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Protective role of mitochondrial uncoupling proteins against age-related oxidative stress in type 2 diabetes mellitus
Maša Čater, Lidija Križančić Bombek, 2022, review article

Abstract: The accumulation of oxidative damage to DNA and other biomolecules plays an important role in the etiology of aging and age-related diseases such as type 2 diabetes mellitus (T2D), atherosclerosis, and neurodegenerative disorders. Mitochondrial DNA (mtDNA) is especially sensitive to oxidative stress. Mitochondrial dysfunction resulting from the accumulation of mtDNA damage impairs normal cellular function and leads to a bioenergetic crisis that accelerates aging and associated diseases. Age-related mitochondrial dysfunction decreases ATP production, which directly affects insulin secretion by pancreatic beta cells and triggers the gradual development of the chronic metabolic dysfunction that characterizes T2D. At the same time, decreased glucose oxidation in skeletal muscle due to mitochondrial damage leads to prolonged postprandial blood glucose rise, which further worsens glucose homeostasis. ROS are not only highly reactive by-products of mitochondrial respiration capable of oxidizing DNA, proteins, and lipids but can also function as signaling and effector molecules in cell membranes mediating signal transduction and inflammation. Mitochondrial uncoupling proteins (UCPs) located in the inner mitochondrial membrane of various tissues can be activated by ROS to protect cells from mitochondrial damage. Mitochondrial UCPs facilitate the reflux of protons from the mitochondrial intermembrane space into the matrix, thereby dissipating the proton gradient required for oxidative phosphorylation. There are five known isoforms (UCP1-UCP5) of mitochondrial UCPs. UCP1 can indirectly reduce ROS formation by increasing glutathione levels, thermogenesis, and energy expenditure. In contrast, UCP2 and UCP3 regulate fatty acid metabolism and insulin secretion by beta cells and modulate insulin sensitivity. Understanding the functions of UCPs may play a critical role in developing pharmacological strategies to combat T2D. This review summarizes the current knowledge on the protective role of various UCP homologs against age-related oxidative stress in T2D.
Keywords: uncoupling proteins, reactive oxygen species, aging, age-related diseases, diabetes
Published in DKUM: 23.08.2023; Views: 321; Downloads: 23
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Salivary gland adaptation to dietary inclusion of hydrolysable tannins in boars
Maša Mavri, Marjeta Čandek-Potokar, Gregor Fazarinc, Martin Škrlep, Catrin Rutland, Božidar Potočnik, Nina Batorek Lukač, Valentina Kubale, 2022, original scientific article

Abstract: The ingestion of hydrolysable tannins as a potential nutrient to reduce boar odor in entire males results in the significant enlargement of parotid glands (parotidomegaly). The objective of this study was to characterize the effects of different levels of hydrolysable tannins in the diet of fattening boars (n = 24) on salivary gland morphology and proline-rich protein (PRP) expression at the histological level. Four treatment groups of pigs (n = 6 per group) were fed either a control (T0) or experimental diet, where the T0 diet was supplemented with 1% (T1), 2% (T2), or 3% (T3) of the hydrolysable tannin-rich extract Farmatan®. After slaughter, the parotid and mandibular glands of the experimental pigs were harvested and dissected for staining using Goldner’s Trichrome method, and immunohistochemical studies with antibodies against PRPs. Morphometric analysis was performed on microtome sections of both salivary glands, to measure the acinar area, the lobular area, the area of the secretory ductal cells, and the sizes of glandular cells and their nuclei. Histological assessment revealed that significant parotidomegaly was only present in the T3 group, based on the presence of larger glandular lobules, acinar areas, and their higher nucleus to cytoplasm ratio. The immunohistochemical method, supported by color intensity measurements, indicated significant increases in basic PRPs (PRB2) in the T3 and acidic PRPs (PRH1/2) in the T1 groups. Tannin upplementation did not affect the histo-morphological properties of the mandibular gland. This study confirms that pigs can adapt to a tannin-rich diet by making structural changes in their parotid salivary gland, indicating its higher functional activity.
Keywords: pigs, dietary supplements, tannins, parotid gland, mandibular gland, immunohistochemistry, histology, proline-rich proteins (PRP), swine
Published in DKUM: 07.07.2023; Views: 355; Downloads: 24
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Production of enzymes from medicinal mushrooms : master's thesis
Gonzalo Herranz Gómez, 2022, master's thesis

