TY  - JOUR
AU  - Marjanović, Đorđe
AU  - Zdravković, Nemanja
AU  - Milovanović, Mirjana
AU  - Nedeljković-Trailović, Jelena
AU  - Robertson, Alan P.
AU  - Todorović, Zoran
AU  - Trailović, Saša
PY  - 2020
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1807
AB  - The neuromuscular system of parasitic nematodes has proven to be an efficient pharmacological target for antihelmintics. Some of the most frequently used antiparasitic drugs are agonists or antagonists of nicotinic acetylcholine receptors (nAChRs). The antinematodal mechanism of action of carvacrol involves the inhibition of parasite muscle contraction. We have examined the interaction of carvacrol with antinematodal drugs that are agonists of different subtypes of nAChRs and monepantel, which is a non-competitive antagonist of this receptor in A. suum. Additionally, we investigated the effect of carvacrol on the muscle type of nAChRs in the mammalian host. As orthosteric agonists of nAChR, pyrantel, morantel and befinijum lead to dose-dependent contractions of the neuromuscular preparation of Ascaris suum. Carvacrol 100 mu M decreased the E-max of pyrantel, morantel and bephenium by 29%, 39% and 12 %, 39 % and 12 % respectively. The EC50 ratio was 3.43, 2.95 and 2.47 for pyrantel, morantel and bephinium, respectively. Carvacrol 300 u mu M reduces the E-max of pyrantel, morantel and bephenium by 71%, 80% and 75 %, 80 % and 75 % respectively. The EC50 ratio for pyrantel, morantel and bephenium was 3.88, 3.19 and 4.83 respectively. Furthermore, carvacrol enhances the inhibitory effect of monepantel on A. mum contractions, which may have an effective clinical application. On the other hand, tested concentrations of carvacrol did not significantly affect the EFS-induced contractions of the rat diaphragm, indicating a lack of interaction with the postsynaptic nAChR at the muscle end plate in mammals, but the highest concentration (300 mu M) caused a clear tetanic fade. Carvacrol exhibited a time and dose-dependent effect on the Rota-rod performances of rats with a high value of the ED50 (421.6 mg/kg). In our research, carvacrol dominantly exhibited characteristics of a non-competitive antagonist of nAChR in A. suum, and enhances the inhibitory effect of monepantel. The combination of monepantel and carvacrol may be clinically very effective, and the carvacrol molecule itself can be used as a promising platform for the development of new anthelmintic drugs.
PB  - Elsevier, Amsterdam
T2  - Veterinary Parasitology
T1  - Carvacrol acts as a potent selective antagonist of different types of nicotinic acetylcholine receptors and enhances the effect of monepantel in the parasitic nematode Ascaris suum
VL  - 278
SP  - UNSP 109031
DO  - 10.1016/j.vetpar.2020.109031
ER  - 

@article{
author = "Marjanović, Đorđe and Zdravković, Nemanja and Milovanović, Mirjana and Nedeljković-Trailović, Jelena and Robertson, Alan P. and Todorović, Zoran and Trailović, Saša",
year = "2020",
abstract = "The neuromuscular system of parasitic nematodes has proven to be an efficient pharmacological target for antihelmintics. Some of the most frequently used antiparasitic drugs are agonists or antagonists of nicotinic acetylcholine receptors (nAChRs). The antinematodal mechanism of action of carvacrol involves the inhibition of parasite muscle contraction. We have examined the interaction of carvacrol with antinematodal drugs that are agonists of different subtypes of nAChRs and monepantel, which is a non-competitive antagonist of this receptor in A. suum. Additionally, we investigated the effect of carvacrol on the muscle type of nAChRs in the mammalian host. As orthosteric agonists of nAChR, pyrantel, morantel and befinijum lead to dose-dependent contractions of the neuromuscular preparation of Ascaris suum. Carvacrol 100 mu M decreased the E-max of pyrantel, morantel and bephenium by 29%, 39% and 12 %, 39 % and 12 % respectively. The EC50 ratio was 3.43, 2.95 and 2.47 for pyrantel, morantel and bephinium, respectively. Carvacrol 300 u mu M reduces the E-max of pyrantel, morantel and bephenium by 71%, 80% and 75 %, 80 % and 75 % respectively. The EC50 ratio for pyrantel, morantel and bephenium was 3.88, 3.19 and 4.83 respectively. Furthermore, carvacrol enhances the inhibitory effect of monepantel on A. mum contractions, which may have an effective clinical application. On the other hand, tested concentrations of carvacrol did not significantly affect the EFS-induced contractions of the rat diaphragm, indicating a lack of interaction with the postsynaptic nAChR at the muscle end plate in mammals, but the highest concentration (300 mu M) caused a clear tetanic fade. Carvacrol exhibited a time and dose-dependent effect on the Rota-rod performances of rats with a high value of the ED50 (421.6 mg/kg). In our research, carvacrol dominantly exhibited characteristics of a non-competitive antagonist of nAChR in A. suum, and enhances the inhibitory effect of monepantel. The combination of monepantel and carvacrol may be clinically very effective, and the carvacrol molecule itself can be used as a promising platform for the development of new anthelmintic drugs.",
publisher = "Elsevier, Amsterdam",
journal = "Veterinary Parasitology",
title = "Carvacrol acts as a potent selective antagonist of different types of nicotinic acetylcholine receptors and enhances the effect of monepantel in the parasitic nematode Ascaris suum",
volume = "278",
pages = "UNSP 109031",
doi = "10.1016/j.vetpar.2020.109031"
}

Marjanović, Đ., Zdravković, N., Milovanović, M., Nedeljković-Trailović, J., Robertson, A. P., Todorović, Z.,& Trailović, S.. (2020). Carvacrol acts as a potent selective antagonist of different types of nicotinic acetylcholine receptors and enhances the effect of monepantel in the parasitic nematode Ascaris suum. in Veterinary Parasitology
Elsevier, Amsterdam., 278, UNSP 109031.
https://doi.org/10.1016/j.vetpar.2020.109031

Marjanović Đ, Zdravković N, Milovanović M, Nedeljković-Trailović J, Robertson AP, Todorović Z, Trailović S. Carvacrol acts as a potent selective antagonist of different types of nicotinic acetylcholine receptors and enhances the effect of monepantel in the parasitic nematode Ascaris suum. in Veterinary Parasitology. 2020;278:UNSP 109031.
doi:10.1016/j.vetpar.2020.109031 .

Marjanović, Đorđe, Zdravković, Nemanja, Milovanović, Mirjana, Nedeljković-Trailović, Jelena, Robertson, Alan P., Todorović, Zoran, Trailović, Saša, "Carvacrol acts as a potent selective antagonist of different types of nicotinic acetylcholine receptors and enhances the effect of monepantel in the parasitic nematode Ascaris suum" in Veterinary Parasitology, 278 (2020):UNSP 109031,
https://doi.org/10.1016/j.vetpar.2020.109031 . .

