In mouse SCNT embryo cultures, 50 μM procyanidin B1 reduced ROS-probe signals at the eight-cell and blastocyst stages. Caspase-3 protein did not change significantly despite decreased transcript expression. Higher concentrations (80–150 μM) reduced blastocyst formation, showing that the developmental response depended on dose. [2]
Kv10.1 current inhibition and tested-channel selectivity
Cell-based study · Human Kv10.1 expressed in HEK293 cells; Kir2.1, hERG and KCNQ1/KCNE1 comparators in transfected HEK cells. [1]
Procyanidin B1 inhibited Kv10.1 currents in whole-cell recordings. At 100 μM it did not significantly affect the three comparator currents under the reported conditions. This establishes a limited channel panel, not universal target specificity.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0, 0.1, 1, 10, 100 and 500 μM for Kv10.1; 100 μM for comparator channels.
- Method
- Whole-cell patch-clamp recordings at room temperature (22–25 °C); Kv10.1 concentration-response fitted at +100 mV. Astemizole (10 μM) was the Kv10.1 positive control; channel-specific blockers served as comparator controls.
- Co-treatment
- Single-compound B1 exposures; positive-control blockers tested separately/sequentially as shown in the figures.
- Reported limits
- An exposure duration and a B1 solvent/vehicle specifically for patch-clamp assays were not specified in the read electrophysiology section. DMSO reported for proliferation is not transferred to this assay.
Methods and source locations
Compound reported in the paper: Procyanidin B1; Compound label in the paper: Procyanidin B1
Evidence concerns the material named procyanidin B1 in this paper; no Adooq lot validation or equivalence to other procyanidin isomers, extracts or salts is established.
Sections 2.1, 2.2, 2.5, 2.11, 3.2 and 3.5; Figures 1 and 4; Table 1
Inhibition of Kv10.1 current at +100 mV: Whole-cell patch clamp; Hill fit; n=5; mean ± SE per Section 2.11. This is an assay endpoint, not an animal dose. — Figure 1c and caption; Section 3.2; Section 2.11
Kir2.1, hERG and KCNQ1/KCNE1 current responses: Whole-cell recordings, n=7 in Figure 4 caption; comparator blockers Ba2+, dofetilide and TEA reduced corresponding currents. No exact percentage inferred from bars. — Figure 4a–f and caption; Section 3.5
Dose-dependent effects in mouse SCNT embryo cultures
Cell-based study · Mouse SCNT embryos derived from B6D2F1 female oocytes, cultured in vitro in KSOM; no systemic B1 administration. [2]
At 50 μM, B1 reduced ROS probe signals at the eight-cell and blastocyst stages. Caspase-3 transcript decreased, whereas its measured protein signal did not change significantly. Higher tested concentrations (80–150 μM) reduced blastocyst formation, limiting any general developmental-benefit claim.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0, 20, 50, 80, 100, 120 and 150 μM for developmental dose comparison; 0 versus 50 μM for redox and apoptosis-marker assays.
- Exposure duration
- Embryos followed through blastocyst formation at 72–84 h after SCNT; two-cell, four-cell and eight-cell observations at 24 h, 48 h and 48–60 h, respectively.
- Method
- B1 supplementation of in vitro KSOM culture; fluorescence-probe readouts and immunofluorescence/qPCR compared with unsupplemented KSOM.
- Vehicle
- KSOM medium; B1 dissolved in KSOM per Section 4.3.
- Co-treatment
- No second pharmacological treatment in the adopted B1-versus-control comparison.
- Reported limits
- Paper-specific purity, lot and CAS were not reported in the read reagent/treatment sections. Embryo numbers in figures do not establish independence of each embryo across the three repeated experiments.
Methods and source locations
Compound reported in the paper: Procyanidin B1; Compound label in the paper: PB1
Evidence concerns the material named procyanidin B1 in this paper; no Adooq lot validation or equivalence to other procyanidin isomers, extracts or salts is established.
Results 2.1, 2.1.1 and 2.1.3; Figures 1, 2 and 4; Methods 4.2, 4.3, 4.6–4.8 and 4.10
ROS-sensitive H2DCFDA fluorescence at eight-cell stage: Mean ± SD, experiment repeated three times; Figure 2C labels n=20 embryos in each group. Signal intensity is a probe endpoint. — Results 2.1.1; Figure 2A–C and caption; Method 4.8
ROS-sensitive H2DCFDA fluorescence at blastocyst stage: Mean ± SD, experiment repeated three times; Figure 2C labels control n=18 and treated n=16 embryos. No exact values read from bars. — Results 2.1.1; Figure 2A–C and caption; Method 4.8
Caspase-3 protein and transcript in blastocysts: Immunofluorescence and qPCR; Figure 4D labels n=12 control and n=10 treated for protein; caption describes three independent experiments. Protein result is retained despite transcript decrease. — Results 2.1.3; Figure 4A, D and E and caption; Method 4.6
Blastocyst formation at higher B1 concentrations: Viewed Figure 1D shows reductions and significance markers at these concentrations; exact percentages are not reconstructed. Caption reports mean ± SD of three independent experiments; Dunnett test in Method 4.10. — Figure 1D and caption; Results 2.1; Discussion; Methods 4.3 and 4.10
