TY  - JOUR
AU  - Pajic, Tanja
AU  - Stevanović, Katarina
AU  - Todorović, Nataša
AU  - Krmpot, Aleksandar J.
AU  - Živić, Miroslav
AU  - Savić-Šević, Svetlana
AU  - Lević, Steva M.
AU  - Stanić, Marina
AU  - Pantelić, Dejan
AU  - Jelenković, Brana
AU  - Rabasović, Mihailo
PY  - 2024
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/3213
AB  - Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for a micron-sized patch-clamp pipette for pA current recordings due to the rigid chitinous cell wall. Here, we report the first femtosecond IR laser nanosurgery of the cell wall of the filamentous fungi, which enabled patch-clamp measurements on protoplasts released from hyphae. A reproducible and highly precise (diffraction-limited, submicron resolution) method for obtaining viable released protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in the cell wall was achieved from different regions of the hyphae. The plasma membrane of the obtained protoplasts formed tight and high-resistance (GΩ) contacts with the recording pipette. The entire nanosurgical procedure followed by the patch-clamp technique could be completed in less than 1 hour. Compared to previous studies using heterologously expressed channels, this technique provides the opportunity to identify new ionic currents and to study the properties of the ion channels in the protoplasts of filamentous fungi in their native environment.
PB  - Springer Nature
T2  - Microsystems & Nanoengineering
T1  - In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi
SP  - 47
VL  - 10
DO  - 10.1038/s41378-024-00664-x
ER  - 

@article{
author = "Pajic, Tanja and Stevanović, Katarina and Todorović, Nataša and Krmpot, Aleksandar J. and Živić, Miroslav and Savić-Šević, Svetlana and Lević, Steva M. and Stanić, Marina and Pantelić, Dejan and Jelenković, Brana and Rabasović, Mihailo",
year = "2024",
abstract = "Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for a micron-sized patch-clamp pipette for pA current recordings due to the rigid chitinous cell wall. Here, we report the first femtosecond IR laser nanosurgery of the cell wall of the filamentous fungi, which enabled patch-clamp measurements on protoplasts released from hyphae. A reproducible and highly precise (diffraction-limited, submicron resolution) method for obtaining viable released protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in the cell wall was achieved from different regions of the hyphae. The plasma membrane of the obtained protoplasts formed tight and high-resistance (GΩ) contacts with the recording pipette. The entire nanosurgical procedure followed by the patch-clamp technique could be completed in less than 1 hour. Compared to previous studies using heterologously expressed channels, this technique provides the opportunity to identify new ionic currents and to study the properties of the ion channels in the protoplasts of filamentous fungi in their native environment.",
publisher = "Springer Nature",
journal = "Microsystems & Nanoengineering",
title = "In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi",
pages = "47",
volume = "10",
doi = "10.1038/s41378-024-00664-x"
}

Pajic, T., Stevanović, K., Todorović, N., Krmpot, A. J., Živić, M., Savić-Šević, S., Lević, S. M., Stanić, M., Pantelić, D., Jelenković, B.,& Rabasović, M.. (2024). In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi. in Microsystems & Nanoengineering
Springer Nature., 10, 47.
https://doi.org/10.1038/s41378-024-00664-x

Pajic T, Stevanović K, Todorović N, Krmpot AJ, Živić M, Savić-Šević S, Lević SM, Stanić M, Pantelić D, Jelenković B, Rabasović M. In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi. in Microsystems & Nanoengineering. 2024;10:47.
doi:10.1038/s41378-024-00664-x .

Pajic, Tanja, Stevanović, Katarina, Todorović, Nataša, Krmpot, Aleksandar J., Živić, Miroslav, Savić-Šević, Svetlana, Lević, Steva M., Stanić, Marina, Pantelić, Dejan, Jelenković, Brana, Rabasović, Mihailo, "In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi" in Microsystems & Nanoengineering, 10 (2024):47,
https://doi.org/10.1038/s41378-024-00664-x . .

