Journal: Insect molecular biology

Article Title: Brief exposures of human body lice to sub-lethal amounts of ivermectin over transcribes detoxification genes involved in tolerance

doi: 10.1111/j.1365-2583.2011.01097.x

Figure Lengend Snippet: Phylogenetic trees of cytochrome P450 monooxygenases (P450) and ABC transporters from deduced amino acid sequences of genes from female body lice that were significantly induced following a brief exposure to a sub-lethal amount of ivermectin. Louse genes over-transcribed following both immersion and contact exposures and that are highly expressed are given in bold red font. Louse genes over-transcribed following both immersion and contact exposures that are modestly expressed are given in normal red font. Louse genes that were over-transcribed by only one of the two exposure means and/or were either modestly or weakly expressed are given in normal black font. Non-louse genes used for comparisons are given in bold black font with the organism’s name.

Article Snippet: Total RNA was extracted with Trizol (MRC, Cincinnati, OH) from ivermectin-treated and untreated lice, treated with DNAase l and cDNA template for qPCR synthesized from 5 μg total RNA with SuperScript III Reverse Transcriptase (Invitrogen, Carlsbad, CA). qPCR was performed in a 20 μl reaction mixture containing 5 μl of 1/50-diluted cDNA, 5 pmol forward and reverse primers and 2× DyNAmo HS SYBR Green qPCR master mix (Finnzyme, Espoo, Finland) using Chromo4 thermal cycler (Bio-Rad, Hercules, CA).

Techniques:

Journal: Insect molecular biology

Article Title: Brief exposures of human body lice to sub-lethal amounts of ivermectin over transcribes detoxification genes involved in tolerance

doi: 10.1111/j.1365-2583.2011.01097.x

Figure Lengend Snippet: Comparative tolerance to lethal contact concentrations of ivermectin (1 %, w/v, IVM in panel A or 5 %, w/v, IVM in panel B) using female body lice pretreated by immersion with a sub-lethal concentration of ivermectin (10 −9 M) in ethanol. Asterisks (*) indicate that pretreated regressions are significantly different from ethanol (EtOH)-treated control regressions using the maximum log-likelihood ratio test ( P

Article Snippet: Total RNA was extracted with Trizol (MRC, Cincinnati, OH) from ivermectin-treated and untreated lice, treated with DNAase l and cDNA template for qPCR synthesized from 5 μg total RNA with SuperScript III Reverse Transcriptase (Invitrogen, Carlsbad, CA). qPCR was performed in a 20 μl reaction mixture containing 5 μl of 1/50-diluted cDNA, 5 pmol forward and reverse primers and 2× DyNAmo HS SYBR Green qPCR master mix (Finnzyme, Espoo, Finland) using Chromo4 thermal cycler (Bio-Rad, Hercules, CA).

Techniques: Concentration Assay

Journal: Insect molecular biology

Article Title: Brief exposures of human body lice to sub-lethal amounts of ivermectin over transcribes detoxification genes involved in tolerance

doi: 10.1111/j.1365-2583.2011.01097.x

Figure Lengend Snippet: Relative increases in the transcript levels of cytochrome P450 monooxygenase (P450) genes determined by qPCR following 2 (panels A and B) and 4 (panels C and D) h post ivermectin (IVM) treatments of female lice, where normalized fold-increase = normalized basal transcript level × [relative fold-increase of transcript level following IVM treatment (10 −6 M for direct contact (panels A and C); 10 −9 M IVM in ethanol for immersion (panels B and D)) − 1]. Asterisks (*) indicate a significant increase in transcript levels over respective controls using Student’s t-test ( P

Article Snippet: Total RNA was extracted with Trizol (MRC, Cincinnati, OH) from ivermectin-treated and untreated lice, treated with DNAase l and cDNA template for qPCR synthesized from 5 μg total RNA with SuperScript III Reverse Transcriptase (Invitrogen, Carlsbad, CA). qPCR was performed in a 20 μl reaction mixture containing 5 μl of 1/50-diluted cDNA, 5 pmol forward and reverse primers and 2× DyNAmo HS SYBR Green qPCR master mix (Finnzyme, Espoo, Finland) using Chromo4 thermal cycler (Bio-Rad, Hercules, CA).

Techniques: Real-time Polymerase Chain Reaction

Journal: Insect molecular biology

Article Title: Brief exposures of human body lice to sub-lethal amounts of ivermectin over transcribes detoxification genes involved in tolerance

doi: 10.1111/j.1365-2583.2011.01097.x

Figure Lengend Snippet: Relative increases in the transcript levels of ABC transporter genes determined by qPCR following 2 (panels A and B) and 4 (panels C and D) h post ivermectin (IVM) treatments of female lice, where normalized fold-increase = normalized basal transcript level × [relative fold-increase of transcript level following IVM treatment (10 −6 M for contact (panels A and C); 10 −9 M IVM for immersion (panels B and D)) − 1]. Asterisks (*) indicate a significant increase in transcript levels over respective controls using Student’s t-test ( P

Article Snippet: Total RNA was extracted with Trizol (MRC, Cincinnati, OH) from ivermectin-treated and untreated lice, treated with DNAase l and cDNA template for qPCR synthesized from 5 μg total RNA with SuperScript III Reverse Transcriptase (Invitrogen, Carlsbad, CA). qPCR was performed in a 20 μl reaction mixture containing 5 μl of 1/50-diluted cDNA, 5 pmol forward and reverse primers and 2× DyNAmo HS SYBR Green qPCR master mix (Finnzyme, Espoo, Finland) using Chromo4 thermal cycler (Bio-Rad, Hercules, CA).

Techniques: Real-time Polymerase Chain Reaction

Journal: Insect molecular biology

Article Title: Brief exposures of human body lice to sub-lethal amounts of ivermectin over transcribes detoxification genes involved in tolerance

doi: 10.1111/j.1365-2583.2011.01097.x

Figure Lengend Snippet: Log time versus logit mortality regression analyses of a lethal contact concentration of 1 % (w/v) ivermectin (IVM) using female body lice pretreated by immersion with either a sub-lethal concentration of verapamil (VRP; 1 %, w/v) or dexamethasone (DEX; 1 %, w/v) in ethanol. Asterisks (*) indicate that pretreated regressions are significantly different from ethanol (EtOH)-treated control regressions using the maximum log-likelihood ratio test ( P

Article Snippet: Total RNA was extracted with Trizol (MRC, Cincinnati, OH) from ivermectin-treated and untreated lice, treated with DNAase l and cDNA template for qPCR synthesized from 5 μg total RNA with SuperScript III Reverse Transcriptase (Invitrogen, Carlsbad, CA). qPCR was performed in a 20 μl reaction mixture containing 5 μl of 1/50-diluted cDNA, 5 pmol forward and reverse primers and 2× DyNAmo HS SYBR Green qPCR master mix (Finnzyme, Espoo, Finland) using Chromo4 thermal cycler (Bio-Rad, Hercules, CA).

Techniques: Concentration Assay