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  - Pajić, Tanja
AU  - Stevanović, Katarina
AU  - Todorović, Nataša
AU  - Lević, Steva
AU  - Savić Šević, Svetlana
AU  - Pantelić, Dejan
AU  - Živić, Miroslav
AU  - Rabasović, Mihailo D
AU  - Krmpot, Aleksandar J
PY  - 2023
UR  - https://www.eventclass.org/contxt_emim2023/online-program/session?s=PW36#e609
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6289
AB  - Introduction
Electrophysiology studies of ion channels, in live filamentous fungi by patch clamp method are notpossible due to presence of rigid chitinous cell wall that prevents patch clamp pipette to access theplasma membrane. We present laser nano-surgery of the fungal cell wall that enables patch clampelectrophysiology studies. Similar approaches as one-time reports utilizing nanosecond laser pulseslong time ago were not pursued further [1,2]. Here, we demonstrate reproducible method usingfemtosecond lasers accompanied by two-photon excitation fluorescence (TPEF) imaging of hyphae.
Methods
A wild-type strain of filamentous fungus
Phycomyces blakesleeanus
(Burgeff ) [NRRL 1555(-)] were grownon glass coverslips with hand-etched grid, coated with a thin layer of 50% collagen type I as animmobilizer. Home built nonlinear laser scanning microscope [3,4] utilizing Ti:Sa tunable fs laser wasused for TPEF imaging of hyphae and the cell surgery. The latter is enabled with the custom made add-on in soft ware. Coverslip with hyphae is transferred to another microscope setup for patch clamp,consisting of micromanipulators and precise electronics for pA current measurements. The surgicalincisions and released protoplasts were additionally imaged by scanning electron microscopy for whichtreated hyphae had to undergo critical point drying procedure.
Results/Discussion
Hyphae were stained by Calcofluor White and treated with an exocytosis inhibitor (brefeldin A) and arespiration inhibitor (sodium azide) to prevent cell wall regeneration. Since the cell wall and the plasmamembrane are in the close contact [4] hyphae were kept in hyperosmotic solution to retract thecytoplasm from the cell wall. Surgical spot-wise pattern was precisely positioned at TPEF image ofselected hypha at the place where the plasma membrane was retracted. The dwell time (1s) and thelaser power (4-15mW) were set with fixed repetition rate (76MHz), pulse duration (160fs) and laserwavelength (730nm). Upon the surgery, hyphae were gently deplasmolysed. A protoplast with plasmamembrane accessible for the patch clamp pipette was released through the surgical incision (Fig 1). The>GΩ seal resistance was achieved. Numerous ion channels are recorded in diff erent configurations (oncell, inside-out, whole cell and out-out) (Fig 2).
Conclusions
The whole process (cell surgery + patch clamping) is rather complex and specific steps have to be strictlyfollowed for high success rate and reproducibility. Also, chemicals concentrations, solutions osmolarity,timing and cutting parameters have to be kept in the specified narrow range. Obtained currentrecordings provide valuable information on fungal cell membrane ionic channels.
PB  - European Society for Molecular Imaging
C3  - European Molecular Imaging Meeting: 18th Annual Meeting of the European Society for Molecular Imaging: EMIM 2023; 2023 Mar 14-17; Saltzburg, Austria
T1  - Laser nano-surgery of fungal cell wall to enable patch clamping
SP  - 1095
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6289
ER  - 

@conference{
author = "Pajić, Tanja and Stevanović, Katarina and Todorović, Nataša and Lević, Steva and Savić Šević, Svetlana and Pantelić, Dejan and Živić, Miroslav and Rabasović, Mihailo D and Krmpot, Aleksandar J",
year = "2023",
abstract = "Introduction
Electrophysiology studies of ion channels, in live filamentous fungi by patch clamp method are notpossible due to presence of rigid chitinous cell wall that prevents patch clamp pipette to access theplasma membrane. We present laser nano-surgery of the fungal cell wall that enables patch clampelectrophysiology studies. Similar approaches as one-time reports utilizing nanosecond laser pulseslong time ago were not pursued further [1,2]. Here, we demonstrate reproducible method usingfemtosecond lasers accompanied by two-photon excitation fluorescence (TPEF) imaging of hyphae.
Methods
A wild-type strain of filamentous fungus
Phycomyces blakesleeanus
(Burgeff ) [NRRL 1555(-)] were grownon glass coverslips with hand-etched grid, coated with a thin layer of 50% collagen type I as animmobilizer. Home built nonlinear laser scanning microscope [3,4] utilizing Ti:Sa tunable fs laser wasused for TPEF imaging of hyphae and the cell surgery. The latter is enabled with the custom made add-on in soft ware. Coverslip with hyphae is transferred to another microscope setup for patch clamp,consisting of micromanipulators and precise electronics for pA current measurements. The surgicalincisions and released protoplasts were additionally imaged by scanning electron microscopy for whichtreated hyphae had to undergo critical point drying procedure.
Results/Discussion
Hyphae were stained by Calcofluor White and treated with an exocytosis inhibitor (brefeldin A) and arespiration inhibitor (sodium azide) to prevent cell wall regeneration. Since the cell wall and the plasmamembrane are in the close contact [4] hyphae were kept in hyperosmotic solution to retract thecytoplasm from the cell wall. Surgical spot-wise pattern was precisely positioned at TPEF image ofselected hypha at the place where the plasma membrane was retracted. The dwell time (1s) and thelaser power (4-15mW) were set with fixed repetition rate (76MHz), pulse duration (160fs) and laserwavelength (730nm). Upon the surgery, hyphae were gently deplasmolysed. A protoplast with plasmamembrane accessible for the patch clamp pipette was released through the surgical incision (Fig 1). The>GΩ seal resistance was achieved. Numerous ion channels are recorded in diff erent configurations (oncell, inside-out, whole cell and out-out) (Fig 2).
Conclusions
The whole process (cell surgery + patch clamping) is rather complex and specific steps have to be strictlyfollowed for high success rate and reproducibility. Also, chemicals concentrations, solutions osmolarity,timing and cutting parameters have to be kept in the specified narrow range. Obtained currentrecordings provide valuable information on fungal cell membrane ionic channels.",
publisher = "European Society for Molecular Imaging",
journal = "European Molecular Imaging Meeting: 18th Annual Meeting of the European Society for Molecular Imaging: EMIM 2023; 2023 Mar 14-17; Saltzburg, Austria",
title = "Laser nano-surgery of fungal cell wall to enable patch clamping",
pages = "1095",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6289"
}

Pajić, T., Stevanović, K., Todorović, N., Lević, S., Savić Šević, S., Pantelić, D., Živić, M., Rabasović, M. D.,& Krmpot, A. J.. (2023). Laser nano-surgery of fungal cell wall to enable patch clamping. in European Molecular Imaging Meeting: 18th Annual Meeting of the European Society for Molecular Imaging: EMIM 2023; 2023 Mar 14-17; Saltzburg, Austria
European Society for Molecular Imaging., 1095.
https://hdl.handle.net/21.15107/rcub_ibiss_6289

Pajić T, Stevanović K, Todorović N, Lević S, Savić Šević S, Pantelić D, Živić M, Rabasović MD, Krmpot AJ. Laser nano-surgery of fungal cell wall to enable patch clamping. in European Molecular Imaging Meeting: 18th Annual Meeting of the European Society for Molecular Imaging: EMIM 2023; 2023 Mar 14-17; Saltzburg, Austria. 2023;:1095.
https://hdl.handle.net/21.15107/rcub_ibiss_6289 .

Pajić, Tanja, Stevanović, Katarina, Todorović, Nataša, Lević, Steva, Savić Šević, Svetlana, Pantelić, Dejan, Živić, Miroslav, Rabasović, Mihailo D, Krmpot, Aleksandar J, "Laser nano-surgery of fungal cell wall to enable patch clamping" in European Molecular Imaging Meeting: 18th Annual Meeting of the European Society for Molecular Imaging: EMIM 2023; 2023 Mar 14-17; Saltzburg, Austria (2023):1095,
https://hdl.handle.net/21.15107/rcub_ibiss_6289 .

TY  - JOUR
AU  - Stevanović, Katarina
AU  - Čepkenović, Bogdana
AU  - Križak, Strahinja
AU  - Živić, Miroslav
AU  - Todorović, Nataša
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5978
AB  - Studies of ion currents in filamentous fungi are a prerequisite for forming a complete
understanding of their physiology. Cytoplasmic droplets (CDs), obtained from sporangiophores of
Phycomyces blakesleeanus, are a model system that enables the characterization of ion currents in
the native membrane, including the currents mediated by the channels not yet molecularly identified.
Osmotically activated anionic current with outward rectification (ORIC) is a dominant current in
the membrane of cytoplasmic droplets under the conditions of hypoosmotic stimulation. We have
previously reported remarkable functional similarities of ORIC with the vertebrate volume regulated
anionic current (VRAC), such as dose-dependent activation by osmotic difference, ion selectivity
sequence, and time and voltage dependent profile of the current. Using the patch clamp method on
the CD membrane, we further resolve VRAC-like ORIC characteristics in this paper. We examine
the inhibition by extracellular ATP and carbenoxolone, the permeation of glutamate in presence of
chloride, selectivity for nitrates, and activation by GTP, and we show its single channel behavior
in excised membrane. We propose that ORIC is a functional counterpart of vertebrate VRAC in
filamentous fungi, possibly with a similar essential role in anion efflux during cell volume regulation.
PB  - Basel: MDPI
T2  - Journal of Fungi
T1  - Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current
IS  - 6
VL  - 9
DO  - 10.3390/jof9060637
SP  - 637
ER  - 

@article{
author = "Stevanović, Katarina and Čepkenović, Bogdana and Križak, Strahinja and Živić, Miroslav and Todorović, Nataša",
year = "2023",
abstract = "Studies of ion currents in filamentous fungi are a prerequisite for forming a complete
understanding of their physiology. Cytoplasmic droplets (CDs), obtained from sporangiophores of
Phycomyces blakesleeanus, are a model system that enables the characterization of ion currents in
the native membrane, including the currents mediated by the channels not yet molecularly identified.
Osmotically activated anionic current with outward rectification (ORIC) is a dominant current in
the membrane of cytoplasmic droplets under the conditions of hypoosmotic stimulation. We have
previously reported remarkable functional similarities of ORIC with the vertebrate volume regulated
anionic current (VRAC), such as dose-dependent activation by osmotic difference, ion selectivity
sequence, and time and voltage dependent profile of the current. Using the patch clamp method on
the CD membrane, we further resolve VRAC-like ORIC characteristics in this paper. We examine
the inhibition by extracellular ATP and carbenoxolone, the permeation of glutamate in presence of
chloride, selectivity for nitrates, and activation by GTP, and we show its single channel behavior
in excised membrane. We propose that ORIC is a functional counterpart of vertebrate VRAC in
filamentous fungi, possibly with a similar essential role in anion efflux during cell volume regulation.",
publisher = "Basel: MDPI",
journal = "Journal of Fungi",
title = "Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current",
number = "6",
volume = "9",
doi = "10.3390/jof9060637",
pages = "637"
}

Stevanović, K., Čepkenović, B., Križak, S., Živić, M.,& Todorović, N.. (2023). Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current. in Journal of Fungi
Basel: MDPI., 9(6), 637.
https://doi.org/10.3390/jof9060637

Stevanović K, Čepkenović B, Križak S, Živić M, Todorović N. Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current. in Journal of Fungi. 2023;9(6):637.
doi:10.3390/jof9060637 .

Stevanović, Katarina, Čepkenović, Bogdana, Križak, Strahinja, Živić, Miroslav, Todorović, Nataša, "Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current" in Journal of Fungi, 9, no. 6 (2023):637,
https://doi.org/10.3390/jof9060637 . .

TY  - JOUR
AU  - Pajić, Tanja
AU  - Todorović, Nataša
AU  - Živić, Miroslav
AU  - Nikolić, Stanko N
AU  - Rabasović, Mihailo D
AU  - Clayton, Andrew H A
AU  - Krmpot, Aleksandar J
PY  - 2022
UR  - http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=PMC9637149
UR  - https://www.nature.com/articles/s41598-022-23502-4
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5235
AB  - We report the utilization of Third-Harmonic Generation microscopy for label-free live cell imaging of lipid droplets in the hypha of filamentous fungus Phycomyces blakesleeanus . THG microscopy images showed bright spherical features dispersed throughout the hypha cytoplasm in control conditions and a transient increase in the number of bright features after complete nitrogen starvation. Colocalization analysis of THG and lipid-counterstained images disclosed that the cytoplasmic particles were lipid droplets. Particle Size Analysis and Image Correlation Spectroscopy were used to quantify the number density and size of lipid droplets. The two analysis methods both revealed an increase from 16 × 10 −3 to 23 × 10 −3 lipid droplets/µm 2 after nitrogen starvation and a decrease in the average size of the droplets (range: 0.5–0.8 µm diameter). In conclusion, THG imaging, followed by PSA and ICS, can be reliably used for filamentous fungi for the in vivo quantification of lipid droplets without the need for labeling and/or fixation. In addition, it has been demonstrated that ICS is suitable for THG microscopy.
PB  - Berlin: Nature Portfolio
T2  - Scientific Reports
T1  - Label-free third harmonic generation imaging and quantification of lipid droplets in live filamentous fungi
IS  - 1
VL  - 12
DO  - 10.1038/s41598-022-23502-4
SP  - 18760
ER  - 

@article{
author = "Pajić, Tanja and Todorović, Nataša and Živić, Miroslav and Nikolić, Stanko N and Rabasović, Mihailo D and Clayton, Andrew H A and Krmpot, Aleksandar J",
year = "2022",
abstract = "We report the utilization of Third-Harmonic Generation microscopy for label-free live cell imaging of lipid droplets in the hypha of filamentous fungus Phycomyces blakesleeanus . THG microscopy images showed bright spherical features dispersed throughout the hypha cytoplasm in control conditions and a transient increase in the number of bright features after complete nitrogen starvation. Colocalization analysis of THG and lipid-counterstained images disclosed that the cytoplasmic particles were lipid droplets. Particle Size Analysis and Image Correlation Spectroscopy were used to quantify the number density and size of lipid droplets. The two analysis methods both revealed an increase from 16 × 10 −3 to 23 × 10 −3 lipid droplets/µm 2 after nitrogen starvation and a decrease in the average size of the droplets (range: 0.5–0.8 µm diameter). In conclusion, THG imaging, followed by PSA and ICS, can be reliably used for filamentous fungi for the in vivo quantification of lipid droplets without the need for labeling and/or fixation. In addition, it has been demonstrated that ICS is suitable for THG microscopy.",
publisher = "Berlin: Nature Portfolio",
journal = "Scientific Reports",
title = "Label-free third harmonic generation imaging and quantification of lipid droplets in live filamentous fungi",
number = "1",
volume = "12",
doi = "10.1038/s41598-022-23502-4",
pages = "18760"
}

Pajić, T., Todorović, N., Živić, M., Nikolić, S. N., Rabasović, M. D., Clayton, A. H. A.,& Krmpot, A. J.. (2022). Label-free third harmonic generation imaging and quantification of lipid droplets in live filamentous fungi. in Scientific Reports
Berlin: Nature Portfolio., 12(1), 18760.
https://doi.org/10.1038/s41598-022-23502-4

Pajić T, Todorović N, Živić M, Nikolić SN, Rabasović MD, Clayton AHA, Krmpot AJ. Label-free third harmonic generation imaging and quantification of lipid droplets in live filamentous fungi. in Scientific Reports. 2022;12(1):18760.
doi:10.1038/s41598-022-23502-4 .

Pajić, Tanja, Todorović, Nataša, Živić, Miroslav, Nikolić, Stanko N, Rabasović, Mihailo D, Clayton, Andrew H A, Krmpot, Aleksandar J, "Label-free third harmonic generation imaging and quantification of lipid droplets in live filamentous fungi" in Scientific Reports, 12, no. 1 (2022):18760,
https://doi.org/10.1038/s41598-022-23502-4 . .

