Supplementary data for article : Kolarević, S.; Milovanović, D.; Kračun-Kolarević, M.; Kostić, J.; Sunjog, K.; Martinović, R.; Đorđević, J.; Novaković, I.; Sladić, D.; Vuković-Gačić, B. Evaluation of Genotoxic Potential of Avarol, Avarone, and Its Methoxy and Methylamino Derivatives in Prokaryotic and Eukaryotic Test Models. Drug and Chemical Toxicology 2019, 42 (2), 130–139. https://doi.org/10.1080/01480545.2017.1413108

Supplementary material for: [https://doi.org/10.1080/01480545.2017.1413108]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/322

Effects of 5-Fluorouracil, Etoposide and CdCl2 in Aquatic Oligochaeta Limnodrilus udekemianus Claparede (Tubificidae) Measured by Comet Assay

Genotoxicity of 5-fluorouracil (5-FU), etoposide (ET) and cadmium chloride (CdCl2) was evaluated in Limnodrilus udekemianus, cosmopolitan tubificid species, by alkaline single-cell gel electrophoresis (comet assay). Groups of 50 individuals were exposed in vivo in water-only short-term (96 h) tests to 5-FU (0.004, 0.04, 0.4, 4 and 40 mu M), ET (0.004. 0.04, 0.4 and 4 mu M) and CdCl2 (0.004, 0.04, 0.4, 4 and 40 mu M). Mortality of worms was observed only for CdCl2 (4 and 40 mu M). Cell viability lower than 70 \% was detected for 5-FU (0.4, 4 and 40 mu M), ET (4 mu M) and CdCl2 (0.4 and 4 mu M). All tested substances induced significant increase of DNA damage except 0.004 mu M of ET. L. udekemianus being sensitive to all tested substances indicates that it can be used in ecogenotoxicology studies. Concern should be raised to cytostatics, especially to 5-FU, since concentration of 0.004 mu M induced DNA damage is similar to ones detected in wastewaters.Water Air and Soil Pollution (2015), 226(242

Supplementary data for article : Kolarević, S.; Milovanović, D.; Kračun-Kolarević, M.; Kostić, J.; Sunjog, K.; Martinović, R.; Đorđević, J.; Novaković, I.; Sladić, D.; Vuković-Gačić, B. Evaluation of Genotoxic Potential of Avarol, Avarone, and Its Methoxy and Methylamino Derivatives in Prokaryotic and Eukaryotic Test Models. Drug and Chemical Toxicology 2019, 42 (2), 130–139. https://doi.org/10.1080/01480545.2017.1413108

Supplementary material for: [https://doi.org/10.1080/01480545.2017.1413108]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/322

Antitumor activity of Lamiaceae plants frequently used in Serbian folk medicine and cuisine.

Recently, cancer research has focused on searching for new and more effective antitumor agents of natural origin that can activate multiple defence mechanisms and selectively damage transformed cells. The goal of this research was to assess different antitumor mechanisms of ethanolic extracts of 18 Lamiaceae species traditionally used in Serbian folk medicine and cuisine, as well as their genotoxic potential towards HCT-116 (colorectal cancer) cells. The viability of treated HCT-116 cells was assessed by MTT assay; the production of reactive oxygen species (ROS) by treated HCT- 116 cells was determined using NBT assay, while their production of nitric oxide (NO) was evaluated using Griess assay. The genotoxic activity of the extracts on HCT-116 cells was tested in Comet assay, using etoposide as a positive control. The results indicated that lavender, basil, and rosemary inhibited the proliferation of these cells, significantly lowering their viability. Moreover, lavender and thyme extracts displayed a significant increase in ROS production, whereas ground- ivy, hyssop, lemon balm, peppermint, basil, rosemary, sage, and winter savory have significantly lowered their production. The results of the Griess assay suggested that lavender, motherwort, peppermint, basil, rosemary, sage, winter savory, ironwort, and thyme have significantly increased the production of NO. Furthermore, Comet assay results pointed out that motherwort, peppermint, basil, oregano, marjoram, winter savory, ironwort, wild thyme, thyme, and mountain germander possess genotoxic potential towards HCT-116 cells, while only basil had genotoxic activity statistically similar to etoposide. The obtained results are in accordance with our previous findings, which indicated that these extracts have antigenotoxic and genoprotective activities towards normal cells. Finally, it can be concluded that these traditionally valued plants might act as potent antitumor agents by modulating the proliferation and production of ROS and NO by cancer cells, as well as by expressing significant genotoxic properties towards cancer cells

