1. Initial glutathione depletion during short-term bed rest : pinpointing synthesis and degradation checkpoints in the γ-glutamyl cycleFilippo Giorgio Di Girolamo, Filippo Mearelli, Mariella Sturma, Nicola Fiotti, Kaja Teraž, Alja Ivetac, Alessio Nunnari, Pierandrea Vinci, Boštjan Šimunič, Rado Pišot, Gianni Biolo, 2024, izvirni znanstveni članek Povzetek: Hypokinesia triggers oxidative stress and accelerates the turnover of the glutathione system via the γ-glutamyl cycle. Our study aimed to identify the regulatory checkpoints controlling intracellular glutathione levels. We measured the intermediate substrates of the γ-glutamyl cycle in erythrocytes from 19 healthy young male volunteers before and during a 10-day experimental bed rest. Additionally, we tracked changes in glutathione levels and specific metabolite ratios up to 21 days of bed rest. Using gas chromatography-mass spectrometry and the internal standard technique, we observed a 9 ± 9% decrease in glutathione levels during the first 5 days of bed rest, followed by an 11 ± 9% increase from the 5th to the 10th day, nearly returning to baseline ambulatory levels. The cysteinyl-glycine-to-glutathione ratio, reflecting γ-glutamyl cyclotransferase activity (a key enzyme in glutathione breakdown), rose by 14 ± 22% in the first 5 days and then fell by 10 ± 14% over the subsequent 5 days, again approaching baseline levels. Additionally, the γ-glutamyl cysteine-to-cysteine ratio, indicative of γ-glutamyl cysteine synthetase activity (crucial for glutathione synthesis), increased by 12 ± 30% on day 5 and by 29 ± 41% on day 10 of bed rest. The results observed on day 21 of bed rest confirm those seen on day 10. By calculating the ratio of product concentration to precursor concentration, we assessed the efficiency of these key enzymes in glutathione turnover. These results were corroborated by directly measuring glutathione synthesis and degradation rates in vivo using stable isotope techniques. Our findings reveal significant changes in glutathione kinetics during the initial days of bed rest and identify potential therapeutic targets for maintaining glutathione levels.
Ključne besede: antioxidant, muscle unloading, glutathion turnover, γ-glutamyl cycle, gamma-glutamyl cycle
Objavljeno v DiRROS: 21.11.2024; Ogledov: 115; Prenosov: 1102
 Celotno besedilo (1,84 MB)
Gradivo ima več datotek! Več... |
2. Environmental and economic impacts of hydroxyapatite mineralized wood : LCA and LCC analysisMatic Sitar, Manja Kitek Kuzman, Leon Oblak, Katarina Remic, 2024, izvirni znanstveni članek Povzetek: Wood is considered a promising raw material for the circular bioeconomy and has the ability to store biogenic carbon, and this is one reason why we want to extend the service life of the wood. In order to consider the influence of durability in our study, we used two wood species with different lifespans. Beech (Fagus sylvatica L.) belongs to the group of very sensitive wood species, as the durability of the untreated wood is estimated to be around 5 years; meanwhile, pine (Pinus sylvestris L.) belongs to the group of moderately resistant wood species, where the durability of the untreated wood is estimated to be up to 15 years. While toxic chemicals are often used for wood preservation, hydroxyapatite offers an environmentally friendly solution for wood mineralization. This study presents life cycle assessment (LCA) and life cycle cost (LCC) analyses comparing a novel hydroxyapatite (HAp) mineralization method with a service life of 50 years to a non-mineralized reference alternative. LCA was based on EN ISO 14040 and EN ISO 14044, while LCC was adapted from the European Commission’s LCC tool for public procurement. The results of the LCA show that mineralized wood has a lower overall impact on the environment than surface-treated beech wood but a higher impact than surface-treated pine wood. Most impact categories were determined by electricity consumption with the exception of stratospheric ozone depletion, water consumption, and land use. Water consumption proved to be the category where the mineralization process was problematic due to water consumption during the leaching process. The LCC showed that mineralized wood is the most cost-effective solution for the exterior façade, as all costs, but especially investment costs, were lower. The differences in the LCA and LCC results are mainly due to the different lifetimes of the two alternatives. It can be concluded that if energy-intensive processes and chemicals are used in the production of the material, the extended lifetime must be sufficient to account for the additional impacts that occur during the production phase.