TY  - JOUR
AU  - Vranković, Jelena
AU  - Božanić, Milenka
AU  - Živić, Miroslav
AU  - Marković, Zoran
AU  - Marjanović, Stefan
AU  - Golubović, Vukosav
AU  - Živić, Ivana
PY  - 2022
UR  - https://linkinghub.elsevier.com/retrieve/pii/S2352513422003830
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5176
AB  - A 102-day feeding trial was conducted to evaluate the effects of four different fish feeding patterns on carp pond water quality and antioxidant biomarkers [superoxide dismutase (SOD), Catalase (CAT), Glutathione peroxidase (GPx), Glutathione reductase (GR), Glutathione S-transferase (GST) and Thiol groups (SH)], protein content and biomass of the midge Chironomus plumosus. Farmed fish were fed two commercial diets: Soprofish 25/7 Standard (containing 25% protein and 7 % fat) and Soprofish 32/7 Profi Effect (containing 32 % protein and 7 % fat). These feeds were combined during a feeding trial in four different feeding patterns, designated as A, B, C and D. In feeding pattern A, the fish received Soprofish 25/7 Standard throughout the experiment and in feeding pattern D, Soprofish 32/7 Profi Effect. During feeding patterns B and C, a mixed feeding pattern was used, alternating between lower and higher protein diets. The study revealed no significant effects (P > 0.05) of the feeding regimens on water quality, so their effects on C. plumosus larvae can be evaluated independently. The activities of the chironomid enzymes CAT and GR showed a clear statistically significant dependence on the feeding pattern (P < 0.05), which increased with increasing protein content. On the other hand, the activity of SOD as well as larval biomass were noticeably affected by water quality, the former increasing with its deterioration and the latter with its improvement. The activities of GPx and GST were correlated with each other, but no statistically significant relationship was found with any of the measured abiotic factors (P > 0.05). Overall, our results suggest that long-term supplementation with a highly concentrated protein diet may have preventive effects against oxidative stress and support the use of C. plumosus as a model for assessing the effects of organic pollution on pond zoobenthic fauna.
PB  - Elsevier B.V.
T2  - Aquaculture Reports
T1  - Antioxidant biomarker profile of chironomid larvae from carp ponds: Evaluation of the effects of different fish feeding patterns
VL  - 27
DO  - 10.1016/j.aqrep.2022.101387
SP  - 101387
ER  - 

@article{
author = "Vranković, Jelena and Božanić, Milenka and Živić, Miroslav and Marković, Zoran and Marjanović, Stefan and Golubović, Vukosav and Živić, Ivana",
year = "2022",
abstract = "A 102-day feeding trial was conducted to evaluate the effects of four different fish feeding patterns on carp pond water quality and antioxidant biomarkers [superoxide dismutase (SOD), Catalase (CAT), Glutathione peroxidase (GPx), Glutathione reductase (GR), Glutathione S-transferase (GST) and Thiol groups (SH)], protein content and biomass of the midge Chironomus plumosus. Farmed fish were fed two commercial diets: Soprofish 25/7 Standard (containing 25% protein and 7 % fat) and Soprofish 32/7 Profi Effect (containing 32 % protein and 7 % fat). These feeds were combined during a feeding trial in four different feeding patterns, designated as A, B, C and D. In feeding pattern A, the fish received Soprofish 25/7 Standard throughout the experiment and in feeding pattern D, Soprofish 32/7 Profi Effect. During feeding patterns B and C, a mixed feeding pattern was used, alternating between lower and higher protein diets. The study revealed no significant effects (P > 0.05) of the feeding regimens on water quality, so their effects on C. plumosus larvae can be evaluated independently. The activities of the chironomid enzymes CAT and GR showed a clear statistically significant dependence on the feeding pattern (P < 0.05), which increased with increasing protein content. On the other hand, the activity of SOD as well as larval biomass were noticeably affected by water quality, the former increasing with its deterioration and the latter with its improvement. The activities of GPx and GST were correlated with each other, but no statistically significant relationship was found with any of the measured abiotic factors (P > 0.05). Overall, our results suggest that long-term supplementation with a highly concentrated protein diet may have preventive effects against oxidative stress and support the use of C. plumosus as a model for assessing the effects of organic pollution on pond zoobenthic fauna.",
publisher = "Elsevier B.V.",
journal = "Aquaculture Reports",
title = "Antioxidant biomarker profile of chironomid larvae from carp ponds: Evaluation of the effects of different fish feeding patterns",
volume = "27",
doi = "10.1016/j.aqrep.2022.101387",
pages = "101387"
}

Vranković, J., Božanić, M., Živić, M., Marković, Z., Marjanović, S., Golubović, V.,& Živić, I.. (2022). Antioxidant biomarker profile of chironomid larvae from carp ponds: Evaluation of the effects of different fish feeding patterns. in Aquaculture Reports
Elsevier B.V.., 27, 101387.
https://doi.org/10.1016/j.aqrep.2022.101387

Vranković J, Božanić M, Živić M, Marković Z, Marjanović S, Golubović V, Živić I. Antioxidant biomarker profile of chironomid larvae from carp ponds: Evaluation of the effects of different fish feeding patterns. in Aquaculture Reports. 2022;27:101387.
doi:10.1016/j.aqrep.2022.101387 .

Vranković, Jelena, Božanić, Milenka, Živić, Miroslav, Marković, Zoran, Marjanović, Stefan, Golubović, Vukosav, Živić, Ivana, "Antioxidant biomarker profile of chironomid larvae from carp ponds: Evaluation of the effects of different fish feeding patterns" in Aquaculture Reports, 27 (2022):101387,
https://doi.org/10.1016/j.aqrep.2022.101387 . .

TY  - CONF
AU  - Stevanović, Katarina
AU  - Pajić, Tanja
AU  - Živić, Miroslav
AU  - Krmpot, Aleksandar
AU  - Rabasović, Mihailo
AU  - Todorović, Nataša
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5742
AB  - Prisustvo i aktivni karakter ćelijskog zida koji prekriva plazma membranu filamentoznih gljiva su do sad bili nesavladive prepreke da joj se pristupi elektrofiziološkim metodama na način koji bi omogućio kvalitetno registrovanje jonskih struja. Nedavno, mi smo razvili i optimizovali protokol koji omogućava kvalitetno snimanje struja, metodom nametnute voltaže na deliću membrane, sa plazma membrane protoplasta gljive oslobođenih od zida subćelijskom mikrohirurgijom korišćenjem femtosekundnog Ti:Sa lasera.1,2 Među registrovanim plazmamembranskim strujama protoplasta gljive dominantne su struje nošene različitim anjonskim vrstama. Na osnovu potencijala reverzije u asimetričnim jonskim uslovima identifikovani tipovi jonskih struja su, u najvećem broju slučajeva, bili slabo (42%) ili izrazito (35%) selektivni za hlor u prisustvu K, NO3, glutamatnog jona i visokih koncentracija Ca+2. Među nekoliko različitih struja slabije selektivnosti za hlor, daleko najzastupljenija je struja, konduktivnosti 21 ± 1 pS, prisutna u 42% svih kontakata registracije. Analiza aktivnosti pojedinačnih kanala ove, najzastupljenije, struje pokazuje da verovatnoća otvorenosti kanala slabo raste sa depolarizacijom membrane (Po(-80) = 0,3; Po(50) = 0,45) i opada na više depolarisanim vrednostima (Po(80) = 0,2). Učestalost paketića aktivnosti je najveća na hiperpolarisanim potencijalima (f(-80 mV) > 200 Hz) i dvostruko manja na depolarisanim (f(80 mV) < 100 Hz).
AB  - Присуство и активни карактер ћелијског зида који прекрива плазма мембрану
филаментозних гљива су до сад били несавладиве препреке да јој се приступи
електрофизиолошким методама на начин који би омогућио квалитетно
регистровање јонских струја. Недавно, ми смо развили и оптимизовали протокол
који омогућава квалитетно снимање струја, методом наметнуте волтаже на делићу
мембране, са плазма мембране протопласта гљиве ослобођених од зида
субћелијском микрохирургијом коришћењем фемтосекундног Ti:Sa ласера.1,2 Међу
регистрованим плазмамембранским струјама протопласта гљиве доминантне су
струје ношене различитим анјонским врстама. На основу потенцијала реверзије у
асиметричним јонским условима идентификовани типови јонских струја су, у
највећем броју случајева, били слабо (42%) или изразито (35%) селективни за хлор
у присуству K, NO3, глутаматног јона и високих концентрација Ca+2. Међу
неколико различитих струја слабије селективности за хлор, далеко најзаступљенија
је струја, кондуктивности 21 ± 1 pS, присутна у 42% свих контаката регистрације.
Анализа активности појединачних канала ове, најзаступљеније, струје показује да
вероватноћа отворености канала слабо расте са деполаризацијом мембране (Po(-80) =
0,3; Po(50) = 0,45) и опада на више деполарисаним вредностима (Po(80) = 0,2).
Учесталост пакетића активности је највећа на хиперполарисаним потенцијалима
(f(-80 mV) > 200 Hz) и двоструко мања на деполарисаним (f(80 mV) < 100 Hz).
PB  - Belgrade: Serbian Biological Society
C3  - Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
T1  - Dominantne struje plazma membrane gljive Phycomyces blakesleeanus registrovane sa protoplasta dobijenih mikrohirurgijom ćelijskog zida femtosekundnim laserom
T1  - Доминантне струје плазма мембране гљиве Phycomyces blakesleeanus регистроване са протопласта добијених микрохирургијом ћелијског зида фемтосекундним ласером
SP  - 24
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5742
ER  - 

@conference{
author = "Stevanović, Katarina and Pajić, Tanja and Živić, Miroslav and Krmpot, Aleksandar and Rabasović, Mihailo and Todorović, Nataša",
year = "2022",
abstract = "Prisustvo i aktivni karakter ćelijskog zida koji prekriva plazma membranu filamentoznih gljiva su do sad bili nesavladive prepreke da joj se pristupi elektrofiziološkim metodama na način koji bi omogućio kvalitetno registrovanje jonskih struja. Nedavno, mi smo razvili i optimizovali protokol koji omogućava kvalitetno snimanje struja, metodom nametnute voltaže na deliću membrane, sa plazma membrane protoplasta gljive oslobođenih od zida subćelijskom mikrohirurgijom korišćenjem femtosekundnog Ti:Sa lasera.1,2 Među registrovanim plazmamembranskim strujama protoplasta gljive dominantne su struje nošene različitim anjonskim vrstama. Na osnovu potencijala reverzije u asimetričnim jonskim uslovima identifikovani tipovi jonskih struja su, u najvećem broju slučajeva, bili slabo (42%) ili izrazito (35%) selektivni za hlor u prisustvu K, NO3, glutamatnog jona i visokih koncentracija Ca+2. Među nekoliko različitih struja slabije selektivnosti za hlor, daleko najzastupljenija je struja, konduktivnosti 21 ± 1 pS, prisutna u 42% svih kontakata registracije. Analiza aktivnosti pojedinačnih kanala ove, najzastupljenije, struje pokazuje da verovatnoća otvorenosti kanala slabo raste sa depolarizacijom membrane (Po(-80) = 0,3; Po(50) = 0,45) i opada na više depolarisanim vrednostima (Po(80) = 0,2). Učestalost paketića aktivnosti je najveća na hiperpolarisanim potencijalima (f(-80 mV) > 200 Hz) i dvostruko manja na depolarisanim (f(80 mV) < 100 Hz)., Присуство и активни карактер ћелијског зида који прекрива плазма мембрану
филаментозних гљива су до сад били несавладиве препреке да јој се приступи
електрофизиолошким методама на начин који би омогућио квалитетно
регистровање јонских струја. Недавно, ми смо развили и оптимизовали протокол
који омогућава квалитетно снимање струја, методом наметнуте волтаже на делићу
мембране, са плазма мембране протопласта гљиве ослобођених од зида
субћелијском микрохирургијом коришћењем фемтосекундног Ti:Sa ласера.1,2 Међу
регистрованим плазмамембранским струјама протопласта гљиве доминантне су
струје ношене различитим анјонским врстама. На основу потенцијала реверзије у
асиметричним јонским условима идентификовани типови јонских струја су, у
највећем броју случајева, били слабо (42%) или изразито (35%) селективни за хлор
у присуству K, NO3, глутаматног јона и високих концентрација Ca+2. Међу
неколико различитих струја слабије селективности за хлор, далеко најзаступљенија
је струја, кондуктивности 21 ± 1 pS, присутна у 42% свих контаката регистрације.
Анализа активности појединачних канала ове, најзаступљеније, струје показује да
вероватноћа отворености канала слабо расте са деполаризацијом мембране (Po(-80) =
0,3; Po(50) = 0,45) и опада на више деполарисаним вредностима (Po(80) = 0,2).
Учесталост пакетића активности је највећа на хиперполарисаним потенцијалима
(f(-80 mV) > 200 Hz) и двоструко мања на деполарисаним (f(80 mV) < 100 Hz).",
publisher = "Belgrade: Serbian Biological Society",
journal = "Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia",
title = "Dominantne struje plazma membrane gljive Phycomyces blakesleeanus registrovane sa protoplasta dobijenih mikrohirurgijom ćelijskog zida femtosekundnim laserom, Доминантне струје плазма мембране гљиве Phycomyces blakesleeanus регистроване са протопласта добијених микрохирургијом ћелијског зида фемтосекундним ласером",
pages = "24",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5742"
}

Stevanović, K., Pajić, T., Živić, M., Krmpot, A., Rabasović, M.,& Todorović, N.. (2022). Dominantne struje plazma membrane gljive Phycomyces blakesleeanus registrovane sa protoplasta dobijenih mikrohirurgijom ćelijskog zida femtosekundnim laserom. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
Belgrade: Serbian Biological Society., 24.
https://hdl.handle.net/21.15107/rcub_ibiss_5742

Stevanović K, Pajić T, Živić M, Krmpot A, Rabasović M, Todorović N. Dominantne struje plazma membrane gljive Phycomyces blakesleeanus registrovane sa protoplasta dobijenih mikrohirurgijom ćelijskog zida femtosekundnim laserom. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia. 2022;:24.
https://hdl.handle.net/21.15107/rcub_ibiss_5742 .

Stevanović, Katarina, Pajić, Tanja, Živić, Miroslav, Krmpot, Aleksandar, Rabasović, Mihailo, Todorović, Nataša, "Dominantne struje plazma membrane gljive Phycomyces blakesleeanus registrovane sa protoplasta dobijenih mikrohirurgijom ćelijskog zida femtosekundnim laserom" in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia (2022):24,
https://hdl.handle.net/21.15107/rcub_ibiss_5742 .