Corrigendum to "European contribution to the study of ROS:A summary of the findings and prospects for the future from the COST action BM1203 (EU-ROS)" [Redox Biol. 13 (2017) 94-162]

The European Cooperation in Science and Technology (COST) provides an ideal framework to establish multi-disciplinary research networks. COST Action BM1203 (EU-ROS) represents a consortium of researchers from different disciplines who are dedicated to providing new insights and tools for better understanding redox biology and medicine and, in the long run, to finding new therapeutic strategies to target dysregulated redox processes in various diseases. This report highlights the major achievements of EU-ROS as well as research updates and new perspectives arising from its members. The EU-ROS consortium comprised more than 140 active members who worked together for four years on the topics briefly described below. The formation of reactive oxygen and nitrogen species (RONS) is an established hallmark of our aerobic environment and metabolism but RONS also act as messengers via redox regulation of essential cellular processes. The fact that many diseases have been found to be associated with oxidative stress established the theory of oxidative stress as a trigger of diseases that can be corrected by antioxidant therapy. However, while experimental studies support this thesis, clinical studies still generate controversial results, due to complex pathophysiology of oxidative stress in humans. For future improvement of antioxidant therapy and better understanding of redox-associated disease progression detailed knowledge on the sources and targets of RONS formation and discrimination of their detrimental or beneficial roles is required. In order to advance this important area of biology and medicine, highly synergistic approaches combining a variety of diverse and contrasting disciplines are needed

The unfolded protein response controls induction and activation of ADAM17/TACE by severe hypoxia and ER stress.

The family of ADAM (a disintegrin and metalloproteinase) proteins has been implicated in tumor initiation and progression. ADAM17/tumor necrosis factor-α (TNFα)-converting enzyme (TACE) has been initially recognized to release TNFα as well as its receptors (TNFRs) from the membrane. ADAM17, TNFα and TNFR have been found upregulated in cancer patients, although the underlying mechanisms remain largely unknown. As hypoxia is a hallmark of cancer that can lead to severe stress conditions accumulating in endoplasmic reticulum (ER) stress and the unfolded protein response (UPR), we investigated the role of these stress conditions in the regulation of ADAM17 and release of TNFR1.We found that severe hypoxia induced ADAM17 expression and activity. Although hypoxia-inducible factor 1α (HIF1α) was important to maintain basal ADAM17 mRNA levels during moderate hypoxia, it was not sufficient to induce ADAM17 levels under severe hypoxia. Instead, we found that ADAM17 induction by severe hypoxia can be mimicked by ER stressors such as Thapsigargin and occurs as a consequence of the activation of the PERK/eIF2α/ATF4 and activating transcription factor 6 (ATF6) arms of UPR in several tumor cell lines. ADAM17 expression was also increased in xenografts displaying ER stress because of treatment with the vascular endothelial growth factor (VEGF) inhibitory antibody Bevacizumab. Additionally, severe hypoxia and ER stress activated ADAM17 and ectodomain shedding of TNFR1 involving mitogen-activated protein (MAP) kinases and reactive oxygen species (ROS). Collectively, these results show that ADAM17 is a novel UPR-regulated gene in response to severe hypoxia and ER stress, which is actively involved in the release of TNFR1 under these conditions. These data provide a novel link between severe hypoxic stress conditions and inflammation in the tumor environment

