Genotoxic effects of inorganic molecule dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH), a promising new therapeutic for the epidermal changes treatment, have been evaluated. In vitro analysis included evaluation of genotoxic and cytotoxic potential of K2(B3O3F4OH) in concentrations of 0.01, 0.02, 0.05 and 0.06 mg/mL applying cytokinesis-block micronucleus cytome assay in human lymphocyte culture. With the increase of concentration the frequency of micronuclei elevated but the differences were not significant. Also, there were no significant differences among the frequencies of nuclear buds and nucleoplasmic bridges between controls and treated cultures. Nuclear division index and nuclear division cytotoxycity index values did not reveal significant cytotoxic effect of K2(B3O3F4OH). In vivo genotoxic effects were analyzed on BALB/c mice applying reticulocytes micronucleus assay. K2(B3O3F4OH) was administrated intraperitoneally in final concentrations of 10, 20, 50 and 55 mg/kg. Significant decrease of reticulocytes ratio and increase of micronuclei frequencies against pre-treatments were found for both sampling periods of 48 and 72 hours of the highest applied concentration. This study confirmed that K2(B3O3F4OH) is not genotoxic in tested concentrations in vitro as well as in concentrations lower than 55 mg/kg in vivo. This study presents a reliable basis for further pre-clinical and potential clinical investigations.

s submitted to the 41st FEBS Congress, which was planned for Kuşadası, Turkey from 3rd to 8th September 2016, and accepted by the Congress Organizing Committee are published in this Special Issue of The FEBS Journal. Unfortunately, the Congress was cancelled by FEBS after the excellent scientific programme was compromised by an insufficient number of confirmed speakers, and so the authors of these abstracts were not able to present their work at the event*. Late-breaking abstracts and abstracts withdrawn after Congress cancellation are not included in this issue. About these abstracts Abstracts submitted to the Congress are not peer-reviewed. In addition, abstracts are published as submitted and are not copyedited prior to publication. We are unable to make corrections of any kind to the abstracts once they are published. Indexing Abstracts published in The FEBS Journal Special Issue for the 41st FEBS Congress will be included individually in the Conference Proceedings Citation Index published by Web of Science. How to cite these abstracts AuthorOne, A., AuthorTwo, B. (2016). Abstract title. FEBS J, 283: Abstract number**. doi:10.1111/febs.13903 * An optional closed online presentation opportunity of short duration on the Congress website was offered after Congress cancellation and may be taken up by some abstract authors. ** Each abstract has been given a unique number beginning with either the letters P or ST; the next part relates to the session in which the speed talk or poster will be presented. 01.02 RNA biology, biogenesis and processing 02.03 Autophagy: Regulation mechanisms 02.04 Mechanisms and regulation of protein translocation 02.05 Intracellular organization 02.06 Human microbiome (microbiota) 02.07 Extracellular matrix and metalloproteinases 09.04 Chemical and biochemical aspects of oxidative stress Mis Miscellaneous 428 The FEBS Journal 283 (Suppl. 1) (2016) 428 DOI: 10.1111/febs.13903 © 2016 The Authors. The FEBS Journal © 2016 FEBS This addendum corrects errors in the Speed Talks and Poster sessions of the Supplement S1. The following abstracts were omitted by mistake from the original line-up. In addition, the abstracts P-02.03.3-002 and P-09.04.4-104 were truncated in the original document and are reproduced in full here. FEBS_v283_s1_13903.indd 1 10/12/2016 2:00:46 PM OMITTED POSTER SESSIONS Tuesday 6 September 12:30–14:30 Autophagy: Regulation mechanisms P-02.03.3-002 Apoptotic and necrotic effects of low dose bisphenol A in SHSY5Y neuroblastoma cells B. Ayazg€ok, T. T€ uyl€ u K€ uc € ukkilinc Hacettepe University Faculty of Pharmacy, Ankara, Turkey Bisphenol A (BPA) is a commonly used chemical in industry to make plastics. “Low-dose” term has been expressed for the first time in studies with BPA in 2001. The value of low dose was received as <1 lM for BPA in in vitro studies. Nowadays majority of the population as a result of today’s lifestyle exposured to low doses BPA chronically, thus importance of low-dose toxicity studies is revealed. In this study we aimed to examine cytotoxicity composed by low dose BPA in SHSY5Y cells in terms of appoptotic and necrotic effects.SHSY5Y cells was seeded at 300.000 cells per well in 6-well plates and cultured in DMEM at 37°C with 5% CO2.SH-SY5Y cells were treated with low dose(1 pM, 1 nM) of BPA. Plate was incubated for 24 and 48 hours. After the incubation period, samples were pooled then washed with PBS in two times and 100 ll aliquot of cells from each sample was centrifuged at 12000 g, +4 °C for 2 minutes and resuspended in 100 ll annexin binding buffer added to 5 ll Annexin V and incubated at room temperature in the dark for 20 minutes. Then, samples were centrifuged and resuspended again in 100 ll of the same buffer and added with 1 ll PI at room temperature for 1–4 minutes and analysed at Tali Image-Based Cytometer. Reducing cell viability of low dose BPA in SHSY5Y neuroblastoma cells is revealed by MTT by our group in our previous studies. Cytotoxicity studies are conducted for 1 pm and 1 nm of BPA in 48 hours by taking into account this result. It is obtained that 1pM reduced cell viability to % 63 and 1 nM BPA decreased cell viability to %78. It is clearly ocuured via examinate of necrotic effects that group treated with 1 pM and 1 nM BPA was significiantly different from control group. It has observed group treated with 1 pM and 1 nM BPA has importantly difference compared control group. The findings obtained from this study explain the cytotoxicity of BPA in SHSY5Y cells through necrotic and late apoptotic pathways. P-02.03.3-005 Lack of Atg5 expression diminished apoptotic potential of cdk inhibitors due to increased Bcl-2 expression in MEF cells A. E. Nezir, E. D. Arisan, A. Coker-Gurkan, P. Obakan,