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  - CONF
AU  - Pajić, Tanja
AU  - Kozakijević, Suzana
AU  - Lukičić, Jovana
AU  - Živić, Miroslav
AU  - Krmpot, Aleksandar
AU  - Rabasović, Mihailo
AU  - Todorović, Nataša
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5744
AB  - Biološka aktivnost selena zavisi od njegove doze i hemijskog oblika. U mikrotragovima povećava antioksidativni kapacitet, dok je u višim koncentracijama toksičan zbog stvaranja reaktivnih vrsta kiseonika i potencijalnog oštećenja mitohondrijalne membrane. Končaste gljive imaju sposobnost koncentrisanja selena u micelijumu i predstavljaju jedan od glavnih puteva za njegov ulazak u ekosisteme. U ovom radu je ispitivan uticaj hroničnog i akutnog dejstva natrijum selenita (uobičajen oblik selena u ishrani) na morfologiju i zastupljenost mitohondrija u živim ćelijama končaste gljive Phycomyces blakesleeanus tokom eksponencijalne faze rasta (EFR). Za in vivo oslikavanje morfologije mitohondrija i njihove dinamike u odgovoru na različite koncentracije selenita korišćena je multifotonska mikroskopija koja omogućava trodimenzionalno oslikavanje u visokoj rezoluciji i smanjeno fotooštećenje i fotoizbeljivanje uzorka zahvaljujući upotrebi infracrvenih ultrabrzih impulsnih lasera. Hronični tretmani selenitom su doveli do uočljivog trenda smanjenja zastupljenosti mitohondrija sa povećanjem koncentracije selenita. Izloženost hifa selenitu, tokom akutnih i hroničnih tretmana, rezultiralo je povećanjem zastupljenosti tubularne morfologije mitohondrija, koja je u akutnom tretmanu većim koncentracijama selenita (0,75 i 1 mM) bila povećana za 50% u odnosu na kontrolu. Kod mlađih hifa EFR izloženih hroničnom delovanju selenita (0,5 do 1 mM) tubularne mitohondrije su bile višestruko duže od onih u starijoj populaciji.
AB  - Биолошка активност селена зависи од његове дозе и хемијског облика. У
микротраговима повећава антиоксидативни капацитет, док је у вишим
концентрацијама токсичан због стварања реактивних врста кисеоника и
потенцијалног оштећења митохондријалне мембране. Кончасте гљиве имају
способност концентрисања селена у мицелијуму и представљају један од главних
путева за његов улазак у екосистеме. У овом раду је испитиван утицај хроничног и
акутног дејства натријум селенита (уобичајен облик селена у исхрани) на
морфологију и заступљеност митохондрија у живим ћелијама кончасте гљиве
Phycomyces blakesleeanus током експоненцијалне фазе раста (ЕФР). За in vivo
осликавање морфологије митохондрија и њихове динамике у одговору на
различите концентрације селенита коришћена је мултифотонска микроскопија која
омогућава тродимензионално осликавање у високој резолуцији и смањено
фотооштећење и фотоизбељивање узорка захваљујући употреби инфрацрвених
ултрабрзих импулсних ласера. Хронични третмани селенитом су довели до
уочљивог тренда смањења заступљености митохондрија са повећањем
концентрације селенита. Изложеност хифа селениту, током акутних и хроничних
третмана, резултирало је повећањем заступљености тубуларне морфологије
митохондрија, која је у акутном третману већим концентрацијама селенита (0,75 и
1 mМ) била повећана за 50% у односу на контролу. Код млађих хифа ЕФР
изложених хроничном деловању селенита (0,5 до 1 mМ) тубуларне митохондрије
су биле вишеструко дуже од оних у старијој популацији.
PB  - Belgrade: Serbian Biological Society
C3  - Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
T1  - In vivo oslikavanje pojedinačnih hifa Phycomyces blakesleeanus metodom multifotonske mikroskopije: uticaj selenita na morfologiju i funkciju mitohondrija
T1  - In vivo осликавање појединачних хифа Phycomyces blakesleeanus методом мултифотонске микроскопије: утицај селенита на морфологију и функцију митохондрија
SP  - 28
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5744
ER  - 

@conference{
author = "Pajić, Tanja and Kozakijević, Suzana and Lukičić, Jovana and Živić, Miroslav and Krmpot, Aleksandar and Rabasović, Mihailo and Todorović, Nataša",
year = "2022",
abstract = "Biološka aktivnost selena zavisi od njegove doze i hemijskog oblika. U mikrotragovima povećava antioksidativni kapacitet, dok je u višim koncentracijama toksičan zbog stvaranja reaktivnih vrsta kiseonika i potencijalnog oštećenja mitohondrijalne membrane. Končaste gljive imaju sposobnost koncentrisanja selena u micelijumu i predstavljaju jedan od glavnih puteva za njegov ulazak u ekosisteme. U ovom radu je ispitivan uticaj hroničnog i akutnog dejstva natrijum selenita (uobičajen oblik selena u ishrani) na morfologiju i zastupljenost mitohondrija u živim ćelijama končaste gljive Phycomyces blakesleeanus tokom eksponencijalne faze rasta (EFR). Za in vivo oslikavanje morfologije mitohondrija i njihove dinamike u odgovoru na različite koncentracije selenita korišćena je multifotonska mikroskopija koja omogućava trodimenzionalno oslikavanje u visokoj rezoluciji i smanjeno fotooštećenje i fotoizbeljivanje uzorka zahvaljujući upotrebi infracrvenih ultrabrzih impulsnih lasera. Hronični tretmani selenitom su doveli do uočljivog trenda smanjenja zastupljenosti mitohondrija sa povećanjem koncentracije selenita. Izloženost hifa selenitu, tokom akutnih i hroničnih tretmana, rezultiralo je povećanjem zastupljenosti tubularne morfologije mitohondrija, koja je u akutnom tretmanu većim koncentracijama selenita (0,75 i 1 mM) bila povećana za 50% u odnosu na kontrolu. Kod mlađih hifa EFR izloženih hroničnom delovanju selenita (0,5 do 1 mM) tubularne mitohondrije su bile višestruko duže od onih u starijoj populaciji., Биолошка активност селена зависи од његове дозе и хемијског облика. У
микротраговима повећава антиоксидативни капацитет, док је у вишим
концентрацијама токсичан због стварања реактивних врста кисеоника и
потенцијалног оштећења митохондријалне мембране. Кончасте гљиве имају
способност концентрисања селена у мицелијуму и представљају један од главних
путева за његов улазак у екосистеме. У овом раду је испитиван утицај хроничног и
акутног дејства натријум селенита (уобичајен облик селена у исхрани) на
морфологију и заступљеност митохондрија у живим ћелијама кончасте гљиве
Phycomyces blakesleeanus током експоненцијалне фазе раста (ЕФР). За in vivo
осликавање морфологије митохондрија и њихове динамике у одговору на
различите концентрације селенита коришћена је мултифотонска микроскопија која
омогућава тродимензионално осликавање у високој резолуцији и смањено
фотооштећење и фотоизбељивање узорка захваљујући употреби инфрацрвених
ултрабрзих импулсних ласера. Хронични третмани селенитом су довели до
уочљивог тренда смањења заступљености митохондрија са повећањем
концентрације селенита. Изложеност хифа селениту, током акутних и хроничних
третмана, резултирало је повећањем заступљености тубуларне морфологије
митохондрија, која је у акутном третману већим концентрацијама селенита (0,75 и
1 mМ) била повећана за 50% у односу на контролу. Код млађих хифа ЕФР
изложених хроничном деловању селенита (0,5 до 1 mМ) тубуларне митохондрије
су биле вишеструко дуже од оних у старијој популацији.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia",
title = "In vivo oslikavanje pojedinačnih hifa Phycomyces blakesleeanus metodom multifotonske mikroskopije: uticaj selenita na morfologiju i funkciju mitohondrija, In vivo осликавање појединачних хифа Phycomyces blakesleeanus методом мултифотонске микроскопије: утицај селенита на морфологију и функцију митохондрија",
pages = "28",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5744"
}

Pajić, T., Kozakijević, S., Lukičić, J., Živić, M., Krmpot, A., Rabasović, M.,& Todorović, N.. (2022). In vivo oslikavanje pojedinačnih hifa Phycomyces blakesleeanus metodom multifotonske mikroskopije: uticaj selenita na morfologiju i funkciju mitohondrija. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
Belgrade: Serbian Biological Society., 28.
https://hdl.handle.net/21.15107/rcub_ibiss_5744

Pajić T, Kozakijević S, Lukičić J, Živić M, Krmpot A, Rabasović M, Todorović N. In vivo oslikavanje pojedinačnih hifa Phycomyces blakesleeanus metodom multifotonske mikroskopije: uticaj selenita na morfologiju i funkciju mitohondrija. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia. 2022;:28.
https://hdl.handle.net/21.15107/rcub_ibiss_5744 .

Pajić, Tanja, Kozakijević, Suzana, Lukičić, Jovana, Živić, Miroslav, Krmpot, Aleksandar, Rabasović, Mihailo, Todorović, Nataša, "In vivo oslikavanje pojedinačnih hifa Phycomyces blakesleeanus metodom multifotonske mikroskopije: uticaj selenita na morfologiju i funkciju mitohondrija" in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia (2022):28,
https://hdl.handle.net/21.15107/rcub_ibiss_5744 .

TY  - CONF
AU  - Bošnjaković, Tamara
AU  - Todorović, Nataša
AU  - Milošević, Milena
AU  - Živić, Miroslav
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5743
AB  - Prihvaćeni model lučenja insulina, po stimulaciji glukozom β ćelija pankreasa, čine: (1) transport glukoze u ćeliju; (2) KATP kanali čija blokada ATP-om dovodi do depolarizacije membrane; (3) voltažno-zavisni kalcijumski kanali koji se aktiviraju depolarizacijom, dozvoljavajući ulazak Ca2+; (4) egzocitoza insulinskih granula pokrenuta Ca2+ jonima.1 Pošto za dostizanje praga aktivacije nije dovoljno samo zatvaranje KATP kanala, jasno je da je uključena i aktivnost drugih jonskih kanala. Ima podataka da je ovaj proces posredovan kanalom VRAC (Volume-Regulated Anion Channel) koji provodi jone hlora i osmolite i aktivira se u hipotoničnoj sredini.2 Mi smo koristili metodu nametnute voltaže na deliću membrane uz oslikavanje signala Ca2+ indikatora, kako bi ispitali prisustvo i ulogu VRAC u električnoj aktivnosti RIN – 5f ćelija, u cilju utvrđivanja potencijalnih ograničenja i optimizacije eksperimentalnog pristupa izučavanja strujnog odgovora β ćelija. U prisustvu cezijuma (Cs+ pip), kojim se blokiraju K kanali, snimljene su familije struja u izotoničnim i hipotoničnim uslovima, kao i spontana i depolarizacijom indukovana aktivnost (akcioni potencijali i depolarizacija u odgovoru na stimulaciju glukozom). Testiran je efekat jedinog postojećeg farmakološkog alata za blokadu VRAC, DCPIB na membranske struje i na Ca2+ odgovor ćelija. Rezultati ukazuju da: (1) u RIN – 5f ćelijama nema inaktivirajućeg oblika VRAC i da hipotonični uslovi, u odsustvu glukoze, ne dovode do aktivacije struja; (2) u izotoničnim uslovima, DCPIB pored očekivanog delimičnog bloka struja ima i stimulativni efekat na druge izlazne struje; (3) preliminarna oslikavanja Ca2+ signala ukazuju da DCPIB inhibira a njegovo uklanjanje povećava aktivnost u prisustvu 0-15 mM glukoze.
AB  - Прихваћени модел лучења инсулина, по стимулацији глукозом β ћелија панкреаса,
чине: (1) транспорт глукозе у ћелију; (2) KATP канали чија блокада ATP-ом доводи
до деполаризације мембране; (3) волтажно-зависни калцијумски канали који се
активирају деполаризацијом, дозвољавајући улазак Ca2+; (4) егзоцитоза
инсулинских гранула покренута Ca2+ јонима.1 Пошто за достизање прага
активације није довољно само затварање KATP канала, јасно је да је укључена и
активност других јонских канала. Има података да је овај процес посредован
каналом VRAC (Volume-Regulated Anion Channel) који проводи јоне хлора и
осмолите и активира се у хипотоничној средини.2 Ми смо користили методу
наметнуте волтаже на делићу мембране уз осликавање сигнала Ca2+ индикатора,
како би испитали присуство и улогу VRAC у електричној активности RIN – 5f
ћелија, у циљу утврђивања потенцијалних ограничења и оптимизације
експерименталног приступа изучавања струјног одговора β ћелија. У присуству
цезијума (Cs+pip), којим се блокирају K канали, снимљене су фамилије струја у
изотоничним и хипотоничним условима, као и спонтана и деполаризацијом
индукована активност (акциони потенцијали и деполаризација у одговору на
стимулацију глукозом). Тестиран је ефекат јединог постојећег фармаколошког
алата за блокаду VRAC, DCPIB на мембранске струје и на Ca2+ одговор ћелија.
Резултати указују да: (1) у RIN – 5f ћелијама нема инактивирајућег облика VRAC и
да хипотонични услови, у одсуству глукозе, не доводе до активације струја; (2) у
изотоничним условима, DCPIB поред очекиваног делимичног блока струја има и
стимулативни ефекат на друге излазне струје; (3) прелиминарна осликавања Ca2+
сигнала указују да DCPIB инхибира а његово уклањање повећава активност у
присуству 0-15 mM глукозе.
PB  - Belgrade: Serbian Biological Society
C3  - Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
T1  - Elektrofiziološka karakterizacija β ćelija pankreasa na ćelijskoj liniji RIN – 5f
T1  - Електрофизиолошка карактеризација β ћелија панкреаса на ћелијској линији RIN – 5f
SP  - 27
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5743
ER  - 

@conference{
author = "Bošnjaković, Tamara and Todorović, Nataša and Milošević, Milena and Živić, Miroslav",
year = "2022",
abstract = "Prihvaćeni model lučenja insulina, po stimulaciji glukozom β ćelija pankreasa, čine: (1) transport glukoze u ćeliju; (2) KATP kanali čija blokada ATP-om dovodi do depolarizacije membrane; (3) voltažno-zavisni kalcijumski kanali koji se aktiviraju depolarizacijom, dozvoljavajući ulazak Ca2+; (4) egzocitoza insulinskih granula pokrenuta Ca2+ jonima.1 Pošto za dostizanje praga aktivacije nije dovoljno samo zatvaranje KATP kanala, jasno je da je uključena i aktivnost drugih jonskih kanala. Ima podataka da je ovaj proces posredovan kanalom VRAC (Volume-Regulated Anion Channel) koji provodi jone hlora i osmolite i aktivira se u hipotoničnoj sredini.2 Mi smo koristili metodu nametnute voltaže na deliću membrane uz oslikavanje signala Ca2+ indikatora, kako bi ispitali prisustvo i ulogu VRAC u električnoj aktivnosti RIN – 5f ćelija, u cilju utvrđivanja potencijalnih ograničenja i optimizacije eksperimentalnog pristupa izučavanja strujnog odgovora β ćelija. U prisustvu cezijuma (Cs+ pip), kojim se blokiraju K kanali, snimljene su familije struja u izotoničnim i hipotoničnim uslovima, kao i spontana i depolarizacijom indukovana aktivnost (akcioni potencijali i depolarizacija u odgovoru na stimulaciju glukozom). Testiran je efekat jedinog postojećeg farmakološkog alata za blokadu VRAC, DCPIB na membranske struje i na Ca2+ odgovor ćelija. Rezultati ukazuju da: (1) u RIN – 5f ćelijama nema inaktivirajućeg oblika VRAC i da hipotonični uslovi, u odsustvu glukoze, ne dovode do aktivacije struja; (2) u izotoničnim uslovima, DCPIB pored očekivanog delimičnog bloka struja ima i stimulativni efekat na druge izlazne struje; (3) preliminarna oslikavanja Ca2+ signala ukazuju da DCPIB inhibira a njegovo uklanjanje povećava aktivnost u prisustvu 0-15 mM glukoze., Прихваћени модел лучења инсулина, по стимулацији глукозом β ћелија панкреаса,
чине: (1) транспорт глукозе у ћелију; (2) KATP канали чија блокада ATP-ом доводи
до деполаризације мембране; (3) волтажно-зависни калцијумски канали који се
активирају деполаризацијом, дозвољавајући улазак Ca2+; (4) егзоцитоза
инсулинских гранула покренута Ca2+ јонима.1 Пошто за достизање прага
активације није довољно само затварање KATP канала, јасно је да је укључена и
активност других јонских канала. Има података да је овај процес посредован
каналом VRAC (Volume-Regulated Anion Channel) који проводи јоне хлора и
осмолите и активира се у хипотоничној средини.2 Ми смо користили методу
наметнуте волтаже на делићу мембране уз осликавање сигнала Ca2+ индикатора,
како би испитали присуство и улогу VRAC у електричној активности RIN – 5f
ћелија, у циљу утврђивања потенцијалних ограничења и оптимизације
експерименталног приступа изучавања струјног одговора β ћелија. У присуству
цезијума (Cs+pip), којим се блокирају K канали, снимљене су фамилије струја у
изотоничним и хипотоничним условима, као и спонтана и деполаризацијом
индукована активност (акциони потенцијали и деполаризација у одговору на
стимулацију глукозом). Тестиран је ефекат јединог постојећег фармаколошког
алата за блокаду VRAC, DCPIB на мембранске струје и на Ca2+ одговор ћелија.
Резултати указују да: (1) у RIN – 5f ћелијама нема инактивирајућег облика VRAC и
да хипотонични услови, у одсуству глукозе, не доводе до активације струја; (2) у
изотоничним условима, DCPIB поред очекиваног делимичног блока струја има и
стимулативни ефекат на друге излазне струје; (3) прелиминарна осликавања Ca2+
сигнала указују да DCPIB инхибира а његово уклањање повећава активност у
присуству 0-15 mM глукозе.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia",
title = "Elektrofiziološka karakterizacija β ćelija pankreasa na ćelijskoj liniji RIN – 5f, Електрофизиолошка карактеризација β ћелија панкреаса на ћелијској линији RIN – 5f",
pages = "27",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5743"
}

Bošnjaković, T., Todorović, N., Milošević, M.,& Živić, M.. (2022). Elektrofiziološka karakterizacija β ćelija pankreasa na ćelijskoj liniji RIN – 5f. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
Belgrade: Serbian Biological Society., 27.
https://hdl.handle.net/21.15107/rcub_ibiss_5743

Bošnjaković T, Todorović N, Milošević M, Živić M. Elektrofiziološka karakterizacija β ćelija pankreasa na ćelijskoj liniji RIN – 5f. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia. 2022;:27.
https://hdl.handle.net/21.15107/rcub_ibiss_5743 .

