Journal: Pesticide biochemistry and physiology

Article Title: High-content phenotypic screening identifies novel chemistries that disrupt mosquito activity and development.

doi: 10.1016/j.pestbp.2022.105037

Figure Lengend Snippet: Fig. 5. Chemical structure of chemistries selected for dose-response, behavioral, and developmental assays. A. Rotenone; B. Ivermectin; C. Nimesulide; D. 6- Methoxy-1,2,3,4-tetrahydro-9H-pyrido[3,4b] indole; E. Metergoline; F. Ritodrine hydrochloride; G. (−)-Quinpirole hydrochloride; H. R(+)-3PPP hydrochloride.

Article Snippet: Nimesulide, Metergoline, Ritodrine hydrochloride, (− )-Quinpirole hydrochloride, 6-Methoxy-1,2,3,4-tetrahydro-9H-pyrido[3,4b] indole, and R(+)-3-PPP hydrochloride, plus positive controls Amitriptyline, Ivermectin and Rotenone were sourced from Tocris Pharmaceuticals (Minneapolis, MN) and Sigma-Aldrich (St. Louis, MO).

Techniques:

Journal: Pesticide biochemistry and physiology

Article Title: High-content phenotypic screening identifies novel chemistries that disrupt mosquito activity and development.

doi: 10.1016/j.pestbp.2022.105037

Figure Lengend Snippet: Fig. 6. Dose-response assays showing toxicity of primary screen hits to L3 larvae of Aedes aegypti at 3, 24, and 48 h post exposure. The parasiticides rotenone and ivermectin, identified/confirmed in the primary/secondary screen, were included as positive controls. Nimesulide and metergoline were selected as examples of toxic chemistries identified by screening. 6-Methoxy-1,2,3,4-tetrahydro-9H-pyrido[3,4b] indole, (−)-Quinpirole hydrochloride, R(+)-3PPP hydrochloride and Ritodrine hydrochloride were selected as examples of non-toxic GPCR-targeting chemistries associated with a range of atypical morphological phenotypes (see Table 2 and Figure 4). The data represent minimum n = 3 biological replicates.

Article Snippet: Nimesulide, Metergoline, Ritodrine hydrochloride, (− )-Quinpirole hydrochloride, 6-Methoxy-1,2,3,4-tetrahydro-9H-pyrido[3,4b] indole, and R(+)-3-PPP hydrochloride, plus positive controls Amitriptyline, Ivermectin and Rotenone were sourced from Tocris Pharmaceuticals (Minneapolis, MN) and Sigma-Aldrich (St. Louis, MO).

Techniques:

Journal: Pharmaceutics

Article Title: Enhanced Intradermal Delivery of Nanosuspensions of Antifilariasis Drugs Using Dissolving Microneedles: A Proof of Concept Study

doi: 10.3390/pharmaceutics11070346

Figure Lengend Snippet: ( a ) Representative response surface plots describing the effect of dependent variables on the particle size, polydispersity indexes (PDI), and zeta potential of doxycycline (DOX) NS. ( b ) Representative response surface plots describing the effect of dependent variables on the particle size, PDI, and zeta potential of albendazole sulfoxide (ABZ-OX) NS. ( c ) Representative response surface plots describing the effect of dependent variables on the particle size, PDI, and zeta potential of ivermectin (IVM) NS.

Article Snippet: Doxycycline monohydrate (DOX) (purity, ≥98%) and ivermectin (purity, ≥98%) of analytical grade were purchased from Alfa Aesar (Lancashire, UK), while chloroform, citric acid, dimethylsulfoxide (DMSO), albendazole sulfoxide (ABZ-OX) (purity, ≥99.9%), hydroxyl propyl methylcellulose (HPMC), poly (vinyl alcohol) (PVA) (MW 9–10 kDa), poly (vinyl alcohol) (PVA) (31–50 kDa), sodium carboxymethylcellulose (NaCMC), sodium hydroxide, and sodium lauryl sulphate (SLS) were purchased from Sigma-Aldrich (Dorset, UK).

Techniques: Zeta Potential Analyzer

Journal: Journal of biomedical science

Article Title: Targeting the G-quadruplex as a novel strategy for developing antibiotics against hypervirulent drug-resistant Staphylococcus aureus.

doi: 10.1186/s12929-024-01109-3

Figure Lengend Snippet: Fig. 7 NMM stabilizes the G4-motif in PmraZ promoter to inhibit transcription/translation. A, B Three constructs showing promoter-less mCherry empty vector (EV) (A I), the recombinant fusion protein, rMraZ-His6-mCherry expression under PgapA and PmraZ promoters (A II–III) were transformed in E. coli DH5α showing the differential expression of rMraZ-His6-mCherry fusion protein (B I–III). C, D Qualitative and quantitative assessment of differential expression of rMraZ-His6-mCherry fusion protein without or with 5 µM NMM using qualitative confocal fluorescence microscopy (C I–III); and quantitative fluorometric estimation of red fluorescent signal intensity of rMraZ-His6-mCherry fusion protein showing that the NMM inhibits the rMraZ-His6-mCherry fusion protein expression due to the inhibition of bacterial coupled transcription/translation (D). E DNA templates for IVT reactions showing T7 promoter with G4-motif (WT PmraZ_G4_3) (lane 1) and T7 promoter without G4-motif (lane 2) tagged with mraZ-his6-stop. F, G Western blot and immunodetection of recombinant MraZ-His6 protein using Anti-His antibody showed the NMM concentration-dependent inhibition of coupled transcription/translation in case of T7 promoter with G4-motif (WT PmraZ_G4_3) (F), while such inhibition was absent where T7 promoter is devoid of G4-motif (G)

Article Snippet: Immunodetection of recombinant MraZ-His6 protein of IVT reactions without or with NMM was carried out with a primary mouse-monoclonal anti-His antibody (Santa Cruz Biotechnology, USA).