Ključne besede: environmental impacts, hydroxyapatite, life cycle assessment (LCA), life cycle cost analysis (LCC), wood mineralization
Objavljeno v DiRROS: 09.09.2024; Ogledov: 292; Prenosov: 2596
Celotno besedilo (2,60 MB)
Gradivo ima več datotek! Več... |
3. Environmental life cycle assessment of railway bridge materials using UHPFRCKarmen Fifer Bizjak, Aljoša Šajna, Katja Slanc, Friderik Knez, 2016, izvirni znanstveni članek Povzetek: The railway infrastructure is a very important component of the world’s total transportation network. Investment in its construction and maintenance is significant on a global scale. Previously published life cycle assessment (LCA) studies performed on road and rail systems very seldom included infrastructures in detail, mainly choosing to focus on vehicle manufacturing and fuel consumption. This article presents results from an environmental study for railway steel bridge materials for the demonstration case of the Buna Bridge in Croatia. The goal of these analyses was to compare two different types of remediation works for railway bridges with different materials and construction types. In the first part, the environmental impact of the classical concrete bridge construction was calculated, whereas in the second one, an alternative new solution, namely, the strengthening of the old steel bridge with ultra-high-performance fibre-reinforced concrete (UHPFRC) deck, was studied. The results of the LCA show that the new solution with UHPFRC deck gives much better environmental performance. Up to now, results of LCA of railway open lines, railway bridges and tunnels have been published, but detailed analyses of the new solution with UHPFRC deck above the old bridge have not previously been performed.
Ključne besede: railway, materials, ife cycle assessment, bridge, environmental
Objavljeno v DiRROS: 23.08.2024; Ogledov: 310; Prenosov: 274
Celotno besedilo (2,33 MB)
Gradivo ima več datotek! Več... |
4. Double strand breaks and cell-cycle arrest induced by the cyanobacterial toxin cylindrospermopsin in HepG2 cellsAlja Štern, Metka Filipič, Matjaž Novak, Bojana Žegura, 2013, izvirni znanstveni članek Povzetek: The newly emerging cyanobacterial cytotoxin cylindrospermopsin (CYN) is increasingly found in surface freshwaters, worldwide. It poses a potential threat to humans after chronic exposure as it was shown to be genotoxic in a range of test systems and is potentially carcinogenic. However, the mechanisms of CYN toxicity and genotoxicity are not well understood. In the present study CYN induced formation of DNA double strand breaks (DSBs), after prolonged exposure (72 h), in human hepatoma cells, HepG2. CYN (0.1–0.5 µg/mL, 24–96 h) induced morphological changes and reduced cell viability in a dose and time dependent manner. No significant increase in lactate dehydrogenase (LDH) leakage could be observed after CYN exposure, indicating that the reduction in cell number was due to decreased cell proliferation and not due to cytotoxicity. This was confirmed by imunocytochemical analysis of the cell-proliferation marker Ki67. Analysis of the cell-cycle using flow-cytometry showed that CYN has an impact on the cell cycle, indicating G0/G1 arrest after 24 h and S-phase arrest after longer exposure (72 and 96 h). Our results provide new evidence that CYN is a direct acting genotoxin, causing DSBs, and these facts need to be considered in the human health risk assessment.
Ključne besede: cylindrospermopsin, cell-cycle, cell-proliferation, double-strand breaks, HepG2 cells
Objavljeno v DiRROS: 02.08.2024; Ogledov: 378; Prenosov: 262
Celotno besedilo (689,29 KB)
Gradivo ima več datotek! Več... |
5. RECQ1 helicase silencing decreases the tumour growth rate of U87 glioblastoma cell xenografts in zebrafish embryosMiloš Vittori, Barbara Breznik, Katja Hrovat, Saša Kenig, Tamara Lah Turnšek, 2017, izvirni znanstveni članek Povzetek: RECQ1 helicase has multiple roles in DNA replication, including restoration of the replication fork and DNA repair, and plays an important role in tumour progression. Its expression is highly elevated in glioblastoma as compared to healthy brain tissue. We studied the effects of small hairpin RNA (shRNA)-induced silencing of RECQ1 helicase on the increase in cell number and the invasion of U87 glioblastoma cells. RECQ1 silencing reduced the rate of increase in the number of U87 cells by 30%. This corresponded with a 40% reduction of the percentage of cells in the G2 phase of the cell cycle, and an accumulation of cells in the G1 phase. These effects were confirmed in vivo, in the brain of zebrafish (Danio rerio) embryos, by implanting DsRed-labelled RECQ1 helicase-silenced and control U87 cells. The growth of resulting tumours was quantified by monitoring the increase in xenograft fluorescence intensity during a three-day period with fluorescence microscopy. The reduced rate of tumour growth, by approximately 30% in RECQ1 helicase-silenced cells, was in line with in vitro measurements of the increase in cell number upon RECQ1 helicase silencing. However, RECQ1 helicase silencing did not affect invasive behaviour of U87 cells in the zebrafish brain. This is the first in vivo confirmation that RECQ1 helicase is a promising molecular target in the treatment of glioblastoma.