Bošnjaković, Tamara, Todorović, Nataša, Milošević, Milena, Živić, Miroslav, "Elektrofiziološka karakterizacija β ćelija pankreasa na ćelijskoj liniji RIN – 5f" in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia (2022):27,
https://hdl.handle.net/21.15107/rcub_ibiss_5743 .

TY  - CONF
AU  - Stevanović, Katarina
AU  - Živić, Miroslav
AU  - Todorović, Nataša
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5741
AB  - Izlazno-rektifikujuća inaktivirajuća struja (IRIS) je dominantna anjonska struja u uslovima osmotskog stresa na membrani poreklom iz sporangiofora gljive Phycomyces blakesleeanus.1 Jedna od njenih upečatljivih karakteristika je obrazac inaktivacije na depolarišućim potencijalima.1 Sa gubitkom adenozin trifosfata (ATF) sa unutarćelijske strane tokom dijalize pipetnim rastvorom, inaktivacija se vremenom ubrzava i dovodi do nepovratnog pada amplitude ukupne struje, dok se sa ATF-om prisutnim u pipetnom rastvoru amplituda struje i konstanta inaktivacije τ održavaju u vremenu.2 Naši najskoriji rezultati ukazuju da je, u prisustvu unutarćelijskog ATF-a oporavak od inaktivacije izazvane depolarizacijom voltažno- i vremenski-zavisan: (1) izlaganje hiperpolarizaciji je neophodno za dostizanje oporavka ≥ 90%; (2) procenat oporavka pri minimalnom vremenu odmora raste sa dubinom hiperpolarizacije; (3) brzina eksponencijalnog pada amplitude pri skraćenju vremena oporavka opada sa dubinom hiperpolarizacije. U uslovima smanjene koncentracije ATF-a koja je postignuta preinkubacijom natrijum-azidom, već u prvom minutu po ulasku u konfiguraciju „cela ćelija“ uočena je ubrzana inaktivacija.2 Dalja analiza voltažne-zavisnosti τ inaktivacije IRIS u različitim uslovima jasno pokazuje pomeraj ka manjim vrednostima τ sa opadanjem očekivane koncentracije ATF-a dok je voltažna zavisnost neznatno izmenjena. IRIS je identifikovana i u istrgnutoj konfiguraciji („spolja spolja“), pri čemu je obrazac oporavka od inaktivacije poslužio kao dodatna potvrda da su registrovana pojedinačna otvaranja upravo kanali u osnovi IRIS.
AB  - Излазно-ректификујућа инактивирајућа струја (ИРИС) је доминантна анјонска
струја у условима осмотског стреса на мембрани пореклом из спорангиофора
гљиве Phycomyces blakesleeanus.1 Једна од њених упечатљивих карактеристика је
образац инактивације на деполаришућим потенцијалима.1 Са губитком аденозин
трифосфата (АТФ) са унутарћелијске стране током дијализе пипетним раствором,
инактивација се временом убрзава и доводи до неповратног пада амплитуде укупне
струје, док се са АТФ-ом присутним у пипетном раствору амплитуда струје и
константа инактивације τ одржавају у времену.2 Наши најскорији резултати указују
да је, у присуству унутарћелијског АТФ-а опоравак од инактивације изазване
деполаризацијом волтажно- и временски-зависан: (1) излагање хиперполаризацији
је неопходно за достизање опоравка ≥ 90%; (2) проценат опоравка при минималном
времену одмора расте са дубином хиперполаризације; (3) брзина експоненцијалног
пада амплитуде при скраћењу времена опоравка опада са дубином
хиперполаризације. У условима смањене концентрације АТФ-а која је постигнута
преинкубацијом натријум-азидом, већ у првом минуту по уласку у конфигурацију
„цела ћелија“ уочена је убрзана инактивација.2 Даља анализа волтажне-зависности
τ инактивације ИРИС у различитим условима јасно показује померај ка мањим
вредностима τ са опадањем очекиване концентрације АТФ-а док је волтажна
зависност незнатно измењена. ИРИС је идентификована и у истргнутој
конфигурацији („споља споља“), при чему је образац опоравка од инактивације
послужио као додатна потврда да су регистрована појединачна отварања управо
канали у основи ИРИС.
PB  - Belgrade: Serbian Biological Society
C3  - Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
T1  - Biofizički profil procesa inaktivacije izlazno-rektifikujuće inaktivirajuće struje gljive Phycomyces blakesleeanus
T1  - Биофизички профил процеса инактивације излазно-ректификујуће инактивирајуће струје гљиве Phycomyces blakesleeanus
SP  - 23
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5741
ER  - 

@conference{
author = "Stevanović, Katarina and Živić, Miroslav and Todorović, Nataša",
year = "2022",
abstract = "Izlazno-rektifikujuća inaktivirajuća struja (IRIS) je dominantna anjonska struja u uslovima osmotskog stresa na membrani poreklom iz sporangiofora gljive Phycomyces blakesleeanus.1 Jedna od njenih upečatljivih karakteristika je obrazac inaktivacije na depolarišućim potencijalima.1 Sa gubitkom adenozin trifosfata (ATF) sa unutarćelijske strane tokom dijalize pipetnim rastvorom, inaktivacija se vremenom ubrzava i dovodi do nepovratnog pada amplitude ukupne struje, dok se sa ATF-om prisutnim u pipetnom rastvoru amplituda struje i konstanta inaktivacije τ održavaju u vremenu.2 Naši najskoriji rezultati ukazuju da je, u prisustvu unutarćelijskog ATF-a oporavak od inaktivacije izazvane depolarizacijom voltažno- i vremenski-zavisan: (1) izlaganje hiperpolarizaciji je neophodno za dostizanje oporavka ≥ 90%; (2) procenat oporavka pri minimalnom vremenu odmora raste sa dubinom hiperpolarizacije; (3) brzina eksponencijalnog pada amplitude pri skraćenju vremena oporavka opada sa dubinom hiperpolarizacije. U uslovima smanjene koncentracije ATF-a koja je postignuta preinkubacijom natrijum-azidom, već u prvom minutu po ulasku u konfiguraciju „cela ćelija“ uočena je ubrzana inaktivacija.2 Dalja analiza voltažne-zavisnosti τ inaktivacije IRIS u različitim uslovima jasno pokazuje pomeraj ka manjim vrednostima τ sa opadanjem očekivane koncentracije ATF-a dok je voltažna zavisnost neznatno izmenjena. IRIS je identifikovana i u istrgnutoj konfiguraciji („spolja spolja“), pri čemu je obrazac oporavka od inaktivacije poslužio kao dodatna potvrda da su registrovana pojedinačna otvaranja upravo kanali u osnovi IRIS., Излазно-ректификујућа инактивирајућа струја (ИРИС) је доминантна анјонска
струја у условима осмотског стреса на мембрани пореклом из спорангиофора
гљиве Phycomyces blakesleeanus.1 Једна од њених упечатљивих карактеристика је
образац инактивације на деполаришућим потенцијалима.1 Са губитком аденозин
трифосфата (АТФ) са унутарћелијске стране током дијализе пипетним раствором,
инактивација се временом убрзава и доводи до неповратног пада амплитуде укупне
струје, док се са АТФ-ом присутним у пипетном раствору амплитуда струје и
константа инактивације τ одржавају у времену.2 Наши најскорији резултати указују
да је, у присуству унутарћелијског АТФ-а опоравак од инактивације изазване
деполаризацијом волтажно- и временски-зависан: (1) излагање хиперполаризацији
је неопходно за достизање опоравка ≥ 90%; (2) проценат опоравка при минималном
времену одмора расте са дубином хиперполаризације; (3) брзина експоненцијалног
пада амплитуде при скраћењу времена опоравка опада са дубином
хиперполаризације. У условима смањене концентрације АТФ-а која је постигнута
преинкубацијом натријум-азидом, већ у првом минуту по уласку у конфигурацију
„цела ћелија“ уочена је убрзана инактивација.2 Даља анализа волтажне-зависности
τ инактивације ИРИС у различитим условима јасно показује померај ка мањим
вредностима τ са опадањем очекиване концентрације АТФ-а док је волтажна
зависност незнатно измењена. ИРИС је идентификована и у истргнутој
конфигурацији („споља споља“), при чему је образац опоравка од инактивације
послужио као додатна потврда да су регистрована појединачна отварања управо
канали у основи ИРИС.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia",
title = "Biofizički profil procesa inaktivacije izlazno-rektifikujuće inaktivirajuće struje gljive Phycomyces blakesleeanus, Биофизички профил процеса инактивације излазно-ректификујуће инактивирајуће струје гљиве Phycomyces blakesleeanus",
pages = "23",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5741"
}

Stevanović, K., Živić, M.,& Todorović, N.. (2022). Biofizički profil procesa inaktivacije izlazno-rektifikujuće inaktivirajuće struje gljive Phycomyces blakesleeanus. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
Belgrade: Serbian Biological Society., 23.
https://hdl.handle.net/21.15107/rcub_ibiss_5741

Stevanović K, Živić M, Todorović N. Biofizički profil procesa inaktivacije izlazno-rektifikujuće inaktivirajuće struje gljive Phycomyces blakesleeanus. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia. 2022;:23.
https://hdl.handle.net/21.15107/rcub_ibiss_5741 .

Stevanović, Katarina, Živić, Miroslav, Todorović, Nataša, "Biofizički profil procesa inaktivacije izlazno-rektifikujuće inaktivirajuće struje gljive Phycomyces blakesleeanus" in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia (2022):23,
https://hdl.handle.net/21.15107/rcub_ibiss_5741 .

TY  - CONF
AU  - Pajić, Tanja
AU  - Krmpot, Aleksandar J.
AU  - Todorović, Nataša
AU  - Živić, Miroslav
AU  - Stevanović, Katarina
AU  - Rabasović, Mihailo D.
PY  - 2021
UR  - https://iupab2020.sbbq.org.br/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4719
AB  - Nonlinear laser scanning microscopy (NLSM), is an advanced optical technique that utilizes ultrashort laser pulses 
for structural and functional imaging, as well as laser manipulation of live organisms and cells. Two modalities of 
NLSM, two photon excitation fluorescence (TPEF) and third harmonic generation (THG) were applied for in vivo and 
label-free study of oxidative and lipid metabolism of individual cells of filamentous fungus Phycomyces 
blakesleeanus. Cell membranes and lipid droplets (LDs) are major sources of THG signal. TPEF allows us to 
determine the redox ratio (reflecting metabolic activity of cells) of the metabolic cofactors FAD and NAD(P)H 
autofluorescence. In addition, slight modifications of the experimental setup, mostly on software, enabled utilization 
of femtosecond laser pulses for precise cell microsurgery of hyphal cell wall. The optimized microsurgery procedure 
we than utilized to obtain protoplasts suitable for patch-clamp electrophysiological recording. Cell surgery of 
filamentous fungus Phycomyces blakesleeanus, were performed by ultrafast Ti:Sa laser (160 fs pulses). The same 
laser was used for in vivo autoTPEF imaging of NAD(P)H and FAD at different wavelengths. For in vivo THG imaging 
of label-free hyphae, we used 1040 nm, 200 fs pulses from Yb KGW laser. In vivo and label-free application of THG 
imaging enabled, accurately and reliably, detection of changes in distribution, total number, and size of LDs in control 
and treatment group of cells. Two-photon microscopy made it possible to obtain a redox ratio using autofluorescences 
of NAD(P)H and FAD in the same regions of live hyphae. The cell microsurgery procedure has been optimized and 
developed, which enabled the subsequent registration of currents on otherwise unaccessible membrane.
PB  - São Paulo: Sociedade Brasileira de Bioquímica e Biologia Molecular
C3  - 20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq
T1  - In vivo metabolic imaging and micromanipulation of individual filamentous fungus cells using different nonlinear laser scanning microscopy modalities
SP  - 358
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4719
ER  - 

@conference{
author = "Pajić, Tanja and Krmpot, Aleksandar J. and Todorović, Nataša and Živić, Miroslav and Stevanović, Katarina and Rabasović, Mihailo D.",
year = "2021",
abstract = "Nonlinear laser scanning microscopy (NLSM), is an advanced optical technique that utilizes ultrashort laser pulses 
for structural and functional imaging, as well as laser manipulation of live organisms and cells. Two modalities of 
NLSM, two photon excitation fluorescence (TPEF) and third harmonic generation (THG) were applied for in vivo and 
label-free study of oxidative and lipid metabolism of individual cells of filamentous fungus Phycomyces 
blakesleeanus. Cell membranes and lipid droplets (LDs) are major sources of THG signal. TPEF allows us to 
determine the redox ratio (reflecting metabolic activity of cells) of the metabolic cofactors FAD and NAD(P)H 
autofluorescence. In addition, slight modifications of the experimental setup, mostly on software, enabled utilization 
of femtosecond laser pulses for precise cell microsurgery of hyphal cell wall. The optimized microsurgery procedure 
we than utilized to obtain protoplasts suitable for patch-clamp electrophysiological recording. Cell surgery of 
filamentous fungus Phycomyces blakesleeanus, were performed by ultrafast Ti:Sa laser (160 fs pulses). The same 
laser was used for in vivo autoTPEF imaging of NAD(P)H and FAD at different wavelengths. For in vivo THG imaging 
of label-free hyphae, we used 1040 nm, 200 fs pulses from Yb KGW laser. In vivo and label-free application of THG 
imaging enabled, accurately and reliably, detection of changes in distribution, total number, and size of LDs in control 
and treatment group of cells. Two-photon microscopy made it possible to obtain a redox ratio using autofluorescences 
of NAD(P)H and FAD in the same regions of live hyphae. The cell microsurgery procedure has been optimized and 
developed, which enabled the subsequent registration of currents on otherwise unaccessible membrane.",
publisher = "São Paulo: Sociedade Brasileira de Bioquímica e Biologia Molecular",
journal = "20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq",
title = "In vivo metabolic imaging and micromanipulation of individual filamentous fungus cells using different nonlinear laser scanning microscopy modalities",
pages = "358",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4719"
}

Pajić, T., Krmpot, A. J., Todorović, N., Živić, M., Stevanović, K.,& Rabasović, M. D.. (2021). In vivo metabolic imaging and micromanipulation of individual filamentous fungus cells using different nonlinear laser scanning microscopy modalities. in 20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq
São Paulo: Sociedade Brasileira de Bioquímica e Biologia Molecular., 358.
https://hdl.handle.net/21.15107/rcub_ibiss_4719

Pajić T, Krmpot AJ, Todorović N, Živić M, Stevanović K, Rabasović MD. In vivo metabolic imaging and micromanipulation of individual filamentous fungus cells using different nonlinear laser scanning microscopy modalities. in 20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq. 2021;:358.
https://hdl.handle.net/21.15107/rcub_ibiss_4719 .

