Effect of Intranasally Administered Insulin on Metabolic Parameters and Inflammation Factors in Control and Diabetic Rats under Conditions of Cerebral Ischemia and Reperfusion

Cover Page

Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription Access

Abstract

The search for natural biologically active substances that have a neuroprotective effect on cerebral ischemia-reperfusion is one of the urgent problems of modern neuroscience and medicine. Intranasally administered insulin (IAI) has a pronounced restorative effect on various neurodegenerative diseases, but the mechanisms of its action and therapeutic effects in cerebral ischemia have not been studied well, including in type 2 diabetes mellitus (DM2), which increases the risk of cerebrovascular dysfunction. The aim of the work was to study the effect of IAI on metabolic parameters and inflammatory factors in male rats with DM2 subjected to the two-vessel ischemia and prolonged forebrain reperfusion, in comparison with non-diabetic animals. A long-term high-fat diet with an injection of a low dose of streptozotocin (25 mg/kg) to rats was used to induce DM2, and a model of the global forebrain two-vessel ischemia induced by occlusion of both common carotids with prolonged reperfusion (IR) for 7 days was used to study cerebral ischemia. Two hours after the end of ischemia, rats were treated with IAI at a dose of 0.5 or 2.0 IU/rat, after which the drug was administered in the same doses daily for 7 subsequent days. It was found that IAI prevents body weight loss in both nondiabetic and diabetic rats that underwent IR, and also increases the total cholesterol level and the proportion of epididymal fat in rats without DM2 after IR. In DM2 rats that underwent IR, IAI in the explored doses reduces the level of postprandial glucose and insulin content in the blood, which indicates an improvement of glucose tolerance, and also reduces the levels of inflammatory factors in the blood – C-reactive protein (at a dose of 0.5 IU/rat/day) and tumor necrosis factor-α (in a dose of 2 IU/rat/day), which reveals its anti-inflammatory potential. Thus, the course treatment with IAI after induction of cerebral ischemia followed by reperfusion leads to an improvement of metabolic parameters and weakens inflammatory reactions in rats with DM2, which may be in demand in the correction of ischemic stroke in patients with DM2.

Full Text

Restricted Access

About the authors

I. I. Zorina

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Author for correspondence.
Email: zorina.inna.spb@gmail.com
Russian Federation, Saint-Petersburg

A. S. Pechalnova

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: zorina.inna.spb@gmail.com
Russian Federation, Saint-Petersburg

E. E. Chernenko

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: zorina.inna.spb@gmail.com
Russian Federation, Saint-Petersburg

K. V. Derkach

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: zorina.inna.spb@gmail.com
Russian Federation, Saint-Petersburg

A. O. Shpakov

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: zorina.inna.spb@gmail.com
Russian Federation, Saint-Petersburg