TY  - JOUR
AU  - Choudhary, Shivani
AU  - Marjanović, Đorđe
AU  - Wong, Colin R.
AU  - Zhang, Xiaoyu
AU  - Abongwa, Melanie
AU  - Coats, Joel R.
AU  - Trailović, Saša
AU  - Martin, Richard J.
AU  - Robertson, Alan P.
PY  - 2019
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1730
AB  - The ongoing and widespread emergence of resistance to the existing anti-nematodal pharmacopeia has made it imperative to develop new anthelminthic agents. Historically, plants have been important sources of therapeutic compounds and offer an alternative to synthetic drugs. Monoterpenoids are phytochemicals that have been shown to produce acute toxic effects in insects and nematodes. Previous studies have shown nicotinic acetylcholine receptors (nAChRs) to be possible targets for naturally occurring plant metabolites such as carvacrol and carveol. In this study we examined the effects of monoterpenoid compounds on a levamisole sensitive nAChR from Oesophagostomum dentatum and a nicotine sensitive nAChR from Ascaris suum. We expressed the receptors in Xenopus laevis oocytes and used two-electrode voltage-clamp to characterize the effect of various compounds on these cys-loop receptors. At 100 mu M the majority of these compounds acted as antagonists. Interestingly, further experiments revealed that both 0.1 mu M and 10 mu M menthol potentiated acetylcholine and levamisole responses in the levamisole sensitive receptor but not the nicotine sensitive receptor. We also investigated the effects of 0.1 mu M menthol on the contractility of A. suum somatic muscle strips. Menthol produced significant potentiation of peak contractions at each concentration of acetylcholine. The positive allosteric modulatory effects of menthol in both in vivo and in vitro experiments suggests menthol as a promising candidate for combination therapy with cholinergic anthelmintics.
PB  - Elsevier Sci Ltd, Oxford
T2  - International Journal for Parasitology-Drugs and Drug Resistance
T1  - Menthol acts as a positive allosteric modulator on nematode levamisole sensitive nicotinic acetylcholine receptors
VL  - 9
SP  - 44
EP  - 53
DO  - 10.1016/j.ijpddr.2018.12.005
ER  - 

@article{
author = "Choudhary, Shivani and Marjanović, Đorđe and Wong, Colin R. and Zhang, Xiaoyu and Abongwa, Melanie and Coats, Joel R. and Trailović, Saša and Martin, Richard J. and Robertson, Alan P.",
year = "2019",
abstract = "The ongoing and widespread emergence of resistance to the existing anti-nematodal pharmacopeia has made it imperative to develop new anthelminthic agents. Historically, plants have been important sources of therapeutic compounds and offer an alternative to synthetic drugs. Monoterpenoids are phytochemicals that have been shown to produce acute toxic effects in insects and nematodes. Previous studies have shown nicotinic acetylcholine receptors (nAChRs) to be possible targets for naturally occurring plant metabolites such as carvacrol and carveol. In this study we examined the effects of monoterpenoid compounds on a levamisole sensitive nAChR from Oesophagostomum dentatum and a nicotine sensitive nAChR from Ascaris suum. We expressed the receptors in Xenopus laevis oocytes and used two-electrode voltage-clamp to characterize the effect of various compounds on these cys-loop receptors. At 100 mu M the majority of these compounds acted as antagonists. Interestingly, further experiments revealed that both 0.1 mu M and 10 mu M menthol potentiated acetylcholine and levamisole responses in the levamisole sensitive receptor but not the nicotine sensitive receptor. We also investigated the effects of 0.1 mu M menthol on the contractility of A. suum somatic muscle strips. Menthol produced significant potentiation of peak contractions at each concentration of acetylcholine. The positive allosteric modulatory effects of menthol in both in vivo and in vitro experiments suggests menthol as a promising candidate for combination therapy with cholinergic anthelmintics.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "International Journal for Parasitology-Drugs and Drug Resistance",
title = "Menthol acts as a positive allosteric modulator on nematode levamisole sensitive nicotinic acetylcholine receptors",
volume = "9",
pages = "44-53",
doi = "10.1016/j.ijpddr.2018.12.005"
}

Choudhary, S., Marjanović, Đ., Wong, C. R., Zhang, X., Abongwa, M., Coats, J. R., Trailović, S., Martin, R. J.,& Robertson, A. P.. (2019). Menthol acts as a positive allosteric modulator on nematode levamisole sensitive nicotinic acetylcholine receptors. in International Journal for Parasitology-Drugs and Drug Resistance
Elsevier Sci Ltd, Oxford., 9, 44-53.
https://doi.org/10.1016/j.ijpddr.2018.12.005

Choudhary S, Marjanović Đ, Wong CR, Zhang X, Abongwa M, Coats JR, Trailović S, Martin RJ, Robertson AP. Menthol acts as a positive allosteric modulator on nematode levamisole sensitive nicotinic acetylcholine receptors. in International Journal for Parasitology-Drugs and Drug Resistance. 2019;9:44-53.
doi:10.1016/j.ijpddr.2018.12.005 .

Choudhary, Shivani, Marjanović, Đorđe, Wong, Colin R., Zhang, Xiaoyu, Abongwa, Melanie, Coats, Joel R., Trailović, Saša, Martin, Richard J., Robertson, Alan P., "Menthol acts as a positive allosteric modulator on nematode levamisole sensitive nicotinic acetylcholine receptors" in International Journal for Parasitology-Drugs and Drug Resistance, 9 (2019):44-53,
https://doi.org/10.1016/j.ijpddr.2018.12.005 . .

TY  - JOUR
AU  - Abongwa, Melanie
AU  - Marjanović, Đorđe
AU  - Tipton, James G.
AU  - Zheng, Fudan
AU  - Martin, Richard J.
AU  - Trailović, Saša
AU  - Robertson, Alan P.
PY  - 2018
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1622
AB  - Zolvix (R) is a recently introduced anthelmintic drench containing monepantel as the active ingredient. Monepantel is a positive allosteric modulator of DEG-3/DES-2 type nicotinic acetylcholine receptors (nAChRs) in several nematode species. The drug has been reported to produce hypercontraction of Caenorhabditis elegans and Haemonchus contortus somatic muscle. We investigated the effects of monepantel on nAChRs from Ascaris suum and Oesophagostomum dentatum heterologously expressed in Xenopus laevis oocytes. Using two-electrode voltagec-lamp electrophysiology, we studied the effects of monepantel on a nicotine preferring homomeric nAChR subtype from A. suum comprising of ACR-16; a pyrantel/tribendimidine preferring heteromeric subtype from O. dentatum comprising UNC-29, UNC-38 and UNC-63 subunits; and a levamisole preferring subtype (O. dentatum) comprising UNC-29, UNC-38, UNC-63 and ACR-8 subunits. For each subtype tested, monepantel applied in isolation produced no measurable currents thereby ruling out an agonist action. When monepantel was continuously applied, it reduced the amplitude of acetylcholine induced currents in a concentration-dependent manner. In all three subtypes, monepantel acted as a non-competitive antagonist on the expressed receptors. ACR-16 from A. suum was particularly sensitive to monepantel inhibition (IC50 values: 1.6 +/- 3.1 nM and 0.2 +/- 2.3 mu M). We also investigated the effects of monepantel on muscle flaps isolated from adult A. suum. The drug did not significantly increase baseline tension when applied on its own. As with acetylcholine induced currents in the heterologously expressed receptors, contractions induced by acetylcholine were antagonized by monepantel. Further investigation revealed that the inhibition was a mixture of competitive and non-competitive antagonism. Our findings suggest that monepantel is active on multiple nAChR subtypes.
PB  - Elsevier Sci Ltd, Oxford
T2  - International Journal for Parasitology-Drugs and Drug Resistance
T1  - Monepantel is a non-competitive antagonist of nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum
VL  - 8
IS  - 1
SP  - 36
EP  - 42
DO  - 10.1016/j.ijpddr.2017.12.001
ER  - 