Techniques: Construct, Plasmid Preparation, Recombinant, Expressing, Transformation Assay, Quantitative Proteomics, Fluorescence, Microscopy, Inhibition, Western Blot, Immunodetection, Concentration Assay

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: Larval development assays were undertaken for all lines, at pre- and post-treatment timepoints, in the third generation of selection. (A) Control (CTL) lines had no treatment applied but sampling was time-matched with the selected lines ((B) IVM = ivermectin and (C) MOX = moxidectin). Y axis shows the proportion of the population in each well that developed to L3.

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Selection, Control, Sampling

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: (A–C) Each sample from each generation is compared with the F0 parental sample and 10 kb window F st values plotted along the chromosomes (I-X). For each of the ivermectin selected (IVM) and moxidectin selected (MOX) lines, a line was plotted indicating five standard deviations above the genome-wide mean of the F3 generations (dashed and dotted respectively). CTL = control lines. Note that no sequence data was available for sample IVM1. (D) Chromosome V pairwise comparisons of third generation ivermectin and moxidectin samples to each other. Horizontal lines are five standard deviations above the mean F st value for the F3 generations – the same as in panels A-C, but with both ivermectin and moxidectin lines overlaid on each faceted plot.

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Genome Wide, Control, Sequencing

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: (A–C) Ratio of nucleotide diversity in three generations (F1 to F3) of sub-therapeutic selection in comparison to F0 sample population. Key: C = control, I = ivermectin, M = moxidectin. L1 to L3 indicate biological replicate selection lines. Pi ratio = Normalised Sample θπ/ Normalised F0 θπ. Nucleotide diversity for each sample window was normalised by dividing by the genome-wide median diversity. Colour indicates the rank within the treatment group, where blue indicates a higher diversity than F0, and red a lower diversity. Note that the rank indicates the genome-wide ranking. Note that no sequence data was available for sample F1_I_L1. (D) Nucleotide diversity (θπ) of the F0 sample along Chromosome V. More negative values indicate lower diversity.

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Selection, Comparison, Control, Genome Wide, Sequencing

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: (A) Ivermectin selected lines, (B) moxidectin selected lines, (C) control lines, (D) F0 sample. Note that no sequence data was available for sample F1_I_L1. More negative values (red) correlate to more rare alleles than expected for the population size, while more positive values (blue) indicate fewer rare alleles than expected and either very few alleles at all (highly conserved), or balancing selection of alleles. If the population is neutrally evolving, Tajima’s D is close to zero (mix of rare and common alleles).

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Control, Sequencing, Selection

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: Negative values indicate a reduction in the reference allele frequency at that site from F0 to F3. Each point is a SNP; red points are those which are significantly different to controls following Holm’s correction of the p-values, and where the corrected p-value is < 0.001. In other words: if red, the reference allele is changing in treated lines differently to control lines, and if negative it is reducing in the treated lines. (A) ivermectin vs control lines (slope coefficient plotted for ivermectin lines), (B) moxidectin vs control lines (slope coefficient plotted for moxidectin lines), (C) selected (ivermectin and moxidectin) vs control lines (slope coefficient plotted for combined selected lines).

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Control

Journal: PLOS Pathogens

Article Title: Analyses of emerging macrocyclic lactone resistance: Speed and signature of ivermectin and moxidectin selection and evidence of a shared genetic locus

doi: 10.1371/journal.ppat.1013578

Figure Lengend Snippet: PCA plots for male (A) and female (B) datasets. Genome-wide differential gene expression visualised as karyoplots (C) with i. ivermectin selected males versus control males, ii. ivermectin selected females versus control females, iii. moxidectin selected males versus control males, iv. moxidectin selected females versus control females. Upregulated genes are in yellow, downregulated genes are in blue, with adjusted P < 0.01 for significance.

Article Snippet: To make the drug plates, stock solutions of ivermectin aglycone (Santa Cruz Animal Health, 202189), thiabendazole (Sigma, T8904), and tetramisole hydrochloride (Sigma, L9756) were prepared and used for doubling dilutions as follows: ivermectin aglycone (stock: 100 μM in DMSO, dilutions: 50 μM to 0.0485 μM), thiabendazole (stock: 1 mM in DMSO, dilutions: 500 μM to 0.06125 μM), tetramisole hydrochloride (stock: 2.5 mM in water, dilutions: 1250 μM to 1.287 μM).

Techniques: Genome Wide, Gene Expression, Control