Journal: Current pharmaceutical biotechnology

Article Title: Oral fluid vs. Urine Analysis to Monitor Synthetic Cannabinoids and Classic Drugs Recent Exposure

doi: 10.2174/1389201018666171122113934

Figure Lengend Snippet: Total ion chromatogram of 11 synthetic cannabinoids and 13 metabolites in urine sample at 15 ng/mL. 1, AB-PINACA 5-Pentanoic acid; 2, A-796260; 3, AB-FUBINACA; 4, JWH-250 4-Hydroxypentyl; 5, JWH-073 4-Butanoic acid; 6, AB-PINACA; 7, JWH-018 5-Pentanoic acid; 8, AM2201 4-Hydroxypentyl; 9, MAM2201 4-Hydroxypentyl; 10, JWH-019 5-Hydroxyhexyl; 11, UR-144 5-Pentanoic acid; 12, JWH-122 4-Hydroxypentyl; 13, XLR-11 4-Hydroxypentyl; 14, UR-144 4-Hydroxypentyl; 15, 5-Fluoro PB-22; 16, JWH-210 4-Hydroxypentyl; 17, APINACA (AKB-48) 5-Hydroxypentyl; 18, PB-22; 19, MAM2201; 20, JWH-073; 21, XLR-11; 22, JWH-018; 23, UR-144; 24, APINACA (AKB-48).

Article Snippet: Synthetic cannabinoids (A-796260, AB-FUBINACA, AB-PINACA, APINACA (AKB-48), JWH-018, JWH-073, MAM2201, PB-22, 5-Fluoro PB-22, UR-144, XLR-11) and metabolites standards (AB-PINACA 5-Pentanoic acid, APINACA (AKB-48) 5-Hydroxypentyl, AM2201 4-Hydroxypentyl, JWH-018 5-Pentanoic acid, JWH-019 5-Hydroxyhexyl, JWH-073 4-Butanoic acid, JWH-122 4-Hydroxypentyl, JWH-210 4-Hydroxypentyl, JWH-250 4-Hydroxypentyl, MAM2201 4-Hydroxypentyl, UR-144 4-Hydroxypentyl, UR-144 5-Pentanoic acid, XLR-11 4-Hydroxypentyl) and internal standards (IS) (JWH-250 4-Hydroxypentyl-d 5 , AM2201 4-Hydroxypentyl-d 5 , and JWH-210 4-Hydroxypentyl-d 5 ) were purchased from Cerilliant (Round Rock, TX) at 100 μg/mL in methanol or acetonitrile.

Techniques:

Journal: Current pharmaceutical biotechnology

Article Title: Oral fluid vs. Urine Analysis to Monitor Synthetic Cannabinoids and Classic Drugs Recent Exposure

doi: 10.2174/1389201018666171122113934

Figure Lengend Snippet: Total ion chromatogram of 11 synthetic cannabinoids in oral fluid sample at 2.5 ng/mL. 1, A-796260; 2, AB-FUBINACA; 3, AB-PINACA; 4, 5-Fluoro PB-22; 5, PB-22; 6, MAM2201; 7, JWH-073; 8, XLR-11; 9, JWH-018; 10, UR-144; 11, APINACA (AKB-48).

Article Snippet: Synthetic cannabinoids (A-796260, AB-FUBINACA, AB-PINACA, APINACA (AKB-48), JWH-018, JWH-073, MAM2201, PB-22, 5-Fluoro PB-22, UR-144, XLR-11) and metabolites standards (AB-PINACA 5-Pentanoic acid, APINACA (AKB-48) 5-Hydroxypentyl, AM2201 4-Hydroxypentyl, JWH-018 5-Pentanoic acid, JWH-019 5-Hydroxyhexyl, JWH-073 4-Butanoic acid, JWH-122 4-Hydroxypentyl, JWH-210 4-Hydroxypentyl, JWH-250 4-Hydroxypentyl, MAM2201 4-Hydroxypentyl, UR-144 4-Hydroxypentyl, UR-144 5-Pentanoic acid, XLR-11 4-Hydroxypentyl) and internal standards (IS) (JWH-250 4-Hydroxypentyl-d 5 , AM2201 4-Hydroxypentyl-d 5 , and JWH-210 4-Hydroxypentyl-d 5 ) were purchased from Cerilliant (Round Rock, TX) at 100 μg/mL in methanol or acetonitrile.