Pajić, Tanja, Krmpot, Aleksandar J., Todorović, Nataša, Živić, Miroslav, Stevanović, Katarina, Rabasović, Mihailo D., "In vivo metabolic imaging and micromanipulation of individual filamentous fungus cells using different nonlinear laser scanning microscopy modalities" in 20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq (2021):358,
https://hdl.handle.net/21.15107/rcub_ibiss_4719 .

TY  - CONF
AU  - Stevanović, Katarina
AU  - Živić, Miroslav
AU  - Todorović, Nataša
PY  - 2021
UR  - https://iupab2020.sbbq.org.br/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4710
AB  - Outwardly rectifying, inactivating current (ORIC), recorded in cytoplasmatic droplets of P.blakesleeanus is 
characterized by a mild outward rectification, selectivity for anions, and a characteristic inactivation on 
depolarizing potentials. In absence of ATP, ORIC has a fast rundown with increasing speed of inactivation, while both processes are delayed by ATPpip (for at least 8min) and the non-hydrolysable AM-PCP (minimum of 5min). Extracellularly applied, ATP has a blocking effect. Flavonoid querzetin leads to a gradual loss of current despite the presence of intracellular ATP. Our research group has shown azide to be the potent respiration inhibitor of P. blakesleeanus, leaving the cells viable but significantly depleted from ATP. We are presenting data recorded in whole cell configuration, showing that: 1.ORIC can be activated despite the depletion of ATP in the cytoplasmatic droplet. However, its inactivation profile is significantly different comparing to normal conditions with or without ATP in the pipette in the first minute of recording, with reduced τ on lower depolarizing potentials. The fraction of current blocked by querzetin with AM-PCP in pipette solution is greater comparing to experiments with ATPpip, even with the 5-fold increase of AM-PCPpip concentration. The time required for current to recover from depolarizing steps is shorter at more hyperpolarized potentials. We have also started to look for the first insights into ORIC single channel activity in out-out configuration, and recorded, upon stimulation of ORIC in whole cell, and patch excision, the step-like pattern of inactivation that strongly resembles one of ORIC. Based on current data, we can conclude that ATP has a significant role in the inactivation process, with a more potent effect comparing to AM-PCP, whether by the means of greater affinity for the binding site or an additional mechanism involving kinase activity.
PB  - São Paulo: Sociedade Brasileira de Bioquímica e Biologia Molecular
C3  - 20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq
T1  - Inactivation properties of ORIC, VRAC-like current of filamentous fungus P. blakesleeanus: the role of ATP and the first glimpse of the single channel behavior
SP  - 143
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4710
ER  - 

@conference{
author = "Stevanović, Katarina and Živić, Miroslav and Todorović, Nataša",
year = "2021",
abstract = "Outwardly rectifying, inactivating current (ORIC), recorded in cytoplasmatic droplets of P.blakesleeanus is 
characterized by a mild outward rectification, selectivity for anions, and a characteristic inactivation on 
depolarizing potentials. In absence of ATP, ORIC has a fast rundown with increasing speed of inactivation, while both processes are delayed by ATPpip (for at least 8min) and the non-hydrolysable AM-PCP (minimum of 5min). Extracellularly applied, ATP has a blocking effect. Flavonoid querzetin leads to a gradual loss of current despite the presence of intracellular ATP. Our research group has shown azide to be the potent respiration inhibitor of P. blakesleeanus, leaving the cells viable but significantly depleted from ATP. We are presenting data recorded in whole cell configuration, showing that: 1.ORIC can be activated despite the depletion of ATP in the cytoplasmatic droplet. However, its inactivation profile is significantly different comparing to normal conditions with or without ATP in the pipette in the first minute of recording, with reduced τ on lower depolarizing potentials. The fraction of current blocked by querzetin with AM-PCP in pipette solution is greater comparing to experiments with ATPpip, even with the 5-fold increase of AM-PCPpip concentration. The time required for current to recover from depolarizing steps is shorter at more hyperpolarized potentials. We have also started to look for the first insights into ORIC single channel activity in out-out configuration, and recorded, upon stimulation of ORIC in whole cell, and patch excision, the step-like pattern of inactivation that strongly resembles one of ORIC. Based on current data, we can conclude that ATP has a significant role in the inactivation process, with a more potent effect comparing to AM-PCP, whether by the means of greater affinity for the binding site or an additional mechanism involving kinase activity.",
publisher = "São Paulo: Sociedade Brasileira de Bioquímica e Biologia Molecular",
journal = "20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq",
title = "Inactivation properties of ORIC, VRAC-like current of filamentous fungus P. blakesleeanus: the role of ATP and the first glimpse of the single channel behavior",
pages = "143",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4710"
}

Stevanović, K., Živić, M.,& Todorović, N.. (2021). Inactivation properties of ORIC, VRAC-like current of filamentous fungus P. blakesleeanus: the role of ATP and the first glimpse of the single channel behavior. in 20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq
São Paulo: Sociedade Brasileira de Bioquímica e Biologia Molecular., 143.
https://hdl.handle.net/21.15107/rcub_ibiss_4710

Stevanović K, Živić M, Todorović N. Inactivation properties of ORIC, VRAC-like current of filamentous fungus P. blakesleeanus: the role of ATP and the first glimpse of the single channel behavior. in 20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq. 2021;:143.
https://hdl.handle.net/21.15107/rcub_ibiss_4710 .

Stevanović, Katarina, Živić, Miroslav, Todorović, Nataša, "Inactivation properties of ORIC, VRAC-like current of filamentous fungus P. blakesleeanus: the role of ATP and the first glimpse of the single channel behavior" in 20th IUPAB Congress, 45th SBBf Congress and 50th Annual Meeting of SBBq (2021):143,
https://hdl.handle.net/21.15107/rcub_ibiss_4710 .

TY  - CONF
AU  - Pajić, Tanja
AU  - Todorović, Nataša
AU  - Živić, Miroslav
AU  - Rabasović, Mihailo D.
AU  - Clayton, Andrew H. A.
AU  - Krmpot, Aleksandar J.
PY  - 2021
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4712
AB  - Oleaginous fungi can accumulate significant amounts of lipids in their mycelium (up to 80% of their biomass), primarily in the form of lipid droplets (LDs). LDs have optical properties that differ from the surrounding aquatic environment, which causes sudden changes in the refractive index. Here, we present in vivo and label-free imaging of individual hyphae of the oleaginous filamentous fungus 
Phycomyces blakesleeanus by Third Harmonic Generation (THG) microscopy method, where LDs 
are the main source of contrast. The LDs quantification from THG images was performed by two image analysis techniques: Image Correlation Spectroscopy (ICS) and software particle counting – Particle Size Analysis (PSA). ICS measures the spatial variation of fluorescence intensity fluctuations in the images, which can then be related to particle density and aggregation state. In order to test and compare the two methods, we used hyphae that undergo nitrogen starvation, which is known to cause alterations in lipid metabolism and the increase in LDs number. For in vivo THG imaging of label-free, > 24-hour old hyphae, we used 1040 nm, 200 fs pulses from Yb KGW laser. Detection was performed in the transmission arm by PMT through Hoya glass UV filter with peak transmission at 340nm. The laser beam was focused with the Zeiss Plan Neofluar 40x1.3 objective lens. For the imaging, the fungi were placed between two cover glasses of 0.17 μm thickness to match the objective lens requirements and for better transmission of the THG signal. An increased number of LDs under nitrogen starvation was observed in THG images and their 
number and size were analyzed using two quantification methods. The comparison of LDs number and 
size obtained by ICS and PSA shows that the number of LDs is approximately the same on average, but 
that ICS consistently detects slightly larger LD number in older group. The mean ICS measured diameter was slightly lower. Using the THG method in vivo and label-free, we can accurately and reliably, over time, detect changes in the localization, total number, and size of LDs in hyphae of the oleaginous filamentous fungus Phycomyces blakesleeanus.
PB  - Belgrade : Institute of Physics
C3  - VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop
T1  - Label-free third harmonic generation imaging of lipid droplets in live filamentous fungi
SP  - 115
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4712
ER  - 

@conference{
author = "Pajić, Tanja and Todorović, Nataša and Živić, Miroslav and Rabasović, Mihailo D. and Clayton, Andrew H. A. and Krmpot, Aleksandar J.",
year = "2021",
abstract = "Oleaginous fungi can accumulate significant amounts of lipids in their mycelium (up to 80% of their biomass), primarily in the form of lipid droplets (LDs). LDs have optical properties that differ from the surrounding aquatic environment, which causes sudden changes in the refractive index. Here, we present in vivo and label-free imaging of individual hyphae of the oleaginous filamentous fungus 
Phycomyces blakesleeanus by Third Harmonic Generation (THG) microscopy method, where LDs 
are the main source of contrast. The LDs quantification from THG images was performed by two image analysis techniques: Image Correlation Spectroscopy (ICS) and software particle counting – Particle Size Analysis (PSA). ICS measures the spatial variation of fluorescence intensity fluctuations in the images, which can then be related to particle density and aggregation state. In order to test and compare the two methods, we used hyphae that undergo nitrogen starvation, which is known to cause alterations in lipid metabolism and the increase in LDs number. For in vivo THG imaging of label-free, > 24-hour old hyphae, we used 1040 nm, 200 fs pulses from Yb KGW laser. Detection was performed in the transmission arm by PMT through Hoya glass UV filter with peak transmission at 340nm. The laser beam was focused with the Zeiss Plan Neofluar 40x1.3 objective lens. For the imaging, the fungi were placed between two cover glasses of 0.17 μm thickness to match the objective lens requirements and for better transmission of the THG signal. An increased number of LDs under nitrogen starvation was observed in THG images and their 
number and size were analyzed using two quantification methods. The comparison of LDs number and 
size obtained by ICS and PSA shows that the number of LDs is approximately the same on average, but 
that ICS consistently detects slightly larger LD number in older group. The mean ICS measured diameter was slightly lower. Using the THG method in vivo and label-free, we can accurately and reliably, over time, detect changes in the localization, total number, and size of LDs in hyphae of the oleaginous filamentous fungus Phycomyces blakesleeanus.",
publisher = "Belgrade : Institute of Physics",
journal = "VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop",
title = "Label-free third harmonic generation imaging of lipid droplets in live filamentous fungi",
pages = "115",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4712"
}

Pajić, T., Todorović, N., Živić, M., Rabasović, M. D., Clayton, A. H. A.,& Krmpot, A. J.. (2021). Label-free third harmonic generation imaging of lipid droplets in live filamentous fungi. in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop
Belgrade : Institute of Physics., 115.
https://hdl.handle.net/21.15107/rcub_ibiss_4712

Pajić T, Todorović N, Živić M, Rabasović MD, Clayton AHA, Krmpot AJ. Label-free third harmonic generation imaging of lipid droplets in live filamentous fungi. in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop. 2021;:115.
https://hdl.handle.net/21.15107/rcub_ibiss_4712 .

Pajić, Tanja, Todorović, Nataša, Živić, Miroslav, Rabasović, Mihailo D., Clayton, Andrew H. A., Krmpot, Aleksandar J., "Label-free third harmonic generation imaging of lipid droplets in live filamentous fungi" in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop (2021):115,
https://hdl.handle.net/21.15107/rcub_ibiss_4712 .

TY  - CONF
AU  - Živić, Miroslav
AU  - Stevanović, Katarina
AU  - Pajić, Tanja
AU  - Rabasović, Mihailo D.
AU  - Krmpot, Aleksandar J.
AU  - Todorović, Nataša
PY  - 2021
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4713
AB  - Electrophysiology of cell membrane ion channels in filamentous fungi, unlike that in yeast, plant and 
animal cells is still in its infancy. The only two reports on single channel patch clamp recordings from 
native cell membrane of filamentous fungi were made on protoplasts obtained after cell wall 
microsurgery with UV pulsed laser more than 20 years ago. Both pioneering papers reported 
fairly high success rates of obtaining gigaohm contacts, but neither was followed by more studies. 
Advanced imaging techniques that enable more controllable surgery process and utilizing the 
femtosecond pulses, after optimizing the protocol, could result in minimally damaging cell wall 
microsurgery. The end result would be reproducibly high quality membrane of “de-walled” protoplasts. 
The membrane quality, property that is of utmost importance for application of patch clamp method, is 
a term describing not only mechanical integrity and cleanliness of the membrane, but the physiological 
fitness of the cell as well, as cells about to enter apoptosis or necrosis, or that were subjected to oxidative 
stress do not have it. We are presenting here, to the best of our knowledge, the first electrophysiological snapshot obtained on filamentous fungi protoplast after cell wall removal by a femtosecond laser microsurgery. Utilization 
of the Ti:Sa femtosecond laser with optimizations of the cell wall microsurgery protocol explained in this conference resulted in protoplasts that were prone to form contacts of high electrical resistance (GΩ) with a patch pipette. Ti:Sa laser operating at 730 nm (76 MHz, 160 fs pulse duration) combined with homemade nonlinear laser scanning microscope, physiological 40x 1.0 NA objective was employed for microsurgery and imaging. Standard patch-clamp set up was used for electrophysiology. In single channel recordings from more than 30 patches, 11 different channel types were distinguished, based on the reversal potential in asymmetric ionic conditions and on the conductance. By far, the most frequent types of conductance were anionic. We have found four groups of ion channel currents, based 
on ion selectivity: 1. Unselective anion currents (not discriminating between chloride and glutamate) 44% of all recorded currents. 2. Anionic currents selective for chloride (carried exclusively or mostly by chloride) 35%. 3. Organic acid permeable anionic currents (discriminating for glutamate over chloride) 17%; 4. Calcium cationic current was recorded once. The range of conductance size (g) was variable, with unselective anionic currents encompassing the
smallest (5 pS) and largest (160 pS) recorded conductances. Calcium conductance was small (6 pS), while organic acid conductances and Cl-selective conductances had similar ranges (10-60 pS). Most of the conductances displayed linear current-voltage relationships.
PB  - Belgrade : Institute of Physics
C3  - VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop
T1  - First glance at a multitude of ion currents on filamentous fungus P. blakesleeanus protoplasts obtained by femtosecond laser microsurgery
SP  - 111
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4713
ER  - 

