Journal: Heliyon

Article Title: An increase in urinary primaquine and a reduction in urinary primaquine-5,6-orthoquinone in the Thai population with CYP2D6 reduced enzyme function

doi: 10.1016/j.heliyon.2024.e24351

Figure Lengend Snippet: Baseline characteristics of the participants in the pharmacokinetic study (n = 44) according to the predicted CYP2D6 phenotypes and CYP2D6 activity scores.

Article Snippet: Two different pairs of primers (Hs00010001_cn and Human RNase P) were used to identify the presence or deletion of the entire CYP2D6 gene versus baseline.

Techniques: Activity Assay

Journal: Heliyon

Article Title: An increase in urinary primaquine and a reduction in urinary primaquine-5,6-orthoquinone in the Thai population with CYP2D6 reduced enzyme function

doi: 10.1016/j.heliyon.2024.e24351

Figure Lengend Snippet: Frequency of the CYP2D6 alleles, genotypes, phenotypes and activity scores.

Article Snippet: Two different pairs of primers (Hs00010001_cn and Human RNase P) were used to identify the presence or deletion of the entire CYP2D6 gene versus baseline.

Techniques: Activity Assay

Journal: Heliyon

Article Title: An increase in urinary primaquine and a reduction in urinary primaquine-5,6-orthoquinone in the Thai population with CYP2D6 reduced enzyme function

doi: 10.1016/j.heliyon.2024.e24351

Figure Lengend Snippet: Pharmacokinetic parameters of primaquine and 5,6-orthoquinone primaquine in plasma and urine according to the predicted CYP2D6 phenotypes.

Article Snippet: Two different pairs of primers (Hs00010001_cn and Human RNase P) were used to identify the presence or deletion of the entire CYP2D6 gene versus baseline.

Techniques: Clinical Proteomics

Journal: Heliyon

Article Title: An increase in urinary primaquine and a reduction in urinary primaquine-5,6-orthoquinone in the Thai population with CYP2D6 reduced enzyme function

doi: 10.1016/j.heliyon.2024.e24351

Figure Lengend Snippet: Plasma primaquine concentration time profiles according to the predicted CYP2D6 phenotypes (A) and the CYP2D6 activity scores (B) Data are presented in median ± median absolute deviation (MAD). MAD was calculated as median ( | X i - X ‾ | ) where X ‾ = median ( X i ). AS: activity score; IM: intermediate metabolizers; NM: normal metabolizers.

Article Snippet: Two different pairs of primers (Hs00010001_cn and Human RNase P) were used to identify the presence or deletion of the entire CYP2D6 gene versus baseline.

Techniques: Clinical Proteomics, Concentration Assay, Activity Assay

Journal: Heliyon

Article Title: An increase in urinary primaquine and a reduction in urinary primaquine-5,6-orthoquinone in the Thai population with CYP2D6 reduced enzyme function

doi: 10.1016/j.heliyon.2024.e24351

Figure Lengend Snippet: The cumulative amount-time profile of primaquine (A) and primaquine 5,6-orthoquinone (B) in urine according to the predicted CYP2D6 phenotypes.

Article Snippet: Two different pairs of primers (Hs00010001_cn and Human RNase P) were used to identify the presence or deletion of the entire CYP2D6 gene versus baseline.

Techniques:

Journal: Heliyon

Article Title: An increase in urinary primaquine and a reduction in urinary primaquine-5,6-orthoquinone in the Thai population with CYP2D6 reduced enzyme function

doi: 10.1016/j.heliyon.2024.e24351

Figure Lengend Snippet: The cumulative amount excreted (CAE) in urine of primaquine (A) and primaquine-5,6-orthoquinone (B) and CAE ratio (C) according to the predicted CYP2D6 phenotypes Data are presented as median. p value for the Mann-Whitney U test between normal metabolizers and intermediate metabolizers. CAE: cumulative amount excreted (CAE) in urine; POQ: primaquine-5,6-orthoquinone.

Article Snippet: Two different pairs of primers (Hs00010001_cn and Human RNase P) were used to identify the presence or deletion of the entire CYP2D6 gene versus baseline.

