Journal: Pharmacognosy Research

Article Title: Effects of mitragynine and 7-hydroxymitragynine (the alkaloids of Mitragyna speciosa Korth) on 4-methylumbelliferone glucuronidation in rat and human liver microsomes and recombinant human uridine 5’-diphospho-glucuronosyltransferase isoforms

doi: 10.4103/0974-8490.159580

Figure Lengend Snippet: Inhibitory effects of mitragynine, 7-hydroxymitragynine, ketamine, buprenorphine, and diclofenac on 4-methylumbelliferone glucuronide formation by rat liver microsomes. Panels to the right show inhibitor concentrations. Each bar represents the mean percentage activity relative to control ± standard error of mean for three independent measurements ( n = 3). Statistical analysis was conducted using one-way ANOVA and Dunnett test. * P < 0.05 versus control (no inhibitor)

Article Snippet: Ketamine hydrochloride and buprenorphine hydrochloride were purchased from Toronto Research Chemicals Inc. (North York, ON, Canada).

Techniques: Activity Assay, Control

Journal: Pharmacognosy Research

Article Title: Effects of mitragynine and 7-hydroxymitragynine (the alkaloids of Mitragyna speciosa Korth) on 4-methylumbelliferone glucuronidation in rat and human liver microsomes and recombinant human uridine 5’-diphospho-glucuronosyltransferase isoforms

doi: 10.4103/0974-8490.159580

Figure Lengend Snippet: Inhibitory effects of mitragynine, 7-hydroxymitragynine, ketamine, buprenorphine, and diclofenac on 4-methylumbelliferone glucuronide formation by recombinant human UGT2B7 isoform. Panels to the right show inhibitor concentrations. Each bar represents the mean percentage activity relative to control ± standard error of mean for three independent measurements ( n = 3). Statistical analysis was conducted using one-way ANOVA and Dunnett test. * P < 0.05 versus control (no inhibitor)

Article Snippet: Ketamine hydrochloride and buprenorphine hydrochloride were purchased from Toronto Research Chemicals Inc. (North York, ON, Canada).

Techniques: Recombinant, Activity Assay, Control

Journal: Pharmacognosy Research

Article Title: Effects of mitragynine and 7-hydroxymitragynine (the alkaloids of Mitragyna speciosa Korth) on 4-methylumbelliferone glucuronidation in rat and human liver microsomes and recombinant human uridine 5’-diphospho-glucuronosyltransferase isoforms

doi: 10.4103/0974-8490.159580

Figure Lengend Snippet: Inhibitory effects of mitragynine, 7-hydroxymitragynine, ketamine, buprenorphine, and diclofenac on 4-methylumbelliferone glucuronide formation by human liver microsomes. Panels to the right show inhibitor concentrations. Each bar represents the mean percentage activity relative to control ± standard error of mean for three independent measurements ( n = 3). Statistical analysis was conducted using one-way ANOVA and Dunnett test. * P < 0.05 versus control (no inhibitor)

Article Snippet: Ketamine hydrochloride and buprenorphine hydrochloride were purchased from Toronto Research Chemicals Inc. (North York, ON, Canada).

Techniques: Activity Assay, Control

Journal: Pharmacognosy Research

Article Title: Effects of mitragynine and 7-hydroxymitragynine (the alkaloids of Mitragyna speciosa Korth) on 4-methylumbelliferone glucuronidation in rat and human liver microsomes and recombinant human uridine 5’-diphospho-glucuronosyltransferase isoforms

doi: 10.4103/0974-8490.159580

Figure Lengend Snippet: Inhibitory effects of mitragynine, 7-hydroxymitragynine, ketamine, buprenorphine, and diclofenac on 4-methylumbelliferone glucuronide formation by recombinant human UGT1A1 isoform. Panels to the right show inhibitor concentrations. Each bar represents the mean percentage activity relative to control ± standard error of mean for three independent measurements ( n = 3). Statistical analysis was conducted using one-way ANOVA and Dunnett test. * P < 0.05 versus control (no inhibitor)

Article Snippet: Ketamine hydrochloride and buprenorphine hydrochloride were purchased from Toronto Research Chemicals Inc. (North York, ON, Canada).

Techniques: Recombinant, Activity Assay, Control

Journal: Analytical and bioanalytical chemistry

Article Title: High-throughput simultaneous analysis of buprenorphine, methadone, cocaine, opiates, nicotine, and metabolites in oral fluid by liquid chromatography tandem mass spectrometry

doi: 10.1007/s00216-010-3903-5

Figure Lengend Snippet: Liquid chromatography tandem mass spectrometry chromatograms of an oral fluid sample spiked at the low quality control concentration, 1.5 μg/L for codeine, 6-acetylmorphine (6AM), 6-acetylcodeine (6AC), heroin, benzoylecgonine (BE), cocaine, cotinine, trans-3′-hydroxycotinine (OH-cotinine), and buprenorphine (BUP), and 3 μg/L for morphine, ecgonine methyl ester (EME), anhydroecgonine methyl ester (AEME), nicotine, norbuprenorphine (NBUP), methadone, and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP). Quantitation transitions of analytes monitored in the first (a), second (b), and third (c) acquisition windows are included