Introduction: Bioflavonoids delphinidin (2-(3,4,5-Trihydroxyphenyl)chromenylium-3,5,7-triol) and luteolin (2-(3,4-Dihydroxyphenyl)-5,7-dihydroxy-4-chromenone) have been recognized as promising antioxidants and anticancer substances. Due to their extensive use, the goal of the research was to determine whether they have any genotoxic potential in vitro.Methods: Analysis of genotoxic potential was performed applying chromosome aberrations test in human lymphocyte culture, as this kind of research was not conducted abundantly for these two bioflavonoids. Delphinidin and luteolin were dissolved in DMSO and added to cultures in final concentrations of 25, 50 and 100 μM.Results: In human lymphocytes cultures Delphinidin induced PCDs in all treatments, potentially affecting the cell cycle and topoisomerase II activity. In concentration of 50 μM luteolin showed strong genotoxic effects and caused significant reduction of cell proliferation.Conclusion: Luteolin exhibited certain genotoxic and cytostatic potential. Delphinidin was not considered genotoxic, however its impact on mitosis, especially topoisomerase II activity, was revealed.

Abstract The genotoxicity of halogenated boroxine [K2(B3O3F4OH)], a novel compound with the potential for prevention and/or treatment of various skin changes, has been confirmed in human lymphocytes. The potential of luteolin and delphinidin in inhibition of the genotoxic and cytotoxic effects of halogenated boroxine in vitro was analyzed applying the chromosome aberration analysis and the cytokinesis-block micronucleus cytome assay in human lymphocyte cultures. The in vitro treatments included addition of boroxine and flavonoids independently, and combined treatments of boroxine with luteolin or delphinidin. In the concentration of 50 μM, luteolin significantly decreased the frequency of micronuclei and nuclear buds. Delphinidin suppressed the occurrence of aberrant cells in the presence of the halogenated boroxine, but also affected their induction in respective delphinidin treatment. Detection of endoreduplications in combined treatments indicated that these flavonoids are potential inhibitors of cell cycle or topoisomerase II activity. The obtained results have confirmed antigenotoxic activity of selected bioflavonoids in vitro. The side effects of potential therapeutic applications of halogenated boroxine may be inhibited in the presence of bioflavonoids in appropriate dosage.

