{"paper_id":"2f5cb5d5-c59c-486f-8046-20edb7fc57f9","body_text":"Stage-Specific Changes in Irisin, Nesfatin-1, and Oxidative Stress Markers in Brucellosis | 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 Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Stage-Specific Changes in Irisin, Nesfatin-1, and Oxidative Stress Markers in Brucellosis Sermin Algul, Feray Ferda Senol, Pinar Oner, Ayse Seker, Oguz Ozcelik This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7884325/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 Background Brucellosis is a globally prevalent foodborne zoonotic disease transmitted primarily through the ingestion of unpasteurized dairy products or undercooked meat. While its infectious etiology is well recognized, the metabolic and oxidative stress-related alterations associated with the disease remain insufficiently characterized. Objectives This study aimed to investigate stage-specific changes in serum irisin and nesfatin-1 levels, alongside markers of oxidative stress, in patients with acute, subacute, and chronic brucellosis. Methods A total of 90 patients diagnosed with brucellosis at different clinical stages and 30 healthy controls were included. Serum levels of irisin, nesfatin-1, total oxidant status (TOS), total antioxidant status (TAS), and the oxidative stress index (OSI; TOS/TAS ratio) were determined using enzyme-linked immunosorbent assay (ELISA). Intergroup comparisons were performed using one-way ANOVA and paired t-tests. Results Serum irisin and nesfatin-1 levels were significantly reduced in brucellosis patients compared to controls, with variations dependent on disease stage (p < 0.05). Concurrently, a significant increase in TOS and OSI values and a decrease in TAS levels were observed across all stages of brucellosis (p < 0.05). Conclusions Brucellosis induces distinct alterations in energy-regulating hormones and redox balance, extending beyond its infectious profile. Reduced levels of irisin and nesfatin-1, in parallel with elevated oxidative stress, suggest potential roles for these hormones as biomarkers in disease staging and metabolic monitoring. These findings underscore the need to incorporate hormonal and oxidative profiling into the clinical evaluation of brucellosis. To our knowledge, this is the first study to evaluate irisin and nesfatin-1 responses across different clinical stages of brucellosis. Brucellosis foodborne zoonosis irisin nesfatin-1 oxidative stress metabolic dysregulation disease staging ELISA Figures Figure 1 Figure 2 Figure 3 Introduction Brucellosis is a globally prevalent foodborne zoonotic infection, primarily transmitted through the consumption of unpasteurized dairy products or undercooked meat from infected animals such as cattle, sheep, goats, and pigs [ 1 , 2 ]. The disease remains endemic in several regions, including the Mediterranean, Middle East, South Asia, and Latin America, with particularly high incidence reported in the southeastern regions of Turkey [ 3 – 6 ]. In many low- and middle-income countries, the burden of brucellosis is compounded by limited food safety enforcement and continued consumption of unprocessed animal products. Clinically, brucellosis exhibits a wide spectrum ranging from acute febrile illness to severe complications such as arthritis, endocarditis, osteomyelitis, and neurobrucellosis [ 7 , 8 ]. While the infectious etiology of brucellosis is well characterized, its systemic consequences—particularly those involving oxidative stress and metabolic regulation—are less well understood. The host response in brucellosis involves chronic immune activation, oxidative imbalance, and potential disruption of hormonal homeostasis [ 9 – 11 ]. Elevated levels of reactive oxygen species (ROS), accompanied by reduced antioxidant defenses, contribute to disease progression and tissue damage [ 12 , 13 ]. Previous studies have consistently reported increased oxidative stress and impaired antioxidant capacity in patients with brucellosis [ 14 , 15 ]. In recent years, the metabolic hormones irisin and nesfatin-1 have gained attention for their dual roles in energy homeostasis and redox regulation. Irisin, secreted by skeletal muscle and adipose tissue, enhances mitochondrial function, glucose uptake, and thermogenesis, and has demonstrated antioxidant effects in both experimental and clinical settings [ 16 , 17 ]. Similarly, nesfatin-1, a hypothalamic peptide with peripheral expression, was initially identified for its anorexigenic and insulin-sensitizing effects but is now also recognized for its involvement in anti-inflammatory and antioxidant pathways [ 18 – 21 ]. Despite increasing evidence of their relevance in metabolic and inflammatory diseases, the roles of irisin and nesfatin-1 in chronic infectious diseases such as brucellosis remain unclear. Given the oxidative and inflammatory burden of brucellosis, these hormones may serve as biomarkers for disease staging and systemic stress. The present study aimed to investigate the stage-specific alterations in serum irisin and nesfatin-1 levels, along with oxidative stress markers—total oxidant status (TOS), total antioxidant status (TAS), and the oxidative stress index (OSI)—in patients with acute, subacute, and chronic brucellosis. We hypothesize that disease progression is associated with distinct hormonal and oxidative patterns, potentially offering novel insights for clinical evaluation and monitoring. To date, no previous study has examined the stage-dependent alterations in these redox-sensitive hormones within the context of brucellosis, highlighting a critical gap that this work aims to address. Materials and Methods Study Design and Participants This cross-sectional study included 120 participants, comprising 90 patients diagnosed with brucellosis and 30 healthy controls. Patients were categorized into three groups according to disease duration: acute (<8 weeks), subacute (8–52 weeks), and chronic (>52 weeks), based on clinical definitions [22]. All patients were recruited from the Infectious Diseases outpatient clinic at Van Yüzüncü Yıl University Faculty of Medicine. -Acute group (n = 30): Mean age 40.4 ± 13.0 years; height 169 ± 6 cm; weight 65.8 ± 6.4 kg -Subacute group (n = 30): Mean age 42.9 ± 16.0 years; height 166 ± 7 cm; weight 68.1 ± 7.6 kg -Chronic group (n = 30): Mean age 46.7 ± 10.0 years; height 168 ± 9 cm; weight 70.4 ± 8.2 kg -Control group (n = 30): Healthy individuals with no history of brucellosis or systemic illness within the past 6 months; mean age 37.5 ± 14.0 years; height 167 ± 5 cm; weight 67.8 ± 6.5 kg Ethical Approval Ethical approval was obtained from the Firat University Non-Invasive Research Ethics Committee (Decision No: 2017/07-13). All participants provided written informed consent, and the study complied with the Declaration of Helsinki. Inclusion and Exclusion Criteria Participants underwent medical screening including clinical history, ECG, physical examination, and routine laboratory tests. Exclusion criteria included pregnancy, BMI >30 kg/m², chronic metabolic or inflammatory diseases, cancer, recent use of supplements, alcohol, tobacco, or hormonal therapy. Diagnostic Criteria Brucellosis diagnosis was confirmed by clinical presentation and serological testing using the Rose-Bengal Plate Test (RBPT) and the Standard Tube Agglutination Test (STAT). A titer ≥1:160 was considered diagnostic [23–25]. Biochemical Analysis Sample