PI and FDA uptake correlates with schistosomula survival.

<p>Schistosomula were incubated with serial dilutions of GA and PI and FDA fluorescence signals in treated schistosomula were measured. Left axis: FDA raw data for each concentration (n = 5) normalized between 50 μM GA-treated control (0% survival) and DMSO treated schistosomula (100% survival). Right axis: PI raw data for each concentration (n = 5) normalized between 50 μM GA-treated control (100% death) and DMSO treated schistosomula (0% death). The error bars represent the standard error, while the dotted lines are the 95% confidence interval of the fitted sigmoid curve. The calculated LD₅₀ was 3.5 μM. The percentage of live and dead parasites, calculated as described in Materials and Methods, are shown.</p

Expression of human tau in the optic nerve of mice transgenic for human mutant P301S tau.

<p>(A–C), Staining for human tau (A, green) and βIII tubulin (B, red) showed co-localisation in the axons of the optic nerve (C, overlay image of A and B; example from 1 month old mouse). (D–F), Staining for tau phosphorylated at S202/T205 (D, green) and βIII tubulin (E, red) showed co-localisation in the axons of the optic nerve (F, overlay image of D and E; example from 5 month old mouse). Arrows indicate examples of co-localisation. Scale bar, 20 µm.</p

Effect of different drugs on viability and phenotype of adult <i>S. mansoni</i> worms <i>in vitro</i>.

<p>Adult worms were incubated with the indicated compounds and phenotypes were scored as described under Materials and Methods. (A) Viability curves of adult schistosomes cultured for 5 days following overnight treatment in the presence of 10 μM hit compounds (mean ± SE, three independent experiments). Different phenotypes of adult schistosomes 5 days after overnight exposure to DMSO alone (B), PZQ (C), GA (D), parthenolide (E), disulfiram (F), menadione (G), thonzonium bromide (H) and sanguinarine chloride hydrate (I). Scale bar = 0.2 cm.</p

Retrograde axonal transport is reduced in optic nerve of mice transgenic for human mutant P301S tau.

<p>Fluorescent cholera toxin B was injected bilaterally into the superior colliculus and the amount transported measured in 5-month-old (A) and 3-month-old (C) P301S tau transgenic and C57/Bl6 control mice. Fluorescence intensity was lower along the length of the optic nerve in transgenic mice at all ages, compared to C57/Bl6 controls. Statistical analysis of the area under the fluorescence intensity curve for each individual showed a significant reduction in retrograde axonal transport in optic nerves from P301S tau transgenic mice at 5 months (B) and at 3 months (D), compared to controls. Data are presented as mean±SEM.</p

Reduced axonal transport in optic nerve of mice transgenic for human mutant P301S tau increases neuronal susceptibility to injury.

<p>Retinal ganglion cell (RGC) survival was quantified following a mild unilateral excitotoxic injury of the left eye (LE) by counting NeuN-positive nuclei in the RGC layer of the whole-mounted retina of 1, 3 and 5 month old mice (A–D). Percentage RGC loss was calculated by comparing the number of surviving RGCs in injured retinas to that of the uninjured right eye (RE). Representative images of NeuN-positive nuclei in injured P301S tau transgenic (A) and C57/Bl6 (C) retinas from 5 month old animals, compared to uninjured contralateral retinas (B and D), are shown as an example. Statistical analysis revealed a significant increase in RGC death following mild excitotoxic injury in P301S tau transgenic retinas, compared to C57/Bl6 control retinas (E), at both 3 months and 5 months of age; however no difference in RGC excitotoxic death was found at 1 month of age between control and transgenic retinas. N-methyl-D-aspartic acid (NMDA) was used as the excitotoxin. Scale bar, 100 µm.</p

ATP quantitation to assess schistosomula survival with known anti-schistosomal drugs.

<p>Schistosomula (100/well) were incubated in triplicate with serial dilutions of the indicated compounds. Data evaluated at 24 and 72 hours post treatment are normalized between 50 μM GA-treated control (0% survival) and DMSO treated schistosomula (100% survival) at each time point. The error bars represent the standard error.</p

Robust penetration of the CTG reagent with preservation of the overall schistosomula morphology.

<p>Representative fluorescence (A), bright field (B) and merge (C) confocal laser microscopy images of schistosomula treated with DMSO and incubated with CTG and the membrane-impermeant DNA dye CellTox green are shown. Scale bar = 25 μm.</p

LD₅₀ of hit compounds on schistosomula.

<p>LD₅₀ of hit compounds on schistosomula.</p

ATP luminescent signal correlates with schistosomula viability.

<p>Schistosomula ranging from 25–200 were incubated with serial dilutions of GA and ATP luminescence signals were measured. Raw data (RLU) for each concentration (n = 5) are reported on the y-axis. The error bars represent the standard error. The table reports the calculated LD₅₀ at different parasite concentrations.</p

ATP quantitation correlates with the number of schistosomula.

<p>Correlation between the number of schistosomula (X-axis, logarithmic scale) and the ATP signals (Y-axis, linear scale). A semi-log plot was used to better visualize the data; a linear correlation between schistosomula numbers and ATP signals is represented by the portion of curve comprised between dotted vertical lines. The LLoQ (Lower Limit of Quantification) was defined as a signal greater than three times the background signal. The ULoQ (Upper Limit of Quantification) was defined as the highest signal lying on the linear correlation. RLU = Relative Luminescent Units.</p