20.500.12556/DiRROS-17760 Exploration of macromolecular phenotype of human skeletal muscle in diabetes using infrared spectroscopy Introduction: The global burden of diabetes mellitus is escalating, and more efficient investigative strategies are needed for a deeper understanding of underlying pathophysiological mechanisms. The crucial role of skeletal muscle in carbohydrate and lipid metabolism makes it one of the most susceptible tissues to diabetes-related metabolic disorders. In tissue studies, conventional histochemical methods have several technical limitations and have been shown to inadequately characterise the biomolecular phenotype of skeletal muscle to provide a holistic view of the pathologically altered proportions of macromolecular constituents. Materials and methods: In this pilot study, we examined the composition of five different human skeletal muscles from male donors diagnosed with type 2 diabetes and non-diabetic controls. We analysed the lipid, glycogen, and collagen content in the muscles in a traditional manner with histochemical assays using different staining techniques. This served as a reference for comparison with the unconventional analysis of tissue composition using Fourier-transform infrared spectroscopy as an alternative methodological approach. Results: A thorough chemometric post-processing of the infrared spectra using a multi-stage spectral decomposition allowed the simultaneous identification of various compositional details from a vibrational spectrum measured in a single experiment. We obtained multifaceted information about the proportions of the different macromolecular constituents of skeletal muscle, which even allowed us to distinguish protein constituents with different structural properties. The most important methodological steps for a comprehensive insight into muscle composition have thus been set and parameters identified that can be used for the comparison between healthy and diabetic muscles. Conclusion: We have established a methodological framework based on vibrational spectroscopy for the detailed macromolecular analysis of human skeletal muscle that can effectively complement or may even serve as an alternative to histochemical assays. As this is a pilot study with relatively small sample sets, we remain cautious at this stage in drawing definitive conclusions about diabetes-related changes in skeletal muscle composition. However, the main focus and contribution of our work has been to provide an alternative, simple and efficient approach for this purpose. We are confident that we have achieved this goal and have brought our methodology to a level from which it can be successfully transferred to a large-scale study that allows the effects of diabetes on skeletal muscle composition and the interrelationships between the macromolecular tissue alterations due to diabetes to be investigated. diabetes mellitus skeletal muscle metabolism macromulecular composition infrared spectroscopy multivariate analysis histochemical assays sladkorna bolezen skeletne mišice metabolizem makromolekulska sestava infrardeča spektroskopija multivariatna analiza histokemični testi true false true Frontiers Research Foundation Frontiers in endocrinology Angleški jezik Slovenski jezik © 2023 Zupančič, Ugwoke, Abdelmonaem, Alibegović, Cvetko, Grdadolnik, Šerbec and Umek. Neznano 2024-01-11 12:04:24 2024-01-11 12:44:39 2024-01-12 03:39:10 0000-00-00 00:00:00 2023 0 0 Soavtorji: Chiedozie Kenneth Ugwoke, Mohamed Elwy Abdelhamed Abdelmonaem, Armin Alibegović, Erika Cvetko, Jože Grdadolnik, Anja Šerbec, Nejc Umek; Nasl. z nasl. zaslona; Opis vira z dne 28. 12. 2023; str. 1-17 Vol. 14 2023 0000-00-00 Zaloznikova Objavljeno apc 0000-00-00 0000-00-00 2023-01-01 616.4 1664-2392 10.3389/fendo.2023.1308373 178901251 https://www.frontiersin.org/articles/10.3389/fendo.2023.1308373/full 1 3102ae7c-b071-11ee-b0d4-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=24110 RAZ_Zupancic_Barbara_2023.pdf RAZ_Zupancic_Barbara_2023.pdf 1 ADCF204E76693E65604F37D80132B35B 775128b31ac4f3802d24d1f98626ebd6ff897bf89d4a257715360e9b18fce324 9f987b13-b076-11ee-b0d4-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=24111 Kemijski inštitut 0 0 0