Article Snippet: Morphine, codeine, 6-acetylmorphine (6AM), heroin, 6-acetylcodeine (6AC), cocaine, benzoylecgonine (BE), ecgonine methyl ester (EME), anhydroecgonine methyl ester (AEME), nicotine, cotinine, BUP, norbuprenorphine (NBUP), methadone, and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) standards and morphine- d 6 , 6AM- d 6 , heroin- d 9 , cocaine- d 3 , BE- d 8 , EME- d 3 , nicotine d 4 , cotinine- d 3 , BUP- d 4 , NBUP- d 3 , methadone- d 3 , and EDDP- d 3 internal standards (IStd) (1 mg/mL or 100 μg/mL ampoules) were obtained from Cerilliant (Round Rock, TX, USA).

Techniques: Liquid Chromatography, Mass Spectrometry, Concentration Assay, Quantitation Assay

Journal: International Journal of Molecular Sciences

Article Title: Opioid-Modulated Receptor Localization and Erk1/2 Phosphorylation in Cells Coexpressing μ-Opioid and Nociceptin Receptors

doi: 10.3390/ijms24021048

Figure Lengend Snippet: HEK 293 cells transiently transfected with MOP-CFP and NOP-YFP were serum-starved overnight; vehicle (ddH 2 O), morphine (1 μM), or buprenorphine (1 μM) was added to cells immediately before FRET/SPT measurements. ( A – C ) present the results obtained under the conditions of pre-treatment, post-bup, and post-mor, respectively. The images of MOP-CPF (shown in cyan), NOP-YFP (shown in yellow), FRET ratio (%), and a magnified view from a white box in the respective FRET ratio image are shown for each condition from left to right. ( D ) is the control image taken from the MOP-CFP-channel at the pretreatment condition, which is used to confirm whether the receptors are properly expressed on the cell surface. ( E ) The histograms of the FRET ratio obtained from each condition were fitted to Gaussian distributions, indicating the peak value with a standard deviation. Scale bar = 20 μm. The bin size of the histogram = 0.01. Twelve cells were used for statistical analysis in each condition.

Article Snippet: Morphine (1 μM, Toronto Research Chemicals, North York, ON, Canada) or buprenorphine (1 μM, Toronto Research Chemicals) was added to cells shortly before starting FRET/SPT measurements and present in the serum-free medium throughout the whole experiment.

Techniques: Transfection, Control, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Opioid-Modulated Receptor Localization and Erk1/2 Phosphorylation in Cells Coexpressing μ-Opioid and Nociceptin Receptors

doi: 10.3390/ijms24021048

Figure Lengend Snippet: Representative confocal images ( A ) and colocalization rates of MOP and NOP on lipid rafts (red) ( B ) from HEK 293 cells stably expressing HA-tagged MOP (green) and/or myc-tagged NOP (magenta) after exposure to morphine (1 μM) or buprenorphine (1 μM) for 20 min at 4 °C. The inset panels display higher magnification images highlighting the colocalization. Scale bars are equal to 10 μm. Each bar represents the mean ± SE value of the percentage of ROI colocalization derived from five independent experiments with 4–6 individual cell images in each group.

Article Snippet: Morphine (1 μM, Toronto Research Chemicals, North York, ON, Canada) or buprenorphine (1 μM, Toronto Research Chemicals) was added to cells shortly before starting FRET/SPT measurements and present in the serum-free medium throughout the whole experiment.

Techniques: Stable Transfection, Expressing, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Opioid-Modulated Receptor Localization and Erk1/2 Phosphorylation in Cells Coexpressing μ-Opioid and Nociceptin Receptors

doi: 10.3390/ijms24021048

Figure Lengend Snippet: Representative immunoblots of Erk1/2 (p44/p42 MAPK) phosphorylation after stimulation with ( A ) morphine and ( B ) buprenorphine in cells stably expressing MOP, NOP, and MOP+NOP receptors. Immunoblotting using anti-phospho-p44/p42 MAPK (Erk1/2) rabbit polyclonal and anti-p44/42 MAPK (Erk1/2) rabbit monoclonal antibodies was performed to visualize opioid-induced phosphorylation of Erk1/2 (p44 MAPK-P and p42 MAPK-P) and total Erk1/2 (p44 MAPK and p42 MAPK), respectively. ( C ) Dose–response curves of the immunoblots. The density of phospho-p44/p42 MAPK versus the density of p44/42 MAPK in control cells (with vehicle treatment) was considered to be 1. All the experiments were repeated no less than four times, and the values represent the average ± SE.

Article Snippet: Morphine (1 μM, Toronto Research Chemicals, North York, ON, Canada) or buprenorphine (1 μM, Toronto Research Chemicals) was added to cells shortly before starting FRET/SPT measurements and present in the serum-free medium throughout the whole experiment.

Techniques: Western Blot, Phospho-proteomics, Stable Transfection, Expressing, Bioprocessing, Control