@conference{
author = "Živić, Miroslav and Stevanović, Katarina and Pajić, Tanja and Rabasović, Mihailo D. and Krmpot, Aleksandar J. and Todorović, Nataša",
year = "2021",
abstract = "Electrophysiology of cell membrane ion channels in filamentous fungi, unlike that in yeast, plant and 
animal cells is still in its infancy. The only two reports on single channel patch clamp recordings from 
native cell membrane of filamentous fungi were made on protoplasts obtained after cell wall 
microsurgery with UV pulsed laser more than 20 years ago. Both pioneering papers reported 
fairly high success rates of obtaining gigaohm contacts, but neither was followed by more studies. 
Advanced imaging techniques that enable more controllable surgery process and utilizing the 
femtosecond pulses, after optimizing the protocol, could result in minimally damaging cell wall 
microsurgery. The end result would be reproducibly high quality membrane of “de-walled” protoplasts. 
The membrane quality, property that is of utmost importance for application of patch clamp method, is 
a term describing not only mechanical integrity and cleanliness of the membrane, but the physiological 
fitness of the cell as well, as cells about to enter apoptosis or necrosis, or that were subjected to oxidative 
stress do not have it. We are presenting here, to the best of our knowledge, the first electrophysiological snapshot obtained on filamentous fungi protoplast after cell wall removal by a femtosecond laser microsurgery. Utilization 
of the Ti:Sa femtosecond laser with optimizations of the cell wall microsurgery protocol explained in this conference resulted in protoplasts that were prone to form contacts of high electrical resistance (GΩ) with a patch pipette. Ti:Sa laser operating at 730 nm (76 MHz, 160 fs pulse duration) combined with homemade nonlinear laser scanning microscope, physiological 40x 1.0 NA objective was employed for microsurgery and imaging. Standard patch-clamp set up was used for electrophysiology. In single channel recordings from more than 30 patches, 11 different channel types were distinguished, based on the reversal potential in asymmetric ionic conditions and on the conductance. By far, the most frequent types of conductance were anionic. We have found four groups of ion channel currents, based 
on ion selectivity: 1. Unselective anion currents (not discriminating between chloride and glutamate) 44% of all recorded currents. 2. Anionic currents selective for chloride (carried exclusively or mostly by chloride) 35%. 3. Organic acid permeable anionic currents (discriminating for glutamate over chloride) 17%; 4. Calcium cationic current was recorded once. The range of conductance size (g) was variable, with unselective anionic currents encompassing the
smallest (5 pS) and largest (160 pS) recorded conductances. Calcium conductance was small (6 pS), while organic acid conductances and Cl-selective conductances had similar ranges (10-60 pS). Most of the conductances displayed linear current-voltage relationships.",
publisher = "Belgrade : Institute of Physics",
journal = "VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop",
title = "First glance at a multitude of ion currents on filamentous fungus P. blakesleeanus protoplasts obtained by femtosecond laser microsurgery",
pages = "111",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4713"
}

Živić, M., Stevanović, K., Pajić, T., Rabasović, M. D., Krmpot, A. J.,& Todorović, N.. (2021). First glance at a multitude of ion currents on filamentous fungus P. blakesleeanus protoplasts obtained by femtosecond laser microsurgery. in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop
Belgrade : Institute of Physics., 111.
https://hdl.handle.net/21.15107/rcub_ibiss_4713

Živić M, Stevanović K, Pajić T, Rabasović MD, Krmpot AJ, Todorović N. First glance at a multitude of ion currents on filamentous fungus P. blakesleeanus protoplasts obtained by femtosecond laser microsurgery. in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop. 2021;:111.
https://hdl.handle.net/21.15107/rcub_ibiss_4713 .

Živić, Miroslav, Stevanović, Katarina, Pajić, Tanja, Rabasović, Mihailo D., Krmpot, Aleksandar J., Todorović, Nataša, "First glance at a multitude of ion currents on filamentous fungus P. blakesleeanus protoplasts obtained by femtosecond laser microsurgery" in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop (2021):111,
https://hdl.handle.net/21.15107/rcub_ibiss_4713 .

TY  - CONF
AU  - AlJamal, Kamal
AU  - Stojadinović, Gordana
AU  - Živić, Miroslav
AU  - Todorović, Nataša
PY  - 2021
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4711
AB  - The chloride channel VRAC has been implicated in apoptosis, proliferation, transport of organic acids and regulatory volume decrease. Regulated by RAS/cGMP and by the ionic strength, it mediates efflux of anions (Cl-, glutamate, gluthatione, taurine, cGAMP). Knowing that chloride transport activity, mediated by NKCC, is indispensable for axon growth, we hypothesized that VRAC could have a role in neurite elongation mechanism. Experimental treatments compared to control were: VRAC selective inhibitor DCPIB (10 µM); BpV (phen) (80 nM), an inhibitor of PTEN. Axon growth was measured on cultured 
dissociated dorsal root ganglia neonatal neurons fixed 24h after treatment and immunostained with anti-neurofilament H (NF-H) phosphorylated antibody. FIJI Simple Neurite Tracer was used for morphometry of individual neurons. 24h post treatment, total neurite length compared to control was decreased by BpV (p=0.0058) and to a lesser extent by DCPIB. In all measured parameters related to the degree of branching, BpV group neurons had small dispersion of values. In addition, DCPIB treated cells had 
larger fractal dimension (<0.05) than control neurons. The effect of DCPIB is mostly related to an increase in Fractal Dimension parameter, according to Principal Component Analysis. The increase in the number of primary branches per increment of cable length is significantly smaller in DCPIB group than in control and BpV -treated neurons. The DCPIB effect demonstrates that an anion current, presumably VRAC, has 
a role in the process of proper branching and directional elongation of neonatal DRG neurites.
C3  - EMBO Workshop Axons 2021: Structure and function
T1  - Morphometric comparison of neurite outgrowth in rat DRG cultures treated with DCPIB and BpV
SP  - 74
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4711
ER  - 

@conference{
author = "AlJamal, Kamal and Stojadinović, Gordana and Živić, Miroslav and Todorović, Nataša",
year = "2021",
abstract = "The chloride channel VRAC has been implicated in apoptosis, proliferation, transport of organic acids and regulatory volume decrease. Regulated by RAS/cGMP and by the ionic strength, it mediates efflux of anions (Cl-, glutamate, gluthatione, taurine, cGAMP). Knowing that chloride transport activity, mediated by NKCC, is indispensable for axon growth, we hypothesized that VRAC could have a role in neurite elongation mechanism. Experimental treatments compared to control were: VRAC selective inhibitor DCPIB (10 µM); BpV (phen) (80 nM), an inhibitor of PTEN. Axon growth was measured on cultured 
dissociated dorsal root ganglia neonatal neurons fixed 24h after treatment and immunostained with anti-neurofilament H (NF-H) phosphorylated antibody. FIJI Simple Neurite Tracer was used for morphometry of individual neurons. 24h post treatment, total neurite length compared to control was decreased by BpV (p=0.0058) and to a lesser extent by DCPIB. In all measured parameters related to the degree of branching, BpV group neurons had small dispersion of values. In addition, DCPIB treated cells had 
larger fractal dimension (<0.05) than control neurons. The effect of DCPIB is mostly related to an increase in Fractal Dimension parameter, according to Principal Component Analysis. The increase in the number of primary branches per increment of cable length is significantly smaller in DCPIB group than in control and BpV -treated neurons. The DCPIB effect demonstrates that an anion current, presumably VRAC, has 
a role in the process of proper branching and directional elongation of neonatal DRG neurites.",
journal = "EMBO Workshop Axons 2021: Structure and function",
title = "Morphometric comparison of neurite outgrowth in rat DRG cultures treated with DCPIB and BpV",
pages = "74",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4711"
}

AlJamal, K., Stojadinović, G., Živić, M.,& Todorović, N.. (2021). Morphometric comparison of neurite outgrowth in rat DRG cultures treated with DCPIB and BpV. in EMBO Workshop Axons 2021: Structure and function, 74.
https://hdl.handle.net/21.15107/rcub_ibiss_4711

AlJamal K, Stojadinović G, Živić M, Todorović N. Morphometric comparison of neurite outgrowth in rat DRG cultures treated with DCPIB and BpV. in EMBO Workshop Axons 2021: Structure and function. 2021;:74.
https://hdl.handle.net/21.15107/rcub_ibiss_4711 .

AlJamal, Kamal, Stojadinović, Gordana, Živić, Miroslav, Todorović, Nataša, "Morphometric comparison of neurite outgrowth in rat DRG cultures treated with DCPIB and BpV" in EMBO Workshop Axons 2021: Structure and function (2021):74,
https://hdl.handle.net/21.15107/rcub_ibiss_4711 .

TY  - CONF
AU  - Stevanović, Katarina
AU  - Pajić, Tanja
AU  - Todorović, Nataša
AU  - Krmpot, Aleksandar J.
AU  - Živić, Miroslav
AU  - Rabasović, Mihailo D.
PY  - 2021
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4714
AB  - We have developed an advanced protocol for laser cell surgery to obtain protoplasts of filamentous 
fungi, suitable for investigation of ion channels, relying on few attempts already made in the past. Among obtained protoplasts, 32% were shown to be “patchable”, meaning that formation of the 
gigaseal by the micropipette was possible. Protoplasts were produced by first making an incision on the 
cell wall of plasmolysed hyphae by a tightly focused femtosecond laser beam. Cell surgery 
is followed by a reduction of solution osmolarity to promote extrusion of protoplast (or more often, a 
part of it) through the cut. The two key points- cell surgery parameters and the proper sequence of the 
solutions used – were subjected to variations to gain insight into parameters that contribute to protoplast 
production and stability. The proper selection of the pipette size and shape with respect to the protoplast 
size was also of great importance. Cell surgery and hyphae imaging was performed by a nonlinear laser scanning microscope. Ti:Sa laser was operating at 730nm, with 76MHz repetition rate and 160fs pulse duration. Water dipping objective lens (40 x 1.0, Zeiss W Plan-Apochromat) on the upright non-linear microscope system made possible to perform laser surgery and patch clamp on two separate systems, while working in the same microscopic chamber. Prior to microsurgery, two-photon excitation fluorescence was used to scan the hyphae cell wall stained with Calcofluor white dye, using the same wavelength as for the surgery. Two-step plasmolysis, with increased concentration of calcium in the more hyperosmotic solution was both efficient for microsurgery to be performed and had a stabilizing effect on protoplasts. Subtle deplasmolysis prior to the patch clamping effective enough to stimulate protoplasts to exit, without 
making the membrane overstretched to interact with the pipette was employed. Optimized concentration 
and type of chemical agens for inhibition of the cell wall production was continuously present in all 
solutions, as an indispensable factor for success.
PB  - Belgrade : Institute of Physics
C3  - VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop
T1  - Laser microsurgery of filamentous fungi: The latest protocol enabling patch-clamp amenable protoplasts
SP  - 104
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4714
ER  - 

@conference{
author = "Stevanović, Katarina and Pajić, Tanja and Todorović, Nataša and Krmpot, Aleksandar J. and Živić, Miroslav and Rabasović, Mihailo D.",
year = "2021",
abstract = "We have developed an advanced protocol for laser cell surgery to obtain protoplasts of filamentous 
fungi, suitable for investigation of ion channels, relying on few attempts already made in the past. Among obtained protoplasts, 32% were shown to be “patchable”, meaning that formation of the 
gigaseal by the micropipette was possible. Protoplasts were produced by first making an incision on the 
cell wall of plasmolysed hyphae by a tightly focused femtosecond laser beam. Cell surgery 
is followed by a reduction of solution osmolarity to promote extrusion of protoplast (or more often, a 
part of it) through the cut. The two key points- cell surgery parameters and the proper sequence of the 
solutions used – were subjected to variations to gain insight into parameters that contribute to protoplast 
production and stability. The proper selection of the pipette size and shape with respect to the protoplast 
size was also of great importance. Cell surgery and hyphae imaging was performed by a nonlinear laser scanning microscope. Ti:Sa laser was operating at 730nm, with 76MHz repetition rate and 160fs pulse duration. Water dipping objective lens (40 x 1.0, Zeiss W Plan-Apochromat) on the upright non-linear microscope system made possible to perform laser surgery and patch clamp on two separate systems, while working in the same microscopic chamber. Prior to microsurgery, two-photon excitation fluorescence was used to scan the hyphae cell wall stained with Calcofluor white dye, using the same wavelength as for the surgery. Two-step plasmolysis, with increased concentration of calcium in the more hyperosmotic solution was both efficient for microsurgery to be performed and had a stabilizing effect on protoplasts. Subtle deplasmolysis prior to the patch clamping effective enough to stimulate protoplasts to exit, without 
making the membrane overstretched to interact with the pipette was employed. Optimized concentration 
and type of chemical agens for inhibition of the cell wall production was continuously present in all 
solutions, as an indispensable factor for success.",
publisher = "Belgrade : Institute of Physics",
journal = "VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop",
title = "Laser microsurgery of filamentous fungi: The latest protocol enabling patch-clamp amenable protoplasts",
pages = "104",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4714"
}

Stevanović, K., Pajić, T., Todorović, N., Krmpot, A. J., Živić, M.,& Rabasović, M. D.. (2021). Laser microsurgery of filamentous fungi: The latest protocol enabling patch-clamp amenable protoplasts. in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop
Belgrade : Institute of Physics., 104.
https://hdl.handle.net/21.15107/rcub_ibiss_4714

Stevanović K, Pajić T, Todorović N, Krmpot AJ, Živić M, Rabasović MD. Laser microsurgery of filamentous fungi: The latest protocol enabling patch-clamp amenable protoplasts. in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop. 2021;:104.
https://hdl.handle.net/21.15107/rcub_ibiss_4714 .

Stevanović, Katarina, Pajić, Tanja, Todorović, Nataša, Krmpot, Aleksandar J., Živić, Miroslav, Rabasović, Mihailo D., "Laser microsurgery of filamentous fungi: The latest protocol enabling patch-clamp amenable protoplasts" in VIII International School and Conference on Photonics PHOTONICA2021 & HEMMAGINERO workshop (2021):104,
https://hdl.handle.net/21.15107/rcub_ibiss_4714 .