@article{
author = "Abongwa, Melanie and Marjanović, Đorđe and Tipton, James G. and Zheng, Fudan and Martin, Richard J. and Trailović, Saša and Robertson, Alan P.",
year = "2018",
abstract = "Zolvix (R) is a recently introduced anthelmintic drench containing monepantel as the active ingredient. Monepantel is a positive allosteric modulator of DEG-3/DES-2 type nicotinic acetylcholine receptors (nAChRs) in several nematode species. The drug has been reported to produce hypercontraction of Caenorhabditis elegans and Haemonchus contortus somatic muscle. We investigated the effects of monepantel on nAChRs from Ascaris suum and Oesophagostomum dentatum heterologously expressed in Xenopus laevis oocytes. Using two-electrode voltagec-lamp electrophysiology, we studied the effects of monepantel on a nicotine preferring homomeric nAChR subtype from A. suum comprising of ACR-16; a pyrantel/tribendimidine preferring heteromeric subtype from O. dentatum comprising UNC-29, UNC-38 and UNC-63 subunits; and a levamisole preferring subtype (O. dentatum) comprising UNC-29, UNC-38, UNC-63 and ACR-8 subunits. For each subtype tested, monepantel applied in isolation produced no measurable currents thereby ruling out an agonist action. When monepantel was continuously applied, it reduced the amplitude of acetylcholine induced currents in a concentration-dependent manner. In all three subtypes, monepantel acted as a non-competitive antagonist on the expressed receptors. ACR-16 from A. suum was particularly sensitive to monepantel inhibition (IC50 values: 1.6 +/- 3.1 nM and 0.2 +/- 2.3 mu M). We also investigated the effects of monepantel on muscle flaps isolated from adult A. suum. The drug did not significantly increase baseline tension when applied on its own. As with acetylcholine induced currents in the heterologously expressed receptors, contractions induced by acetylcholine were antagonized by monepantel. Further investigation revealed that the inhibition was a mixture of competitive and non-competitive antagonism. Our findings suggest that monepantel is active on multiple nAChR subtypes.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "International Journal for Parasitology-Drugs and Drug Resistance",
title = "Monepantel is a non-competitive antagonist of nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum",
volume = "8",
number = "1",
pages = "36-42",
doi = "10.1016/j.ijpddr.2017.12.001"
}

Abongwa, M., Marjanović, Đ., Tipton, J. G., Zheng, F., Martin, R. J., Trailović, S.,& Robertson, A. P.. (2018). Monepantel is a non-competitive antagonist of nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum. in International Journal for Parasitology-Drugs and Drug Resistance
Elsevier Sci Ltd, Oxford., 8(1), 36-42.
https://doi.org/10.1016/j.ijpddr.2017.12.001

Abongwa M, Marjanović Đ, Tipton JG, Zheng F, Martin RJ, Trailović S, Robertson AP. Monepantel is a non-competitive antagonist of nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum. in International Journal for Parasitology-Drugs and Drug Resistance. 2018;8(1):36-42.
doi:10.1016/j.ijpddr.2017.12.001 .

Abongwa, Melanie, Marjanović, Đorđe, Tipton, James G., Zheng, Fudan, Martin, Richard J., Trailović, Saša, Robertson, Alan P., "Monepantel is a non-competitive antagonist of nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum" in International Journal for Parasitology-Drugs and Drug Resistance, 8, no. 1 (2018):36-42,
https://doi.org/10.1016/j.ijpddr.2017.12.001 . .

TY  - CONF
AU  - Trailović, Saša
AU  - Marjanović, Đorđe
AU  - Tipton, James M.
AU  - Abongwa, Melanie
AU  - Zheng, Fudan
AU  - Choudhary, Shivani
AU  - Martin, Richard J.
AU  - Robertson, Alan P.
PY  - 2016
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/3346
AB  - Zolvix ® is a recently introduced anthelmintic drench containing monepantel as the active ingredient. 
Monepantel has been demonstrated to act on deg-3/des-2 containing nAChRs in several nematodes as 
a positive allosteric modulator. The drug produces hypercontraction of nematode muscle. We 
investigated the effects of monepantel on muscle strips isolated form adult Ascaris suum. The drug did 
not significantly increase baseline tension when applied on its own. Contractions induced by 
acetylcholine and several cholininomimetic anthelmintics where antagonized by monepantel. Further 
investigation revealed that the antagonism was non-competitive in nature. We also investigated the 
effect of monepantel on nAChRs from O. dentatum heterologously expressed in Xenopus laevis 
oocytes. We investigated the effects of monepantel on OD3, a levamisole preferring nAChR subtype, 
OD4, a pyrantel preferring subtype and ACR-16 a nicotine preferring subtype. For each subtype tested 
monepantel applied in isolation produced no measureable currents, nor did it produce currents in the 
presence of choline. When monepantel was continuously applied it reduced the amplitude of 
acetylcholine induced currents in a concentration dependent manner for the three nAChR subtypes 
tested. In all three cases monepantel acted as a non-competitive antagonist on the expressed 
receptors. ACR-16 was particularly sensitive to monepantel inhibition. Our findings suggest that the 
mode of action of monepantel is more complex than previously described.
PB  - Society of Toxicology
C3  - Antihelmintics : From discovery to resistance II, Captain’s Room: Marina Village Center: San Diego, CA- Feb. 9 - 12, 2016
T1  - Effects of monepantel on nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum
SP  - 11
EP  - 11
UR  - https://hdl.handle.net/21.15107/rcub_veterinar_3346
ER  - 

@conference{
author = "Trailović, Saša and Marjanović, Đorđe and Tipton, James M. and Abongwa, Melanie and Zheng, Fudan and Choudhary, Shivani and Martin, Richard J. and Robertson, Alan P.",
year = "2016",
abstract = "Zolvix ® is a recently introduced anthelmintic drench containing monepantel as the active ingredient. 
Monepantel has been demonstrated to act on deg-3/des-2 containing nAChRs in several nematodes as 
a positive allosteric modulator. The drug produces hypercontraction of nematode muscle. We 
investigated the effects of monepantel on muscle strips isolated form adult Ascaris suum. The drug did 
not significantly increase baseline tension when applied on its own. Contractions induced by 
acetylcholine and several cholininomimetic anthelmintics where antagonized by monepantel. Further 
investigation revealed that the antagonism was non-competitive in nature. We also investigated the 
effect of monepantel on nAChRs from O. dentatum heterologously expressed in Xenopus laevis 
oocytes. We investigated the effects of monepantel on OD3, a levamisole preferring nAChR subtype, 
OD4, a pyrantel preferring subtype and ACR-16 a nicotine preferring subtype. For each subtype tested 
monepantel applied in isolation produced no measureable currents, nor did it produce currents in the 
presence of choline. When monepantel was continuously applied it reduced the amplitude of 
acetylcholine induced currents in a concentration dependent manner for the three nAChR subtypes 
tested. In all three cases monepantel acted as a non-competitive antagonist on the expressed 
receptors. ACR-16 was particularly sensitive to monepantel inhibition. Our findings suggest that the 
mode of action of monepantel is more complex than previously described.",
publisher = "Society of Toxicology",
journal = "Antihelmintics : From discovery to resistance II, Captain’s Room: Marina Village Center: San Diego, CA- Feb. 9 - 12, 2016",
title = "Effects of monepantel on nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum",
pages = "11-11",
url = "https://hdl.handle.net/21.15107/rcub_veterinar_3346"
}

Trailović, S., Marjanović, Đ., Tipton, J. M., Abongwa, M., Zheng, F., Choudhary, S., Martin, R. J.,& Robertson, A. P.. (2016). Effects of monepantel on nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum. in Antihelmintics : From discovery to resistance II, Captain’s Room: Marina Village Center: San Diego, CA- Feb. 9 - 12, 2016
Society of Toxicology., 11-11.
https://hdl.handle.net/21.15107/rcub_veterinar_3346

Trailović S, Marjanović Đ, Tipton JM, Abongwa M, Zheng F, Choudhary S, Martin RJ, Robertson AP. Effects of monepantel on nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum. in Antihelmintics : From discovery to resistance II, Captain’s Room: Marina Village Center: San Diego, CA- Feb. 9 - 12, 2016. 2016;:11-11.
https://hdl.handle.net/21.15107/rcub_veterinar_3346 .