References

  1. Какорина ЕП, Никитина СЮ (2019) Особенности структуры смертности в Российской Федерации. Пробл соц защиты населения в истории медицины 27: 822–826. [Kakorina EP, Nikitina SY (2019) Features of the structure of mortality in Russian Federation. Probl Sots Gig Zdravookhran Istor Med 27: 822–826. (In Russ)]. https://doi.org/10.32687/0869–866X-2019–27–5–822–826
  2. Vorotnikov AV, Stafeev IS, Menshikov MY, Shestakova MV, Parfyonova YV (2019) Latent Inflammation and Defect in Adipocyte Renewal as a Mechanism of Obesity-Associated Insulin Resistance. Biochemistry (Mosc) 84: 1329–1345. https://doi.org/10.1134/S0006297919110099
  3. Kim H, Lee H (2023) Risk of Stroke and Cardiovascular Disease According to Diabetes Mellitus Status. West J Nurs Res 45: 520–527. https://doi.org/10.1177/01939459231158212
  4. Tsalamandris S, Antonopoulos AS, Oikonomou E, Papamikroulis GA, Vogiatzi G, Papaioannou S, Deftereos S, Tousoulis D (2019) The Role of Inflammation in Diabetes: Current Concepts and Future Perspectives. Eur Cardiol 14: 50–59. https://doi.org/10.15420/ecr.2018.33.1
  5. Акжигитов РГ (ред) (2021) Федеральные клинические рекомендации по ведению больных с ишемическим инсультом и транзиторной ишемической атакой у взрослых. М. Атомиздат. [Akzhigitov RG (ed) (2021) Federal recommendations for the management of patients with ischemic stroke and transient ischemic attack in adults. M. Atomizdat. (In Russ)].
  6. Powers WJ, Rabinstein AA, Ackerson T, Adeoye OM, Bambakidis NC, Becker K, Biller J, Brown M, Demaerschalk BM, Hoh B, Jauch EC, Kidwell CS, Leslie-Mazwi TM, Ovbiagele B, Scott PA, Sheth KN, Southerland AM, Summers DV, Tirschwell DL (2019) Guidelines for the Early Management of Patients With Acute Ischemic Stroke: 2019 Update to the 2018 Guidelines for the Early Management of Acute Ischemic Stroke: A Guideline for Healthcare Professionals From the American Heart Association/American Stroke Association. Stroke 50: e344–e418. https://doi.org/10.1161/STR.0000000000000211
  7. Hallschmid M (2021) Intranasal insulin. J Neuroendocrinol 33: e12934. https://doi.org/10.1111/jne.12934
  8. Zorina II, Avrova NF, Zakharova IO, Shpakov AO (2023) Prospects for the Use of Intranasally Administered Insulin and Insulin-Like Growth Factor-1 in Cerebral Ischemia. Biochemistry (Mosc) 88: 374–391. https://doi.org/10.1134/S0006297923030070
  9. White MF, Kahn CR (2021) Insulin action at a molecular level – 100 years of progress. Mol Metab 52: 101304. https://doi.org/10.1016/j.molmet.2021.101304
  10. Novak V, Mantzoros CS, Novak P, McGlinchey R, Dai W, Lioutas V, Buss S, Fortier CB, Khan F, Aponte Becerra L, Ngo LH (2022) MemAID: Memory advancement with intranasal insulin vs. placebo in type 2 diabetes and control participants: a randomized clinical trial. J Neurol 269: 4817–4835. https://doi.org/10.1007/s00415–022–11119–6
  11. Shpakov AO, Zorina II, Derkach KV (2022) Hot Spots for the Use of Intranasal Insulin: Cerebral Ischemia, Brain Injury, Diabetes Mellitus, Endocrine Disorders and Postoperative Delirium. Int J Mol Sci 24: 3278. https://doi.org/10.3390/ijms24043278
  12. Picone P, Sabatino MA, Ditta LA, Amato A, San Biagio PL, Mulè F, Giacomazza D, Dispenza C, Di Carlo M (2018) Nose-to-brain delivery of insulin enhanced by a nanogel carrier. J Control Release 270: 23–36. https://doi.org/10.1016/j.jconrel.2017.11.040
  13. Fan LW, Carter K, Bhatt A, Pang Y (2019) Rapid transport of insulin to the brain following intranasal administration in rats. Neural Regen Res 14: 1046–1051. https://doi.org/10.4103/1673–5374.250624
  14. Nedelcovych MT, Gadiano AJ, Wu Y, Manning AA, Thomas AG, Khuder SS, Yoo SW, Xu J, McArthur JC, Haughey NJ, Volsky DJ, Rais R, Slusher BS (2018) Pharmacokinetics of Intranasal versus Subcutaneous Insulin in the Mouse. ACS Chem Neurosci 9: 809–816. https://doi.org/10.1021/acschemneuro.7b00434
  15. Xu LB, Huang HD, Zhao M, Zhu GC, Xu Z (2021) Intranasal Insulin Treatment Attenuates Metabolic Distress and Early Brain Injury After Subarachnoid Hemorrhage in Mice. Neurocrit Care 2021 34: 154–166. https://doi.org/10.1007/s12028–020–01011–4