Despite their known toxic properties, various Helleborus species are used as medicaments in folk medicine to treat some diseases and health conditions. As the main mechanism of many cytostatic drugs is based on their cytotoxic activity, there is potential for the toxicity of hellebore to be used in anticancer therapy. This study tested the geno- and cytotoxic effects of extracts of three hellebore taxa (Helleborus odorus, Helleborus multifidus and Helleborus hercegovinus) on meristemic onion (Alliumcepa L.) cells and human lymphocytes. Treatments with Helleborus extracts induced cytotoxic and cytostatic effects in meristemic onion cells as well as in cultivated cytokinesis-blocked human lymphocytes. Cytokinesis-block micronucleus cytome assay indicated that treatments with hellebore extracts induce genotoxic effects in human lymphocytes, and that the significant mechanism of their antiproliferative activity is apoptosis induction.

Despite the fact that Asplenium scolopendrium L. is a wide spread fern which has been used as a human remedy for centuries, there are very poor or no data about activity and genotoxicity of A. scolopendrium extracts. In this work, vacuum dried water and ethanol extracts of A. scolopendrium fronds were tested for their antimicrobial, cytotoxic and genotoxic potential. Antimicrobial activity was evaluated by disk diffusion assay (concentration of extracts 35 mg/ml, 7 mg/ml and 1,4 mg/ml) and there was no inhibition zone for all extracts and for all microorganisms examined (Escherichia coli, Staphylococcus aureus and Candida albicans). Cytotoxic and genotoxic potential of extracts (70 mg/ml, 7 mg/ml and 0,7 mg/ml) was tested using cytokinesis-block micronucleus cytome assay in human lymphocyte cultures. Ethanol blocked division of cells in negative control so only water extracts were analyzed. Extracts didn’t show genotoxic properties but they showed weak cytotoxic properties. NDI (nuclear division index) decreased with increasing concentration of extracts, but there was no statistical significance when compared to negative control (p=0,055).

Ionising radiation is known to cause chromosomal instability, which is observed as increased frequency of chromosomal aberration and micronuclei. These are listed as reliable criteria in biological dosimetry. Numerous experiments conducted on both animal and plant models demonstrated that increase in radiation dosage is followed by increased mutation frequency, and that mutations occur even at the lowest exposure. We used horse blood in vitro irradiated by low doses of ionizing radiation. Cultivation of peripheral blood lymphocytes and micronucleus test were used as biomarkers of genetic damage. The observed aberrations were recorded and classified in accordance with the International System of Cytogenetic Nomenclature. Micronuclei were identified on the basis of criteria proposed by Fenech et al . (8). Analysis of chromosomal aberration showed increased frequency of aberrations in blood cultures exposed to 0,1 Gy and 0,2 Gy compared to the controls. Microscopic analysis of chromosomal damage in in vitro micronucleus test revealed that the applied radiation dose induced micronuclei while no binucleated cells with micronuclei were found in lymphocytes that were not irradiated. In this paper we analysed the influence of low dose ionising radiation on frequency of chromosomal aberration and micronuclei in horse lymphocytes following in vitro exposure to X-rays (0,1 Gy and 0,2 Gy). Key words : chromosomal aberrations, micronuclei, ionising radiation, horse lymphocytes