Trailović, Saša, Marjanović, Đorđe, Tipton, James M., Abongwa, Melanie, Zheng, Fudan, Choudhary, Shivani, Martin, Richard J., Robertson, Alan P., "Effects of monepantel on nicotinic acetylcholine receptors from Ascaris suum and Oesophagostomum dentatum" in Antihelmintics : From discovery to resistance II, Captain’s Room: Marina Village Center: San Diego, CA- Feb. 9 - 12, 2016 (2016):11-11,
https://hdl.handle.net/21.15107/rcub_veterinar_3346 .

TY  - JOUR
AU  - Trailović, Saša
AU  - Marjanović, Đorđe
AU  - Nedeljković-Trailović, Jelena
AU  - Robertson, Alan P.
AU  - Martin, Richard J.
PY  - 2015
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1290
AB  - Essential plant oils (or their active principles) are safe to use and a potentially attractive alternative to current antiparasitic drugs. In the present study, we tested the effects of carvacrol on the isolated tissues of Ascaris suum and investigated potential interactions with other antiparasitic drugs. We used somatic muscle flaps for contraction assays, as well as for electrophysiological investigations. Carvacrol 300 mu M highly significantly inhibited contractions caused by 1, 3, 10, 30, and 100 mu M of ACh (p = 0.0023, p = 0.0002, p = 0.0002, p < 0.0001, and p < 0.0001). The control EC50 for acetylcholine was 8.87 mu M (log EC50 = 0.95 +/- 0.26), while R (max) was 2.53 +/- 0.24 g. The EC50 of acetylcholine in the presence of 300 mu M of carvacrol was 27.71 mu M (log EC50 = 1.44 +/- 0.28) and the R (max) decreased to 1.63 +/- 0.32 g. Furthermore, carvacrol highly significant potentiates inhibitory effect of GABA and piperazine on the contractions induced by ACh. However, carvacrol (100 and 300 mu M), did not produce any changes in the membrane potential or conductance of the A. suum muscle cell. While, 300 mu M of carvacrol showed a significant inhibitory effect on ACh-induced depolarization response. The mean control depolarization was 13.58 +/- 0.66 mV and decreased in presence of carvacrol to 4.50 +/- 1.02 mV (p < 0.0001). Mean control Delta g was 0.168 +/- 0.017 mu S, while in the presence of 300 mu M of carvacrol, Delta g significantly decreased to 0.060 +/- 0.018 Delta S (p = 0.0017). The inhibitory effect on contractions may be the explanation of the antinematodal potential of carvacrol. Moreover, inhibition of depolarizations caused by ACh and reduction of conductance changes directly points to an interaction with the nAChR in A. suum.
PB  - Springer, New York
T2  - Parasitology Research
T1  - Interaction of carvacrol with the Ascaris suum nicotinic acetylcholine receptors and gamma-aminobutyric acid receptors, potential mechanism of antinematodal action
VL  - 114
IS  - 8
SP  - 3059
EP  - 3068
DO  - 10.1007/s00436-015-4508-x
ER  - 

@article{
author = "Trailović, Saša and Marjanović, Đorđe and Nedeljković-Trailović, Jelena and Robertson, Alan P. and Martin, Richard J.",
year = "2015",
abstract = "Essential plant oils (or their active principles) are safe to use and a potentially attractive alternative to current antiparasitic drugs. In the present study, we tested the effects of carvacrol on the isolated tissues of Ascaris suum and investigated potential interactions with other antiparasitic drugs. We used somatic muscle flaps for contraction assays, as well as for electrophysiological investigations. Carvacrol 300 mu M highly significantly inhibited contractions caused by 1, 3, 10, 30, and 100 mu M of ACh (p = 0.0023, p = 0.0002, p = 0.0002, p < 0.0001, and p < 0.0001). The control EC50 for acetylcholine was 8.87 mu M (log EC50 = 0.95 +/- 0.26), while R (max) was 2.53 +/- 0.24 g. The EC50 of acetylcholine in the presence of 300 mu M of carvacrol was 27.71 mu M (log EC50 = 1.44 +/- 0.28) and the R (max) decreased to 1.63 +/- 0.32 g. Furthermore, carvacrol highly significant potentiates inhibitory effect of GABA and piperazine on the contractions induced by ACh. However, carvacrol (100 and 300 mu M), did not produce any changes in the membrane potential or conductance of the A. suum muscle cell. While, 300 mu M of carvacrol showed a significant inhibitory effect on ACh-induced depolarization response. The mean control depolarization was 13.58 +/- 0.66 mV and decreased in presence of carvacrol to 4.50 +/- 1.02 mV (p < 0.0001). Mean control Delta g was 0.168 +/- 0.017 mu S, while in the presence of 300 mu M of carvacrol, Delta g significantly decreased to 0.060 +/- 0.018 Delta S (p = 0.0017). The inhibitory effect on contractions may be the explanation of the antinematodal potential of carvacrol. Moreover, inhibition of depolarizations caused by ACh and reduction of conductance changes directly points to an interaction with the nAChR in A. suum.",
publisher = "Springer, New York",
journal = "Parasitology Research",
title = "Interaction of carvacrol with the Ascaris suum nicotinic acetylcholine receptors and gamma-aminobutyric acid receptors, potential mechanism of antinematodal action",
volume = "114",
number = "8",
pages = "3059-3068",
doi = "10.1007/s00436-015-4508-x"
}

Trailović, S., Marjanović, Đ., Nedeljković-Trailović, J., Robertson, A. P.,& Martin, R. J.. (2015). Interaction of carvacrol with the Ascaris suum nicotinic acetylcholine receptors and gamma-aminobutyric acid receptors, potential mechanism of antinematodal action. in Parasitology Research
Springer, New York., 114(8), 3059-3068.
https://doi.org/10.1007/s00436-015-4508-x

Trailović S, Marjanović Đ, Nedeljković-Trailović J, Robertson AP, Martin RJ. Interaction of carvacrol with the Ascaris suum nicotinic acetylcholine receptors and gamma-aminobutyric acid receptors, potential mechanism of antinematodal action. in Parasitology Research. 2015;114(8):3059-3068.
doi:10.1007/s00436-015-4508-x .

Trailović, Saša, Marjanović, Đorđe, Nedeljković-Trailović, Jelena, Robertson, Alan P., Martin, Richard J., "Interaction of carvacrol with the Ascaris suum nicotinic acetylcholine receptors and gamma-aminobutyric acid receptors, potential mechanism of antinematodal action" in Parasitology Research, 114, no. 8 (2015):3059-3068,
https://doi.org/10.1007/s00436-015-4508-x . .

