AUTOMATED ASSAY OF Caenorhabditis elegans WILD-TYPE AND CYSTATIN MUTANTS THRASHING BEHAVIOUR IN THE PRESENCE OR ABSENCE OF PLANT DERIVED CYSTEINE PROTEINASES (CPs)
Asian Journal of Advances in Medical Science,
Page 30-37
Abstract
The intensive use and over dependence on synthetic anthelmintics for the treatment of nematode infection on only a few drugs with similar mode of action has put pressure on such drug candidates with resultant loss of potency due to development of resistance by target nematodes. Plant materials with promising quality and efficacy to substitute for current anthelmintics include the plant derived cysteine proteinases (CPs). Motility is an important indication of the effectivenes of a drug and is a characteristic of phenotype useful for high thoroughput screening of chemical and theraputic agents. This study determined the effect of cysteine proteinases on motility of C. elegans strains (wild type and cystatin null mutants) using the worm watcher device. Results show that motility of C. elegans was affected differently in PLS or papain. The effect of CP on motility of C. elegans strains was dependent on CP type, time of incubation and concentration of CP. Generally there was no significant difference (P>0.05) between mean motility of WT, cpi-1 and cpi-2 null mutant C. elegans in PLS when compared with PLS+E64 (control). There was a statistically significant (P ˂0.05) effect of papain dose on all the strains. Enzyme specificity on cuticle structural proteins might be responsible for difference in pattern of attack observed between papain and PLS. CP has potency for use as effective anthelminthic.
Keywords:
- Cysteine proteinases
- anthelmintics
- papain
- cystatin
- potency
How to Cite
NJOM, V. S. (2022). AUTOMATED ASSAY OF Caenorhabditis elegans WILD-TYPE AND CYSTATIN MUTANTS THRASHING BEHAVIOUR IN THE PRESENCE OR ABSENCE OF PLANT DERIVED CYSTEINE PROTEINASES (CPs). Asian Journal of Advances in Medical Science, 4(3), 30-37. Retrieved from https://www.mbimph.com/index.php/AJOAIMS/article/view/2976
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Stepek G, et al. Natural plant cysteine proteinases as anthelmintics? Trends Parasitol. 2004;20(7):322-7.
Behnke J, et al. Developing novel anthelmintics from plant cysteine proteinases. Parasites & Vectors. 2008;1(1):29.
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Geerts S, Gryseels B. Anthelmintic resistance in human helminths: A review. Trop Med Int Health. 2001;6(11):915-21.
Shalaby HA. Anthelmintics resistance; How to overcome it? Iranian Journal of Parasitology. 2013;8(1):18-32.
Stepek G, et al. In vitro and in vivo anthelmintic efficacy of plant cysteine proteinases against the rodent gastrointestinal nematode, Trichuris muris. Parasitology. 2006; 132:681-689.
Buttle D, et al. Oral dosing with papaya latex is an effective anthelmintic treatment for sheep infected with Haemonchus contortus. Parasites & Vectors. 2011;4(1):36.
Levecke B, et al. Cysteine proteinases from papaya (Carica papaya) in the treatment of experimental Trichuris suis infection in pigs: two randomized controlled trials. Parasites & Vectors. 2014;7(1):255.
Wink M. Current understanding of modes of action of multicomponent bioactive phytochemicals: potential for nutraceuticals and antimicrobials. Annual Review of Food Science and Technology. 2022;13:337-359.
Malandraki-Miller S, Riley PR. Use of artificial intelligence to enhance phenotypic drug discovery. Drug Discovery Today. 2021; 26(4):887-901.
Culetto E, Sattelle DB. A role for Caenorhabditis elegans in understanding the function and interactions of human disease genes. Hum. Mol. Genet. 2000;9:869-877.
Buckingham S, Sattelle D. Fast, automated measurement of nematode swimming (thrashing) without morphometry. BMC Neuroscience. 2009;10(1):84.
Ha NM, et al. Caenorhabditis elegans as a powerful tool in natural product bioactivity research. Applied Biological Chemistry. 2022; 65(1):1-18.
Devi G. Biological model: Caenorhabditis elegans. Int. J. Curr. Microbiol. App. Sci. 2021;10(05):230-235.