Techniques:

Journal: Current pharmaceutical biotechnology

Article Title: Oral fluid vs. Urine Analysis to Monitor Synthetic Cannabinoids and Classic Drugs Recent Exposure

doi: 10.2174/1389201018666171122113934

Figure Lengend Snippet: Urine samples (n=70) positive results for cannabis, cocaine, opiates, benzodiazepines, amphetamines and synthetic cannabinoids (AB-FUBINACA, PB-22, 5-Fluoro-PB-22, UR-144 metabolites). Cutoffs in urine were 1 ng/mL for synthetic cannabinoids, 50 ng/mL for cannabis, 200 ng/mL for benzodiazepines and 300 ng/mL for cocaine, opiates and amphetamines.

Article Snippet: Synthetic cannabinoids (A-796260, AB-FUBINACA, AB-PINACA, APINACA (AKB-48), JWH-018, JWH-073, MAM2201, PB-22, 5-Fluoro PB-22, UR-144, XLR-11) and metabolites standards (AB-PINACA 5-Pentanoic acid, APINACA (AKB-48) 5-Hydroxypentyl, AM2201 4-Hydroxypentyl, JWH-018 5-Pentanoic acid, JWH-019 5-Hydroxyhexyl, JWH-073 4-Butanoic acid, JWH-122 4-Hydroxypentyl, JWH-210 4-Hydroxypentyl, JWH-250 4-Hydroxypentyl, MAM2201 4-Hydroxypentyl, UR-144 4-Hydroxypentyl, UR-144 5-Pentanoic acid, XLR-11 4-Hydroxypentyl) and internal standards (IS) (JWH-250 4-Hydroxypentyl-d 5 , AM2201 4-Hydroxypentyl-d 5 , and JWH-210 4-Hydroxypentyl-d 5 ) were purchased from Cerilliant (Round Rock, TX) at 100 μg/mL in methanol or acetonitrile.

Techniques:

Journal: Frontiers in Endocrinology

Article Title: VHL-P138R and VHL-L163R Novel Variants: Mechanisms of VHL Pathogenicity Involving HIF-Dependent and HIF-Independent Actions

doi: 10.3389/fendo.2022.854365

Figure Lengend Snippet: Differences in mRNA fibronectin expression for P138R and L163R pVHL variants with similar disrupted deposition patterns. (A) Fibronectin mRNA expression of 786-O, WT, P138R, and L163R cell lines. Results are presented as fold change compared to WT cells. Values are expressed as ± SD of three independent experiments performed in triplicate. ns, not significant; *p = 0.0011, **p = 0.0030, one-way ANOVA and Tukey’s posttest. (B) Cell lines were cultured on coverslips to assess fibronectin deposition with anti-fibronectin Cy5 conjugated (in red) by immunofluorescence. Nuclei were dyed with 5 μg/ml Hoechst as shown in blue. Images were taken at ×40 on a Carl-Zeiss AxioScope A1 microscope.

Article Snippet: Nuclei were dyed with Hoechst (5 µg/ml), and pictures were taken on a Carl-Zeiss AxioScope A1 microscope.

Techniques: Expressing, Cell Culture, Immunofluorescence, Microscopy

Journal: Oncology Letters

Article Title: Inhibition of circular RNA_0000285 prevents cell proliferation and induces apoptosis in thyroid cancer by sponging microRNA-654-3p

doi: 10.3892/ol.2021.12934

Figure Lengend Snippet: miR-654-3p is a direct target of circRNA_0000285. (A) Schematic diagram of the miR-654-3p binding site in the 3′-untranslated region of circRNA_0000285. (B) The association between miR-654-3p and circRNA_0000285 was verified via the dual-luciferase reporter assay. Relative circRNA_0000285 expression in (C) TPC-1 and (D) FTC133 lysates pulled down by circRNA_0000285 probe or oligo probe. (E) Relative miR-654-3p expression in TPC-1 and FTC133 lysates pulled down by circRNA_0000285 probe or oligo probe. (F and G) The RNA immunoprecipitation assay was performed using Ago2 and IgG antibodies to immunoprecipitate. The expression levels of circRNA_0000285 and miR-654-3p in TPC-1 and FTC133 cells were detected via reverse transcription-quantitative PCR analysis. **P<0.01 vs. mimic control; # P<0.05 vs. Oligo probe; ## P<0.01 vs. Oligo probe; && P<0.01 vs. Anti-IgG. miR, microRNA; circRNA, circular RNA; WT, wild-type; MUT, mutant.

