TY  - CONF
AU  - Stevanović, Katarina
AU  - Pajić, Tanja
AU  - Krmpot, Aleksandar
AU  - Rabasović, Mihailo
AU  - Žižić, Milan
AU  - Živić, Miroslav
AU  - Todorović, Nataša
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6246
AB  - The success of patch-clamp giga-ohm seal formation on filamentous fungi Phycomyces blakesleeanus protoplasts was investigated to evaluate their usefulness in ion channel studies on filamentous fungi. Protoplasts were obtained by laser-mediated nanosurgery of the cell wall stained with Calcofluor White. To enable a successful seal formation, it is critical to prevent cell wall regeneration. Since wall integrity responses in fungi involve kinase-dependent pathways, we aimed to sufficiently reduce intracellular ATP availability using the respiratory inhibitor azide. The effect of azide on phosphate metabolites of Phycomyces blakesleeanus, as determined by 31P NMR spectroscopy, was a reduction in intracellular ATP accompanied by a decrease in long chain polyphosphates. Subsequently, all seal formation measurements were performed in the presence of azide, and protoplast viability was confirmed by cytoplasmic streaming. The success of seal formation depends on the size of the protoplasts, as larger protoplasts are more prone to successful seal formation. It was also found that the laser power used in nanosurgery could influence the success of seal formation, as higher values were associated with a lower success rate. The protoplasts released by laser nanosurgery produced by our newly developed method are a good model system for patch clamp on filamentous fungal membranes because their plasma membrane can easily form high-quality seals with the patch pipette.
PB  - Kragujevac: Institute for Information Technologies, University of Belgrade
C3  - Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia
T1  - Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts
DO  - 10.46793/ICCBI23.221S
SP  - 221
EP  - 224
ER  - 

@conference{
author = "Stevanović, Katarina and Pajić, Tanja and Krmpot, Aleksandar and Rabasović, Mihailo and Žižić, Milan and Živić, Miroslav and Todorović, Nataša",
year = "2023",
abstract = "The success of patch-clamp giga-ohm seal formation on filamentous fungi Phycomyces blakesleeanus protoplasts was investigated to evaluate their usefulness in ion channel studies on filamentous fungi. Protoplasts were obtained by laser-mediated nanosurgery of the cell wall stained with Calcofluor White. To enable a successful seal formation, it is critical to prevent cell wall regeneration. Since wall integrity responses in fungi involve kinase-dependent pathways, we aimed to sufficiently reduce intracellular ATP availability using the respiratory inhibitor azide. The effect of azide on phosphate metabolites of Phycomyces blakesleeanus, as determined by 31P NMR spectroscopy, was a reduction in intracellular ATP accompanied by a decrease in long chain polyphosphates. Subsequently, all seal formation measurements were performed in the presence of azide, and protoplast viability was confirmed by cytoplasmic streaming. The success of seal formation depends on the size of the protoplasts, as larger protoplasts are more prone to successful seal formation. It was also found that the laser power used in nanosurgery could influence the success of seal formation, as higher values were associated with a lower success rate. The protoplasts released by laser nanosurgery produced by our newly developed method are a good model system for patch clamp on filamentous fungal membranes because their plasma membrane can easily form high-quality seals with the patch pipette.",
publisher = "Kragujevac: Institute for Information Technologies, University of Belgrade",
journal = "Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia",
title = "Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts",
doi = "10.46793/ICCBI23.221S",
pages = "221-224"
}

Stevanović, K., Pajić, T., Krmpot, A., Rabasović, M., Žižić, M., Živić, M.,& Todorović, N.. (2023). Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts. in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia
Kragujevac: Institute for Information Technologies, University of Belgrade., 221-224.
https://doi.org/10.46793/ICCBI23.221S

Stevanović K, Pajić T, Krmpot A, Rabasović M, Žižić M, Živić M, Todorović N. Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts. in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia. 2023;:221-224.
doi:10.46793/ICCBI23.221S .