Ključne besede: cancer, cell cycle, DNA damage, intravital imaging, RNA interference, theranostics
Objavljeno v DiRROS: 25.07.2024; Ogledov: 333; Prenosov: 223
Celotno besedilo (2,73 MB)
Gradivo ima več datotek! Več... |
6. Microscopy of crustacean cuticle : formation of a flexible extracellular matrix in moulting sea slaters Ligia pallasiiJasna Štrus, Magda Tušek-Žnidarič, Urška Repnik, Andrej Blejec, Adam P. Summers, 2019, izvirni znanstveni članek Povzetek: Structural and functional properties of exoskeleton in moulting sea slaters Ligia pallasii from the Eastern Pacific coast were investigated with CT scanning and electron microscopy. Ultrastructure of preecdysial and postecdysial cuticular layers was described in premoult, intramoult and postmoult animals. Cuticle is a flexible extracellular matrix connected to the epidermal cells through pore channels. During premoult epicuticle and exocuticle are formed and during intramoult and postmoult endocuticular lamellae are deposited and the cuticle is progressively constructed by thickening and mineralization. Cuticle permeability, flexibility and waterproofing capacity change accordingly. Elaboration of epicuticular scales connected to an extensive network of nanotubules, establish its anti-adhesive and hydrophobic properties. Labelling with gold conjugated WGA lectins on Tokuyashu thawed cryosections exposes differences in chitin content between exocuticle and endocuticle. Histochemical staining of cuticle shows presence of acidic carbohydrates/glycoconjugates and lipoproteins in epicuticular layer. Chitin microfibrils are formed at the microvillar border of epidermal cells with abundant Golgi apparatus and secretory vesicles. Numerous spherules associated with nanotubules were observed in the ecdysial space in intramoult animals. The mineral component of the cuticle as visualized with CT scanning indicates progressive mineral resorption from the posterior to the anterior half of the body in premoult animals, its translocation from the anterior to posterior part during intramoult and its progressive deposition in the posterior and anterior exoskeleton during postmoult. Cuticle of sea slaters is a unique biocomposite and biodynamic material constantly reconstructed during frequent moults, and adapted to specific physical and biotic conditions of the high intertidal rocky zone.
Ključne besede: cuticle ultrastructure, micro CT scanning, moult cycle, SEM, TEM, terrestrial isopods
Objavljeno v DiRROS: 23.07.2024; Ogledov: 414; Prenosov: 337
Celotno besedilo (1,06 MB)
Gradivo ima več datotek! Več... |
7. Scaphoideus titanus up-to-the-minute : biology, ecology, and role as a vectorElena Gonella, Giovanni Benelli, Nathalie Arricau-Bouvery, Domenico Bosco, Meta Virant-Doberlet, 2024, pregledni znanstveni članek Povzetek: Native to the Nearctic region, Scaphoideus titanus Ball (Hemiptera: Cicadellidae) has become a major threat for grapevine production after being unintentionally introduced into Europe, where it became the main vector of flavescence dorée phytoplasma, being mainly associated with the genus Vitis. Scaphoideus titanus is a highly efficient vector of the most important phytoplasma affecting grapevine. For this reason, compulsory insecticide treatments have been introduced against this pest in many European countries. Moreover, the continuous expansion of its geographical distribution makes this leafhopper a serious threat for several non-European Countries. In this article, we review the current knowledge about its taxonomy, morphology, biology, ecology, and its role as a vector. Finally, we point out the main challenges for research aimed at reducing S. titanus and flavescence dorée expansion across Europe and avoiding spread of the disease outside the Old World.