  16. Zhu Y, Huang Y, Yang J, Tu R, Zhang X, He WW, Hou CY, Wang XM, Yu JM, Jiang GH (2022) Intranasal insulin ameliorates neurological impairment after intracerebral hemorrhage in mice. Neural Regen Res 17: 210–216. https://doi.org/10.4103/1673–5374.314320
  17. Zorina II, Zakharova IO, Bayunova LV, Avrova NF (2018) Insulin Administration Prevents Accumulation of Conjugated Dienes and Trienes and Inactivation of Na+, K+-ATPase in the Rat Cerebral Cortex during Two-Vessel Forebrain Ischemia and Reperfusion. J Evol Biochem Phys 54: 246–249. https://doi.org/10.1134/S0022093018030109
  18. Zorina II, Galkina OV, Bayunova LV, Zakharova IO (2019) Effect of Insulin on Lipid Peroxidation and Glutathione Levels in a Two-Vessel Occlusion Model of Rat Forebrain Ischemia Followed by Reperfusion. J Evol Biochem Phys 55: 333–335. https://doi.org/10.1134/S0022093019040094
  19. Zakharova IO, Bayunova LV, Zorina II, Sokolova TV, Shpakov AO, Avrova NF (2021) Insulin and α-Tocopherol Enhance the Protective Effect of Each Other on Brain Cortical Neurons under Oxidative Stress Conditions and in Rat Two-Vessel Forebrain Ischemia/Reperfusion Injury. Int J Mol Sci 22: 11768. https://doi.org/10.3390/ijms222111768
  20. Захарова ИО, Баюнова ЛВ, Зорина ИИ, Шпаков АО, Аврова НФ (2022) Инсулин и ганглиозиды мозга предотвращают нарушения метаболизма, вызванные активацией свободнорадикальных реакций, при двухсосудистой ишемии переднего мозга крыс и реперфузии. Рос физиол журн им ИМ Сеченова 108(2): 262–278. [Zakharova IO, Bayunova LV, Zorina II, Shpakov AO, Avrova NF (2022) Insulin and Brain Gangliosides Prevent Metabolic Disorders Caused by Activation of Free Radical Reactions after Two-Vessel Ischemia–Reperfusion Injury to the Rat Forebrain. Rus Phiziol J 108(2): 262–278. (In Russ)]. https://doi.org/10.31857/S086981392202011X
  21. Smith CJ, Sims S-K, Nguyen S, Williams A, McLeod T, Sims-Robinson C (2023) Intranasal insulin helps overcome brain insulin deficiency and improves survival and post-stroke cognitive impairment in male mice. J Neurosci Res 101: 1757–1769. https://doi.org/10.1002/jnr.25237
  22. Bakhtyukov AA, Derkach KV, Sorokoumov VN, Stepochkina AM, Romanova IV, Morina IY, Zakharova IO, Bayunova LV, Shpakov AO (2021) The Effects of Separate and Combined Treatment of Male Rats with Type 2 Diabetes with Metformin and Orthosteric and Allosteric Agonists of Luteinizing Hormone Receptor on Steroidogenesis and Spermatogenesis. Int J Mol Sci 23: 198. https://doi.org/10.3390/ijms23010198
  23. Raval AP, Liu C, Hu BR (2009) Rat Model of Global Cerebral Ischemia: The Two-Vessel Occlusion (2VO) Model of Forebrain Ischemia. In: Chen J, Xu ZC, Xu XM, Zhang JH (eds) Animal Models of Acute Neurological Injuries. Springer Protocols Handbooks. Hum Press. https://doi.org/10.1007/978–1–60327–185–1_7
  24. Qin C, Yang S, Chu YH, Zhang H, Pang XW, Chen L, Zhou LQ, Chen M, Tian DS, Wang W (2022) Signaling pathways involved in ischemic stroke: molecular mechanisms and therapeutic interventions. Signal Transduct Target Ther 7: 215. https://doi.org/10.1038/s41392–022–01064–1
  25. Khoshnam SE, Winlow W, Farzaneh M, Farbood Y, Moghaddam HF (2017) Pathogenic mechanisms following ischemic stroke. Neurol Sci 38: 1167–1186. https://doi.org/10.1007/s10072–017–2938–1
  26. Sidorov EV, Rout M, Xu C, Jordan L, Fields E, Apple B, Smith K, Gordon D, Chainakul J, Sanghera DK (2023) Difference in acute and chronic stage ischemic stroke metabolic markers with controls. J Stroke Cerebrovasc Dis 32: 107211. https://doi.org/10.1016/j.jstrokecerebrovasdis.2023.107211
  27. Haley MJ, White CS, Roberts D, O'Toole K, Cunningham CJ, Rivers-Auty J, O'Boyle C, Lane C, Heaney O, Allan SM, Lawrence CB (2020) Stroke Induces Prolonged Changes in Lipid Metabolism, the Liver and Body Composition in Mice. Transl Stroke Res 11: 837–850. https://doi.org/10.1007/s12975–019–00763–2