Collection Venous blood samples (5 mL) were collected between 08:00–09:00 a.m. after overnight fasting. Samples were centrifuged at 4500 rpm for 5 minutes at +4°C and stored at –80°C until analysis. Hormonal and Oxidative Stress Parameters Serum irisin, nesfatin-1, total oxidant status (TOS), and total antioxidant status (TAS) levels were measured using commercial ELISA kits (SunRed Biological Technology Co., Ltd., Shanghai, China). The technical characteristics of the ELISA kits are summarized in Table 1. Table 1: Technical specifications of commercial ELISA kits used for the measurement of irisin, nesfatin-1, total oxidant status (TOS), and total antioxidant status (TAS) in serum samples. Assay sensitivity, detection ranges, and intra-/inter-assay coefficients of variation (CV) are provided as reported by the manufacturer (SunRed Biological Technology Co., Ltd., Shanghai, China). Parameter Kit Catalog No. Detection Range Sensitivity Intra-/Inter-assay CV Irisin 201-12-5328 0.2–60 ng/mL 0.157 ng/mL <10% / <12% Nesfatin-1 201-12-4341 0.2–35 mmol/L 0.113 mmol/L <10% / <12% TOS 201-12-5807 1.5–400 U/mL 1.362 U/mL <8% / <11% TAS 201-12-7412 0.5–120 U/mL 0.411 U/mL <9% / <11% The Oxidative Stress Index (OSI) was calculated as the ratio of TOS to TAS, providing an integrated measure of oxidative stress burden [26]. Statistical Analysis All data were expressed as mean ± standard error (SE). Normality was assessed using the Kolmogorov–Smirnov test. One-way ANOVA was used to compare intergroup differences, followed by post hoc multiple comparisons. A p-value of <0.05 was considered statistically significant. Results This study demonstrated significant alterations in the levels of irisin, nesfatin-1, total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) across different stages of brucellosis compared to healthy controls. Hormonal Parameters: Irisin and Nesfatin-1 Serum irisin levels were significantly decreased in all brucellosis groups relative to the control group (38.31 ± 1.57 ng/mL). Specifically, levels were 28.47 ± 1.09 ng/mL in the subacute group (p < 0.0001), 24.12 ± 1.48 ng/mL in the acute group (p < 0.0001), and 23.18 ± 0.97 ng/mL in the chronic group (p < 0.0001) (Figure 1). Moreover, the subacute group exhibited significantly higher irisin levels compared to both the acute and chronic groups (p < 0.05), although no significant difference was observed between the acute and chronic groups. Similarly, serum nesfatin-1 levels were significantly lower in all brucellosis groups when compared to the control group (2.063 ± 0.09 mmol/L), with levels of 1.735 ± 0.07 mmol/L in the subacute group (p = 0.01), 1.540 ± 0.06 mmol/L in the acute group (p = 0.001), and 1.427 ± 0.06 mmol/L in the chronic group (p < 0.0001). The subacute group also showed significantly higher nesfatin-1 levels compared to both the acute and chronic groups. However, there were no significant differences in nesfatin-1 between the acute and chronic groups (Figure 1). Oxidative Stress Markers: TAS, TOS, and OSI A progressive decline in TAS was observed with advancing disease stage. The control group had a TAS level of 2.948 ± 0.08 U/mL, which significantly declined to 2.504 ± 0.06 U/mL in the subacute group, 2.085 ± 0.05 U/mL in the acute group, and 1.893 ± 0.07 U/mL in the chronic group (p < 0.05 for all comparisons). Conversely, TOS levels were significantly elevated in the patient groups compared to controls (27.23 ± 1.27 U/mL), with measured values of 39.80 ± 2.01 U/mL (subacute), 45.78 ± 1.96 U/mL (acute), and 47.61 ± 1.53 U/mL (chronic) (p < 0.05 for all) (Figure 1). The OSI values showed a marked elevation in all patient groups relative to controls. Specifically, OSI increased by 68% in the subacute group, 286% in the acute group, and 380% in the chronic group compared to the control group (p < 0.0001 for all comparisons) (Figure 2). Correlation Analysis A significant negative correlation was observed between OSI and serum irisin levels in both the control (R = –0.8133, p < 0.0001) and chronic brucellosis (R = –0.5285, p = 0.01) groups (Figure 3), suggesting that rising oxidative stress is accompanied by a decrease in protective myokine levels. Discussion This study investigated the stage-dependent alterations in oxidative stress markers and the circulating levels of the energy-regulating hormones irisin and nesfatin-1 in patients with brucellosis. Our findings demonstrate that brucellosis—beyond its infectious nature—induces significant disruptions in redox homeostasis and metabolic hormone profiles, particularly as the disease progresses from acute to chronic stages. Oxidative stress is a hallmark of chronic infections and has been increasingly recognized as a contributor to the systemic pathology of brucellosis. We observed elevated levels of total oxidant status (TOS), decreased total antioxidant status (TAS), and a significant increase in oxidative stress index (OSI) across all brucellosis groups compared to healthy controls. These findings are in agreement with previous reports indicating an imbalance between pro-oxidant and antioxidant mechanisms in brucellosis patients, contributing to cellular and tissue damage through lipid peroxidation and protein oxidation [27–31]. Importantly, the current study identifies a clear decline in serum irisin and nesfatin-1 levels in brucellosis patients, with the most pronounced reductions observed in the chronic stage. Both irisin and nesfatin-1 are pleiotropic hormones implicated in energy metabolism, inflammatory regulation, and oxidative stress response. Irisin, primarily secreted by skeletal muscle, has been shown to enhance antioxidant defense through mitochondrial biogenesis and modulation of reactive oxygen species [32, 33]. Similarly, nesfatin-1 exhibits cytoprotective effects via free radical scavenging and anti-inflammatory signaling pathways [34-36]. The observed inverse correlation between OSI and irisin in the chronic brucellosis group highlights the potential of irisin as a redox-sensitive biomarker. These findings align with data from other inflammatory and metabolic conditions where irisin levels reflect oxidative burden [32, 37, 38]. The decline in nesfatin-1 may also indicate impaired stress adaptation mechanisms during prolonged infection and inflammation [34, 35]. Interestingly, some conflicting results in the literature—such as reports of elevated TAS or reduced TOS in brucellosis—may stem from heterogeneity in disease staging, treatment exposure, or population characteristics [25, 29, 30]. Nonetheless, our consistent findings across acute, subacute, and chronic stages underscore the progressive metabolic and oxidative burden imposed by brucellosis. The clinical relevance of these hormonal changes should not be underestimated. Given the central roles of irisin and nesfatin-1 in appetite regulation, glucose homeostasis, and mitochondrial function, their suppression may contribute to fatigue, weight loss, and altered energy metabolism often observed in chronic brucellosis. Moreover, these hormones may serve as adjunctive biomarkers to aid in disease staging or therapeutic monitoring, particularly in resource-limited settings where advanced diagnostic modalities may be lacking. From a public health perspective, our findings reinforce the need to consider brucellosis not only as an infectious disease but also as a metabolic stressor with potential long-term systemic consequences. Future research should