Njom VS. Mechanism of attack and molecular target (s) for plant cysteine proteinases on cuticles of parasitic nematodes and C. elegans. 2016, University of Sheffield.
Marcellino C, et al. WormAssay: a novel computer application for whole-plate motion-based screening of macroscopic parasites. PLoS Negl Trop Dis. 2012;6(1):31.
Buckingham S, Sattelle D. Strategies for automated analysis of C. elegans locomotion. Invert Neurosci. 2008;8(3):121-131.
Stepek G, et al. Natural plant cysteine proteinases as anthelmintics? Trends in Parasitol. 2004;20:322-327.
Stepek G, et al. The anthelmintic efficacy of plant-derived cysteine proteinases against the rodent gastrointestinal nematode, Heligmosomoides polygyrus, in vivo. Parasitology. 2007;134:1409-1419.
Stepek G, et al. Anthelmintic action of plant cysteine proteinases against the rodent stomach nematode, Protospirura muricola, in vitro and in vivo. Parasitology. 2007;134:103-112.
Luoga W, et al. The anthelmintic efficacy of papaya latex in a rodent–nematode model is not dependent on fasting before treatment. Journal of Helminthology. 2012;86(03):311-316.
Lalruatfela B, et al. Nematocidal effects of tobacco infusion (tuibur) against intestinal helminth parasites of chicken. Journal of Environmental Biology. 2020;41(4):840- 844.
Mansur F, et al. The anthelmintic efficacy of natural plant cysteine proteinases against two rodent cestodes Hymenolepis diminuta and Hymenolepis microstoma in vitro. Vet Parasitol.
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Njom VS, et al. The effects of plant cysteine proteinases on the nematode cuticle. Parasites & Vectors. 2021;14(1):1-11.
Stepek G, et al. Assessment of the anthelmintic effect of natural plant cysteine proteinases against the gastrointestinal nematode, Heligmosomoides polygyrus, in vitro. Parasitology. 2005;130(Pt 2):203-11.
Grote A, et al. Cysteine proteases during larval migration and development of helminths in their final host. PLoS Neglected Tropical Diseases. 2018;12(8):e0005919.
Maizels RM, et al. Immune evasion genes from filarial nematodes. International Journal for Parasitology. 2001;31(9):889-898.
Murray J, et al. Bm-CPI-2, a cystatin from Brugia malayi nematode parasites, differs from Caenorhabditis elegans cystatins in a specific site mediating inhibition of the antigen-processing enzyme AEP. Mol Biochem Parasitol. 2005;139(2):197-203.
Gregory WF, Maizels RM. Cystatins from filarial parasites: evolution, adaptation and function in the host-parasite relationship. Int J Biochem Cell Biol. 2008;40(6-7):1389-98.
Page AP, Johnstone IL. The cuticle. WormBook. 2007;19:1-15.
Barrett AJ, et al. Introduction, in Handbook of Proteolytic Enzymes. 2013;Academic Press. li-liv.
Buckingham SD, et al. Automated phenotyping of mosquito larvae enables high-throughput screening for novel larvicides and offers potential for smartphone-based detection of larval insecticide resistance. PLoS Neglected Tropical Diseases. 2021;15(6): e0008639.
Buttle DJ, et al. The preparation of fully active chymopapain free of contaminating proteinases. Biol Chem Hoppe Seyler. 1990; 371(11):1083-8.
Zucker S, et al. Proteolytic activities of papain, chymopapain and papaya proteinase III. Biochim Biophys Acta. 1985;828:196-204.
Buttle D, et al. The preparation of fully active chymopapain free of contaminating proteinases. Biol Chem Hoppe-Seyler. 1990; 371:1083-1088.
Rawlings N, et al. MEROPS: The peptidase database. Nucl Acids Res. 2008;36:D320- D325.
Rawlings ND, Salvesen G. Handbook of Proteolytic Enzymes. Elsevier Science; 2012.
Rawlings ND, Barrett AJ, Finn R. Twenty years of the MEROPS database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Research. 2016;44(D1):D343-D350.
Cronin C, et al. An automated system for measuring parameters of nematode sinusoidal movement. BMC Genet. 2005;6(1):5.
Cronin C, Feng Z, Schafer W. Automated imaging of C. elegans behavior. Methods Mol Biol. 2006;351:241-251.