Techniques: MANN-WHITNEY

Journal: Plant Physiology

Article Title: mTERF8, a Member of the Mitochondrial Transcription Termination Factor Family, Is Involved in the Transcription Termination of Chloroplast Gene psbJ ^{1 [OPEN]}

doi: 10.1104/pp.19.00906

Figure Lengend Snippet: Expression pattern of the chloroplast polycistron psbJ-psbL-psbF-psbE in the knockout line of mTERF8. A, RT-qPCR analysis of chloroplast gene expression in wild type (WT) and the mterf8 mutant. TUBLIN4 was used as an internal control. Relative expression patterns of plastid-encoded genes, including psbA, rbcL, psbJ, rpoA, rrna16, and accD of wild type and mterf8 are shown. B, Schematic illustration of the chloroplast polycistron psbJ-psbL-psbF-psbE in Arabidopsis, and relative expression levels of the chloroplast genes psbJ, psbI, psbF, and psbE in wild type and mterf8. C, Profile of the psbJ-psbL-psbF-psbE polycistron in wild type and mterf8 by RNA blot analysis. The gray values of the transcripts in wild type and mterf8 mutant are 135.8 and 146.5, respectively. The values were measured by Image J software. D, Expression of the chloroplast genes psbJ and psbE in wild type treated with spectinomycin or not. The tublin4 gene was used as control. For the RT-qPCR results, the data represent the mean of triplicate experiments for each gene. Bars represent standard deviations calculated by ABI7300 system SDS software. Two independent biological replicates showed similar results.

Article Snippet: Total RNA (5 µg) was circularized using T4 RNA ligase (New England Biolabs). cDNA spanning the junction of the 5′ and 3′ ligated ends was synthesized with the corresponding psbJ and psbA reverse gene primers, respectively ( Supplemental Table S1 ), using SuperScript II RNaseH reverse transcriptase (Invitrogen).

Techniques: Expressing, Knock-Out, Quantitative RT-PCR, Mutagenesis, Northern blot, Software

Journal: Plant Physiology

Article Title: mTERF8, a Member of the Mitochondrial Transcription Termination Factor Family, Is Involved in the Transcription Termination of Chloroplast Gene psbJ ^{1 [OPEN]}

doi: 10.1104/pp.19.00906

Figure Lengend Snippet: mTERF8 binds to the chloroplast gene psbJ. A, EMSAs using six DNA probes from psbJ of Arabidopsis. Labeled DNA probes were incubated with recombinant MBP:mTERF8 protein, whereas free biotinylated DNA probes only (Mock) or incubated with empty MBP protein (MBP) served as negative controls. Increasing concentrations of the corresponding competitor probes (competitor 1, 3, 4, 5, 6, and 7) were used, respectively. The terminal region of the psbA gene was used as a competitor probe (competitor 2) in the analysis. The location of the six probes is shown in the schematic of the polycistron. **Bands of free probes; *bands of shifted probes. B, Chloroplast chromatin immunoprecipitation (cpCHIP) analysis of the relative enrichment of the four representative chloroplast genes including psbA, psbJ, rbcL, rpoA, and rrn16. C, cpCHIP analysis of the relative enrichment of the four chloroplast genes psbE, psbF, psbL, and psbJ. D, Distribution of the four different fragments in the psbJ gene. The names of the four fragments (psbJ-1, psbJ-2, psbJ-3, and psbJ-4) are indicated on the right. E, cpCHIP analysis of the relative enrichment of the four different fragments of the psbJ gene. Bars = standard deviations calculated by ABI7300 system SDS software.

Article Snippet: Total RNA (5 µg) was circularized using T4 RNA ligase (New England Biolabs). cDNA spanning the junction of the 5′ and 3′ ligated ends was synthesized with the corresponding psbJ and psbA reverse gene primers, respectively ( Supplemental Table S1 ), using SuperScript II RNaseH reverse transcriptase (Invitrogen).

Techniques: Labeling, Incubation, Recombinant, Chromatin Immunoprecipitation, Software

Journal: Plant Physiology

Article Title: mTERF8, a Member of the Mitochondrial Transcription Termination Factor Family, Is Involved in the Transcription Termination of Chloroplast Gene psbJ ^{1 [OPEN]}

doi: 10.1104/pp.19.00906

Figure Lengend Snippet: Transcription termination activity of MBP:mTERF8 in vitro. A, Transcription termination activity of MBP:mTERF8 in vitro. MBP:mTERF8 was added to the T7 RNA polymerase transcription system in vitro with the Arabidopsis psbJ gene as a template. B to F, Transcription termination activity of MBP:mTERF8 in vitro. MBP:mTERF8 was added to the T7 RNA polymerase transcription system in vitro with the Arabidopsis psbA, rbcL, psaJ, petD, and petA genes as a template. All the above transcripts were separated by electrophoresis on a 2% agarose gel, then stained with ethidium bromide.

Article Snippet: Total RNA (5 µg) was circularized using T4 RNA ligase (New England Biolabs). cDNA spanning the junction of the 5′ and 3′ ligated ends was synthesized with the corresponding psbJ and psbA reverse gene primers, respectively ( Supplemental Table S1 ), using SuperScript II RNaseH reverse transcriptase (Invitrogen).

Techniques: Activity Assay, In Vitro, Electrophoresis, Agarose Gel Electrophoresis, Staining