Article Snippet: Control-small interfering (si)RNA (1 µg, 5′-AAGACAUUGUGUGUCCGCCTT-3′), circRNA_0000285-siRNA (1 µg, 5′-CCCCAGCUAUUCAAGUGUAAA−3′), inhibitor control (50 nM, 5′-AAGUCAGGUGAUGGACAGCAUA-3′) and miR-654-3p inhibitor (50 nM, 5′-AAGGUGAUGGUCAGCAGACAUA-3′) (all purchased from Shanghai GenePharma Co., Ltd.) were transfected into TPC-1 and FTC133 cells using Lipofectamine 2000 ® (Thermo Fisher Scientific, Inc.) at 37°C for 48 h, according to the manufacturer's instructions.

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Immunoprecipitation, Real-time Polymerase Chain Reaction, Mutagenesis

Journal: Oncology Letters

Article Title: Inhibition of circular RNA_0000285 prevents cell proliferation and induces apoptosis in thyroid cancer by sponging microRNA-654-3p

doi: 10.3892/ol.2021.12934

Figure Lengend Snippet: Expression levels of circRNA_0000285 and miR-654-3p in thyroid carcinoma cells. (A) RT-qPCR analysis was performed to detect circRNA_0000285 expression in thyroid cancer cells. (B) RT-qPCR analysis was performed to detect miR-654-3p expression in thyroid cancer cells. ## P<0.01 vs. Nthy-ori 3-1 cells. circRNA, circular RNA; miR, microRNA; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Control-small interfering (si)RNA (1 µg, 5′-AAGACAUUGUGUGUCCGCCTT-3′), circRNA_0000285-siRNA (1 µg, 5′-CCCCAGCUAUUCAAGUGUAAA−3′), inhibitor control (50 nM, 5′-AAGUCAGGUGAUGGACAGCAUA-3′) and miR-654-3p inhibitor (50 nM, 5′-AAGGUGAUGGUCAGCAGACAUA-3′) (all purchased from Shanghai GenePharma Co., Ltd.) were transfected into TPC-1 and FTC133 cells using Lipofectamine 2000 ® (Thermo Fisher Scientific, Inc.) at 37°C for 48 h, according to the manufacturer's instructions.

Techniques: Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Inhibition of circular RNA_0000285 prevents cell proliferation and induces apoptosis in thyroid cancer by sponging microRNA-654-3p

doi: 10.3892/ol.2021.12934

Figure Lengend Snippet: miR-654-3p inhibitor reverses the effects of circRNA_0000285 on miR-654-3p expression. Control-siRNA, circRNA_0000285-siRNA, inhibitor control and miR-654-3p inhibitor sequences were transfected into TPC-1 and FTC133 cells for 48 h. circRNA_0000285 expression was detected via RT-qPCR analysis in (A) TPC-1 and (B) FTC133 cells following transfection with circRNA_0000285-siRNA or control-siRNA. miR-654-3p expression was detected via RT-qPCR analysis in (C) TPC-1 and (D) FTC133 cells following transfection with miR-654-3p inhibitor or inhibitor control. RT-qPCR analysis was performed to detect miR-654-3p expression in (E) TPC-1 and (F) FTC133 cells transfected with circRNA 0000285-siRNA + inhibitor control or circRNA 0000285-siRNA + miR-654-3p inhibitor. **P<0.01 vs. control-siRNA; ## P<0.01 vs. inhibitor control; && P<0.01 vs. si-circ + inhibitor control. miR, microRNA; circRNA, circular RNA; RT-qPCR, reverse transcription-quantitative PCR; si, small interfering.

