Journal: Phytotherapy Research

Article Title: Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes

doi: 10.1002/ptr.6097

Figure Lengend Snippet: Influence of major components of essential oil from Chenopodium ambrosioides L. on the cellular oxygen consumption of Leishmania tarentolae promastigotes (LtP). Oxygen consumption of LtP (72–100 × 10 6 cells/ml) was assessed by a Clark‐type electrode in air‐saturated medium containing 14.6 mM glucose. Increasing concentrations of compounds were added subsequently using DMSO as vehicle. At 1% DMSO (highest final concentration), O 2 consumption of LtP was inhibited by 1.74 ± 9.46%. Data are means ± standard deviation of four independent experiments. Asc = Ascaridole; Car = carvacrol; Caryo = caryophyllene oxide

Article Snippet: Leishmania tarentolae promastigotes (LtP) strain P10 from Jena Bioscience (Germany) was used.

Techniques: Concentration Assay, Standard Deviation

Journal: Phytotherapy Research

Article Title: Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes

doi: 10.1002/ptr.6097

Figure Lengend Snippet: Detection of superoxide radicals in Leishmania tarentolae promastigotes (LtP) and the influence of major compounds of essential oil from Chenopodium ambrosioides L. Superoxide radicals in LtP converted the cyclic hydroxylamine CMH to a stable nitroxyl radical shown in (a). The intensity of the middle peak (Ipp) is proportional to the amount of superoxide formed. (b) The influence of major components of essential oil (100 μM) and antimycin A (AA, 5 μM) on the radical formation in LtP. Buffer (Bu) indicates samples without LtP. The reaction with 400 μM CMH was performed in suspensions containing 5 × 10 8 LtP/ml in phosphate‐buffered saline with 15 mM glucose, 100 μM DFO, and 25 μM DTPA. Data represent mean ± standard deviation of quadruplicate experiments. * Significant differences versus LtP on the level p < .1. Asc = Ascaridole; Car = carvacrol; Caryo = caryophyllene oxide

Article Snippet: Leishmania tarentolae promastigotes (LtP) strain P10 from Jena Bioscience (Germany) was used.

Techniques: Saline, Standard Deviation, Electron Paramagnetic Resonance

Journal: Phytotherapy Research

Article Title: Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes

doi: 10.1002/ptr.6097

Figure Lengend Snippet: Formation of superoxide radicals in Leishmania tarentolae promastigotes (LtP) triggered by ascaridole (Asc) increased with Asc concentration, incubation time, and iron availability. Superoxide radical formation was assessed by reaction of 400 μM CMH in suspensions containing 5 × 10 8 LtP/ml in phosphate‐buffered saline with 15 mM glucose and 100 μM DFO and 25 μM DTPA (white bars). Immediate effects (0‐hr incubation) of Asc in LtP in the presence of chelators (DFO/DTPA). After 1‐hr incubation (light grey) and after 1‐hr incubation in the absence of iron chelators (dark grey). Data represent mean ± standard deviation of quadruplicate experiments. * Significant differences versus LtP on the level p < .05. Bu = buffer controls without LtP

Article Snippet: Leishmania tarentolae promastigotes (LtP) strain P10 from Jena Bioscience (Germany) was used.

Techniques: Concentration Assay, Incubation, Saline, Standard Deviation

Journal: Phytotherapy Research

Article Title: Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes

doi: 10.1002/ptr.6097

Figure Lengend Snippet: Mitochondrial coupling in Leishmania tarentolae promastigotes (LtP). (a) Typical experiment to assess mitochondrial coupling in LtP. LtP cells (110 ± 13 × 10 6 /ml) in Schneider's medium supplemented with 6 μM hemin were equilibrated with air within the reaction chamber of a Clark‐type electrode. After closing the chamber, the basal cellular O 2 consumption was measured. By addition of 5 μM oligomycin (Oligo), an inhibitor of ATP synthase, oxygen consumption coupled to ATP production was blocked. Finally, the uncoupler CCCP (0.5 μM) was added to obtain the capacity of mitochondrial electron transport in LtP at non‐coupled respiration. (b) O 2 consumption rates obtained from the respective slopes in the left graph. Data represent mean ± standard deviation of four independent experiments. * Significant differences versus LtP on the level p < .05

Article Snippet: Leishmania tarentolae promastigotes (LtP) strain P10 from Jena Bioscience (Germany) was used.