  28. Scherbakov N, Pietrock C, Sandek A, Ebner N, Valentova M, Springer J, Schefold JC, von Haehling S, Anker SD, Norman K, Haeusler KG, Doehner W (2019) Body weight changes and incidence of cachexia after stroke. J Cachexia Sarcopenia Muscle 10: 611–620. https://doi.org/10.1002/jcsm.12400
  29. Cai L, Geng X, Hussain M, Liu Z, Gao Z, Liu S, Du H, Ji X, Ding Y (2015) Weight loss: indication of brain damage and effect of combined normobaric oxygen and ethanol therapy after stroke. Neurol Res 37: 441–446. https://doi.org/10.1179/1743132815Y.0000000033
  30. English C, McLennan H, Thoirs K, Coates A, Bernhardt J (2010) Loss of skeletal muscle mass after stroke: a systematic review. Int J Stroke 5: 395–402. https://doi.org/10.1111/j.1747–4949.2010.00467
  31. Gungor L, Arsava EM, Guler A, Togay Isikay C, Aykac O, Batur Caglayan HZ, Kozak HH, Aydingoz U, Topcuoglu MA (2023) MASS investigators. Determinants of in-hospital muscle loss in acute ischemic stroke – Results of the Muscle Assessment in Stroke Study (MASS). Clin Nutr 42: 431–439. https://doi.org/10.1016/j.clnu.2023.01.017
  32. Akhtar N, Singh R, Kamran S, Joseph S, Morgan D, Uy RT, Treit S, Shuaib A (2024) Association between serum triglycerides and stroke type, severity, and prognosis. Analysis in 6558 patients. BMC Neurol 24: 88. https://doi.org/10.1186/s12883–024–03572–9
  33. Ding PF, Zhang HS, Wang J, Gao YY, Mao JN, Hang CH, Li W (2022) Insulin resistance in ischemic stroke: Mechanisms and therapeutic approaches. Front Endocrinol (Lausanne) 13: 1092431. https://doi.org/ 10.3389/fendo.2022.1092431
  34. Grizard J, Dardevet D, Balage M, Larbaud D, Sinaud S, Savary I, Grzelkowska K, Rochon C, Tauveron I, Obled C (1999) Insulin action on skeletal muscle protein metabolism during catabolic states. Reprod Nutr Dev 39: 61–74. https://doi.org/10.1051/rnd:19990104
  35. Ho-Palma AC, Toro P, Rotondo F, Romero MDM, Alemany M, Remesar X, Fernández-López JA (2019) Insulin Controls Triacylglycerol Synthesis through Control of Glycerol Metabolism and Despite Increased Lipogenesis. Nutrients 11: 513. https://doi.org/10.3390/nu11030513
  36. Katsiki N, Mikhailidis DP, Gotzamani-Psarrakou A, Didangelos TP, Yovos JG, Karamitsos DT (2011) Effects of improving glycemic control with insulin on leptin, adiponectin, ghrelin and neuropeptidey levels in patients with type 2 diabetes mellitus: a pilot study. Open Cardiovasc Med J 5: 136–147. https://doi.org/10.2174/1874192401105010136
  37. Hallschmid M, Higgs S, Thienel M, Ott V, Lehnert H (2012) Postprandial administration of intranasal insulin intensifies satiety and reduces intake of palatable snacks in women. Diabetes 61: 782–789. https://doi.org/10.2337/db11–1390
  38. Scherer T, Sakamoto K, Buettner C (2021) Brain insulin signalling in metabolic homeostasis and disease. Nat Rev Endocrinol 17: 468–483. https://doi.org/10.1038/s41574–021–00498-x
  39. Xiao X, Luo Y, Peng D (2022) Updated Understanding of the Crosstalk Between Glucose/Insulin and Cholesterol Metabolism. Front Cardiovasc Med 9: 879355. https://doi.org/10.3389/fcvm.2022.879355
  40. Brabazon F, Wilson CM, Jaiswal S, Reed J, Frey WH, Nd, Byrnes KR (2017) Intranasal insulin treatment of an experimental model of moderate traumatic brain injury. J Cereb Blood Flow Metab 37: 3203–3218. https://doi.org/10.1177/0271678X16685106
  41. Xu LB, Huang HD, Zhao M, Zhu GC, Xu Z (2021) Intranasal Insulin Treatment Attenuates Metabolic Distress and Early Brain Injury After Subarachnoid Hemorrhage inтMice. Neurocrit Care 34: 154–166. https://doi.org/10.1007/s12028–020–01011–4
  42. Xue Y, Zeng X, Tu WJ, Zhao J (2022) Tumor Necrosis Factor-α: The Next Marker of Stroke. Dis Markers 2022: 2395269. https://doi.org/10.1155/2022/2395269
  43. Collino M, Aragno M, Castiglia S, Tomasinelli C, Thiemermann C, Boccuzzi G, Fantozzi R (2009) Insulin reduces cerebral ischemia/reperfusion injury in the hippocampus of diabetic rats: a role for glycogen synthase kinase-3beta. Diabetes 58: 235–242. https://doi.org/10.2337/db08–0691
  44. Lioutas VA, Novak V (2016) Intranasal insulin neuroprotection in ischemic stroke. Neural Regen Res 11: 400–401. https://doi.org/10.4103/1673–5374.179040