TY  - CONF
AU  - Pajić, Tanja
AU  - Todorović, Nataša
AU  - Krmpot, Aleksandar J.
AU  - Rabasović, Mihailo D.
AU  - Jelenković, Branislav
AU  - Živić, Miroslav
PY  - 2020
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4643
AB  - Nonlinear laser scanning microscopy (NLSM) enables non-invasive, label-free visualization of living cell, their internal structures and metabolic state (which is a general marker of cell health). The metabolic activity of the cell can be monitored through the optical redox ratio of endogenous fluorophores in cytoplasm and mitochondria. Optical redox ratio provides relative changes in NAD(P)H and FAD concentrations, i.e. oxidation-reduction state in the cell. Lipid droplets (LDs) also play an important role in cellular energy metabolism. LDs are dynamic organelles that interact with molecules involved in metabolism and change their numbers and size depending on the metabolic state of the cell. In stressed cells, LDs maintain energy and redox homeostasis. Here we combined two powerful modalities of NLSM, Two Photon Excitation fluorescence (TPEF) and Third Harmonic Generation (THG), to optically detect how the molecules and organelles, involved in energy metabolism, are distributed. THG is a nonlinear coherent optical effect in which the incident laser beam interacts with a medium producing the light of exactly three times shorter wavelenght than the incidental one. THG mostly ocuurs at interfaces where  change of refractive index is steep, like water-lipid structures. Therefore, cell membranes and lipid droplets are major sources of THG signal. For observation of a map of the oxidation-reduction ratio in a living cell, how the NADH and FAD ratio changes, we used two photon nonlinear microscopy. For in vivo TPEF imaging of optical redox ratio from the same region on hyphae of filamentous fungus Phycomyces blakesleeanus, we used ultrafast Ti:Sa laser, 160 fs pulses. The autofluorescence of NAD(P)H was excited by laser pulses at 730 nm and signal was collected through 479/40 filter, while for autofluorescence of FAD we used excitation by the same laser pulses at 860 nm, 530/43 filter. For THG imaging, we used 1040 nm, 200 fs pulses from Yb KGW laser, and detection was performed by PMT through Hoya glass UV filter with peak transmission at 340nm. Both laser beams were focused with the same objective lens, Zeiss Plan Neofluar 40x1.3. With our nonlinear microscope and its different modalities, we were able to localize the two metabolic molecules, NAD(P)H and FAD, clearly see the lipid droplets, their distribution, number and size, and to characterize cell metabolism. In stressed cells, we obtained a decrease in the redox ratio and a higher number of lipid droplets under nitrogen starvation.
PB  - Belgrade : Institute of Physics
C3  - Book of Abstracts: 13th Photonics Workshop: Conferenece; 2020 Mar 8-12; Kopaonik, Serbia.
T1  - In vivo metabolic imaging of fungus Phycomyces blakesleeanus using label-free nonlinear laser scanning microscopy
SP  - 23
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4643
ER  - 

@conference{
author = "Pajić, Tanja and Todorović, Nataša and Krmpot, Aleksandar J. and Rabasović, Mihailo D. and Jelenković, Branislav and Živić, Miroslav",
year = "2020",
abstract = "Nonlinear laser scanning microscopy (NLSM) enables non-invasive, label-free visualization of living cell, their internal structures and metabolic state (which is a general marker of cell health). The metabolic activity of the cell can be monitored through the optical redox ratio of endogenous fluorophores in cytoplasm and mitochondria. Optical redox ratio provides relative changes in NAD(P)H and FAD concentrations, i.e. oxidation-reduction state in the cell. Lipid droplets (LDs) also play an important role in cellular energy metabolism. LDs are dynamic organelles that interact with molecules involved in metabolism and change their numbers and size depending on the metabolic state of the cell. In stressed cells, LDs maintain energy and redox homeostasis. Here we combined two powerful modalities of NLSM, Two Photon Excitation fluorescence (TPEF) and Third Harmonic Generation (THG), to optically detect how the molecules and organelles, involved in energy metabolism, are distributed. THG is a nonlinear coherent optical effect in which the incident laser beam interacts with a medium producing the light of exactly three times shorter wavelenght than the incidental one. THG mostly ocuurs at interfaces where  change of refractive index is steep, like water-lipid structures. Therefore, cell membranes and lipid droplets are major sources of THG signal. For observation of a map of the oxidation-reduction ratio in a living cell, how the NADH and FAD ratio changes, we used two photon nonlinear microscopy. For in vivo TPEF imaging of optical redox ratio from the same region on hyphae of filamentous fungus Phycomyces blakesleeanus, we used ultrafast Ti:Sa laser, 160 fs pulses. The autofluorescence of NAD(P)H was excited by laser pulses at 730 nm and signal was collected through 479/40 filter, while for autofluorescence of FAD we used excitation by the same laser pulses at 860 nm, 530/43 filter. For THG imaging, we used 1040 nm, 200 fs pulses from Yb KGW laser, and detection was performed by PMT through Hoya glass UV filter with peak transmission at 340nm. Both laser beams were focused with the same objective lens, Zeiss Plan Neofluar 40x1.3. With our nonlinear microscope and its different modalities, we were able to localize the two metabolic molecules, NAD(P)H and FAD, clearly see the lipid droplets, their distribution, number and size, and to characterize cell metabolism. In stressed cells, we obtained a decrease in the redox ratio and a higher number of lipid droplets under nitrogen starvation.",
publisher = "Belgrade : Institute of Physics",
journal = "Book of Abstracts: 13th Photonics Workshop: Conferenece; 2020 Mar 8-12; Kopaonik, Serbia.",
title = "In vivo metabolic imaging of fungus Phycomyces blakesleeanus using label-free nonlinear laser scanning microscopy",
pages = "23",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4643"
}

Pajić, T., Todorović, N., Krmpot, A. J., Rabasović, M. D., Jelenković, B.,& Živić, M.. (2020). In vivo metabolic imaging of fungus Phycomyces blakesleeanus using label-free nonlinear laser scanning microscopy. in Book of Abstracts: 13th Photonics Workshop: Conferenece; 2020 Mar 8-12; Kopaonik, Serbia.
Belgrade : Institute of Physics., 23.
https://hdl.handle.net/21.15107/rcub_ibiss_4643

Pajić T, Todorović N, Krmpot AJ, Rabasović MD, Jelenković B, Živić M. In vivo metabolic imaging of fungus Phycomyces blakesleeanus using label-free nonlinear laser scanning microscopy. in Book of Abstracts: 13th Photonics Workshop: Conferenece; 2020 Mar 8-12; Kopaonik, Serbia.. 2020;:23.
https://hdl.handle.net/21.15107/rcub_ibiss_4643 .

Pajić, Tanja, Todorović, Nataša, Krmpot, Aleksandar J., Rabasović, Mihailo D., Jelenković, Branislav, Živić, Miroslav, "In vivo metabolic imaging of fungus Phycomyces blakesleeanus using label-free nonlinear laser scanning microscopy" in Book of Abstracts: 13th Photonics Workshop: Conferenece; 2020 Mar 8-12; Kopaonik, Serbia. (2020):23,
https://hdl.handle.net/21.15107/rcub_ibiss_4643 .

TY  - JOUR
AU  - Cvetić Antić, Tijana
AU  - Janošević, Dušica
AU  - Maksimović, Vuk M.
AU  - Živić, Miroslav
AU  - Budimir, Snežana
AU  - Glamočlija, Jasmina
AU  - Mitrović, Aleksandra Lj
PY  - 2020
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3730
AB  - We present changes in Tacitus bellus antioxidative system that specifically correspond to subsequent phases of hemibiotroph Fusarium verticillioides infection revealed by histological analysis. T. bellus response to spore germination 6 h post inoculation (hpi), manifested as first oxidative burst, was characterized by transient decrease in malondialdehyde (MDA) content, transient increase in catalase (CAT), low level of superoxide dismutase (SOD) and peroxidase (POD) activity, as well as with transient decrease in total antioxidant capacity (TAC), total phenol content (TPC) and phenylalanine ammonium lyase activity (PAL), and no changes in polyphenol oxidase (PPO) activity, or phenolic profile. During the biotrophic phase of F. verticillioides infection, characterized by hyphae spread intercellularly in epidermal and mesophyll tissue, the host antioxidative system was suppressed. The transition to necrotrophic phase of F. verticillioides infection (inter- and intracellular colonization and sporulation), occurred 3-4 days post inoculation (dpi). During the necrotrophic phase, 5-7 dpi, slowed progression of colonization of T. bellus mesophyll cells occurred and it coincided with sharp increase in MDA content and CAT, SOD and POD activities, but the drop in TAC, TPC content, and PPO activity, as well as the production of phytotoxin fusaric acid. Presented results add to the knowledge of events and mechanisms related to the transition from biotrophy to necrotrophy in F. verticillioides.
PB  - NLM (Medline)
T2  - Journal of Plant Physiology
T1  - Biochemical and histological characterization of succulent plant Tacitus bellus response to Fusarium verticillioides infection in vitro
VL  - 244
DO  - 10.1016/j.jplph.2019.153086
SP  - 153086
ER  - 

@article{
author = "Cvetić Antić, Tijana and Janošević, Dušica and Maksimović, Vuk M. and Živić, Miroslav and Budimir, Snežana and Glamočlija, Jasmina and Mitrović, Aleksandra Lj",
year = "2020",
abstract = "We present changes in Tacitus bellus antioxidative system that specifically correspond to subsequent phases of hemibiotroph Fusarium verticillioides infection revealed by histological analysis. T. bellus response to spore germination 6 h post inoculation (hpi), manifested as first oxidative burst, was characterized by transient decrease in malondialdehyde (MDA) content, transient increase in catalase (CAT), low level of superoxide dismutase (SOD) and peroxidase (POD) activity, as well as with transient decrease in total antioxidant capacity (TAC), total phenol content (TPC) and phenylalanine ammonium lyase activity (PAL), and no changes in polyphenol oxidase (PPO) activity, or phenolic profile. During the biotrophic phase of F. verticillioides infection, characterized by hyphae spread intercellularly in epidermal and mesophyll tissue, the host antioxidative system was suppressed. The transition to necrotrophic phase of F. verticillioides infection (inter- and intracellular colonization and sporulation), occurred 3-4 days post inoculation (dpi). During the necrotrophic phase, 5-7 dpi, slowed progression of colonization of T. bellus mesophyll cells occurred and it coincided with sharp increase in MDA content and CAT, SOD and POD activities, but the drop in TAC, TPC content, and PPO activity, as well as the production of phytotoxin fusaric acid. Presented results add to the knowledge of events and mechanisms related to the transition from biotrophy to necrotrophy in F. verticillioides.",
publisher = "NLM (Medline)",
journal = "Journal of Plant Physiology",
title = "Biochemical and histological characterization of succulent plant Tacitus bellus response to Fusarium verticillioides infection in vitro",
volume = "244",
doi = "10.1016/j.jplph.2019.153086",
pages = "153086"
}

Cvetić Antić, T., Janošević, D., Maksimović, V. M., Živić, M., Budimir, S., Glamočlija, J.,& Mitrović, A. L.. (2020). Biochemical and histological characterization of succulent plant Tacitus bellus response to Fusarium verticillioides infection in vitro. in Journal of Plant Physiology
NLM (Medline)., 244, 153086.
https://doi.org/10.1016/j.jplph.2019.153086

Cvetić Antić T, Janošević D, Maksimović VM, Živić M, Budimir S, Glamočlija J, Mitrović AL. Biochemical and histological characterization of succulent plant Tacitus bellus response to Fusarium verticillioides infection in vitro. in Journal of Plant Physiology. 2020;244:153086.
doi:10.1016/j.jplph.2019.153086 .

Cvetić Antić, Tijana, Janošević, Dušica, Maksimović, Vuk M., Živić, Miroslav, Budimir, Snežana, Glamočlija, Jasmina, Mitrović, Aleksandra Lj, "Biochemical and histological characterization of succulent plant Tacitus bellus response to Fusarium verticillioides infection in vitro" in Journal of Plant Physiology, 244 (2020):153086,
https://doi.org/10.1016/j.jplph.2019.153086 . .

TY  - JOUR
AU  - Radojević, Anđelina
AU  - Mirčić, Dejan
AU  - Živić, Miroslav
AU  - Perić Mataruga, Vesna
AU  - Božanić, Milenka
AU  - Stojanović, Katarina
AU  - Lukičić, Jovana
AU  - Živić, Ivana
PY  - 2019
UR  - http://www.doiserbia.nb.rs/Article.aspx?ID=0354-46641900004R
UR  - http://www.serbiosoc.org.rs/arch/index.php/abs/article/view/3771
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3399
AB  - The aim of the present study was to establish the extent to which the outlet waters of trout farms affect the ecosystems of the Crnica and Skrapež rivers in Serbia. We monitored selected biomarkers of oxidative stress: superoxide dismutase (SOD), glutathione peroxidase (GPx) and total glutathione (GSH) in larvae of the species Ecdyonurus venosus, and simultaneously analyzed the changes in the physical and chemical parameters. The investigations were carried out in spring at four localities along the Skrapež and Crnica rivers: one upstream (the control localities), and three downstream from the fish farm outlets. On the Skrapež River, the fish farm was clearly visible and was markedly changed by the chemical parameters of the water, manifested as a decreased concentration of dissolved oxygen, increased concentrations of nitrates, nitrites, ammonium ions, total phosphorus and total organic carbon, and increased SOD and GPx activities and decreased GSH concentration in larvae from the first downstream locality as compared to the control locality. On the Crnica River, due to the high values of water flow (around 3 m3/s), effluents from the fish farm had no effect on the chemical parameters of the water or on the tested biomarkers.
PB  - Serbian Biological Society
T2  - Archives of Biological Sciences
T1  - Influence of trout farm effluents on selected oxidative stress biomarkers in larvae of Ecdyonurus venosus (Ephemeroptera, Heptageniidae)
IS  - 2
VL  - 71
DO  - 10.2298/ABS181220004R
SP  - 225
EP  - 233
ER  - 

@article{
author = "Radojević, Anđelina and Mirčić, Dejan and Živić, Miroslav and Perić Mataruga, Vesna and Božanić, Milenka and Stojanović, Katarina and Lukičić, Jovana and Živić, Ivana",
year = "2019",
abstract = "The aim of the present study was to establish the extent to which the outlet waters of trout farms affect the ecosystems of the Crnica and Skrapež rivers in Serbia. We monitored selected biomarkers of oxidative stress: superoxide dismutase (SOD), glutathione peroxidase (GPx) and total glutathione (GSH) in larvae of the species Ecdyonurus venosus, and simultaneously analyzed the changes in the physical and chemical parameters. The investigations were carried out in spring at four localities along the Skrapež and Crnica rivers: one upstream (the control localities), and three downstream from the fish farm outlets. On the Skrapež River, the fish farm was clearly visible and was markedly changed by the chemical parameters of the water, manifested as a decreased concentration of dissolved oxygen, increased concentrations of nitrates, nitrites, ammonium ions, total phosphorus and total organic carbon, and increased SOD and GPx activities and decreased GSH concentration in larvae from the first downstream locality as compared to the control locality. On the Crnica River, due to the high values of water flow (around 3 m3/s), effluents from the fish farm had no effect on the chemical parameters of the water or on the tested biomarkers.",
publisher = "Serbian Biological Society",
journal = "Archives of Biological Sciences",
title = "Influence of trout farm effluents on selected oxidative stress biomarkers in larvae of Ecdyonurus venosus (Ephemeroptera, Heptageniidae)",
number = "2",
volume = "71",
doi = "10.2298/ABS181220004R",
pages = "225-233"
}

Radojević, A., Mirčić, D., Živić, M., Perić Mataruga, V., Božanić, M., Stojanović, K., Lukičić, J.,& Živić, I.. (2019). Influence of trout farm effluents on selected oxidative stress biomarkers in larvae of Ecdyonurus venosus (Ephemeroptera, Heptageniidae). in Archives of Biological Sciences
Serbian Biological Society., 71(2), 225-233.
https://doi.org/10.2298/ABS181220004R

Radojević A, Mirčić D, Živić M, Perić Mataruga V, Božanić M, Stojanović K, Lukičić J, Živić I. Influence of trout farm effluents on selected oxidative stress biomarkers in larvae of Ecdyonurus venosus (Ephemeroptera, Heptageniidae). in Archives of Biological Sciences. 2019;71(2):225-233.
doi:10.2298/ABS181220004R .

Radojević, Anđelina, Mirčić, Dejan, Živić, Miroslav, Perić Mataruga, Vesna, Božanić, Milenka, Stojanović, Katarina, Lukičić, Jovana, Živić, Ivana, "Influence of trout farm effluents on selected oxidative stress biomarkers in larvae of Ecdyonurus venosus (Ephemeroptera, Heptageniidae)" in Archives of Biological Sciences, 71, no. 2 (2019):225-233,
https://doi.org/10.2298/ABS181220004R . .