explore the mechanistic links between Brucella-induced oxidative damage, endocrine dysregulation, and nutritional status. Longitudinal studies are warranted to evaluate whether restoring redox balance and hormonal function—through pharmacologic, dietary, or lifestyle interventions—can improve clinical outcomes in affected individuals. Conclusion This study provides compelling evidence that brucellosis exerts a significant systemic impact beyond its infectious etiology, notably altering oxidative stress parameters and key metabolic hormones such as irisin and nesfatin-1. The progressive decrease in these energy-regulating hormones across disease stages, in conjunction with elevated oxidative stress markers, underscores the nutritional-metabolic burden imposed by this foodborne zoonosis. These findings support the integration of hormonal and redox biomarkers into the clinical evaluation of brucellosis and suggest potential utility for disease staging and therapeutic monitoring. From a public health standpoint, improving food safety practices and considering nutritional support strategies may enhance disease control and patient outcomes in endemic regions. Declarations Funding Statement The authors declare that no specific grants from funding agencies in the public, commercial, or not-for-profit sectors were received for the conduct of this study. Ethical Approval Ethical clearance for this study was obtained from the Firat University Non-Invasive Local Ethics Committee (Decision No: 2017/07-13). All procedures performed were in accordance with the ethical standards of the institutional and national research committees and with the 1964 Helsinki Declaration and its later amendments. Consent to Participate declaration All participants provided written informed consent prior to inclusion in the study. Clinical Trial Number Not Applicable. Conflict of Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Acknowledgements The authors have no acknowledgements to disclose. Author Contributions SA, AS, and OO were responsible for conceptualization, data interpretation, and critical revision of the manuscript. FFS and PO contributed to data acquisition, including blood sampling and clinical documentation. All authors reviewed and approved the final version of the manuscript. Data Availability: The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. References Yumuk Z, O’Callaghan D. Brucellosis in Turkey—an overview. Int J Infect Dis . 2012;16(4):228–35. doi:10.1016/j.ijid.2011.12.011 Galińska EM, Zagórski J. Brucellosis in humans—etiology, diagnostics, clinical forms. Ann Agric Environ Med . 2013;20(2):233–8. Guler S, Kokoglu OF, Ucmak H, et al. Human brucellosis in Turkey: different clinical presentations. J Infect Dev Ctries . 2014;8(5):581–8. doi:10.3855/jidc.3510 Zhang N, Zhou H, Huang DS, Guan P. Brucellosis awareness and knowledge in communities worldwide: a systematic review and meta-analysis of 79 observational studies. PLoS Negl Trop Dis . 2019;13(5):0007366. doi:10.1371/journal.pntd.0007366 Moriyón I, Blasco JM, Letesson JJ, De Massis F, Moreno E. Brucellosis and one health: inherited and future challenges. Microorganisms . 2023;11(8):2070. doi:10.3390/microorganisms11082070 Alqaseer K, Al-Khafajy AAM, Almkhadhree EAK. Serological and molecular detection of human brucellosis in rural areas in Wasit Province, Iraq. Arch Razi Inst . 2023;78(1):369–78. doi:10.22092/ARI.2022.359002.2352 Samadi A, Amiri M, Hailat N. The reasons behind long-term endemicity of brucellosis in low and middle-income countries: challenges and future perspectives. Curr Microbiol . 2024;81(3):82. doi:10.1007/s00284-023-03605-5 Yagupsky P, Morata P, Colmenero JD. Laboratory diagnosis of human brucellosis. Clin Microbiol Rev . 2019;33(1):e00073–19. doi:10.1128/CMR.00073-19 Sen P, Demirdal T, Nemli SA. Predictive value of inflammation markers in brucellosis. Arch Iran Med . 2019;22(11):640–5. Karaagac L, Koruk ST, Koruk I, Aksoy N. Decreasing oxidative stress in response to treatment in patients with brucellosis: could it be used to monitor treatment? Int J Infect Dis . 2011;15(5):346–9. doi:10.1016/j.ijid.2011.01.009 Demirpençe O, Sevim B, Yıldırım M, et al. Serum paraoxonase, TAS, TOS and ceruloplasmin in brucellosis. Int J Clin Exp Med . 2014;7(6):1592–7. Filomeni G, De Zio D, Cecconi F. Oxidative stress and autophagy: the clash between damage and metabolic needs. Cell Death Differ . 2015;22(3):377–88. doi:10.1038/cdd.2014.150 Teleanu DM, Niculescu AG, Lungu II, et al. An overview of oxidative stress, neuroinflammation, and neurodegenerative diseases. Int J Mol Sci . 2022;23(11):5938. doi:10.3390/ijms23115938 Melek IM, Erdogan S, Celik S, et al. Evaluation of oxidative stress and inflammation in long-term Brucella melitensis infection. Mol Cell Biochem . 2006;293(1-2):203–9. doi:10.1007/s11010-006-9243-2 Esen R, Aslan M, Kucukoglu ME, et al. Serum paraoxonase activity, total thiols levels, and oxidative status in patients with acute brucellosis. Wien Klin Wochenschr . 2015;127(11-12):427–33. doi:10.1007/s00508-015-0720-z Boström P, Wu J, Jedrychowski MP, et al. A PGC1-α-dependent myokine that drives brown-fat-like development of white fat and thermogenesis. Nature . 2012;481(7382):463–8. doi:10.1038/nature10777 Ozcelik O, Algul S, Yilmaz B. Nesfatin-1 and irisin levels in response to the soccer matches performed in morning, afternoon and at night in young trained male subjects. Cell Mol Biol (Noisy-le-grand) . 2018;64(13):130–3. Oh-I S, Shimizu H, Satoh T, et al. Identification of nesfatin-1 as a satiety molecule in the hypothalamus. Nature . 2006;443(7112):709–12. doi:10.1038/nature05162 Askari H, Rajani SF, Poorebrahim M, et al. A glance at the therapeutic potential of irisin against diseases involving inflammation, oxidative stress, and apoptosis: an introductory review. Pharmacol Res . 2018;129:44–55. doi:10.1016/j.phrs.2018.01.012 Ayada C, Toru Ü, Genç O, et al. Balanced oxidative status by nesfatin-1 in intestinal ischemia-reperfusion. Int J Clin Exp Med . 2015;8(3):3318–24. Zhang Z, Zhang X, Chen X, et al. Clinical features of human brucellosis and risk factors for focal complications. Int J Gen Med . 2022;15:7373–82. doi:10.2147/IJGM.S380328 Ertek M, Yazgi H, Ozkurt Z, et al. Comparison of the diagnostic value of the standard tube agglutination test and the ELISA IgG and IgM in patients with brucellosis. Turk J Med Sci . 2006;36(3):159–63. Yohannes M, Gill JP, Ghatak S, et al. Comparative evaluation of the Rose Bengal plate test, standard tube agglutination test and complement fixation test. Rev Sci Tech . 2012;31(3):979–84. doi:10.20506/rst.31.3.2175 Andriopoulos P, Tsironi M, Deftereos S, et al. Acute brucellosis: presentation, diagnosis, and treatment of 144 cases. Int J Infect Dis . 2007;11(1):52–7. doi:10.1016/j.ijid.2005.10.011 Karaagac L, Koruk ST, Koruk I, Aksoy N. Decreasing oxidative stress in response to treatment in patients with brucellosis: could it be used to monitor treatment? Int J Infect Dis . 2011;15(5):346–9. doi:10.1016/j.ijid.2011.01.009 Moreno E, Blasco JM, Moriyón I. Facing the human and animal brucellosis conundrums: the forgotten lessons. Microorganisms . 2022;10(5):942. doi:10.3390/microorganisms10050942 Perin G, Bottari NB, Silva AD, et al. Cholinesterase's activities of infected mice by Brucella ovis . Microb Pathog . 2019;132:137–140. doi:10.1016/j.micpath.2019.04.036 Baldi PC, Giambartolomei GH. Immunopathology of Brucella infection. Recent Pat Antiinfect Drug Discov . 