Macroinvertebrate communities along the Velika Morava River

This paper presents the results of a faunistic study of the macrozoobenthos of the Velika Morava River. The investigation was conducted during the summer and autumn months in 2010. A total of 84 macroinvertebrate taxa have been identified, with Insecta (Ephemeroptera) as the most diverse and Oligochaeta as the most abundant groups. A tubificid worm, Limnodrilus hoffmeisteri, was the most important species with regard to relative abundance and frequency of occurrence. Two rare and endangered species, Theodoxus transversalis and Unio crassus, were recorded, as well as 5 alien species. Locality VM4 (Markovac Bridge) is of particular interest as the northernmost locality, as well as having the most abundant population of T. transversalis found. Despite being in the lower stretch of the river, this site is particularly taxa-rich, presumably due to conspicuous microhabitat diversity. Water temperature and pH value were determined to be the most important factors of the 32 environmental variables tested. Multivariate analyses revealed separation of summer samples compared to autumn. The Mann-Whitney test showed significant differences in fauna only in the case of ecoregions, confirming their current delineation and the transitional character of this river.Ministry of Education, Science and Technological Development of the Republic of Serbia {[}TR 37009, OI 173025

Evaluation of genotoxic potential of avarol, avarone, and its methoxy and methylamino derivatives in prokaryotic and eukaryotic test models

In this study, mutagenic and genotoxic potential of anti-tumor compounds avarol, avarone, and its derivatives 3′-methoxyavarone, 4′-(methylamino)avarone and 3′-(methylamino)avarone was evaluated and compared to cytostatics commonly used in chemotherapy (5-fluorouracil, etoposid, and cisplatin). Mutagenic potential of selected hydroquinone and quinones was assessed in prokaryotic model by the SOS/umuC assay in Salmonella typhimurium TA1535/pSK1002. Genotoxic potential was also assessed in eukaryotic models using comet assay in human fetal lung cell line (MRC-5), human adenocarcinoma epithelial cell line (A549), and in human peripheral blood cells (HPBC). The results indicated that avarol and avarone do not exert mutagenic/genotoxic potential. Among the studied avarone derivatives, mutagenic potential was detected by SOS/umuC test for 3′-(methylamino)avarone, but only after metabolic activation. The results of comet assay indicated that 3′-methoxyavarone and 3′-(methylamino)avarone have a significant impact on the level of DNA damage in the MRC-5 cell line. Genotoxic potential was not observed in A549 cells or HPBC probably due to a different uptake rate for the compounds and lower in metabolism rate within these cells.Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3033

Molecular signaling pathways in right ventricular impairment of adult patients after tetralogy of Fallot repair.

Background: Right ventricular impairment (RVI) secondary to altered hemodynamics contributes to morbidity and mortality in adult patients after tetralogy of Fallot (TOF) repair. The goal of this study was to describe signaling pathways contributing to right ventricular (RV) remodeling by analyzing over lifetime alterations of RV gene expression in affected patients. Methods: RV tissue was collected at the time of cardiac surgery in 13 patients with a diagnosis of TOF. RNA was isolated and whole transcriptome sequencing was performed. Gene profiles were compared between a group of 6 adults with signs of RVI undergoing right ventricle to pulmonary artery conduit surgery and a group of 7 infants, undergoing TOF correction. Definition of RVI in adult patients was based on clinical symptoms, evidence of RV hypertrophy, dilation, dysfunction or elevated pressure on echocardiographic, cardiovascular magnetic resonance, or catheterization evaluation. Results: Median age was 34 years in RVI patients and 5 months in infants. Based on P adjusted value <0.01, RNA sequencing of RV specimens identified a total of 3,010 differentially expressed genes in adult patients with TOF and RVI as compared to infant patients with TOF. Gene Ontology and Kyoto Encyclopedia of Genes databases highlighted pathways involved in cellular metabolism, cell-cell communication, cell cycling and cellular contractility to be dysregulated in adults with corrected TOF and chronic RVI. Conclusions: RV transcriptome profiling in adult patients with RVI after TOF repair allows identification of signaling pathways, contributing to pathologic RV remodeling and helps in the discovery of biomarkers for disease progression and of new therapeutic targets