Ključne besede: vector distribution, life cycle, morphology, taxonomy, feeding behaviour, entomology, insects
Objavljeno v DiRROS: 23.07.2024; Ogledov: 372; Prenosov: 187
Celotno besedilo (982,26 KB)
Gradivo ima več datotek! Več... |
8. HepG2 spheroids as a biosensor-like cell-based system for (geno)toxicity assessmentMartina Štampar, Sonja Žabkar, Metka Filipič, Bojana Žegura, 2022, izvirni znanstveni članek Povzetek: 3D spheroids developed from HepG2 cells were used as a biosensor-like system for the detection of (geno)toxic effects induced by chemicals. Benzo(a)pyrene (B(a)P) and amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) with well-known mechanisms of action were used for system validation. HepG2 spheroids grown for 3 days were exposed to BaP and PhIP for 24 and 72 h. The growth and viability of spheroids were monitored by planimetry and Live/Dead staining of cells. Multi-parametric flow cytometric analysis was applied for simultaneous detection of specific end-effects including cell cycle analysis (Hoechst staining), cell proliferation (KI67 marker), and DNA double-strand breaks (ℽH2AX) induced by genotoxic compounds. Depending on the exposure concentration/time, BaP reduced spheroid growth, affected cell proliferation by arresting cells in S and G2 phase and induced DNA double-strand breaks (DSB). Simultaneous staining of ℽH2AX formation and cell cycle analysis revealed that after BaP (10 μM; 24 h) exposure 60% of cells in G0/G1 phase had DNA DSB, while after 72 h only 20% of cells contained DSB indicating efficient repair of DNA lesions. PhIP did not influence the spheroid size whereas accumulation of cells in the G2 phase occurred after both treatment times. The evaluation of DNA damage revealed that at 200 μM PhIP 50% of cells in G0/G1 phase had DNA DSB, which after 72-h exposure dropped to 40%, showing lower repair capacity of PhIP-induced DSB compared to BaP-induced. The developed approach using simultaneous detection of several parameters provides mechanistic data and thus contributes to more reliable genotoxicity assessment of chemicals as a high-content screening tool.
Ključne besede: in vitro 3D cell model, HepG2, flow cytometry, cell cycle, proliferation, DNA strand, breaks
Objavljeno v DiRROS: 16.07.2024; Ogledov: 349; Prenosov: 194
Celotno besedilo (6,21 MB)
Gradivo ima več datotek! Več... |
9. Adverse toxic effects of tyrosine kinase inhibitors on non-target zebrafish liver (ZFL) cellsKatja Kološa, Bojana Žegura, Martina Štampar, Metka Filipič, Matjaž Novak, 2023, izvirni znanstveni članek Ključne besede: zebrafish liver cells, ZFL, tyrosine kinase inhibitors, cytotoxicity, cell cycle, genotoxicity, environmental hazard
Objavljeno v DiRROS: 12.07.2024; Ogledov: 594; Prenosov: 238
Celotno besedilo (3,30 MB)
Gradivo ima več datotek! Več... |
10. Influence of alkylthio and arylthio derivatives of tert-butylquinone on the induction of DNA damage in a human hepatocellular carcinoma cell line (HepG2)Jelena Djordjević, Stoimir Kolarević, Jovana Jovanović Marić, Margareta Kračun-Kolarević, Bojana Žegura, Alja Štern, Dušan M. Sladić, Irena Novaković, Branka Vuković-Gačić, 2024, izvirni znanstveni članek Povzetek: The aim of this study was to investigate the effects of tert-butylquinone (TBQ) and its alkylthio and arylthio
derivatives on DNA in vitro, using acellular and cellular test systems. Direct interaction with DNA was studied
using the plasmid pUC19. Cytotoxic (MTS assay) and genotoxic (comet assay and γH2AX focus assays) effects,
and their influence on the cell cycle were studied in the HepG2 cell line. Our results show that TBQ and its
derivatives did not directly interact with DNA. The strongest cytotoxic effect on the HepG2 cells was observed for
the derivative 2-tert-butyl-5,6-(ethylenedithio)-1,4-benzoquinone (IC50 64.68 and 55.64 μM at 24-h and 48-h
treatment, respectively). The tested derivatives did not significantly influence the cell cycle distribution in the
exposed cellular populations. However, all derivatives showed a genotoxic activity stronger than that of TBQ in
the comet assay, with 2-tert-butyl-5,6-(ethylenedithio)-1,4-benzoquinone producing the strongest effect. The
same derivative also induced DNA double-strand breaks in the γH2AX focus assay.
Ključne besede: TBQ derivatives, HepG2 cell line, comet assay, γH2AX assay, cell cycle analysis, cytotoxicity
Objavljeno v DiRROS: 11.07.2024; Ogledov: 401; Prenosov: 248
 Celotno besedilo (7,34 MB)
Gradivo ima več datotek! Več... |