2013;8(1):18–26. doi:10.2174/1574891X11308010005 Serefhanoglu K, Taskin A, Turan H, et al. Evaluation of oxidative status in patients with brucellosis. Braz J Infect Dis . 2009;13(4):249–51. doi:10.1590/s1413-86702009000400001 Usta M, Aras Z, Tas A. Oxidant and antioxidant parameters in patients with Brucella canis . Clin Biochem . 2012;45(4–5):366–7. doi:10.1016/j.clinbiochem.2011.12.028 Barzilai A, Yamamoto K. DNA damage responses to oxidative stress. DNA Repair (Amst) . 2004;3(8–9):1109–15. doi:10.1016/j.dnarep.2004.03.002 Slate-Romano JJ, Yano N, Zhao TC. Irisin reduces inflammatory signaling pathways in inflammation-mediated metabolic syndrome. Mol Cell Endocrinol . 2022;552:111676. doi:10.1016/j.mce.2022.111676 Algul S, Senol FF, Ugras S, et al. Investigation of irisin level in brucella patients. East J Med . 2021;26(2):135–8. doi:10.5505/ejm.2021.70431 Xu Y, Chen F. Antioxidant, anti-inflammatory and anti-apoptotic activities of nesfatin-1: a review. J Inflamm Res . 2020;13:607–17. doi:10.2147/JIR.S273446 Caylak E. IFN-γ, IL-10, ghrelin and nesfatin-1 levels in acute brucella patients. Ann Clin Anal Med . 2022;13(7):716–20. doi:10.4328/ACAM.21061 Algul S, Ozcelik O. Günün farklı zamanlarında yapılan futbol maçlarında oksidan-antioksidan dengenin incelenmesi. Genel Tıp Derg . 2017;27(4):129–35. Canpolat Erkan RE, Tekin R. Investigation of new inflammatory biomarkers in patients with brucella. PLoS One . 2024;19(2):0297550. doi:10.1371/journal.pone.0297550 Mazur-Bialy AI, Kozlowska K, Pochec E, et al. Myokine irisin-induced protection against oxidative stress in vitro. J Physiol Pharmacol . 2018;69(1):117–25. 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compared to the control group.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7884325/v1/b043f91f47ad467e25715b2e.png\"},{\"id\":96180654,\"identity\":\"8d03995e-e666-4e67-97b7-6584dedd0e1e\",\"added_by\":\"auto\",\"created_at\":\"2025-11-18 12:32:49\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":33287,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eOS index for the control, subacute, acute, and chronic brucellosis groups.\\u003c/p\\u003e\\n\\u003cp\\u003e* reflects statistically significant differences compared to the control group\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026amp; reflects statistically significant differences compared to the subacute group\\u003c/p\\u003e\\n\\u003cp\\u003eΨ reflects statistically significant differences compared to the acute group\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7884325/v1/450a79a2e4a548641a6ea323.png\"},{\"id\":96180656,\"identity\":\"562df8ff-ad3a-437f-afa2-dee37c089ec6\",\"added_by\":\"auto\",\"created_at\":\"2025-11-18 12:32:49\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":33309,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLinear regression showing the correlation between OSI and irisin levels in patients with chronic brucellosis.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7884325/v1/15fb267b26c7d191339f95ac.png\"},{\"id\":98380727,\"identity\":\"2cbafd69-92e6-46e7-a3af-b3da2e1856cf\",\"added_by\":\"auto\",\"created_at\":\"2025-12-17 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class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e]. The disease remains endemic in several regions, including the Mediterranean, Middle East, South Asia, and Latin America, with particularly high incidence reported in the southeastern regions of Turkey [\\u003cspan additionalcitationids=\\\"CR4 CR5\\\" citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e]. In many low- and middle-income countries, the burden of brucellosis is compounded by limited food safety enforcement and continued consumption of unprocessed animal products.\\u003c/p\\u003e\\u003cp\\u003eClinically, brucellosis exhibits a wide spectrum ranging from acute febrile illness to severe complications such as arthritis, endocarditis, osteomyelitis, and neurobrucellosis [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e]. While the infectious etiology of brucellosis is well characterized, its systemic consequences\\u0026mdash;particularly those involving oxidative stress and metabolic regulation\\u0026mdash;are less well understood.\\u003c/p\\u003e\\u003cp\\u003eThe host response in brucellosis involves chronic immune activation, oxidative imbalance, and potential disruption of hormonal homeostasis [\\u003cspan additionalcitationids=\\\"CR10\\\" citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e]. Elevated levels of reactive oxygen species (ROS), accompanied by reduced antioxidant defenses, contribute to disease progression and tissue damage [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e]. Previous studies have consistently reported increased oxidative stress and impaired antioxidant capacity in patients with brucellosis [\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e].\\u003c/p\\u003e\\u003cp\\u003eIn recent years, the metabolic hormones irisin and nesfatin-1 have gained attention for their dual roles in energy homeostasis and redox regulation. Irisin, secreted by skeletal muscle and adipose tissue, enhances mitochondrial function, glucose uptake, and thermogenesis, and has demonstrated antioxidant effects in both experimental and clinical settings [\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e]. Similarly, nesfatin-1, a hypothalamic peptide with peripheral expression, was initially identified for its anorexigenic and insulin-sensitizing effects but is now also recognized for its involvement in anti-inflammatory and antioxidant pathways [\\u003cspan additionalcitationids=\\\"CR19 CR20\\\" citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e].\\u003c/p\\u003e\\u003cp\\u003eDespite increasing evidence of their relevance in metabolic and inflammatory diseases, the roles of irisin and nesfatin-1 in chronic infectious diseases such as brucellosis remain unclear. Given the oxidative and inflammatory burden of brucellosis, these hormones may serve as biomarkers for disease staging and systemic stress.\\u003c/p\\u003e\\u003cp\\u003eThe present study aimed to investigate the stage-specific alterations in serum irisin and nesfatin-1 levels, along with oxidative stress markers\\u0026mdash;total oxidant status (TOS), total antioxidant status (TAS), and the oxidative stress index (OSI)\\u0026mdash;in patients with acute, subacute, and chronic brucellosis. We hypothesize that disease progression is associated with distinct hormonal and oxidative patterns, potentially offering novel insights for clinical evaluation and monitoring. To date, no previous study has examined the stage-dependent alterations in these redox-sensitive hormones within the context of brucellosis, highlighting a critical gap that this work aims to address.\\u003c/p\\u003e\"},{\"header\":\"Materials and Methods\",\"content\":\"\\u003ch4\\u003e\\u003cstrong\\u003eStudy Design and Participants\\u003c/strong\\u003e\\u003c/h4\\u003e\\n\\u003cp\\u003eThis cross-sectional study included 120 participants, comprising 90 patients diagnosed with brucellosis and 30 healthy controls. Patients were categorized into three groups according to disease duration: acute (\\u0026lt;8 weeks), subacute (8\\u0026ndash;52 weeks), and chronic (\\u0026gt;52 weeks), based on clinical definitions [22]. All patients were recruited from the Infectious Diseases outpatient clinic at Van Y\\u0026uuml;z\\u0026uuml;nc\\u0026uuml; Yıl University Faculty of Medicine.