Stevanović, Katarina, Pajić, Tanja, Krmpot, Aleksandar, Rabasović, Mihailo, Žižić, Milan, Živić, Miroslav, Todorović, Nataša, "Patch clamp pipette giga seal forming success on the nanosurgery-obtained filamentous fungi protoplasts" in Book of Proceedings: 2nd International Conference on Chemo and Bioinformatics ICCBIKG 2023; 2023 Sep 28-29; Kragujevac, Serbia (2023):221-224,
https://doi.org/10.46793/ICCBI23.221S . .

TY  - CONF
AU  - Žižić, Milan
AU  - Stanić, Marina
AU  - Aquilanti, Giuliana
AU  - Cvetić Antić, Tijana
AU  - Todorović, Nataša
AU  - Živić, Miroslav
AU  - Zakrzewska, Joanna
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5733
AB  - In this study, mycelium of fungus Phycomyces blakesleeanus was exposed to soluble toxic form of selenium, selenite (Se+4), with the aim of determining the flow and products of its biotransformation. Selenite is reduced to Se0 in the form selenium nanoparticles (SeNPs) and Se-2 in the form of methylated volatile selenides. Low concentrations of Se+4 in the mycelium form RSSeSR type compounds, which could be a step in the process of SeNPs formation, or incorporation of Se into metabolites such as Se-amino acids.
PB  - Belgrade: Society of Physical Chemists of Serbia
C3  - Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia
T1  - Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus
SP  - 93
EP  - 96
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5733
ER  - 

@conference{
author = "Žižić, Milan and Stanić, Marina and Aquilanti, Giuliana and Cvetić Antić, Tijana and Todorović, Nataša and Živić, Miroslav and Zakrzewska, Joanna",
year = "2022",
abstract = "In this study, mycelium of fungus Phycomyces blakesleeanus was exposed to soluble toxic form of selenium, selenite (Se+4), with the aim of determining the flow and products of its biotransformation. Selenite is reduced to Se0 in the form selenium nanoparticles (SeNPs) and Se-2 in the form of methylated volatile selenides. Low concentrations of Se+4 in the mycelium form RSSeSR type compounds, which could be a step in the process of SeNPs formation, or incorporation of Se into metabolites such as Se-amino acids.",
publisher = "Belgrade: Society of Physical Chemists of Serbia",
journal = "Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia",
title = "Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus",
pages = "93-96",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5733"
}

Žižić, M., Stanić, M., Aquilanti, G., Cvetić Antić, T., Todorović, N., Živić, M.,& Zakrzewska, J.. (2022). Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus. in Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia
Belgrade: Society of Physical Chemists of Serbia., 93-96.
https://hdl.handle.net/21.15107/rcub_ibiss_5733

Žižić M, Stanić M, Aquilanti G, Cvetić Antić T, Todorović N, Živić M, Zakrzewska J. Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus. in Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia. 2022;:93-96.
https://hdl.handle.net/21.15107/rcub_ibiss_5733 .

Žižić, Milan, Stanić, Marina, Aquilanti, Giuliana, Cvetić Antić, Tijana, Todorović, Nataša, Živić, Miroslav, Zakrzewska, Joanna, "Selenite metabolism in the mycelium of the fungus Phycomyces blakesleeanus" in Proceedings: 16th International Conference on Fundamental and Applied Aspects of Physical Chemistry: Physical Chemistry 2022, Vol. 1; 2022 Sep 26-30; Belgrade, Serbia (2022):93-96,
https://hdl.handle.net/21.15107/rcub_ibiss_5733 .