Supplementary files

Supplementary Files
Action
1. JATS XML
Download
2. Fig. 1. Body weight loss in rats with and without DM2, who underwent double-vascular ischemia and reperfusion, 3 (a) and 7 (b) days after the start of reperfusion and the effect of intranasally administered insulin on it. The loss of body weight in % of total body weight on the 3rd (a) and 7th (b) days of reperfusion is presented. The designation of the groups: C-SO – falsely operated rats without DM2; C–IR – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion; C–IR-0.5 – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/ day; DM2-SO – falsely operated rats with DM2; DM2-IR – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion; DM2-IR-0.5 – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/day; DM2-IR-2 – rats with DM2, who underwent bicascular forebrain ischemia and reperfusion and received IVI at a dose of 2 IU/rat/day. The data is presented in the form of M ± SEM (n = 6 in

Download (197KB)
Indexing metadata
3. Fig. 2. The proportion of visceral (a) and epididymal (b) fat in rats undergoing forebrain ischemia and 7-day reperfusion, with and without DM2, and the effect of intranasally administered insulin on them. The proportion of adipose tissue in % of total body weight is presented. The designation of the groups: C-SO – falsely operated rats without DM2; C–IR – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion; C–IR-0.5 – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/ day; DM2-SO – falsely operated rats with DM2; DM2-IR – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion; DM2-IR-0.5 – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/day; DM2-IR-2 – rats with DM2, who underwent bicascular forebrain ischemia and reperfusion and received IVI at a dose of 2 IU/rat/day. The data are presented in the form of M ± SEM (n = 6 in all groups).

Download (256KB)
Indexing metadata
4. Fig. 3. Postprandial glucose levels in the blood of rats with and without DM2 who underwent forebrain ischemia and reperfusion, and the effect of intranasally administered insulin on them on the 3rd and 7th days of reperfusion. The designation of the groups: C-SO – falsely operated rats without DM2; C–IR – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion; C–IR-0.5 – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/ day; DM2-SO – falsely operated rats with DM2; DM2-IR – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion; DM2-IR-0.5 – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/day; DM2-IR-2 – rats with DM2, who underwent bicascular forebrain ischemia and reperfusion and received IVI at a dose of 2 IU/rat/day.a – differences are significant compared with C-SO at p < 0.05;b – differences are significant compared with C–IR at p < 0.05;c – differences are significant

Download (219KB)
Indexing metadata
5. Fig. 4. The level of postprandial insulin in the blood of rats with and without DM2 who underwent forebrain ischemia and reperfusion for 7 days, and the effect of intranasally administered insulin on it. The designation of the groups: C-SO – falsely operated rats without DM2; C–IR – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion; C–IR-0.5 – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/ day; DM2-SO – falsely operated rats with DM2; DM2-IR – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion; DM2-IR-0.5 – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/day; DM2-IR-2 – rats with DM2, who underwent bicascular forebrain ischemia and reperfusion and received IVI at a dose of 2 IU/rat/day. The data are presented in the form of M ± SEM (n = 6 in all groups).

Download (170KB)
Indexing metadata
6. Fig. 5. Levels of total cholesterol (a) and triglycerides (b) in the blood of rats with and without DM2 who underwent double-vascular ischemia reperfusion for 7 days, and the effect of intranasally administered insulin on them. The designation of the groups: C-SO – falsely operated rats without DM2; C–IR – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion; C–IR-0.5 – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/ day; DM2-SO – falsely operated rats with DM2; DM2-IR – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion; DM2-IR-0.5 – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/day; DM2-IR-2 – rats with DM2, who underwent bicascular forebrain ischemia and reperfusion and received IVI at a dose of 2 IU/rat/day. The data are presented in the form of M ± SEM (n = 6 in all groups).

Download (246KB)
Indexing metadata
7. Fig. 6. Levels of TNFa (a) and C-reactive protein (CRP) (b) in the blood of rats with and without DM2, who underwent dvuvascular ischemia, on the 3rd day of reperfusion, and the effect of intranasally administered insulin on them. The designation of the groups: C-SO – falsely operated rats without DM2; C–IR – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion; C–IR-0.5 – rats without DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/ day; DM2-SO – falsely operated rats with DM2; DM2-IR – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion; DM2-IR-0.5 – rats with DM2 who underwent bicavascular forebrain ischemia and reperfusion and received IVI at a dose of 0.5 IU/rat/day; DM2-IR-2 – rats with DM2, who underwent bicascular forebrain ischemia and reperfusion and received IVI at a dose of 2 IU/rat/day. The data are presented in the form of M ± SEM (n = 6 in all groups).

Download (265KB)
Indexing metadata

Copyright (c) 2024 Russian Academy of Sciences