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e-Acute group (n = 30):\\u003c/strong\\u003e Mean age 40.4 \\u0026plusmn; 13.0 years; height 169 \\u0026plusmn; 6 cm; weight 65.8 \\u0026plusmn; 6.4 kg\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e-Subacute group (n = 30):\\u003c/strong\\u003e Mean age 42.9 \\u0026plusmn; 16.0 years; height 166 \\u0026plusmn; 7 cm; weight 68.1 \\u0026plusmn; 7.6 kg\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e-Chronic group (n = 30):\\u003c/strong\\u003e Mean age 46.7 \\u0026plusmn; 10.0 years; height 168 \\u0026plusmn; 9 cm; weight 70.4 \\u0026plusmn; 8.2 kg\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e-Control group (n = 30):\\u003c/strong\\u003e Healthy individuals with no history of brucellosis or systemic illness within the past 6 months; mean age 37.5 \\u0026plusmn; 14.0 years; height 167 \\u0026plusmn; 5 cm; weight 67.8 \\u0026plusmn; 6.5 kg\\u003c/p\\u003e\\n\\u003ch4\\u003e\\u003cstrong\\u003eEthical Approval\\u003c/strong\\u003e\\u003c/h4\\u003e\\n\\u003cp\\u003eEthical approval was obtained from the Firat University Non-Invasive Research Ethics Committee (Decision No: 2017/07-13). All participants provided written informed consent, and the study complied with the Declaration of Helsinki.\\u003c/p\\u003e\\n\\u003ch4\\u003e\\u003cstrong\\u003eInclusion and Exclusion Criteria\\u003c/strong\\u003e\\u003c/h4\\u003e\\n\\u003cp\\u003eParticipants underwent medical screening including clinical history, ECG, physical examination, and routine laboratory tests. Exclusion criteria included pregnancy, BMI \\u0026gt;30 kg/m\\u0026sup2;, chronic metabolic or inflammatory diseases, cancer, recent use of supplements, alcohol, tobacco, or hormonal therapy.\\u003c/p\\u003e\\n\\u003ch4\\u003e\\u003cstrong\\u003eDiagnostic Criteria\\u003c/strong\\u003e\\u003c/h4\\u003e\\n\\u003cp\\u003eBrucellosis diagnosis was confirmed by clinical presentation and serological testing using the Rose-Bengal Plate Test (RBPT) and the Standard Tube Agglutination Test (STAT). A titer \\u0026ge;1:160 was considered diagnostic [23\\u0026ndash;25].\\u003c/p\\u003e\\n\\u003ch3\\u003e\\u003cstrong\\u003eBiochemical Analysis\\u003c/strong\\u003e\\u003c/h3\\u003e\\n\\u003ch4\\u003e\\u003cstrong\\u003eSample Collection\\u003c/strong\\u003e\\u003c/h4\\u003e\\n\\u003cp\\u003eVenous blood samples (5 mL) were collected between 08:00\\u0026ndash;09:00 a.m. after overnight fasting. Samples were centrifuged at 4500 rpm for 5 minutes at +4\\u0026deg;C and stored at \\u0026ndash;80\\u0026deg;C until analysis.\\u003c/p\\u003e\\n\\u003ch4\\u003e\\u003cstrong\\u003eHormonal and Oxidative Stress Parameters\\u003c/strong\\u003e\\u003c/h4\\u003e\\n\\u003cp\\u003eSerum irisin, nesfatin-1, total oxidant status (TOS), and total antioxidant status (TAS) levels were measured using commercial ELISA kits (SunRed Biological Technology Co., Ltd., Shanghai, China).\\u0026nbsp;The technical characteristics of the ELISA kits are summarized in Table 1.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 1:\\u003c/strong\\u003e Technical specifications of commercial ELISA kits used for the measurement of irisin, nesfatin-1, total oxidant status (TOS), and total antioxidant status (TAS) in serum samples. Assay sensitivity, detection ranges, and intra-/inter-assay coefficients of variation (CV) are provided as reported by the manufacturer (SunRed Biological Technology Co., Ltd., Shanghai, China).\\u003c/p\\u003e\\n\\u003ctable border=\\\"1\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eParameter\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eKit Catalog No.\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eDetection Range\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eSensitivity\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eIntra-/Inter-assay CV\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eIrisin\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e201-12-5328\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e0.2\\u0026ndash;60 ng/mL\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e0.157 ng/mL\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;10% / \\u0026lt;12%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eNesfatin-1\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e201-12-4341\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e0.2\\u0026ndash;35 mmol/L\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e0.113 mmol/L\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;10% / \\u0026lt;12%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eTOS\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e201-12-5807\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e1.5\\u0026ndash;400 U/mL\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e1.362 U/mL\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;8% / \\u0026lt;11%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eTAS\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e201-12-7412\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e0.5\\u0026ndash;120 U/mL\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e0.411 U/mL\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 121px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;9% / \\u0026lt;11%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003eThe \\u003cstrong\\u003eOxidative Stress Index (OSI)\\u003c/strong\\u003e was calculated as the ratio of TOS to TAS, providing an integrated measure of oxidative stress burden [26].\\u003c/p\\u003e\\n\\u003ch3\\u003e\\u003cstrong\\u003eStatistical Analysis\\u003c/strong\\u003e\\u003c/h3\\u003e\\n\\u003cp\\u003eAll data were expressed as mean \\u0026plusmn; standard error (SE). Normality was assessed using the Kolmogorov\\u0026ndash;Smirnov test. One-way ANOVA was used to compare intergroup differences, followed by post hoc multiple comparisons. A p-value of \\u0026lt;0.05 was considered statistically significant.\\u003c/p\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003eThis study demonstrated significant alterations in the levels of irisin, nesfatin-1, total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) across different stages of brucellosis compared to healthy controls.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eHormonal Parameters: Irisin and Nesfatin-1\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eSerum irisin levels were significantly decreased in all brucellosis groups relative to the control group (38.31 \\u0026plusmn; 1.57 ng/mL). Specifically, levels were 28.47 \\u0026plusmn; 1.09 ng/mL in the subacute group (p \\u0026lt; 0.0001), 24.12 \\u0026plusmn; 1.48 ng/mL in the acute group (p \\u0026lt; 0.0001), and 23.18 \\u0026plusmn; 0.97 ng/mL in the chronic group (p \\u0026lt; 0.0001) (Figure 1). Moreover, the subacute group exhibited significantly higher irisin levels compared to both the acute and chronic groups (p \\u0026lt; 0.05), although no significant difference was observed between the acute and chronic groups.\\u003c/p\\u003e\\n\\u003cp\\u003eSimilarly, serum nesfatin-1 levels were significantly lower in all brucellosis groups when compared to the control group (2.063 \\u0026plusmn; 0.09 mmol/L), with levels of 1.735 \\u0026plusmn; 0.07 mmol/L in the subacute group (p = 0.01), 1.540 \\u0026plusmn; 0.06 mmol/L in the acute group (p = 0.001), and 1.427 \\u0026plusmn; 0.06 mmol/L in the chronic group (p \\u0026lt; 0.0001). The subacute group also showed significantly higher nesfatin-1 levels compared to both the acute and chronic groups. However, there were no significant differences in nesfatin-1 between the acute and chronic groups (Figure 1).