TY  - JOUR
AU  - Puttachary, S.
AU  - Trailović, Saša
AU  - Robertson, Alan P.
AU  - Thompson, D.P.
AU  - Woods, D.J.
AU  - Martin, Richard J.
PY  - 2013
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1068
AB  - Startect® is a novel anthelmintic combination of derquantel and abamectin. It is hypothesized that derquantel and abamectin interact pharmacologically. We investigated the effects of derquantel, abamectin and their combination on somatic muscle nicotinic acetylcholine receptors and pharyngeal muscle glutamate gated chloride receptor channels of Ascaris suum. We used muscle-strips to test the effects of abamectin, derquantel, and abamectin + derquantel together on the contraction responses to different concentrations of acetylcholine. We found that abamectin reduced the response to acetylcholine, as did derquantel. In combination (abamectin + derquantel), inhibition of the higher acetylcholine concentration response was greater than the predicted additive effect. A two-micropipette current-clamp technique was used to study electrophysiological effects of the anthelmintics on: (1) acetylcholine responses in somatic muscle and; (2) on l-glutamate responses in pharyngeal preparations. On somatic muscle, derquantel (0.1-30 μM) produced a potent (IC50 0.22, CI 0.18-0.28 μM) reversible antagonism of acetylcholine depolarizations. Abamectin (0.3 μM) produced a slow onset inhibition of acetylcholine depolarizations. We compared effects of abamectin and derquantel on muscle preparations pretreated for 30 min with these drugs. The effect of the combination was significantly greater than the predicted additive effect of both drugs at higher acetylcholine concentrations. On the pharynx, application of derquantel produced no significant effect by itself or on responses to abamectin and l-glutamate. Abamectin increased the input conductance of the pharynx (EC50 0.42, CI 0.13-1.36 μM). Our study demonstrates that abamectin and derquantel interact at nicotinic acetylcholine receptors on the somatic muscle and suggested synergism can occur.
T2  - Molecular and Biochemical Parasitology
T1  - Derquantel and abamectin: Effects and interactions on isolated tissues of Ascaris suum
VL  - 188
IS  - 2
SP  - 79
EP  - 86
DO  - 10.1016/j.molbiopara.2013.02.004
ER  - 

@article{
author = "Puttachary, S. and Trailović, Saša and Robertson, Alan P. and Thompson, D.P. and Woods, D.J. and Martin, Richard J.",
year = "2013",
abstract = "Startect® is a novel anthelmintic combination of derquantel and abamectin. It is hypothesized that derquantel and abamectin interact pharmacologically. We investigated the effects of derquantel, abamectin and their combination on somatic muscle nicotinic acetylcholine receptors and pharyngeal muscle glutamate gated chloride receptor channels of Ascaris suum. We used muscle-strips to test the effects of abamectin, derquantel, and abamectin + derquantel together on the contraction responses to different concentrations of acetylcholine. We found that abamectin reduced the response to acetylcholine, as did derquantel. In combination (abamectin + derquantel), inhibition of the higher acetylcholine concentration response was greater than the predicted additive effect. A two-micropipette current-clamp technique was used to study electrophysiological effects of the anthelmintics on: (1) acetylcholine responses in somatic muscle and; (2) on l-glutamate responses in pharyngeal preparations. On somatic muscle, derquantel (0.1-30 μM) produced a potent (IC50 0.22, CI 0.18-0.28 μM) reversible antagonism of acetylcholine depolarizations. Abamectin (0.3 μM) produced a slow onset inhibition of acetylcholine depolarizations. We compared effects of abamectin and derquantel on muscle preparations pretreated for 30 min with these drugs. The effect of the combination was significantly greater than the predicted additive effect of both drugs at higher acetylcholine concentrations. On the pharynx, application of derquantel produced no significant effect by itself or on responses to abamectin and l-glutamate. Abamectin increased the input conductance of the pharynx (EC50 0.42, CI 0.13-1.36 μM). Our study demonstrates that abamectin and derquantel interact at nicotinic acetylcholine receptors on the somatic muscle and suggested synergism can occur.",
journal = "Molecular and Biochemical Parasitology",
title = "Derquantel and abamectin: Effects and interactions on isolated tissues of Ascaris suum",
volume = "188",
number = "2",
pages = "79-86",
doi = "10.1016/j.molbiopara.2013.02.004"
}

Puttachary, S., Trailović, S., Robertson, A. P., Thompson, D.P., Woods, D.J.,& Martin, R. J.. (2013). Derquantel and abamectin: Effects and interactions on isolated tissues of Ascaris suum. in Molecular and Biochemical Parasitology, 188(2), 79-86.
https://doi.org/10.1016/j.molbiopara.2013.02.004

Puttachary S, Trailović S, Robertson AP, Thompson D, Woods D, Martin RJ. Derquantel and abamectin: Effects and interactions on isolated tissues of Ascaris suum. in Molecular and Biochemical Parasitology. 2013;188(2):79-86.
doi:10.1016/j.molbiopara.2013.02.004 .

Puttachary, S., Trailović, Saša, Robertson, Alan P., Thompson, D.P., Woods, D.J., Martin, Richard J., "Derquantel and abamectin: Effects and interactions on isolated tissues of Ascaris suum" in Molecular and Biochemical Parasitology, 188, no. 2 (2013):79-86,
https://doi.org/10.1016/j.molbiopara.2013.02.004 . .

TY  - JOUR
AU  - Trailović, Saša
AU  - Ivanović, Saša
AU  - Nedeljković-Trailović, Jelena
AU  - Robertson, Alan P.
PY  - 2010
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/703
AB  - Cholinergic receptors of parasitic nematodes are one of the most important possible sites of action of antiparasitic drugs. This paper presents some of our own results of electrophysiological and pharamcological examinations of nicotinic and muscarinic receptors of nematodes, as well as data from literature on a new class of anthelmintics that act precisely on cholinergic receptors. The nicotinic acetylcholine receptor (nAChR) is located on somatic muscle cells of nematodes and it is responsible for the coordination of parasite movement. Cholinomimetic anthelmintics act on this receptor, as well as acetylcholine, an endogenic neurotransmitter, but they are not sensitive to enzyme acetylcholineesterase which dissolves acetylcholine. As opposed to the nicotinic receptor of vertebra, whose structure has been examined thoroughly, the stoichiometry of the nicotinic receptor of nematodes is not completely known. However, on the grounds of knowledge acquired so far, a model has been constructed recently of the potential composition of a type of nematodes nicotinic receptor, as the site of action of anthelmintics. Based on earlier investigations, it is supposed that a conventional muscarinic receptor exists in nematodes as well, so that it can also be a new pharamocological target for the development of antinematode drugs. The latest class of synthesized anthelmintics, named aminoacetonitriles (AAD), act via the nicotinic receptor. Monepantel is the first drug from the AAD group as a most significant candidate for registration in veterinary medicine. Even though several groups of cholinomimetic anthelmintics (imiodazothiazoles, tetrahydropyrimidines, organophosphat anthelmintics) have been in use in veterinary practice for many years now, it is evident that cholinergic receptors of nematodes still present an attractive place in the examinations and development of new antinematode drugs. .
AB  - Holinergički receptori parazitskih nematoda jedno su od najznačajnijih mogućih mesta delovanja antiparazitskih lekova. U ovom radu prikazani su neki od rezultata elektrofizioloških i farmakoloških ispitivanja nikotinskog i muskarinskog receptora nematoda, kao i literaturni podaci o novoj klasi antihelmintika koji deluju upravo na holinergičke receptore. Nikotinski acetilholinski receptor (nAChR) se nalazi na somatskim mišićnim ćelijama nematoda i odgovoran je za koordinaciju kretanja parazita. Holinomimetički antihelmintici deluju na ovaj receptor kao i acetilholin, endogeni neurotransmiter, ali nisu osetljivi na enzim acetilholin-esterazu koja razlaže acetilholin. Za razliku od nikotinskog receptora vertebrata, čija je struktura dobro ispitana, stohiometrija nikotinskog receptora nematoda nije u potpunosti poznata. Međutim, u skorije vreme na osnovu dosadašnjih saznanja konstruisan je model potencijalnog sastava jedne vrste nikotinskog receptora nematoda, kao mesta delovanja antihelmintika. Na osnovu ranijih istraživanja postoji pretpostavka da klasični muskarinski receptor postoji i kod nematoda, tako da i on može biti novo farmakološko ciljno mesto za razvoj antinematodnih lekova. Najnovija klasa sintetisanih antihelmintika, nazvana aminoacetonitrili (AAD), deluje preko nikotinskog receptora. Monepantel je prvi lek iz grupe AAD kao najznačajniji kandidat za registraciju u veterinarskoj medicini. Iako se u veterinarskoj kliničkoj praksi već duži niz godina primenjuje nekoliko grupa holinomimetičkih antihelmintika (imiodazotiazoli, tetrahidropirimidini, organofosfatni antihelmintici), očigledno je da holinergički receptori nematoda i dalje predstavljaju atraktivno mesto u ispitivanju i razvoju novih antinematodnih lekova.
PB  - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
T2  - Veterinarski Glasnik
T1  - Pharmacological receptors of nematoda as target points for action of antiparasitic drugs
T1  - Farmakološki receptori nematoda kao ciljna mesta delovanja antiparazitskih lekova
VL  - 64
IS  - 3-4
SP  - 253
EP  - 264
DO  - 10.2298/VETGL1004253T
ER  - 