TY  - CONF
AU  - Božanić, Milenka
AU  - Todorović, Dajana
AU  - Radojević, Anđelina
AU  - Perić Mataruga, Vesna
AU  - Živić, Miroslav
AU  - Stojanović, Katarina
AU  - Živić, Ivana
PY  - 2018
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4022
AB  - Za analizu aktivnosti superoksid dismutaze (SOD), pod uticajem ispusnih voda pastrmskog ribnjaka u reci Skrapez, koriscena je larva vrste Ephemera danica kao model organizam. Istrazivanja su obavljena sezonski tokom 2015. i 2016. godine, na cetiri lokaliteta, jedan iznad, (SKI, kontrolni lokalitet) i tri ispod (SK2, SK3 i SK4) pastrmskog ribnjaka. Za analizu podataka koriscena je analiza koinercije (CIA). Od hemijskih i fizickih parametara analizirani su: temperatura, brzina vode, rastvoreni kiseonik, ukupni azot, amonijak, nitriti, nitrati i fosfati. Aktivnost SOD karakterise veca sezonska nego longitudinalna varijabilnost. CIA analiza pokazuje da je Fl osa, osa sezonskih promena, a enzim SOD se nalazi na njenom negativnom kraju. SOD karakterise statisticki znacajna korelacija sa rastvorenim kiseonikom (R=0,714, P=0,002), ukupnim azotom (R=0,592, P=0,016) i temperaturom (R=0,834, P<0,001). U odnosu na kontrolni lokalitet dolazi do opadanja aktivnosti SOD od proleca do zime. Povecana aktivnost ovog enzima u prolece i leto je posledica povisene temperature, sa kojom je SOD korelisan, kao i vece metabolicke aktivnosti organizma. Na aktivnost SOD, snazan uticaj ima promena faktora unutrasnje i spoljasnje sredine uslovljena smenom godisnjih doba, posto su kako u odnosu na amplitudu aktivnosti, tako i na obrazac njihovih sezonskih promena lokaliteti nizvodno od ispusta ribnjaka veoma slicni kontrolnom lokalitetu. Izuzetak je jesen, kada aktivnost SOD dostize maksimum na SK2. To moze biti posledica koncentracije kiseonika koja upravo u jesen na ovom lokalitetu ima najnizu vrednost (8,06 mg/I). Na osnovu dobijenih rezultata, zakljucuje se da ispusne vode pastrmskog ribnjaka izazivaju stres kod vrste E. danica sto se reflektuje kroz izmenjenu aktivnost SOD.
PB  - Belgrade: Serbian Biological Society
C3  - Drugi kongres biologa Srbije
T1  - Aktivnost superoksid dismutaze larve Ephemera danica - biomarker oksidativnog stresa izazvang otpadnim vodama pastrmskih ribnjaka
SP  - 309
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4022
ER  - 

@conference{
author = "Božanić, Milenka and Todorović, Dajana and Radojević, Anđelina and Perić Mataruga, Vesna and Živić, Miroslav and Stojanović, Katarina and Živić, Ivana",
year = "2018",
abstract = "Za analizu aktivnosti superoksid dismutaze (SOD), pod uticajem ispusnih voda pastrmskog ribnjaka u reci Skrapez, koriscena je larva vrste Ephemera danica kao model organizam. Istrazivanja su obavljena sezonski tokom 2015. i 2016. godine, na cetiri lokaliteta, jedan iznad, (SKI, kontrolni lokalitet) i tri ispod (SK2, SK3 i SK4) pastrmskog ribnjaka. Za analizu podataka koriscena je analiza koinercije (CIA). Od hemijskih i fizickih parametara analizirani su: temperatura, brzina vode, rastvoreni kiseonik, ukupni azot, amonijak, nitriti, nitrati i fosfati. Aktivnost SOD karakterise veca sezonska nego longitudinalna varijabilnost. CIA analiza pokazuje da je Fl osa, osa sezonskih promena, a enzim SOD se nalazi na njenom negativnom kraju. SOD karakterise statisticki znacajna korelacija sa rastvorenim kiseonikom (R=0,714, P=0,002), ukupnim azotom (R=0,592, P=0,016) i temperaturom (R=0,834, P<0,001). U odnosu na kontrolni lokalitet dolazi do opadanja aktivnosti SOD od proleca do zime. Povecana aktivnost ovog enzima u prolece i leto je posledica povisene temperature, sa kojom je SOD korelisan, kao i vece metabolicke aktivnosti organizma. Na aktivnost SOD, snazan uticaj ima promena faktora unutrasnje i spoljasnje sredine uslovljena smenom godisnjih doba, posto su kako u odnosu na amplitudu aktivnosti, tako i na obrazac njihovih sezonskih promena lokaliteti nizvodno od ispusta ribnjaka veoma slicni kontrolnom lokalitetu. Izuzetak je jesen, kada aktivnost SOD dostize maksimum na SK2. To moze biti posledica koncentracije kiseonika koja upravo u jesen na ovom lokalitetu ima najnizu vrednost (8,06 mg/I). Na osnovu dobijenih rezultata, zakljucuje se da ispusne vode pastrmskog ribnjaka izazivaju stres kod vrste E. danica sto se reflektuje kroz izmenjenu aktivnost SOD.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Drugi kongres biologa Srbije",
title = "Aktivnost superoksid dismutaze larve Ephemera danica - biomarker oksidativnog stresa izazvang otpadnim vodama pastrmskih ribnjaka",
pages = "309",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4022"
}

Božanić, M., Todorović, D., Radojević, A., Perić Mataruga, V., Živić, M., Stojanović, K.,& Živić, I.. (2018). Aktivnost superoksid dismutaze larve Ephemera danica - biomarker oksidativnog stresa izazvang otpadnim vodama pastrmskih ribnjaka. in Drugi kongres biologa Srbije
Belgrade: Serbian Biological Society., 309.
https://hdl.handle.net/21.15107/rcub_ibiss_4022

Božanić M, Todorović D, Radojević A, Perić Mataruga V, Živić M, Stojanović K, Živić I. Aktivnost superoksid dismutaze larve Ephemera danica - biomarker oksidativnog stresa izazvang otpadnim vodama pastrmskih ribnjaka. in Drugi kongres biologa Srbije. 2018;:309.
https://hdl.handle.net/21.15107/rcub_ibiss_4022 .

Božanić, Milenka, Todorović, Dajana, Radojević, Anđelina, Perić Mataruga, Vesna, Živić, Miroslav, Stojanović, Katarina, Živić, Ivana, "Aktivnost superoksid dismutaze larve Ephemera danica - biomarker oksidativnog stresa izazvang otpadnim vodama pastrmskih ribnjaka" in Drugi kongres biologa Srbije (2018):309,
https://hdl.handle.net/21.15107/rcub_ibiss_4022 .

TY  - JOUR
AU  - Vranković, Jelena
AU  - Živić, Miroslav
AU  - Radojević, Anđelina
AU  - Perić Mataruga, Vesna
AU  - Todorović, Dajana
AU  - Marković, Zoran
AU  - Živić, Ivana
PY  - 2018
UR  - https://www.sciencedirect.com/science/article/pii/S0147651318306560?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3114
AB  - The current study reports a combined seasonal monitoring of the Crnica River watercourse using physico-chemical parameters and biomarkers measured in Gammarus dulensis with the aim of correlating the effects of pollution with the level of oxidative stress biomarkers in this amphipod. The pollution source is identified as a trout farm in the upper part of the Crnica River (Eastern Serbia). Physico-chemical water parameters and sediment trace element concentrations were measured in the Crnica River during three seasons. Three localities downstream [CR2, CR3, and CR4 (20 m, 400 m, and 1.3 km from the trout farm discharge, respectively)] were assessed in comparison with a reference location (CR1), 250 m upstream from the trout aquaculture in the Crnica River. Antioxidant biomarkers including the enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), and glutathione-S-transferase (GST) were determined in pooled samples of G. dulensis. Our findings showed statistically different antioxidant enzyme levels in gammarids from the first downstream locality (CR2) compared to those from other three localities. The results indicate that oxidative stress occurred to some extent despite the assumed antioxidant induction as a result of exposure to elevated levels of some elements and changed water parameters, the effects seeming in part to be mediated by suppression of SOD and CAT (during summer and autumn); and by suppression of GPX and GR (during autumn) at the CR2 locality. The activity of GST in all seasons was higher at the CR2 locality than at the other localities, suggesting the presence of some permanent xenobiotic next to the aquaculture output. Seasonal difference in the physical and chemical water parameters and sediment trace element concentrations were observed. Overall, our results indicate a locality- and season-dependent antioxidant enzymes response in G. dulensis, pointing at progressively reduced deleterious effect of trout farm outputs with movement further downstream.
PB  - Academic Press
T2  - Ecotoxicology and Environmental Safety
T1  - Evaluation of oxidative stress biomarkers in the freshwater gammarid Gammarus dulensis exposed to trout farm outputs
VL  - 163
DO  - 10.1016/J.ECOENV.2018.07.061
SP  - 84
EP  - 95
ER  - 

@article{
author = "Vranković, Jelena and Živić, Miroslav and Radojević, Anđelina and Perić Mataruga, Vesna and Todorović, Dajana and Marković, Zoran and Živić, Ivana",
year = "2018",
abstract = "The current study reports a combined seasonal monitoring of the Crnica River watercourse using physico-chemical parameters and biomarkers measured in Gammarus dulensis with the aim of correlating the effects of pollution with the level of oxidative stress biomarkers in this amphipod. The pollution source is identified as a trout farm in the upper part of the Crnica River (Eastern Serbia). Physico-chemical water parameters and sediment trace element concentrations were measured in the Crnica River during three seasons. Three localities downstream [CR2, CR3, and CR4 (20 m, 400 m, and 1.3 km from the trout farm discharge, respectively)] were assessed in comparison with a reference location (CR1), 250 m upstream from the trout aquaculture in the Crnica River. Antioxidant biomarkers including the enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), and glutathione-S-transferase (GST) were determined in pooled samples of G. dulensis. Our findings showed statistically different antioxidant enzyme levels in gammarids from the first downstream locality (CR2) compared to those from other three localities. The results indicate that oxidative stress occurred to some extent despite the assumed antioxidant induction as a result of exposure to elevated levels of some elements and changed water parameters, the effects seeming in part to be mediated by suppression of SOD and CAT (during summer and autumn); and by suppression of GPX and GR (during autumn) at the CR2 locality. The activity of GST in all seasons was higher at the CR2 locality than at the other localities, suggesting the presence of some permanent xenobiotic next to the aquaculture output. Seasonal difference in the physical and chemical water parameters and sediment trace element concentrations were observed. Overall, our results indicate a locality- and season-dependent antioxidant enzymes response in G. dulensis, pointing at progressively reduced deleterious effect of trout farm outputs with movement further downstream.",
publisher = "Academic Press",
journal = "Ecotoxicology and Environmental Safety",
title = "Evaluation of oxidative stress biomarkers in the freshwater gammarid Gammarus dulensis exposed to trout farm outputs",
volume = "163",
doi = "10.1016/J.ECOENV.2018.07.061",
pages = "84-95"
}

Vranković, J., Živić, M., Radojević, A., Perić Mataruga, V., Todorović, D., Marković, Z.,& Živić, I.. (2018). Evaluation of oxidative stress biomarkers in the freshwater gammarid Gammarus dulensis exposed to trout farm outputs. in Ecotoxicology and Environmental Safety
Academic Press., 163, 84-95.
https://doi.org/10.1016/J.ECOENV.2018.07.061

Vranković J, Živić M, Radojević A, Perić Mataruga V, Todorović D, Marković Z, Živić I. Evaluation of oxidative stress biomarkers in the freshwater gammarid Gammarus dulensis exposed to trout farm outputs. in Ecotoxicology and Environmental Safety. 2018;163:84-95.
doi:10.1016/J.ECOENV.2018.07.061 .

Vranković, Jelena, Živić, Miroslav, Radojević, Anđelina, Perić Mataruga, Vesna, Todorović, Dajana, Marković, Zoran, Živić, Ivana, "Evaluation of oxidative stress biomarkers in the freshwater gammarid Gammarus dulensis exposed to trout farm outputs" in Ecotoxicology and Environmental Safety, 163 (2018):84-95,
https://doi.org/10.1016/J.ECOENV.2018.07.061 . .

TY  - JOUR
AU  - Opačić, Miloš
AU  - Stević, Zorica
AU  - Baščarević, Vladimir
AU  - Živić, Miroslav
AU  - Spasić, Mihajlo
AU  - Spasojević, Ivan
PY  - 2018
UR  - http://www.liebertpub.com/doi/10.1089/ars.2017.7433
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3065
AB  - The monitoring of progression in amyotrophic lateral sclerosis (ALS) relies on clinical outcome measures that take months to interpret, such as revised ALS functional rating scale (ALSFRS-R) score, with no approved biomarkers. A number of clinical studies have documented the involvement of oxidative stress in ALS pathology. Pertinent to this, we propose to evaluate oxidation-reduction potential (ORP) of cerebrospinal fluid (CSF) as a potential indicator of ALS progression. The case-control study included 24 patients with neurological non-neurodegenerative disorders (controls) and 82 ALS patients with different degrees of disease (ALSFRS-R score: 21-47). ORP was significantly higher in ALS patients than controls. It was not dependent on age or gender. A strong negative correlation was found between ORP and ALSFRS-R score for all patients and patients with spinal onset. In other words, ORP increased with ALS progression. No correlation was found for the subset of patients with bulbar onset, most likely because of the physical distance between neurodegenerative loci and the site of CSF collection. These results lead to the hypothesis that ORP of CSF has a potential as monitoring biomarker in ALS, particularly in the cohort of patients with spinal onset. Antioxid. Redox Signal. 28, 1570-1575.
T2  - Antioxidants and Redox Signaling
T1  - Can Oxidation-Reduction Potential of Cerebrospinal Fluid Be a Monitoring Biomarker in Amyotrophic Lateral Sclerosis?
IS  - 17
VL  - 28
DO  - 10.1089/ars.2017.7433
SP  - 1570
EP  - 1575
ER  - 

@article{
author = "Opačić, Miloš and Stević, Zorica and Baščarević, Vladimir and Živić, Miroslav and Spasić, Mihajlo and Spasojević, Ivan",
year = "2018",
abstract = "The monitoring of progression in amyotrophic lateral sclerosis (ALS) relies on clinical outcome measures that take months to interpret, such as revised ALS functional rating scale (ALSFRS-R) score, with no approved biomarkers. A number of clinical studies have documented the involvement of oxidative stress in ALS pathology. Pertinent to this, we propose to evaluate oxidation-reduction potential (ORP) of cerebrospinal fluid (CSF) as a potential indicator of ALS progression. The case-control study included 24 patients with neurological non-neurodegenerative disorders (controls) and 82 ALS patients with different degrees of disease (ALSFRS-R score: 21-47). ORP was significantly higher in ALS patients than controls. It was not dependent on age or gender. A strong negative correlation was found between ORP and ALSFRS-R score for all patients and patients with spinal onset. In other words, ORP increased with ALS progression. No correlation was found for the subset of patients with bulbar onset, most likely because of the physical distance between neurodegenerative loci and the site of CSF collection. These results lead to the hypothesis that ORP of CSF has a potential as monitoring biomarker in ALS, particularly in the cohort of patients with spinal onset. Antioxid. Redox Signal. 28, 1570-1575.",
journal = "Antioxidants and Redox Signaling",
title = "Can Oxidation-Reduction Potential of Cerebrospinal Fluid Be a Monitoring Biomarker in Amyotrophic Lateral Sclerosis?",
number = "17",
volume = "28",
doi = "10.1089/ars.2017.7433",
pages = "1570-1575"
}

Opačić, M., Stević, Z., Baščarević, V., Živić, M., Spasić, M.,& Spasojević, I.. (2018). Can Oxidation-Reduction Potential of Cerebrospinal Fluid Be a Monitoring Biomarker in Amyotrophic Lateral Sclerosis?. in Antioxidants and Redox Signaling, 28(17), 1570-1575.
https://doi.org/10.1089/ars.2017.7433

Opačić M, Stević Z, Baščarević V, Živić M, Spasić M, Spasojević I. Can Oxidation-Reduction Potential of Cerebrospinal Fluid Be a Monitoring Biomarker in Amyotrophic Lateral Sclerosis?. in Antioxidants and Redox Signaling. 2018;28(17):1570-1575.
doi:10.1089/ars.2017.7433 .

Opačić, Miloš, Stević, Zorica, Baščarević, Vladimir, Živić, Miroslav, Spasić, Mihajlo, Spasojević, Ivan, "Can Oxidation-Reduction Potential of Cerebrospinal Fluid Be a Monitoring Biomarker in Amyotrophic Lateral Sclerosis?" in Antioxidants and Redox Signaling, 28, no. 17 (2018):1570-1575,
https://doi.org/10.1089/ars.2017.7433 . .