degree.\n\nOksana V. Bakun et al. \n836\nhave a negative correlation with the duration of \ninfertility, significantly increased IL-1β mRNA gene \nexpression levels in women with primary infertility than \nwith secondary infertility .Therefore, we recommend \nthe proposed preparation for assisted reproductive \ntechnologies with the inclusion of a probiotic.\npathogenesis of endometriosis and inflammation. The \ninclusion of probiotics in a comprehensive regimen \nof preparation for assisted reproductive technologies \nleads to a noticeable improvement in the patient’s \nwell-being and a significant decrease in IL1β mRNA \ngene expression. IL-1β mRNA gene expression levels \n \nFig. 9. Regression line characterizing the dependence of level expression mRNA IL1β before \ntreatment from infertility duration. \n \nFig. 10. Regression \nline characterizing the \ndependence of level \nexpression mRNA IL1β \nafter treatment from \ninfertility duration.\nREFERENCES\n 1.  Gao Y , Shen M, Ma X et al. Seven Hormonal Biomarkers for Diagnosing Endometriosis: Meta-Analysis and Adjusted Indirect Comparison \nof Diagnostic Test Accuracy. J Minim Invasive Gynecol. 2019;26:1026-35. doi: 10.1016/j.jmig.2019.04.004.  DOI\n 2.  Kolanska K, Alijotas-Reig J, Cohen J et al. Endometriosis with infertility: A comprehensive review on the role of immune deregulation \nand immunomodulation therapy. Am J Reprod Immunol. 2021;85(3):e13384. doi: 10.1111/aji.13384.  DOI\n 3.  Pant A, Moar K, K Arora T, Maurya PK. Biomarkers of endometriosis. Clin Chim Acta. 2023;549:117563. doi: 10.1016/j.cca.2023.117563. \nDOI\n 4.  Lamceva J, Uljanovs R, Strumfa I. The Main Theories on the Pathogenesis of Endometriosis. Int J Mol Sci. 2023;24(5):1-13. doi: 10.3390/\nijms24054254.  DOI\n 5.  Bulun SE, Yildiz S, Adli M et al. 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Int J Fertil Steril. 2019;13(3):178-83. doi: 10.22074/ijfs.2019.5584.  \nDOI\n 10.  Jia-Juan Wu, Qin-Yu Zhou, Dong-Mei Liu et al. Evaluation of the safety and probiotic properties of Lactobacillus gasseri LGZ1029 based \non whole genome analysis. Food science & Technology. 2023;187:1-7. doi: 10.1016/j.lwt.2023.114759.  DOI\n 11.  Taylor RN, Berga SL, Zou E et al. Interleukin-1β induces and accelerates human endometrial stromal cell senescence and impairs \ndecidualization via the c-Jun N-terminal kinase pathway. Cell Death Discov. 2024;10:288. doi: 10.1038/s41420-024-02048-6. DOI\n\n837\nCorrelation between MRNA IL1Β and type, duration of infertility in women with endometriosis on the stage preparing...\nEthical approval for this study was obtained from the Medical Ethics Committee of the Bukovinian State Medical Uni-\nversity, Chernivtsi, Ukraine (approval ID: No. 6 from 8.10.2024). \nCONFLICT OF INTEREST\nThe Authors declare no conflict of interest\nCORRESPONDING AUTHOR\nOksana V. Bakun\nBukovinian State Medical University\n129 Golovna St., 58000 Chernivtsi, Ukraine\ne – mail: kupchanko06@gmail.com \nORCID AND CONTRIBUTIONSHIP\nOksana V. Bakun: 0000-0002-4742-2265  \nKristina V. Dyak: 0000-0002-6568-7945  \nOksana V. Kolesnik: 0000-0001-9087-2635  \nTetiana V. Protsak: 0000-0002-9620-3667  \nYevheniia A. Dudka: 0000-0002-8301-500X  \n – Work concept and design,  – Data collection and analysis,  – Responsibility for statistical analysis,  – Writing the article,  – Critical review,  – Final approval of the article\nRECEIVED: 10.12.2024\nACCEPTED: 25.03.2025\n CREATIVE COMMONS 4.0","source_license":"CC0","license_restricted":false}