TY  - CONF
AU  - Opačić, Miloš
AU  - Stevic, Zorica
AU  - Živić, Miroslav
AU  - Spasojević, Ivan
PY  - 2016
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/2957
AB  - Background: ALS is an oxidative stress-related fastpaced
motor neuron disease for which there are no
accurate biomarkers of progression. This largely hampers
the development of therapeutics. Our aim was to test the
applicability of oxidation-reduction potential (ORP) of
cerebrospinal fluid (CSF) in ALS progression follow-up.
Methods: ORP was measured using RedoxSYS (Aytu
BioScience, Inc.) in CSF of 49 ALS patients (mean age
63 1.29 years; range 43-80 years; mean ALSFRSr score
37.47 0.90, range 21-48; m/f = 35/14) and 15 controls
(mean age 48.93 3.68 years, range 21-67 years; m/f =
10/5).
Results: Pearson correlation coefficient (R) between
ALSFRSr score and ORP was 0.27 (p = 0.06). R was
higher ( 0.45) when two outlier values were excluded
from the calculus. Importantly, ALS patients had significantly
higher mean ORP (ALS: 121.83 2.76mV;
controls: 111.14 2.94 mV; p = 0.033). The difference
was also significant for ORP normalized to age. Of note,
ORP is reciprocally proportional to the pro-oxidative
settings in biological samples.
Discussion: Increased ORP values in ALS patients
further confirm the role of oxidative stress in this
neurodegenerative disease. ORP might find application
as a biomarker for ALS progression/prognosis but further
measurements on a larger cohort is warranted.
PB  - Taylor and Francis
C3  - Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration
T1  - Oxidation-reduction potential of cerebrospinal fluid as a potential biomarker for ALS progression
SP  - 183
DO  - 10.1080/21678421.2016.1232061
ER  - 

@conference{
author = "Opačić, Miloš and Stevic, Zorica and Živić, Miroslav and Spasojević, Ivan",
year = "2016",
abstract = "Background: ALS is an oxidative stress-related fastpaced
motor neuron disease for which there are no
accurate biomarkers of progression. This largely hampers
the development of therapeutics. Our aim was to test the
applicability of oxidation-reduction potential (ORP) of
cerebrospinal fluid (CSF) in ALS progression follow-up.
Methods: ORP was measured using RedoxSYS (Aytu
BioScience, Inc.) in CSF of 49 ALS patients (mean age
63 1.29 years; range 43-80 years; mean ALSFRSr score
37.47 0.90, range 21-48; m/f = 35/14) and 15 controls
(mean age 48.93 3.68 years, range 21-67 years; m/f =
10/5).
Results: Pearson correlation coefficient (R) between
ALSFRSr score and ORP was 0.27 (p = 0.06). R was
higher ( 0.45) when two outlier values were excluded
from the calculus. Importantly, ALS patients had significantly
higher mean ORP (ALS: 121.83 2.76mV;
controls: 111.14 2.94 mV; p = 0.033). The difference
was also significant for ORP normalized to age. Of note,
ORP is reciprocally proportional to the pro-oxidative
settings in biological samples.
Discussion: Increased ORP values in ALS patients
further confirm the role of oxidative stress in this
neurodegenerative disease. ORP might find application
as a biomarker for ALS progression/prognosis but further
measurements on a larger cohort is warranted.",
publisher = "Taylor and Francis",
journal = "Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration",
title = "Oxidation-reduction potential of cerebrospinal fluid as a potential biomarker for ALS progression",
pages = "183",
doi = "10.1080/21678421.2016.1232061"
}

Opačić, M., Stevic, Z., Živić, M.,& Spasojević, I.. (2016). Oxidation-reduction potential of cerebrospinal fluid as a potential biomarker for ALS progression. in Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration
Taylor and Francis., 183.
https://doi.org/10.1080/21678421.2016.1232061

Opačić M, Stevic Z, Živić M, Spasojević I. Oxidation-reduction potential of cerebrospinal fluid as a potential biomarker for ALS progression. in Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration. 2016;:183.
doi:10.1080/21678421.2016.1232061 .

Opačić, Miloš, Stevic, Zorica, Živić, Miroslav, Spasojević, Ivan, "Oxidation-reduction potential of cerebrospinal fluid as a potential biomarker for ALS progression" in Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration (2016):183,
https://doi.org/10.1080/21678421.2016.1232061 . .