\\u003c/p\\u003e\\n\\u003ch4\\u003e\\u003cstrong\\u003eOxidative Stress Markers: TAS, TOS, and OSI\\u003c/strong\\u003e\\u003c/h4\\u003e\\n\\u003cp\\u003eA progressive decline in TAS was observed with advancing disease stage. The control group had a TAS level of 2.948 \\u0026plusmn; 0.08 U/mL, which significantly declined to 2.504 \\u0026plusmn; 0.06 U/mL in the subacute group, 2.085 \\u0026plusmn; 0.05 U/mL in the acute group, and 1.893 \\u0026plusmn; 0.07 U/mL in the chronic group (p \\u0026lt; 0.05 for all comparisons).\\u003c/p\\u003e\\n\\u003cp\\u003eConversely, TOS levels were significantly elevated in the patient groups compared to controls (27.23 \\u0026plusmn; 1.27 U/mL), with measured values of 39.80 \\u0026plusmn; 2.01 U/mL (subacute), 45.78 \\u0026plusmn; 1.96 U/mL (acute), and 47.61 \\u0026plusmn; 1.53 U/mL (chronic) (p \\u0026lt; 0.05 for all) (Figure 1).\\u003c/p\\u003e\\n\\u003cp\\u003eThe OSI values showed a marked elevation in all patient groups relative to controls. Specifically, OSI increased by 68% in the subacute group, 286% in the acute group, and 380% in the chronic group compared to the control group (p \\u0026lt; 0.0001 for all comparisons) (Figure 2).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCorrelation Analysis\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eA significant negative correlation was observed between OSI and serum irisin levels in both the control (R = \\u0026ndash;0.8133, p \\u0026lt; 0.0001) and chronic brucellosis (R = \\u0026ndash;0.5285, p = 0.01) groups (Figure 3), suggesting that rising oxidative stress is accompanied by a decrease in protective myokine levels.\\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eThis study investigated the stage-dependent alterations in oxidative stress markers and the circulating levels of the energy-regulating hormones irisin and nesfatin-1 in patients with brucellosis. Our findings demonstrate that brucellosis\\u0026mdash;beyond its infectious nature\\u0026mdash;induces significant disruptions in redox homeostasis and metabolic hormone profiles, particularly as the disease progresses from acute to chronic stages.\\u003c/p\\u003e\\n\\u003cp\\u003eOxidative stress is a hallmark of chronic infections and has been increasingly recognized as a contributor to the systemic pathology of brucellosis. We observed elevated levels of total oxidant status (TOS), decreased total antioxidant status (TAS), and a significant increase in oxidative stress index (OSI) across all brucellosis groups compared to healthy controls. These findings are in agreement with previous reports indicating an imbalance between pro-oxidant and antioxidant mechanisms in brucellosis patients, contributing to cellular and tissue damage through lipid peroxidation and protein oxidation [27\\u0026ndash;31].\\u003c/p\\u003e\\n\\u003cp\\u003eImportantly, the current study identifies a clear decline in serum irisin and nesfatin-1 levels in brucellosis patients, with the most pronounced reductions observed in the chronic stage. Both irisin and nesfatin-1 are pleiotropic hormones implicated in energy metabolism, inflammatory regulation, and oxidative stress response. Irisin, primarily secreted by skeletal muscle, has been shown to enhance antioxidant defense through mitochondrial biogenesis and modulation of reactive oxygen species [32, 33]. Similarly, nesfatin-1 exhibits cytoprotective effects via free radical scavenging and anti-inflammatory signaling pathways [34-36].\\u003c/p\\u003e\\n\\u003cp\\u003eThe observed inverse correlation between OSI and irisin in the chronic brucellosis group highlights the potential of irisin as a redox-sensitive biomarker. These findings align with data from other inflammatory and metabolic conditions where irisin levels reflect oxidative burden [32, 37, 38]. The decline in nesfatin-1 may also indicate impaired stress adaptation mechanisms during prolonged infection and inflammation [34, 35].\\u003c/p\\u003e\\n\\u003cp\\u003eInterestingly, some conflicting results in the literature\\u0026mdash;such as reports of elevated TAS or reduced TOS in brucellosis\\u0026mdash;may stem from heterogeneity in disease staging, treatment exposure, or population characteristics [25, 29, 30]. Nonetheless, our consistent findings across acute, subacute, and chronic stages underscore the progressive metabolic and oxidative burden imposed by brucellosis.\\u003c/p\\u003e\\n\\u003cp\\u003eThe clinical relevance of these hormonal changes should not be underestimated. Given the central roles of irisin and nesfatin-1 in appetite regulation, glucose homeostasis, and mitochondrial function, their suppression may contribute to fatigue, weight loss, and altered energy metabolism often observed in chronic brucellosis. Moreover, these hormones may serve as adjunctive biomarkers to aid in disease staging or therapeutic monitoring, particularly in resource-limited settings where advanced diagnostic modalities may be lacking.\\u003c/p\\u003e\\n\\u003cp\\u003eFrom a public health perspective, our findings reinforce the need to consider brucellosis not only as an infectious disease but also as a metabolic stressor with potential long-term systemic consequences. Future research should explore the mechanistic links between Brucella-induced oxidative damage, endocrine dysregulation, and nutritional status. Longitudinal studies are warranted to evaluate whether restoring redox balance and hormonal function\\u0026mdash;through pharmacologic, dietary, or lifestyle interventions\\u0026mdash;can improve clinical outcomes in affected individuals.\\u003c/p\\u003e\"},{\"header\":\"Conclusion\",\"content\":\"\\u003cp\\u003eThis study provides compelling evidence that brucellosis exerts a significant systemic impact beyond its infectious etiology, notably altering oxidative stress parameters and key metabolic hormones such as irisin and nesfatin-1. The progressive decrease in these energy-regulating hormones across disease stages, in conjunction with elevated oxidative stress markers, underscores the nutritional-metabolic burden imposed by this foodborne zoonosis. These findings support the integration of hormonal and redox biomarkers into the clinical evaluation of brucellosis and suggest potential utility for disease staging and therapeutic monitoring. From a public health standpoint, improving food safety practices and considering nutritional support strategies may enhance disease control and patient outcomes in endemic regions.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cbr\\u003e\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003ch3\\u003e\\u003cstrong\\u003eFunding Statement\\u003c/strong\\u003e\\u003c/h3\\u003e\\n\\u003cp\\u003eThe authors declare that no specific grants from funding agencies in the public, commercial, or not-for-profit sectors were received for the conduct of this study.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eEthical Approval\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eEthical clearance for this study was obtained from the Firat University Non-Invasive Local Ethics Committee (Decision No: 2017/07-13). All procedures performed were in accordance with the ethical standards of the institutional and national research committees and with the 1964 Helsinki Declaration and its later amendments.