Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: An automated high-throughput system for phenotypic screening of chemical libraries on C. elegans and parasitic nematodes

doi: 10.1016/j.ijpddr.2017.11.004

Figure Lengend Snippet: Determination of the effect of acute treatment (3 h) with important anthelmintics on C. elegans adult motility using the INVAPP/Paragon movement index algorithm. Blue line fitted using the 3-parameter log-logistic model. EC 50 values are shown in parentheses, with standard error for this estimate calculated using drc . Each dot indicates movement score for one well. n = 5 wells for each concentration, with each replicate concentration on different plates. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Techniques: Concentration Assay

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: An automated high-throughput system for phenotypic screening of chemical libraries on C. elegans and parasitic nematodes

doi: 10.1016/j.ijpddr.2017.11.004

Figure Lengend Snippet: Determination of the effect of important anthelmintics on C. elegans growth using the INVAPP/Paragon movement index algorithm. Chronic treatment for 5 days from L1 stage. Blue line fitted using the 3-parameter log-logistic model. EC 50 values are shown in parentheses, with standard error for this estimate calculated using drc . Each dot indicates movement score for one well. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Techniques:

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: An automated high-throughput system for phenotypic screening of chemical libraries on C. elegans and parasitic nematodes

doi: 10.1016/j.ijpddr.2017.11.004

Figure Lengend Snippet: Utilising the INVAPP/Paragon system with ensheathed L3 T. circumcincta . (A) INVAPP/Paragon motility thresholding of single wells treated with DMSO or a DMSO solution of ivermectin (assay concentration 100 μM). Black pixels show those scored as being ‘motile’ by the software. (B) Concentration-response curve for treatment with the anthelmintic ivermectin, n = 4. Each replicate was carried out in different experimental sessions, using one batch of L3 T. circumcincta . Curve fitted using the four-parameter log-logistic function. Error range on the EC 50 estimate indicates standard error (delta method).

Article Snippet: Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Techniques: Concentration Assay, Software

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: An automated high-throughput system for phenotypic screening of chemical libraries on C. elegans and parasitic nematodes

doi: 10.1016/j.ijpddr.2017.11.004

Figure Lengend Snippet: Utilising the INVAPP/Paragon system with exsheathed L3 H. contortus . (A) INVAPP/Paragon motility thresholding of single wells treated with DMSO or a DMSO solution of ivermectin (assay concentration 100 μM). Black pixels show those scored as being ‘motile’ by the software. (B) Concentration-response curve for treatment with the anthelmintic ivermectin, n = 4. Each replicate drug treatment was carried out in different experimental sessions, each time freshly exsheathing the worms by bleach treatment, using one batch of ensheathed L3 H. contortus . Curve fitted using the four-parameter log-logistic function. Error range on the EC 50 estimate indicates standard error (delta method).

Article Snippet: Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Techniques: Concentration Assay, Software

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: An automated high-throughput system for phenotypic screening of chemical libraries on C. elegans and parasitic nematodes

doi: 10.1016/j.ijpddr.2017.11.004

Figure Lengend Snippet: Utilising the INVAPP/Paragon system with adult T. muris . (A) Example of section of movie, showing control and levamisole-treated wells. Selected movie frames are shown at 3-s intervals (B) Pixels for these wells that are considered to be motile as determined by the algorithm are shown in red. (C) Concentration-response curves for treatment with the anthelmintic levamisole, n = 4 wells per concentration, each replicate well on different plates using independent batches of T. muris from different mice, over 2 experimental sessions. Curve fitted using the three-parameter log-logistic function. Error range on the EC 50 estimate indicates standard error (delta method). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Techniques: Control, Concentration Assay

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: An automated high-throughput system for phenotypic screening of chemical libraries on C. elegans and parasitic nematodes

doi: 10.1016/j.ijpddr.2017.11.004

Figure Lengend Snippet: Screening the Pathogen Box in the C. elegans growth screen. (A) Volcano plot showing the results of the primary screen (n = 5 replicates per compound, concentration = 10 μM, each replicate was on different plates, over two experimental sessions). Each point represents one compound. Effective size is shown on the x axis, as log 2 -fold change (ratio of the median movement for the repeats of the compound to the median movement of DMSO-only wells). Statistical significance is shown on the y axis as the -log 10 P value in the Mann-Whitney-Wilcoxon test. A location at the top left of this plot indicates anthelmintic activity. (B) Secondary rescreen of hit compounds, using library material, in order of their activity in the primary screen. Statistical significance compared to DMSO-only control calculated by Dunnett's multiple comparison test (n = 5, * indicates P < 0.05, ** indicates P < 0.005, **** indicates P < 0.0005). (C,D,E) Concentration-response curves showing the activity of known anthelmintics – (C) tolfenpyrad, (D) auranofin, (E) isradipine – that were found in the Pathogen box screen retested using solid material (supplied by the Medicines for Malaria Venture) in the C. elegans growth assay. A concentration-response curve for mebendazole in the assay was presented in . Error bars indicate standard deviation, n = 4 wells per concentration per compound. Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Article Snippet: Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Techniques: Concentration Assay, MANN-WHITNEY, Activity Assay, Control, Comparison, Growth Assay, Standard Deviation

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: An automated high-throughput system for phenotypic screening of chemical libraries on C. elegans and parasitic nematodes

doi: 10.1016/j.ijpddr.2017.11.004

Figure Lengend Snippet: Concentration-response relationships for selected hit compounds in the C. elegans growth screen, using solid material supplied by Medicines for Malaria Venture. (A) MMV007920 (B) MMV020152 (C) MMV652003 (D) MMV688372. N = 4 wells per concentration per compound. Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Article Snippet: Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Techniques: Concentration Assay

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: An automated high-throughput system for phenotypic screening of chemical libraries on C. elegans and parasitic nematodes

doi: 10.1016/j.ijpddr.2017.11.004

Figure Lengend Snippet: Benzoxaborole compounds as anthelmintics. Structures of (A) the screen hit MMV652003 and (B) the related benzoxaborole tavaborole, which is approved as an anti-fungal. (C) Concentration-response curve for tavaborole in the C. elegans growth assay. N = 5 wells per concentration. Blue line fitted using three parameter log logistic model with drc . (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Curve fitting was undertaken using three parameter log logistic model (Graphpad Prism).

Techniques: Concentration Assay, Growth Assay