@article{
author = "Trailović, Saša and Ivanović, Saša and Nedeljković-Trailović, Jelena and Robertson, Alan P.",
year = "2010",
abstract = "Cholinergic receptors of parasitic nematodes are one of the most important possible sites of action of antiparasitic drugs. This paper presents some of our own results of electrophysiological and pharamcological examinations of nicotinic and muscarinic receptors of nematodes, as well as data from literature on a new class of anthelmintics that act precisely on cholinergic receptors. The nicotinic acetylcholine receptor (nAChR) is located on somatic muscle cells of nematodes and it is responsible for the coordination of parasite movement. Cholinomimetic anthelmintics act on this receptor, as well as acetylcholine, an endogenic neurotransmitter, but they are not sensitive to enzyme acetylcholineesterase which dissolves acetylcholine. As opposed to the nicotinic receptor of vertebra, whose structure has been examined thoroughly, the stoichiometry of the nicotinic receptor of nematodes is not completely known. However, on the grounds of knowledge acquired so far, a model has been constructed recently of the potential composition of a type of nematodes nicotinic receptor, as the site of action of anthelmintics. Based on earlier investigations, it is supposed that a conventional muscarinic receptor exists in nematodes as well, so that it can also be a new pharamocological target for the development of antinematode drugs. The latest class of synthesized anthelmintics, named aminoacetonitriles (AAD), act via the nicotinic receptor. Monepantel is the first drug from the AAD group as a most significant candidate for registration in veterinary medicine. Even though several groups of cholinomimetic anthelmintics (imiodazothiazoles, tetrahydropyrimidines, organophosphat anthelmintics) have been in use in veterinary practice for many years now, it is evident that cholinergic receptors of nematodes still present an attractive place in the examinations and development of new antinematode drugs. ., Holinergički receptori parazitskih nematoda jedno su od najznačajnijih mogućih mesta delovanja antiparazitskih lekova. U ovom radu prikazani su neki od rezultata elektrofizioloških i farmakoloških ispitivanja nikotinskog i muskarinskog receptora nematoda, kao i literaturni podaci o novoj klasi antihelmintika koji deluju upravo na holinergičke receptore. Nikotinski acetilholinski receptor (nAChR) se nalazi na somatskim mišićnim ćelijama nematoda i odgovoran je za koordinaciju kretanja parazita. Holinomimetički antihelmintici deluju na ovaj receptor kao i acetilholin, endogeni neurotransmiter, ali nisu osetljivi na enzim acetilholin-esterazu koja razlaže acetilholin. Za razliku od nikotinskog receptora vertebrata, čija je struktura dobro ispitana, stohiometrija nikotinskog receptora nematoda nije u potpunosti poznata. Međutim, u skorije vreme na osnovu dosadašnjih saznanja konstruisan je model potencijalnog sastava jedne vrste nikotinskog receptora nematoda, kao mesta delovanja antihelmintika. Na osnovu ranijih istraživanja postoji pretpostavka da klasični muskarinski receptor postoji i kod nematoda, tako da i on može biti novo farmakološko ciljno mesto za razvoj antinematodnih lekova. Najnovija klasa sintetisanih antihelmintika, nazvana aminoacetonitrili (AAD), deluje preko nikotinskog receptora. Monepantel je prvi lek iz grupe AAD kao najznačajniji kandidat za registraciju u veterinarskoj medicini. Iako se u veterinarskoj kliničkoj praksi već duži niz godina primenjuje nekoliko grupa holinomimetičkih antihelmintika (imiodazotiazoli, tetrahidropirimidini, organofosfatni antihelmintici), očigledno je da holinergički receptori nematoda i dalje predstavljaju atraktivno mesto u ispitivanju i razvoju novih antinematodnih lekova.",
publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd",
journal = "Veterinarski Glasnik",
title = "Pharmacological receptors of nematoda as target points for action of antiparasitic drugs, Farmakološki receptori nematoda kao ciljna mesta delovanja antiparazitskih lekova",
volume = "64",
number = "3-4",
pages = "253-264",
doi = "10.2298/VETGL1004253T"
}

Trailović, S., Ivanović, S., Nedeljković-Trailović, J.,& Robertson, A. P.. (2010). Pharmacological receptors of nematoda as target points for action of antiparasitic drugs. in Veterinarski Glasnik
Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 64(3-4), 253-264.
https://doi.org/10.2298/VETGL1004253T

Trailović S, Ivanović S, Nedeljković-Trailović J, Robertson AP. Pharmacological receptors of nematoda as target points for action of antiparasitic drugs. in Veterinarski Glasnik. 2010;64(3-4):253-264.
doi:10.2298/VETGL1004253T .

Trailović, Saša, Ivanović, Saša, Nedeljković-Trailović, Jelena, Robertson, Alan P., "Pharmacological receptors of nematoda as target points for action of antiparasitic drugs" in Veterinarski Glasnik, 64, no. 3-4 (2010):253-264,
https://doi.org/10.2298/VETGL1004253T . .