Article Snippet: Control-small interfering (si)RNA (1 µg, 5′-AAGACAUUGUGUGUCCGCCTT-3′), circRNA_0000285-siRNA (1 µg, 5′-CCCCAGCUAUUCAAGUGUAAA−3′), inhibitor control (50 nM, 5′-AAGUCAGGUGAUGGACAGCAUA-3′) and miR-654-3p inhibitor (50 nM, 5′-AAGGUGAUGGUCAGCAGACAUA-3′) (all purchased from Shanghai GenePharma Co., Ltd.) were transfected into TPC-1 and FTC133 cells using Lipofectamine 2000 ® (Thermo Fisher Scientific, Inc.) at 37°C for 48 h, according to the manufacturer's instructions.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Inhibition of circular RNA_0000285 prevents cell proliferation and induces apoptosis in thyroid cancer by sponging microRNA-654-3p

doi: 10.3892/ol.2021.12934

Figure Lengend Snippet: miR-654-3p inhibitor reverses the effects of circRNA_0000285 on TPC-1 cell proliferation and apoptosis. Control-siRNA, circRNA_0000285-siRNA, inhibitor control and miR-654-3p inhibitor sequences were transfected into TPC-1 cells for 48 h. (A) The viability of TPC-1 cells was assessed via the MTT assay. (B) Detection of apoptotic TPC-1 cells was assessed via flow cytometric analysis. (C) Quantitative analysis of apoptotic cells. (D) Western blot analysis was performed to detect the protein expression levels of Bax and Bcl-2 in the different groups. (E) Bax/GAPDH ratio and (F) Bcl-2/GAPDH ratio. (G) Assessment of the Bax/Bcl-2 ratio. **P<0.01 vs. control-siRNA; ## P<0.01 vs. si-circ + inhibitor control. miR, microRNA; circRNA, circular RNA; si, small interfering; Bcl-2, B-cell lymphoma 2; OD, optical density; PI, propidium iodide.

Article Snippet: Control-small interfering (si)RNA (1 µg, 5′-AAGACAUUGUGUGUCCGCCTT-3′), circRNA_0000285-siRNA (1 µg, 5′-CCCCAGCUAUUCAAGUGUAAA−3′), inhibitor control (50 nM, 5′-AAGUCAGGUGAUGGACAGCAUA-3′) and miR-654-3p inhibitor (50 nM, 5′-AAGGUGAUGGUCAGCAGACAUA-3′) (all purchased from Shanghai GenePharma Co., Ltd.) were transfected into TPC-1 and FTC133 cells using Lipofectamine 2000 ® (Thermo Fisher Scientific, Inc.) at 37°C for 48 h, according to the manufacturer's instructions.

Techniques: Transfection, MTT Assay, Western Blot, Expressing

Journal: Oncology Letters

Article Title: Inhibition of circular RNA_0000285 prevents cell proliferation and induces apoptosis in thyroid cancer by sponging microRNA-654-3p

doi: 10.3892/ol.2021.12934

Figure Lengend Snippet: miR-654-3p inhibitor reverses the effects of circRNA_0000285 on FTC133 cell viability and apoptosis. Control-siRNA, circRNA_0000285-siRNA, inhibitor control and miR-654-3p inhibitor sequences were transfected into FTC133 cells for 48 h. (A) The viability of FTC133 cells was assessed via the MTT assay. (B) Detection of apoptotic FTC133 cells was assessed via flow cytometric analysis. (C) Quantitative analysis of apoptotic cells. (D) Western blot analysis was performed to detect the protein expression levels of Bax and Bcl-2 levels in the different groups. (E) Bax/GAPDH ratio and (F) Bcl-2/GAPDH ratio. (G) Assessment of the Bax/Bcl-2 ratio. **P<0.01 vs. control-siRNA; ## P<0.01 vs. si-circ + inhibitor control. miR, microRNA; circRNA, circular RNA; si, small interfering; Bcl-2, B-cell lymphoma 2; OD, optical density; PI, propidium iodide.

Article Snippet: Control-small interfering (si)RNA (1 µg, 5′-AAGACAUUGUGUGUCCGCCTT-3′), circRNA_0000285-siRNA (1 µg, 5′-CCCCAGCUAUUCAAGUGUAAA−3′), inhibitor control (50 nM, 5′-AAGUCAGGUGAUGGACAGCAUA-3′) and miR-654-3p inhibitor (50 nM, 5′-AAGGUGAUGGUCAGCAGACAUA-3′) (all purchased from Shanghai GenePharma Co., Ltd.) were transfected into TPC-1 and FTC133 cells using Lipofectamine 2000 ® (Thermo Fisher Scientific, Inc.) at 37°C for 48 h, according to the manufacturer's instructions.

Techniques: Transfection, MTT Assay, Western Blot, Expressing