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConsent to Participate declaration\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAll participants provided written informed consent prior to inclusion in the study.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eClinical Trial Number\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot Applicable.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflict of Interest\\u003c/strong\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgements\\u003c/strong\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors have no acknowledgements to disclose.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthor Contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eSA, AS, and OO were responsible for conceptualization, data interpretation, and critical revision of the manuscript. FFS and PO contributed to data acquisition, including blood sampling and clinical documentation. All authors reviewed and approved the final version of the manuscript.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eData Availability:\\u003c/strong\\u003e\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n \\u003cli\\u003eYumuk Z, O\\u0026rsquo;Callaghan D. Brucellosis in Turkey\\u0026mdash;an overview. \\u003cem\\u003eInt J Infect Dis\\u003c/em\\u003e. 2012;16(4):228\\u0026ndash;35. doi:10.1016/j.ijid.2011.12.011\\u003c/li\\u003e\\n \\u003cli\\u003eGalińska EM, Zag\\u0026oacute;rski J. Brucellosis in humans\\u0026mdash;etiology, diagnostics, clinical forms. \\u003cem\\u003eAnn Agric Environ Med\\u003c/em\\u003e. 2013;20(2):233\\u0026ndash;8.\\u003c/li\\u003e\\n \\u003cli\\u003eGuler S, Kokoglu OF, Ucmak H, et al. Human brucellosis in Turkey: different clinical presentations. \\u003cem\\u003eJ Infect Dev Ctries\\u003c/em\\u003e. 2014;8(5):581\\u0026ndash;8. doi:10.3855/jidc.3510\\u003c/li\\u003e\\n \\u003cli\\u003eZhang N, Zhou H, Huang DS, Guan P. Brucellosis awareness and knowledge in communities worldwide: a systematic review and meta-analysis of 79 observational studies. \\u003cem\\u003ePLoS Negl Trop Dis\\u003c/em\\u003e. 2019;13(5):0007366. doi:10.1371/journal.pntd.0007366\\u003c/li\\u003e\\n \\u003cli\\u003eMoriy\\u0026oacute;n I, Blasco JM, Letesson JJ, De Massis F, Moreno E. Brucellosis and one health: inherited and future challenges. \\u003cem\\u003eMicroorganisms\\u003c/em\\u003e. 2023;11(8):2070. doi:10.3390/microorganisms11082070\\u003c/li\\u003e\\n \\u003cli\\u003eAlqaseer K, Al-Khafajy AAM, Almkhadhree EAK. Serological and molecular detection of human brucellosis in rural areas in Wasit Province, Iraq. \\u003cem\\u003eArch Razi Inst\\u003c/em\\u003e. 2023;78(1):369\\u0026ndash;78. doi:10.22092/ARI.2022.359002.2352\\u003c/li\\u003e\\n \\u003cli\\u003eSamadi A, Amiri M, Hailat N. The reasons behind long-term endemicity of brucellosis in low and middle-income countries: challenges and future perspectives. \\u003cem\\u003eCurr Microbiol\\u003c/em\\u003e. 2024;81(3):82. doi:10.1007/s00284-023-03605-5\\u003c/li\\u003e\\n \\u003cli\\u003eYagupsky P, Morata P, Colmenero JD. Laboratory diagnosis of human brucellosis. \\u003cem\\u003eClin Microbiol Rev\\u003c/em\\u003e. 2019;33(1):e00073\\u0026ndash;19. doi:10.1128/CMR.00073-19\\u003c/li\\u003e\\n \\u003cli\\u003eSen P, Demirdal T, Nemli SA. Predictive value of inflammation markers in brucellosis. \\u003cem\\u003eArch Iran Med\\u003c/em\\u003e. 2019;22(11):640\\u0026ndash;5.\\u003c/li\\u003e\\n \\u003cli\\u003eKaraagac L, Koruk ST, Koruk I, Aksoy N. Decreasing oxidative stress in response to treatment in patients with brucellosis: could it be used to monitor treatment? \\u003cem\\u003eInt J Infect Dis\\u003c/em\\u003e. 2011;15(5):346\\u0026ndash;9. doi:10.1016/j.ijid.2011.01.009\\u003c/li\\u003e\\n \\u003cli\\u003eDemirpen\\u0026ccedil;e O, Sevim B, Yıldırım M, et al. Serum paraoxonase, TAS, TOS and ceruloplasmin in brucellosis. \\u003cem\\u003eInt J Clin Exp Med\\u003c/em\\u003e. 2014;7(6):1592\\u0026ndash;7.\\u003c/li\\u003e\\n \\u003cli\\u003e\\u0026nbsp;Filomeni G, De Zio D, Cecconi F. Oxidative stress and autophagy: the clash between damage and metabolic needs. \\u003cem\\u003eCell Death Differ\\u003c/em\\u003e. 2015;22(3):377\\u0026ndash;88. doi:10.1038/cdd.2014.150\\u003c/li\\u003e\\n \\u003cli\\u003eTeleanu DM, Niculescu AG, Lungu II, et al. An overview of oxidative stress, neuroinflammation, and neurodegenerative diseases. \\u003cem\\u003eInt J Mol Sci\\u003c/em\\u003e. 2022;23(11):5938. doi:10.3390/ijms23115938\\u003c/li\\u003e\\n \\u003cli\\u003eMelek IM, Erdogan S, Celik S, et al. Evaluation of oxidative stress and inflammation in long-term \\u003cem\\u003eBrucella melitensis\\u003c/em\\u003e infection. \\u003cem\\u003eMol Cell Biochem\\u003c/em\\u003e. 2006;293(1-2):203\\u0026ndash;9. doi:10.1007/s11010-006-9243-2\\u003c/li\\u003e\\n \\u003cli\\u003eEsen R, Aslan M, Kucukoglu ME, et al. Serum paraoxonase activity, total thiols levels, and oxidative status in patients with acute brucellosis. \\u003cem\\u003eWien Klin Wochenschr\\u003c/em\\u003e. 2015;127(11-12):427\\u0026ndash;33. doi:10.1007/s00508-015-0720-z\\u003c/li\\u003e\\n \\u003cli\\u003eBostr\\u0026ouml;m P, Wu J, Jedrychowski MP, et al. A PGC1-\\u0026alpha;-dependent myokine that drives brown-fat-like development of white fat and thermogenesis. \\u003cem\\u003eNature\\u003c/em\\u003e. 2012;481(7382):463\\u0026ndash;8. doi:10.1038/nature10777\\u003c/li\\u003e\\n \\u003cli\\u003eOzcelik O, Algul S, Yilmaz B. Nesfatin-1 and irisin levels in response to the soccer matches performed in morning, afternoon and at night in young trained male subjects. \\u003cem\\u003eCell Mol Biol (Noisy-le-grand)\\u003c/em\\u003e. 2018;64(13):130\\u0026ndash;3.\\u003c/li\\u003e\\n \\u003cli\\u003eOh-I S, Shimizu H, Satoh T, et al. Identification of nesfatin-1 as a satiety molecule in the hypothalamus. \\u003cem\\u003eNature\\u003c/em\\u003e. 2006;443(7112):709\\u0026ndash;12. doi:10.1038/nature05162\\u003c/li\\u003e\\n \\u003cli\\u003eAskari H, Rajani SF, Poorebrahim M, et al. A glance at the therapeutic potential of irisin against diseases involving inflammation, oxidative stress, and apoptosis: an introductory review. \\u003cem\\u003ePharmacol Res\\u003c/em\\u003e. 2018;129:44\\u0026ndash;55. doi:10.1016/j.phrs.2018.01.012\\u003c/li\\u003e\\n \\u003cli\\u003eAyada C, Toru \\u0026Uuml;, Gen\\u0026ccedil; O, et al. Balanced oxidative status by nesfatin-1 in intestinal ischemia-reperfusion. \\u003cem\\u003eInt J Clin Exp Med\\u003c/em\\u003e. 2015;8(3):3318\\u0026ndash;24.\\u003c/li\\u003e\\n \\u003cli\\u003eZhang Z, Zhang X, Chen X, et al. Clinical features of human brucellosis and risk factors for focal complications. \\u003cem\\u003eInt J Gen Med\\u003c/em\\u003e. 2022;15:7373\\u0026ndash;82. doi:10.2147/IJGM.S380328\\u003c/li\\u003e\\n \\u003cli\\u003eErtek M, Yazgi H, Ozkurt Z, et al. Comparison of the diagnostic value of the standard tube agglutination test and the ELISA IgG and IgM in patients with brucellosis. \\u003cem\\u003eTurk J Med Sci\\u003c/em\\u003e. 2006;36(3):159\\u0026ndash;63.