Abstract: The purpose of this master's thesis was to determine the ability of certain organisms of the kingdom Fungi to produce a series of enzymes in their active form by solid-state fermentation. In this study, two types of fungi were used, Pleurotus ostreatus and Ganoderma lucidum. For this purpose, different growth media, cultivation times (8 and 10 days), extraction procedures (shaking and homogenization) and extraction medium (distilled water, sodium citrate buffer and sodium phosphate buffer) were used. First, for P. ostreatus mushroom, the optimization of the extraction procedure and time for isolation of enzymes in their active form (α-amylase, glucoamylase, cellulase, laccase, and protease) was studied. It was observed that the highest total protein concentration in mycelium extract was obtained by 8 min of homogenization (0.8607 mg/mL, and distilled water). Using the shaking procedure, the highest enzyme activities were achieved for α-amylase (24 h, 8.0413 U/mL, and sodium citrate buffer) and protease (3 h, 0.0040 U/mL, and sodium citrate buffer). With the homogenization process, the highest activities were achieved for the enzymes glucoamylase (10 min, 6.7113 U/mL, and sodium citrate buffer) and laccase (8 min, 12.2500 U/mL, and sodium citate buffer). For the mushroom G. lucidum, the growth medium and the extraction procedure were optimized, using the same extraction medium (sodium citrate buffer). In this case, α-amylase, glucoamylase, cellulase, laccase, protease, catalase, peroxidase, superoxidase dismutase (SOD), and lipase were studied. It was observed that the highest total protein concentration was obtained with 4 min of homogenization (0.0338 mg/mL). Furthermore, using the homogenization process, the highest activities were achieved for α-amylase (4 min, 16.3459 U/mL) and SOD (4 min, 9.2615 U/mL). With the shaking procedure, the highest activities were achieved for cellulase (3 h, 1.6332 U/mL), lipase (3 h, 16.924 U/mL), glucoamylase (3 h, 14.6737 U/mL), peroxidase (3 h, 0.0156 U/mL), protease (3 h, 0.0080 U/mL) and laccase (24 h, 20.7083 U/mL).
Keywords: medicinal mushrooms, Pleurotus ostreatus, Ganoderma lucidum, total proteins, enzymes activities.
Published in DKUM: 06.07.2022; Views: 636; Downloads: 35
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Variations in leaf total protein, phenolic and thiol contents amongst old varieties of mulberry from the Gorizia region
Tina Ugulin, Tamas Bakonyi, Rebeka Lucijana Berčič, Andreja Urbanek Krajnc, 2015, original scientific article

Abstract: Preserving the plant genetic resources of genus Morus is insuff­cient but undoubtedly vital for conservation of the world’s germplasm for our successors. This research was focused on old mulberry varieties from the Gorizia region in Slovenia which were assessed for their contents on crucial metabolites (proteins, phenolics and thiols) in leaves regarding their antioxidant and nutraceutical potentials. Total proteins were measured spectrophotometrically by following the procedure of Bradford, the total phenolic contents were determined using the Folin-Ciocalteu method and thiols were established with monobromobimane fluorescent dye. The presented metabolite screening showed that some of the evaluated genotypes had higher concentrations of glutathione and were superior in contents of proteins and phenolics when compared to the results of other authors and could be propagated as highly recommendable feed for silkworms, and other animals.
Keywords: feeds, glutathione, Morus, phenols, proteins
Published in DKUM: 14.11.2017; Views: 1636; Downloads: 502
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Quality and delivery of milk in Slovenia
Marjan Janžekovič, Maksimiljan Brus, Lidija Mužerlin, 2007, original scientific article

Abstract: Milk production represents the possibility of earning monthly income and assures social security to many farms in Slovenia. In Slovenia the European rules and regulations on the quality of the delivered milk have been introduced, relating to the hygienic quality of milk and the count of somatic cells. The purchase price of the milk depends also on the protein and fat content. The aim of the research was to establish, whether the quality of the delivered milk complies with the prescriptions regulated by the Slovene legislation. The quality of the delivered milk in the area of the Šentjur agricultural cooperative (ACŠ) and in the entire area of Slovenia (SLO) has been studied. The bacteriological data in the individual quality classes, the data on the total count of somatic cels and the data on the fat and protein contents have been analyzed. 36 monthly accounts have been included in statistical processing. It has been established that the delivered milk of the areas in question does not differ in the relevant quality values.
Keywords: cattle, milk quality, fats, proteins, micro-organisms, somatic cells
Published in DKUM: 10.07.2017; Views: 1219; Downloads: 129
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Effect of concentrate exchange on dairy farm
Marjan Janžekovič, Milan Repič, 2009, original scientific article