This study was undertaken in order to evaluate possible antioxidative and antiproliferative activities of three Helleborus taxa. The dry leaves and roots of three Helleborus taxa were extracted with ethanol and water. A phytochemical evaluation of the selected extracts was performed using spectrophotometric methods and a 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity assay was used for measuring the antioxidative activity of extracts. The antiproliferative activity of the three Helleborus taxa was studied using Burkitt's lymphoma B cells (BJAB) cell lines. The phytochemical evaluation showed that the leaves contain high levels of total phenolic and flavonoid content. Results from the DPPH assay indicated that the activity of the ethanol and water extracts of the leaves was higher than that of positive control (thymol). Extracts from the roots of H. odorus also displayed higher antioxidant activity than the positive probe, while H. mulifidus and H. hercegovinus root extracts were less effective. A statistically significant correlation between total phenolic content and antioxidative properties indicates that these compounds contribute to the antioxidant activity. The highest percentage of cell growth inhibition was observed when testing the water root extracts of H. multifidus (50.14%) and H. hercegovinus (49.04%). In contrast, the water leaf extract of H. hercegovinus exhibited the lowest inhibition of cell growth (8.59%), although it showed strong antioxidant activity.

We have examined antiproliferative, cytotoxic, and genotoxic potential of a halogenated boroxine dipotassium trioxohydroxytetrafluorotriborate [K2(B3O3F4OH)]. The impact on cell growth was evaluated by alamarBlue assay in basal cell carcinoma culture. Cytostatic, cytotoxic, and genotoxic potential were evaluated in lymphocytes culture, applying cytokinesis-block micronucleus cytome assay and chromosome aberrations analysis. Tested concentrations (0.05, 0.1, 0.2, and 0.4 mg/mL) were correlated with inhibition of cell growth in basal cell carcinoma culture and with the lymphocytes proliferation. Clastogenic activity has been confirmed, without evidences of aneugenic activity, in human lymphocytes.

Lilium martagon L. var. cattaniae Vis. (Liliaceae) is endemic plant of Dinaridi mountain. In this work we established protocol for fast in vitro propagation and multiplication of Lilium martagon var. cattaniae. The aim was to enable fast production of plant material as potential source of pharmaceutically valuable secondary metabolites. Seeds of L.martagon var. cattaniae were germinated on a Murashige and Skoog basal medium with a supplement of 0.15 mg/l gibberellic acid (GA3), and multiplication was performed on MS medium supplemented with 0.1 mg/l gibberellic acid (GA3), 0.2 mg/l indole-3-butyric acid (IBA) and 0.5 mg/l 6-ben- zylaminopurine (BAP). We used ultrasound assisted extraction to prepare extracts of leaves and bulbs of Lilium martagon var. cattaniae, which were evaluated for their genotoxic potential using Allium test and cytokinesis-block micronucleus test in human lymphocytes culture. There was statistically significant difference between all used concentrations of lilium extracts and control on proliferation of cells of root tip of onion (Allium cepa). In cytokinesis-block micronucleus test no statistically significant difference between frequencies of analyzed parameters in samples treated with tested concentrations of extracts and control was obtained.

Cytogenetic biomarkers, such as micronuclei in peripheral blood or oral mucosa, are widely used for evaluation of exposure to genotoxins or carcinogens. Tobacco is one of the strongest carcinogens, responsible for development of different types of cancers. The aim of this study was to assess the genotoxicity of cigarette consumption in young smokers and to correlate results of cytogenetic analysis in peripheral blood lymphocytes and exfoliated buccal cells. The study was conducted on samples taken from 43 smokers and 44 non-smokers, young individuals from Bosnia and Herzegovina. Significantly higher frequency of micronuclei in peripheral blood lymphocytes was observed in smokers (p < 0.05). No significant correlations were found for age, duration and intensity of smoking, and frequency of micronuclei in lymphocytes. Significantly higher frequency of degenerated (apoptotic) buccal cells was also revealed in smokers (p < 0.05). The frequency of apoptotic cells in smokers was significantly influenced by the age of participants (F = 8.649; p < 0.01) and duration of smoking (F = 5.389; p < 0.05). Results of cytogenetic analysis conducted in peripheral blood and exfoliated buccal cells are in significant positive correlation, indicating complementarities of those analyses.