TY  - JOUR
AU  - Trailović, Saša
AU  - Robertson, Alan P.
AU  - Nedeljković-Trailović, Jelena
PY  - 2009
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/570
AB  - Eugenol, an essential oil of clove oil possesses several different pharmacological properties, including antimicrobial, antifungal, insecticidal and antihelmintic. With regard to the digestive tract, eugenol has shown spasmolitic and relaxant effects. To elucidate some of the mechanisms involved, the effects of eugenol on contractions of isolated rat ileum induced by electrical field stimulation (EFS) were investigated. Eugenol (100 µM) significantly and reversibly reduced EFS induced contractions by approximately 80%. Control contractions were 2.00±0.18 g, while contractions in the presence of eugenol decreased to 0.40±0.02 g (n=7; P=0.0001), respectively. Moreover, eugenol (100 µM) reversibly decreased ileal basal tonus from 0.88±0.04 g to 0.65±0.04 g (n=7; P=0.0002). After incubation with the nitric oxide synthase inhibitor nitro-Larginine methyl ester (L-NAME), eugenol (100 µM) still significantly inhibited ileum contractions (reduction by 75.30%), from 1.70±0.09 g to 0.42±0.08 g (n=6; P=0.0001), respectively. Likewise, incubation with L-NAME did not alter the eugenol relaxant effect on ileal basal tonus. Mean control basal tonus was 0.84±0.07 g and significantly decreased after the addition of eugenol (100 µM), to 0.37±0.09 g (n=6; P=0.0003). After incubation with 300 µM of L-NAME mean basal tonus was 0.85±0.08 g, while eugenol significantly relaxed ileal preparations in the presence of L-NAME to 0.34±0.11 g (n=6; P=0.0003). Our results suggest that eugenol reversibly inhibits contractions caused by EFS and induces relaxation of rat ileum via a mechanism largely independent of NO activity.
AB  - Eugenol, aktivni princip ulja karanfilića, poseduje različita farmakološka svojstva, kao što su antimikrobno, antimikotičko, insekticidno i antihelmintičko. U digestivnom traktu deluje spazmolitički i relaksantno. U cilju bližeg upoznavanja mehanizma dejstva eugenola, ispitan je njegov efekt na kontrakcije iziolovanog ileuma pacova izazvane spoljnom stimulacijom (electrical field stimulation-EFS). Eugenol (100 µM) signifikantno, reverzibilno redukuje kontrakcije ileuma izazvane EFS za oko 80%. Kontrolne kontrakcije iznosile su 2,00 ± 0,18 g, dok su kontrakcije u prisutvu eugenola bile 0,40 ± 0,02 g (n=7; P=0,0001). Eugenol (100 µM), takođe signifikantno smanjuje i bazalni tonus izolovanog ileuma sa 0,88 ± 0,04 g na 0,65 ± 0,04 g (n=7; P=0,0002). Posle inkubacije sa ihibitorom sintaze azotnog oksida L-NAME, eugenol i dalje signifikantno redukuje kontrakcije ileuma (za 75,30%) sa kontrolnih 1,70 ± 0,09 g na 0,42±0,08 g (n=6; P=0,0001). Isto tako, inkubacija sa L-NAME ne utiče na relaksantni efekt eugenola na bazalni tonus ileuma. Kontrolna srednja vrednost bazalnog tonusa bila je 0,84 ± 0,07 g i signifikantno je smanjena posle dodavanja eugenola (100 µM) na 0,37 ± 0,09 g (n=6; P=0,0003). Posle inkubacije sa 300 µML-NAME, srednji bazalni tonus bio je 0,85±0,08 g, dok je eugenol i u prisustvu L-NAME relaksirao izolovani ileum na 0,34 ± 0,11 g (n=6; P=0,0003). Dobijeni rezultati ukazuju da eugenol reverzibilno inhibiše kontrakcije ileuma izazvane spoljnom stimulacijom i dovodi do relaksacije izolovanog ileuma pacova mehanizmom nezavisnim od aktivnosti azotnog oksida.
PB  - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
T2  - Acta Veterinaria-Beograd
T1  - Inhibitory effect of eugenol on rat ileal motility in vitro
T1  - Inhibitorni efekt eugenola na motilitet ileuma pacova in vitro
VL  - 59
IS  - 2-3
SP  - 123
EP  - 131
DO  - 10.2298/AVB0903123T
ER  - 

@article{
author = "Trailović, Saša and Robertson, Alan P. and Nedeljković-Trailović, Jelena",
year = "2009",
abstract = "Eugenol, an essential oil of clove oil possesses several different pharmacological properties, including antimicrobial, antifungal, insecticidal and antihelmintic. With regard to the digestive tract, eugenol has shown spasmolitic and relaxant effects. To elucidate some of the mechanisms involved, the effects of eugenol on contractions of isolated rat ileum induced by electrical field stimulation (EFS) were investigated. Eugenol (100 µM) significantly and reversibly reduced EFS induced contractions by approximately 80%. Control contractions were 2.00±0.18 g, while contractions in the presence of eugenol decreased to 0.40±0.02 g (n=7; P=0.0001), respectively. Moreover, eugenol (100 µM) reversibly decreased ileal basal tonus from 0.88±0.04 g to 0.65±0.04 g (n=7; P=0.0002). After incubation with the nitric oxide synthase inhibitor nitro-Larginine methyl ester (L-NAME), eugenol (100 µM) still significantly inhibited ileum contractions (reduction by 75.30%), from 1.70±0.09 g to 0.42±0.08 g (n=6; P=0.0001), respectively. Likewise, incubation with L-NAME did not alter the eugenol relaxant effect on ileal basal tonus. Mean control basal tonus was 0.84±0.07 g and significantly decreased after the addition of eugenol (100 µM), to 0.37±0.09 g (n=6; P=0.0003). After incubation with 300 µM of L-NAME mean basal tonus was 0.85±0.08 g, while eugenol significantly relaxed ileal preparations in the presence of L-NAME to 0.34±0.11 g (n=6; P=0.0003). Our results suggest that eugenol reversibly inhibits contractions caused by EFS and induces relaxation of rat ileum via a mechanism largely independent of NO activity., Eugenol, aktivni princip ulja karanfilića, poseduje različita farmakološka svojstva, kao što su antimikrobno, antimikotičko, insekticidno i antihelmintičko. U digestivnom traktu deluje spazmolitički i relaksantno. U cilju bližeg upoznavanja mehanizma dejstva eugenola, ispitan je njegov efekt na kontrakcije iziolovanog ileuma pacova izazvane spoljnom stimulacijom (electrical field stimulation-EFS). Eugenol (100 µM) signifikantno, reverzibilno redukuje kontrakcije ileuma izazvane EFS za oko 80%. Kontrolne kontrakcije iznosile su 2,00 ± 0,18 g, dok su kontrakcije u prisutvu eugenola bile 0,40 ± 0,02 g (n=7; P=0,0001). Eugenol (100 µM), takođe signifikantno smanjuje i bazalni tonus izolovanog ileuma sa 0,88 ± 0,04 g na 0,65 ± 0,04 g (n=7; P=0,0002). Posle inkubacije sa ihibitorom sintaze azotnog oksida L-NAME, eugenol i dalje signifikantno redukuje kontrakcije ileuma (za 75,30%) sa kontrolnih 1,70 ± 0,09 g na 0,42±0,08 g (n=6; P=0,0001). Isto tako, inkubacija sa L-NAME ne utiče na relaksantni efekt eugenola na bazalni tonus ileuma. Kontrolna srednja vrednost bazalnog tonusa bila je 0,84 ± 0,07 g i signifikantno je smanjena posle dodavanja eugenola (100 µM) na 0,37 ± 0,09 g (n=6; P=0,0003). Posle inkubacije sa 300 µML-NAME, srednji bazalni tonus bio je 0,85±0,08 g, dok je eugenol i u prisustvu L-NAME relaksirao izolovani ileum na 0,34 ± 0,11 g (n=6; P=0,0003). Dobijeni rezultati ukazuju da eugenol reverzibilno inhibiše kontrakcije ileuma izazvane spoljnom stimulacijom i dovodi do relaksacije izolovanog ileuma pacova mehanizmom nezavisnim od aktivnosti azotnog oksida.",
publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd",
journal = "Acta Veterinaria-Beograd",
title = "Inhibitory effect of eugenol on rat ileal motility in vitro, Inhibitorni efekt eugenola na motilitet ileuma pacova in vitro",
volume = "59",
number = "2-3",
pages = "123-131",
doi = "10.2298/AVB0903123T"
}

Trailović, S., Robertson, A. P.,& Nedeljković-Trailović, J.. (2009). Inhibitory effect of eugenol on rat ileal motility in vitro. in Acta Veterinaria-Beograd
Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 59(2-3), 123-131.
https://doi.org/10.2298/AVB0903123T

Trailović S, Robertson AP, Nedeljković-Trailović J. Inhibitory effect of eugenol on rat ileal motility in vitro. in Acta Veterinaria-Beograd. 2009;59(2-3):123-131.
doi:10.2298/AVB0903123T .