Restif C, Metaxas D. Tracking the swimming motions of C. elegans worms with applications in aging studies. Med Image Comput Comput Assist Interv. 2008;11(Pt 1):35-42.
Anderson GL, Boyd WA, Williams PL. Assessment of sublethal endpoints for toxicity testing with the nematode Caenorhabditis elegans. Environ Toxicol Chem. 2001;20(4): 833-8.
Anderson GL, Cole RD, Williams PL. Assessing behavioral toxicity with Caenorhabditis elegans. Environ Toxicol Chem. 2004;23(5):1235-40.
Behnke J, et al. Developing novel anthelmintics from plant cysteine proteinases. Parasites & Vectors. 2008;1(29).
Luoga W, et al. The anthelmintic efficacy of papaya latex in a rodent- nematode model is not dependent on fasting before treatment. J Helminthol. 2012;86(3): 311-6.
Stepek G, et al. Natural plant cysteine proteinases as anthelmintics? Trends Parasitol. 2004;20(7):322-7.
Behnke J, et al. Developing novel anthelmintics from plant cysteine proteinases. Parasites & Vectors. 2008;1(1):29.
Geerts S, Gryseels B. Drug resistance in human helminths: current situation and lessons from livestock. Clin Microbiol Rev. 2000;13: 202-222.
Geerts S, Gryseels B. Anthelmintic resistance in human helminths: A review. Trop Med Int Health. 2001;6(11):915-21.
Shalaby HA. Anthelmintics resistance; How to overcome it? Iranian Journal of Parasitology. 2013;8(1):18-32.
Stepek G, et al. In vitro and in vivo anthelmintic efficacy of plant cysteine proteinases against the rodent gastrointestinal nematode, Trichuris muris. Parasitology. 2006; 132:681-689.
Buttle D, et al. Oral dosing with papaya latex is an effective anthelmintic treatment for sheep infected with Haemonchus contortus. Parasites & Vectors. 2011;4(1):36.
Levecke B, et al. Cysteine proteinases from papaya (Carica papaya) in the treatment of experimental Trichuris suis infection in pigs: two randomized controlled trials. Parasites & Vectors. 2014;7(1):255.
Wink M. Current understanding of modes of action of multicomponent bioactive phytochemicals: potential for nutraceuticals and antimicrobials. Annual Review of Food Science and Technology. 2022;13:337-359.
Malandraki-Miller S, Riley PR. Use of artificial intelligence to enhance phenotypic drug discovery. Drug Discovery Today. 2021; 26(4):887-901.
Culetto E, Sattelle DB. A role for Caenorhabditis elegans in understanding the function and interactions of human disease genes. Hum. Mol. Genet. 2000;9:869-877.
Buckingham S, Sattelle D. Fast, automated measurement of nematode swimming (thrashing) without morphometry. BMC Neuroscience. 2009;10(1):84.
Ha NM, et al. Caenorhabditis elegans as a powerful tool in natural product bioactivity research. Applied Biological Chemistry. 2022; 65(1):1-18.
Devi G. Biological model: Caenorhabditis elegans. Int. J. Curr. Microbiol. App. Sci. 2021;10(05):230-235.
Njom VS. Mechanism of attack and molecular target (s) for plant cysteine proteinases on cuticles of parasitic nematodes and C. elegans. 2016, University of Sheffield.
Marcellino C, et al. WormAssay: a novel computer application for whole-plate motion-based screening of macroscopic parasites. PLoS Negl Trop Dis. 2012;6(1):31.
Buckingham S, Sattelle D. Strategies for automated analysis of C. elegans locomotion. Invert Neurosci. 2008;8(3):121-131.
Stepek G, et al. Natural plant cysteine proteinases as anthelmintics? Trends in Parasitol. 2004;20:322-327.
Stepek G, et al. The anthelmintic efficacy of plant-derived cysteine proteinases against the rodent gastrointestinal nematode, Heligmosomoides polygyrus, in vivo. Parasitology. 2007;134:1409-1419.
Stepek G, et al. Anthelmintic action of plant cysteine proteinases against the rodent stomach nematode, Protospirura muricola, in vitro and in vivo. Parasitology. 2007;134:103-112.
Luoga W, et al. The anthelmintic efficacy of papaya latex in a rodent–nematode model is not dependent on fasting before treatment. Journal of Helminthology. 2012;86(03):311-316.