TY  - JOUR
AU  - Stanić, Marin
AU  - Križak, Strahinja
AU  - Jovanović, Mirna
AU  - Pajić, Tanja
AU  - Ćirić, Ana
AU  - Žižić, Milan
AU  - Zakrzewska, Joanna
AU  - Cvetić Antić, Tijana
AU  - Todorović, Nataša
AU  - Živić, Miroslav
PY  - 2017
UR  - http://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.000429
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2730
AB  - Increasing resistance of fungal strains to known fungicides has prompted identification of new candidates for fungicides among substances previously used for other purposes. We have tested the effects of known anion channel inhibitors anthracene-9-carboxylic acid (A9C) and niflumic acid (NFA) on growth, energy metabolism and anionic current of mycelium of fungus Phycomyces blakesleeanus. Both inhibitors significantly decreased growth and respiration of mycelium, but complete inhibition was only achieved by 100 and 500 μM NFA for growth and respiration, respectively. A9C had no effect on respiration of human NCI-H460 cell line and very little effect on cucumber root sprout clippings, which nominates this inhibitor for further investigation as a potential new fungicide. Effects of A9C and NFA on respiration of isolated mitochondria of P. blakesleeanus were significantly smaller, which indicates that their inhibitory effect on respiration of mycelium is indirect. NMR spectroscopy showed that both A9C and NFA decrease the levels of ATP and polyphosphates in the mycelium of P. blakesleeanus, but only A9C caused intracellular acidification. Outwardly rectifying, fast inactivating instantaneous anionic current (ORIC) was also reduced to 33±5 and 21±3% of its pre-treatment size by A9C and NFA, respectively, but only in the absence of ATP. It can be assumed from our results that the regulation of ORIC is tightly linked to cellular energy metabolism in P. blakesleeanus, and the decrease in ATP and polyphosphate levels could be a direct cause of growth inhibition.
T2  - Microbiology
T1  - Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites
IS  - 3
VL  - 163
DO  - 10.1099/mic.0.000429
SP  - 364
EP  - 372
ER  - 

@article{
author = "Stanić, Marin and Križak, Strahinja and Jovanović, Mirna and Pajić, Tanja and Ćirić, Ana and Žižić, Milan and Zakrzewska, Joanna and Cvetić Antić, Tijana and Todorović, Nataša and Živić, Miroslav",
year = "2017",
abstract = "Increasing resistance of fungal strains to known fungicides has prompted identification of new candidates for fungicides among substances previously used for other purposes. We have tested the effects of known anion channel inhibitors anthracene-9-carboxylic acid (A9C) and niflumic acid (NFA) on growth, energy metabolism and anionic current of mycelium of fungus Phycomyces blakesleeanus. Both inhibitors significantly decreased growth and respiration of mycelium, but complete inhibition was only achieved by 100 and 500 μM NFA for growth and respiration, respectively. A9C had no effect on respiration of human NCI-H460 cell line and very little effect on cucumber root sprout clippings, which nominates this inhibitor for further investigation as a potential new fungicide. Effects of A9C and NFA on respiration of isolated mitochondria of P. blakesleeanus were significantly smaller, which indicates that their inhibitory effect on respiration of mycelium is indirect. NMR spectroscopy showed that both A9C and NFA decrease the levels of ATP and polyphosphates in the mycelium of P. blakesleeanus, but only A9C caused intracellular acidification. Outwardly rectifying, fast inactivating instantaneous anionic current (ORIC) was also reduced to 33±5 and 21±3% of its pre-treatment size by A9C and NFA, respectively, but only in the absence of ATP. It can be assumed from our results that the regulation of ORIC is tightly linked to cellular energy metabolism in P. blakesleeanus, and the decrease in ATP and polyphosphate levels could be a direct cause of growth inhibition.",
journal = "Microbiology",
title = "Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites",
number = "3",
volume = "163",
doi = "10.1099/mic.0.000429",
pages = "364-372"
}

Stanić, M., Križak, S., Jovanović, M., Pajić, T., Ćirić, A., Žižić, M., Zakrzewska, J., Cvetić Antić, T., Todorović, N.,& Živić, M.. (2017). Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites. in Microbiology, 163(3), 364-372.
https://doi.org/10.1099/mic.0.000429

Stanić M, Križak S, Jovanović M, Pajić T, Ćirić A, Žižić M, Zakrzewska J, Cvetić Antić T, Todorović N, Živić M. Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites. in Microbiology. 2017;163(3):364-372.
doi:10.1099/mic.0.000429 .

Stanić, Marin, Križak, Strahinja, Jovanović, Mirna, Pajić, Tanja, Ćirić, Ana, Žižić, Milan, Zakrzewska, Joanna, Cvetić Antić, Tijana, Todorović, Nataša, Živić, Miroslav, "Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites" in Microbiology, 163, no. 3 (2017):364-372,
https://doi.org/10.1099/mic.0.000429 . .