\\u003c/li\\u003e\\n \\u003cli\\u003eYohannes M, Gill JP, Ghatak S, et al. Comparative evaluation of the Rose Bengal plate test, standard tube agglutination test and complement fixation test. \\u003cem\\u003eRev Sci Tech\\u003c/em\\u003e. 2012;31(3):979\\u0026ndash;84. doi:10.20506/rst.31.3.2175\\u003c/li\\u003e\\n \\u003cli\\u003eAndriopoulos P, Tsironi M, Deftereos S, et al. Acute brucellosis: presentation, diagnosis, and treatment of 144 cases. \\u003cem\\u003eInt J Infect Dis\\u003c/em\\u003e. 2007;11(1):52\\u0026ndash;7. doi:10.1016/j.ijid.2005.10.011\\u003c/li\\u003e\\n \\u003cli\\u003eKaraagac L, Koruk ST, Koruk I, Aksoy N. Decreasing oxidative stress in response to treatment in patients with brucellosis: could it be used to monitor treatment? \\u003cem\\u003eInt J Infect Dis\\u003c/em\\u003e. 2011;15(5):346\\u0026ndash;9. doi:10.1016/j.ijid.2011.01.009\\u003c/li\\u003e\\n \\u003cli\\u003eMoreno E, Blasco JM, Moriy\\u0026oacute;n I. Facing the human and animal brucellosis conundrums: the forgotten lessons. \\u003cem\\u003eMicroorganisms\\u003c/em\\u003e. 2022;10(5):942. doi:10.3390/microorganisms10050942\\u003c/li\\u003e\\n \\u003cli\\u003ePerin G, Bottari NB, Silva AD, et al. Cholinesterase\\u0026apos;s activities of infected mice by \\u003cem\\u003eBrucella ovis\\u003c/em\\u003e. \\u003cem\\u003eMicrob Pathog\\u003c/em\\u003e. 2019;132:137\\u0026ndash;140. doi:10.1016/j.micpath.2019.04.036\\u003c/li\\u003e\\n \\u003cli\\u003eBaldi PC, Giambartolomei GH. Immunopathology of Brucella infection. \\u003cem\\u003eRecent Pat Antiinfect Drug Discov\\u003c/em\\u003e. 2013;8(1):18\\u0026ndash;26. doi:10.2174/1574891X11308010005\\u003c/li\\u003e\\n \\u003cli\\u003eSerefhanoglu K, Taskin A, Turan H, et al. Evaluation of oxidative status in patients with brucellosis. \\u003cem\\u003eBraz J Infect Dis\\u003c/em\\u003e. 2009;13(4):249\\u0026ndash;51. doi:10.1590/s1413-86702009000400001\\u003c/li\\u003e\\n \\u003cli\\u003eUsta M, Aras Z, Tas A. Oxidant and antioxidant parameters in patients with \\u003cem\\u003eBrucella canis\\u003c/em\\u003e. \\u003cem\\u003eClin Biochem\\u003c/em\\u003e. 2012;45(4\\u0026ndash;5):366\\u0026ndash;7. doi:10.1016/j.clinbiochem.2011.12.028\\u003c/li\\u003e\\n \\u003cli\\u003eBarzilai A, Yamamoto K. DNA damage responses to oxidative stress. \\u003cem\\u003eDNA Repair (Amst)\\u003c/em\\u003e. 2004;3(8\\u0026ndash;9):1109\\u0026ndash;15. doi:10.1016/j.dnarep.2004.03.002\\u003c/li\\u003e\\n \\u003cli\\u003eSlate-Romano JJ, Yano N, Zhao TC. Irisin reduces inflammatory signaling pathways in inflammation-mediated metabolic syndrome. \\u003cem\\u003eMol Cell Endocrinol\\u003c/em\\u003e. 2022;552:111676. doi:10.1016/j.mce.2022.111676\\u003c/li\\u003e\\n \\u003cli\\u003eAlgul S, Senol FF, Ugras S, et al. Investigation of irisin level in brucella patients. \\u003cem\\u003eEast J Med\\u003c/em\\u003e. 2021;26(2):135\\u0026ndash;8. doi:10.5505/ejm.2021.70431\\u003c/li\\u003e\\n \\u003cli\\u003eXu Y, Chen F. Antioxidant, anti-inflammatory and anti-apoptotic activities of nesfatin-1: a review. \\u003cem\\u003eJ Inflamm Res\\u003c/em\\u003e. 2020;13:607\\u0026ndash;17. doi:10.2147/JIR.S273446\\u003c/li\\u003e\\n \\u003cli\\u003eCaylak E. IFN-\\u0026gamma;, IL-10, ghrelin and nesfatin-1 levels in acute brucella patients. \\u003cem\\u003eAnn Clin Anal Med\\u003c/em\\u003e. 2022;13(7):716\\u0026ndash;20. doi:10.4328/ACAM.21061\\u003c/li\\u003e\\n \\u003cli\\u003eAlgul S, Ozcelik O. G\\u0026uuml;n\\u0026uuml;n farklı zamanlarında yapılan futbol ma\\u0026ccedil;larında oksidan-antioksidan dengenin incelenmesi. \\u003cem\\u003eGenel Tıp Derg\\u003c/em\\u003e. 2017;27(4):129\\u0026ndash;35.\\u003c/li\\u003e\\n \\u003cli\\u003eCanpolat Erkan RE, Tekin R. Investigation of new inflammatory biomarkers in patients with brucella. \\u003cem\\u003ePLoS One\\u003c/em\\u003e. 2024;19(2):0297550. doi:10.1371/journal.pone.0297550\\u003c/li\\u003e\\n \\u003cli\\u003eMazur-Bialy AI, Kozlowska K, Pochec E, et al. Myokine irisin-induced protection against oxidative stress in vitro. \\u003cem\\u003eJ Physiol Pharmacol\\u003c/em\\u003e. 2018;69(1):117\\u0026ndash;25.\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"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\":\"info@researchsquare.com\",\"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\":\"Brucellosis, foodborne zoonosis, irisin, nesfatin-1, oxidative stress, metabolic dysregulation, disease staging, ELISA\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-7884325/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-7884325/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003ch2\\u003eBackground\\u003c/h2\\u003e\\u003cp\\u003eBrucellosis is a globally prevalent foodborne zoonotic disease transmitted primarily through the ingestion of unpasteurized dairy products or undercooked meat. While its infectious etiology is well recognized, the metabolic and oxidative stress-related alterations associated with the disease remain insufficiently characterized.\\u003c/p\\u003e\\u003ch2\\u003eObjectives\\u003c/h2\\u003e\\u003cp\\u003eThis study aimed to investigate stage-specific changes in serum irisin and nesfatin-1 levels, alongside markers of oxidative stress, in patients with acute, subacute, and chronic brucellosis.\\u003c/p\\u003e\\u003ch2\\u003eMethods\\u003c/h2\\u003e\\u003cp\\u003eA total of 90 patients diagnosed with brucellosis at different clinical stages and 30 healthy controls were included. Serum levels of irisin, nesfatin-1, total oxidant status (TOS), total antioxidant status (TAS), and the oxidative stress index (OSI; TOS/TAS ratio) were determined using enzyme-linked immunosorbent assay (ELISA). Intergroup comparisons were performed using one-way ANOVA and paired t-tests.\\u003c/p\\u003e\\u003ch2\\u003eResults\\u003c/h2\\u003e\\u003cp\\u003eSerum irisin and nesfatin-1 levels were significantly reduced in brucellosis patients compared to controls, with variations dependent on disease stage (p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05). Concurrently, a significant increase in TOS and OSI values and a decrease in TAS levels were observed across all stages of brucellosis (p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05).\\u003c/p\\u003e\\u003ch2\\u003eConclusions\\u003c/h2\\u003e\\u003cp\\u003eBrucellosis induces distinct alterations in energy-regulating hormones and redox balance, extending beyond its infectious profile. Reduced levels of irisin and nesfatin-1, in parallel with elevated oxidative stress, suggest potential roles for these hormones as biomarkers in disease staging and metabolic monitoring. These findings underscore the need to incorporate hormonal and oxidative profiling into the clinical evaluation of brucellosis. To our knowledge, this is the first study to evaluate irisin and nesfatin-1 responses across different clinical stages of brucellosis.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Stage-Specific Changes in Irisin, Nesfatin-1, and Oxidative Stress Markers in Brucellosis\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2025-11-18 12:32:44\",\"doi\":\"10.21203/rs.3.rs-7884325/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"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\":\"5508c8df-f240-4232-a432-003d2e59d856\",\"owner\":[],\"postedDate\":\"November 18th, 2025\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2025-12-17T07:39:30+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2025-11-18 12:32:44\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-7884325\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-7884325\",\"identity\":\"rs-7884325\",\"version\":[\"v1\"]},\"buildId\":\"8U1c8b4HqxoKbykW_rLl7\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}