Trailović, Saša, Robertson, Alan P., Nedeljković-Trailović, Jelena, "Inhibitory effect of eugenol on rat ileal motility in vitro" in Acta Veterinaria-Beograd, 59, no. 2-3 (2009):123-131,
https://doi.org/10.2298/AVB0903123T . .

TY  - JOUR
AU  - Trailović, Saša
AU  - Robertson, Alan P.
AU  - Clark, Cheryl L.
AU  - Martin, Richard J.
PY  - 2002
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/2624
AB  - A two-micropipette current-clamp technique was used
to record electrophysiological responses from the somatic
muscle of Ascaris suum. Levamisole and acetylcholine
were applied to the bag region of the muscle using a
microperfusion system. Depolarizations produced by 10 s
applications of 10 µmol l–1 levamisole or 20 s applications
of 10 µmol l–1 acetylcholine were recorded. The effect on
the peak membrane potential change of the kinase
antagonists H-7, staurosporine, KN-93 and genistein was
observed. H-7 (30 µmol l–1), a non-selective antagonist of
protein kinases A, C and G but which has little effect on
Ca2+/calmodulin-dependent kinase II (CaM kinase II), did
not produce a significant effect on the peak response to
levamisole or acetylcholine. Staurosporine (1 µmol l–1), a
non-selective kinase antagonist that has effects on protein
kinases A, C and G, CaM kinase and tyrosine kinase,
reduced the mean peak membrane potential response to
levamisole from 6.8 mV to 3.9 mV (P<0.0001) and the
mean response to acetylcholine from 5.5 mV to 2.8 mV
(P=0.0016). The difference between the effects of H-7 and
staurosporine suggested the involvement of CaM kinase II
and/or tyrosine kinase. KN-93, a selective CaM kinase II
antagonist, reduced the mean peak response to levamisole
from 6.2 mV to 2.7 mV (P=0.035) and the mean peak
response of acetylcholine from 4.7 mV to 2.0 mV
(P=0.0004). The effects indicated the involvement of CaM
kinase II in the phosphorylation of levamisole and
acetylcholine receptors. The effect of extracellular Ca2+ on
the response to levamisole was assessed by comparing
responses to levamisole in normal and in low-Ca2+ bathing
solutions. The response to levamisole was greater in the
presence of Ca2+, an effect that may be explained by
stimulation of CaM kinase II. Genistein (90 µmol l–1), a
selective tyrosine kinase antagonist, reduced peak
membrane potential responses to levamisole from a mean
of 6.4 mV to 3.3 mV (P=0.001). This effect indicated the
involvement of tyrosine kinase in maintaining the
receptor.
PB  - The Company of Biologists
T2  - Journal of Experimental Biology
T1  - Levamisole receptor phosphorylation: effects of kinase antagonists on membrane potential responses in Ascaris suum suggest that CaM kinase and tyrosine kinase regulate sensitivity to levamisole
VL  - 205
SP  - 3979
EP  - 3988
DO  - 10.1242/jeb.205.24.3979
ER  - 

@article{
author = "Trailović, Saša and Robertson, Alan P. and Clark, Cheryl L. and Martin, Richard J.",
year = "2002",
abstract = "A two-micropipette current-clamp technique was used
to record electrophysiological responses from the somatic
muscle of Ascaris suum. Levamisole and acetylcholine
were applied to the bag region of the muscle using a
microperfusion system. Depolarizations produced by 10 s
applications of 10 µmol l–1 levamisole or 20 s applications
of 10 µmol l–1 acetylcholine were recorded. The effect on
the peak membrane potential change of the kinase
antagonists H-7, staurosporine, KN-93 and genistein was
observed. H-7 (30 µmol l–1), a non-selective antagonist of
protein kinases A, C and G but which has little effect on
Ca2+/calmodulin-dependent kinase II (CaM kinase II), did
not produce a significant effect on the peak response to
levamisole or acetylcholine. Staurosporine (1 µmol l–1), a
non-selective kinase antagonist that has effects on protein
kinases A, C and G, CaM kinase and tyrosine kinase,
reduced the mean peak membrane potential response to
levamisole from 6.8 mV to 3.9 mV (P<0.0001) and the
mean response to acetylcholine from 5.5 mV to 2.8 mV
(P=0.0016). The difference between the effects of H-7 and
staurosporine suggested the involvement of CaM kinase II
and/or tyrosine kinase. KN-93, a selective CaM kinase II
antagonist, reduced the mean peak response to levamisole
from 6.2 mV to 2.7 mV (P=0.035) and the mean peak
response of acetylcholine from 4.7 mV to 2.0 mV
(P=0.0004). The effects indicated the involvement of CaM
kinase II in the phosphorylation of levamisole and
acetylcholine receptors. The effect of extracellular Ca2+ on
the response to levamisole was assessed by comparing
responses to levamisole in normal and in low-Ca2+ bathing
solutions. The response to levamisole was greater in the
presence of Ca2+, an effect that may be explained by
stimulation of CaM kinase II. Genistein (90 µmol l–1), a
selective tyrosine kinase antagonist, reduced peak
membrane potential responses to levamisole from a mean
of 6.4 mV to 3.3 mV (P=0.001). This effect indicated the
involvement of tyrosine kinase in maintaining the
receptor.",
publisher = "The Company of Biologists",
journal = "Journal of Experimental Biology",
title = "Levamisole receptor phosphorylation: effects of kinase antagonists on membrane potential responses in Ascaris suum suggest that CaM kinase and tyrosine kinase regulate sensitivity to levamisole",
volume = "205",
pages = "3979-3988",
doi = "10.1242/jeb.205.24.3979"
}

Trailović, S., Robertson, A. P., Clark, C. L.,& Martin, R. J.. (2002). Levamisole receptor phosphorylation: effects of kinase antagonists on membrane potential responses in Ascaris suum suggest that CaM kinase and tyrosine kinase regulate sensitivity to levamisole. in Journal of Experimental Biology
The Company of Biologists., 205, 3979-3988.
https://doi.org/10.1242/jeb.205.24.3979

Trailović S, Robertson AP, Clark CL, Martin RJ. Levamisole receptor phosphorylation: effects of kinase antagonists on membrane potential responses in Ascaris suum suggest that CaM kinase and tyrosine kinase regulate sensitivity to levamisole. in Journal of Experimental Biology. 2002;205:3979-3988.
doi:10.1242/jeb.205.24.3979 .

Trailović, Saša, Robertson, Alan P., Clark, Cheryl L., Martin, Richard J., "Levamisole receptor phosphorylation: effects of kinase antagonists on membrane potential responses in Ascaris suum suggest that CaM kinase and tyrosine kinase regulate sensitivity to levamisole" in Journal of Experimental Biology, 205 (2002):3979-3988,
https://doi.org/10.1242/jeb.205.24.3979 . .