Lalruatfela B, et al. Nematocidal effects of tobacco infusion (tuibur) against intestinal helminth parasites of chicken. Journal of Environmental Biology. 2020;41(4):840- 844.
Mansur F, et al. The anthelmintic efficacy of natural plant cysteine proteinases against two rodent cestodes Hymenolepis diminuta and Hymenolepis microstoma in vitro. Vet Parasitol.
Mansur F, et al. The anthelmintic efficacy of natural plant cysteine proteinases against the equine tapeworm, Anoplocephala perfoliata in vitro. Journal of Helminthology. 2015; FirstView:1-8.
Njom VS, et al. The effects of plant cysteine proteinases on the nematode cuticle. Parasites & Vectors. 2021;14(1):1-11.
Stepek G, et al. Assessment of the anthelmintic effect of natural plant cysteine proteinases against the gastrointestinal nematode, Heligmosomoides polygyrus, in vitro. Parasitology. 2005;130(Pt 2):203-11.
Grote A, et al. Cysteine proteases during larval migration and development of helminths in their final host. PLoS Neglected Tropical Diseases. 2018;12(8):e0005919.
Maizels RM, et al. Immune evasion genes from filarial nematodes. International Journal for Parasitology. 2001;31(9):889-898.
Murray J, et al. Bm-CPI-2, a cystatin from Brugia malayi nematode parasites, differs from Caenorhabditis elegans cystatins in a specific site mediating inhibition of the antigen-processing enzyme AEP. Mol Biochem Parasitol. 2005;139(2):197-203.
Gregory WF, Maizels RM. Cystatins from filarial parasites: evolution, adaptation and function in the host-parasite relationship. Int J Biochem Cell Biol. 2008;40(6-7):1389-98.
Page AP, Johnstone IL. The cuticle. WormBook. 2007;19:1-15.
Barrett AJ, et al. Introduction, in Handbook of Proteolytic Enzymes. 2013;Academic Press. li-liv.
Buckingham SD, et al. Automated phenotyping of mosquito larvae enables high-throughput screening for novel larvicides and offers potential for smartphone-based detection of larval insecticide resistance. PLoS Neglected Tropical Diseases. 2021;15(6): e0008639.
Buttle DJ, et al. The preparation of fully active chymopapain free of contaminating proteinases. Biol Chem Hoppe Seyler. 1990; 371(11):1083-8.
Zucker S, et al. Proteolytic activities of papain, chymopapain and papaya proteinase III. Biochim Biophys Acta. 1985;828:196-204.
Buttle D, et al. The preparation of fully active chymopapain free of contaminating proteinases. Biol Chem Hoppe-Seyler. 1990; 371:1083-1088.
Rawlings N, et al. MEROPS: The peptidase database. Nucl Acids Res. 2008;36:D320- D325.
Rawlings ND, Salvesen G. Handbook of Proteolytic Enzymes. Elsevier Science; 2012.
Rawlings ND, Barrett AJ, Finn R. Twenty years of the MEROPS database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Research. 2016;44(D1):D343-D350.
Cronin C, et al. An automated system for measuring parameters of nematode sinusoidal movement. BMC Genet. 2005;6(1):5.
Cronin C, Feng Z, Schafer W. Automated imaging of C. elegans behavior. Methods Mol Biol. 2006;351:241-251.
Restif C, Metaxas D. Tracking the swimming motions of C. elegans worms with applications in aging studies. Med Image Comput Comput Assist Interv. 2008;11(Pt 1):35-42.
Anderson GL, Boyd WA, Williams PL. Assessment of sublethal endpoints for toxicity testing with the nematode Caenorhabditis elegans. Environ Toxicol Chem. 2001;20(4): 833-8.
Anderson GL, Cole RD, Williams PL. Assessing behavioral toxicity with Caenorhabditis elegans. Environ Toxicol Chem. 2004;23(5):1235-40.
Behnke J, et al. Developing novel anthelmintics from plant cysteine proteinases. Parasites & Vectors. 2008;1(29).
Luoga W, et al. The anthelmintic efficacy of papaya latex in a rodent- nematode model is not dependent on fasting before treatment. J Helminthol. 2012;86(3): 311-6.
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