Abstract: The purpose of the research was to establish the effect of concentrates A (C-A) and B (C-B) on the quantity and composition of the produced milk of dairy cows. The research included cows of Holstein Friesian and Simmental breeds and crossbreeds with Simmental breed. The test took place in two periods. The first period with the C-A lasted from December 2004 to February 2005 and included 113 cows. During the second period the C-B was fed. The test endured from April to June 2005. The results of monthly controls of milk quantity and protein and fat contents were entered into the Excel programme and processed with the statistical programme SPSS for Windows 12.0. When the concentrate B was fed, the milk quantity amounted to 21.99 L/day and was statistically significantly (P<0.05) greater than the milk quantity in case of feeding the concentrate A, when the milk quantity was 19.12 L/day. No statistically significant differences (P<0.05) of the protein and fat content in the milk were established.
Keywords: milking cows, concentrate, milk quantity, proteins, fats
Published in DKUM: 10.07.2017; Views: 1018; Downloads: 110
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Aquaporin-based biomimetic polymeric membranes
Joachim Habel, Michael R. Hansen, Søren Kynde, Nanna Larsen, Søren Roi Midtgaard, Grethe Vestergaard Jensen, Julie Bomholt, Anayo Ogbonna, Kristoffer Almdal, Alexander Schulz, Claus Hélix-Nielsen, 2015, review article

Abstract: In recent years, aquaporin biomimetic membranes (ABMs) for water separation have gained considerable interest. Although the first ABMs are commercially available, there are still many challenges associated with further ABM development. Here, we discuss the interplay of the main components of ABMs: aquaporin proteins (AQPs), block copolymers for AQP reconstitution, and polymer-based supporting structures. First, we briefly cover challenges and review recent developments in understanding the interplay between AQP and block copolymers. Second, we review some experimental characterization methods for investigating AQP incorporation including freeze-fracture transmission electron microscopy, fluorescence correlation spectroscopy, stopped-flow light scattering, and small-angle X-ray scattering. Third, we focus on recent efforts in embedding reconstituted AQPs in membrane designs that are based on conventional thin film interfacial polymerization techniques. Finally, we describe some new developments in interfacial polymerization using polyhedral oligomeric silsesquioxane cages for increasing the physical and chemical durability of thin film composite membranes.
Keywords: aquaporins, biomimetic membranes, block copolymers, proteopolymersomes, polyhedral oligomeric silsesquioxanes, polyamide layer, microfluidics, membrane proteins, protein-polymer-interactions
Published in DKUM: 21.06.2017; Views: 2087; Downloads: 164
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On the role of anisotropy of membrane constituents on the elastic properties of highly curved lipid membranes
Šarka Perutkova, 2017, doctoral dissertation

Abstract: The primary objective of the thesis covers in the theoretical study the role of anisotropic membrane components in the elasticity of highly curved biological membranes. To show the importance of anisotropy, we focused on one type of non-lamellar membrane self-assembly - the inverted hexagonal phase and the membrane tubular protrusions with attached proteins. These two structures represent excellent examples of highly curved structures in which the anisotropy of molecules or small domains plays an important role. In the first part of the thesis, we developed a theoretical model describing the stability of the inverted hexagonal phase, which considers lipid anisotropy and deviations from the circularity of the pivotal plane cross-section. We applied a wedge-like model of phospholipid molecules, in which the phospholipid molecule is described as a wedge, with the angle of the wedge increasing with temperature. However, we also took into account the average orientation of lipids by including the deviatoric bending energy contribution derived from statistical physics. Theoretical predictions of our model showed that a crosssection of the inverted hexagonal phase is an intermediate between a circle and a hexagon, and that it has lower energy than the circular cross-section. The results were in agreement with observations gathered by the small angle X-ray scattering. By comparing our results with the experiments, we predicted some values of the mean intrinsic curvature and the phospholipid chain stiffness. In the second part of the thesis, we developed a theoretical model, which describes the stabilisation of membrane nanotubes containing attached anisotropic flexible rod-like proteins. We derived the free energy of a vesicle with nanotube taking into account the rotational averaging of the anisotropic attached proteins. We also added the entropy contribution due to the non-homogeneous lateral distribution of proteins. Our theoretical results showed that rod-like attached proteins and membrane domains can stabilise the membrane tubular protrusions if we consider the protein/ domain anisotropy. Our results were also in agreement with experimental results in which isotropic membrane constituents were found on the tips of the nanotube or on the mother vesicle; however, the anisotropic membrane constituents were detected along the nanotubes. Our results showed that rod-like attached proteins and membrane domains can stabilise the membrane tubular protrusions if we consider the protein/domain anisotropy. The anisotropy of membrane constituents can lower the membrane free energy in regions of high curvature. The main aim of the thesis was to show that the anisotropy of membrane constituents can lower the membrane free energy in regions of high curvature and that the rotational averaging of anisotropic membrane components should be considered in the evaluation of the membrane free energy at highly curved membrane structures.
Keywords: Biomembranes, Lipid anisotropy, Inverted hexagonal phase, Rotational averaging, Rod-like proteins, Membrane nanotubes, Membrane protein sorting
Published in DKUM: 25.05.2017; Views: 2076; Downloads: 104
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