Colorimetric quantitation of filarial viability

Comley, J. C. W.; Rees, Mike; Turner, Claire H.; Jenkins, D. C.

doi:10.1016/0020-7519(89)90024-6

Cited by 110 publications

(59 citation statements)

References 3 publications

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“…Furthermore, a parallel between viability and MTT incorporation was observed in inhibitor-treated parasites. The present results further corroborate evidence presented in previous reports (6,7) that the MTT reduction assay could be an effective tool for evaluating the filaricidal activity of therapeutic agents.…”

Section: Materuils and Methodssupporting

confidence: 92%

Brugia malayi and Acanthocheilonema viteae: antifilarial activity of transglutaminase inhibitors in vitro

Rao

Mehta

Subrahmanyam

et al. 1991

Antimicrob Agents Chemother

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The possible involvement of transglutaminase-catalyzed reactions in survival of adult worms, microfilariae (mf), and infective larvae of the Milarial parasite Brugia malayi was studied in vitro by using the specific pseudosubstrate monodansylcadaverine (MDC) and the active-site inhibitors cystamine or iodoacetamide. These inhibitors significantly inhibited parasite mobility in a dose-dependent manner. This inhibition was associated with irreversible biochemical lesions followed by filarial death. A. structurally related, inactive analog of MDC, dimethyldansylcadaverine, did not affect the mobility or survival of the parasites. Adult worms failed to release mf when they were incubated in the presence of MDC or cystamine, and this inhibitory effect on mf release was concentration dependent. Similar embryostatic and macrofilaricidal effects of MDC were observed in Acanthocheilonema viteae adult worms. These studies suggest that transglutaminase-catalyzed reactions may play an important role in the growth, development, and survival of filarial parasites.

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Section: Materuils and Methodssupporting

confidence: 92%

Brugia malayi and Acanthocheilonema viteae: antifilarial activity of transglutaminase inhibitors in vitro

Rao

Mehta

Subrahmanyam

et al. 1991

Antimicrob Agents Chemother

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“…Worms were cultured in the presence of different concentrations of NMMP for 7 d. Motility was scored daily using a scoring system described previously (31), with 0 being immotile and 4 being highly active and motile and the results for each group presented as a percentage of the control group. Viability was evaluated at day 7 using the MTT assay (32,33). The reduction of MTT for each group is presented as a percentage of the control group.…”

Section: Methodsmentioning

confidence: 99%

Interdomain lateral gene transfer of an essential ferrochelatase gene in human parasitic nematodes

Novelli

Jiang

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Lateral gene transfer events between bacteria and animals highlight an avenue for evolutionary genomic loss/gain of function. Herein, we report functional lateral gene transfer in animal parasitic nematodes. Members of the Nematoda are heme auxotrophs, lacking the ability to synthesize heme; however, the human filarial parasite Brugia malayi has acquired a bacterial gene encoding ferrochelatase (BmFeCH), the terminal step in heme biosynthesis. BmFeCH, encoded by a 9-exon gene, is a mitochondrial-targeted, functional ferrochelatase based on enzyme assays, complementation, and inhibitor studies. Homologs have been identified in several filariae and a nonfilarial nematode. RNAi and ex vivo inhibitor experiments indicate that BmFeCH is essential for viability, validating it as a potential target for filariasis control.

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“…Examples are limited to MTT staining of filarial nematodes including Onchocerca volvulus (Comley et al 1989) and SYTOX green (DNA intercalating stain with fluorescent properties similar to EthD and PI) staining of the non-parasitic nematode Caenorhabditis elegans (Gill et al 2003). The lack of studies reporting the use of dyes/stains in distinguishing nematode viability is likely due to the robust nature of the nematode's cuticle, which prevents many dyes/stains from entering the coelomate body plan as well as the fact that phenotype such as motility, as opposed to viability, is the most commonly accepted readout utilised in both academia and industry Pax 1986, Smout et al 2010).…”

Section: Methodologies For Characterising Nematode Viability and Phenmentioning

confidence: 99%

Cross-disciplinary approaches for measuring parasitic helminth viability and phenotype

Peak

Hoffmann

2011

An. Acad. Bras. Ciênc.

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Parasitic worms (helminths) within the Phyla Nematoda and Platyhelminthes are responsible for some of the most debilitating and chronic infectious diseases of human and animal populations across the globe. As no subunit vaccine for any parasitic helminth is close to being developed, the frontline strategy for intervention is administration of therapeutic, anthelmintic drugs. Worryingly, and unsurprising due to co-evolutionary mechanisms, many of these worms are developing resistance to the limited compound classes currently being used. This unfortunate reality has led to a renaissance in next generation anthelmintic discovery within both academic and industrial sectors. However, a major bottleneck in this process is the lack of quantitative methods for screening large numbers of small molecules for their effects on the whole organism. Development of methodologies that can objectively and rapidly distinguish helminth viability or phenotype would be an invaluable tool in the anthelmintic discovery pipeline. Towards this end, we describe how several basic techniques currently used to assess single cell eukaryote viability have been successfully applied to parasitic helminths. We additionally demonstrate how some of these methodologies have been adopted for high-throughput use and further modified for assessing worm phenotype. Continued development in this area is aimed at increasing the rate by which novel anthelmintics are identified and subsequently translated into everyday, practical applications.

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Colorimetric quantitation of filarial viability

Cited by 110 publications

References 3 publications

Brugia malayi and Acanthocheilonema viteae: antifilarial activity of transglutaminase inhibitors in vitro

Brugia malayi and Acanthocheilonema viteae: antifilarial activity of transglutaminase inhibitors in vitro

Interdomain lateral gene transfer of an essential ferrochelatase gene in human parasitic nematodes

Cross-disciplinary approaches for measuring parasitic helminth viability and phenotype

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