Techniques: Standard Deviation

Journal: Phytotherapy Research

Article Title: Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes

doi: 10.1002/ptr.6097

Figure Lengend Snippet: Mitochondrial uncoupling in Leishmania tarentolae promastigotes (LtP) in the presence of major components of essential oil from Chenopodium ambrosioides L. Uncoupling was assessed by stimulation of oxygen consumption in the presence of the ATP synthase inhibitor oligomycin (Oligo). LtP cells (1 × 10 8 /ml) in brain heart infusion medium were supplemented with Oligo (5 μM) and decreasing concentrations of ascaridole (Asc), carvacrol (Car), caryophyllene oxide (Caryo), and the uncoupler CCCP. Oxygen consumption was assessed in 96‐well OxoPlates for 40 min at 26 °C. Oxygen consumption rates were normalized to the respiration of non‐inhibited Leishmania (LtP = 100%). Data represent mean ± standard deviation of 3–4 experiments. * Significant differences versus LtP + Oligo on the level p < .05

Article Snippet: Leishmania tarentolae promastigotes (LtP) strain P10 from Jena Bioscience (Germany) was used.

Techniques: Standard Deviation

Journal: Phytotherapy Research

Article Title: Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes

doi: 10.1002/ptr.6097

Figure Lengend Snippet: Effects of ascaridole (Asc) on mitochondrial coupling in Leishmania tarentolae promastigotes (LtP) upon prolonged incubation. Different cell batches with 1 × 10 8 LtP/ml in Schneider's medium plus 6 μM hemin were incubated in culture tubes either with DMSO (vehicle for Asc; LtP) or with 200 μM Asc (LtP + Asc). From these culture stocks, aliquots were taken for O 2 consumption measurements at 0, 6, and 24 hr. Mean cell counts during measurements were adjusted with medium to approximately 1–2 × 10 8 LtP/ml. Respiratory control ratios were calculated as the ratios of O 2 consumption rates in the presence of 5 μM oligomycin plus 0.5 μM CCCP to oligomycin‐inhibited O 2 consumption rates as shown in Figure . Data represent mean ± standard deviation of four independent experiments. * Significant differences versus LtP on the level p < .05

Article Snippet: Leishmania tarentolae promastigotes (LtP) strain P10 from Jena Bioscience (Germany) was used.

Techniques: Incubation, Control, Standard Deviation

Journal: Phytotherapy Research

Article Title: Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes

doi: 10.1002/ptr.6097

Figure Lengend Snippet: Influence of major components of essential oil from Chenopodium ambrosioides L. on the low molecular thiol status of Leishmania tarentolae promastigotes (LtP) after 5 hr incubation at 26 °C in phosphate‐buffered saline/glucose medium. Low molecular thiol status was assessed by measuring the rate of fluorescence evolution over 1 hr from the conjugation of CMF (arising from 5 μM CMFDA) to low molecular thiols in LtP (1 × 10 7 cells/ml) in phosphate‐buffered saline/glucose. Results represent mean ± standard deviation of three experiments. * Significant differences versus LtP on the level p < .05. Asc = Ascaridole; Bu = buffer; Car = carvacrol; Caryo = caryophyllene oxide

Article Snippet: Leishmania tarentolae promastigotes (LtP) strain P10 from Jena Bioscience (Germany) was used.

Techniques: Incubation, Saline, Fluorescence, Conjugation Assay, Standard Deviation

Journal: Phytotherapy Research

Article Title: Interaction of ascaridole, carvacrol, and caryophyllene oxide from essential oil of Chenopodium ambrosioides L. with mitochondria in Leishmania and other eukaryotes

doi: 10.1002/ptr.6097

Figure Lengend Snippet: Scheme of the mitochondrial effects of major components of essential oil from Chenopodium ambrosioides L. on Leishmania tarentolae promastigotes (LtP). Asc = Ascaridole; Car = carvacrol; Caryo = caryophyllene oxide; IMM, inner mitochondrial membrane

Article Snippet: Leishmania tarentolae promastigotes (LtP) strain P10 from Jena Bioscience (Germany) was used.

Techniques: Membrane

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Paeoniflorin Attenuates Dexamethasone-Induced Apoptosis of Osteoblast Cells and Promotes Bone Formation via Regulating AKT/mTOR/Autophagy Signaling Pathway

doi: 10.1155/2021/6623464

Figure Lengend Snippet: Effect of paeoniflorin on DEX-induced apoptosis in MC3T3-E1 cells. (a, b) The extent of apoptosis of MC3T3-E1 cells was detected by flow cytometry after Annexin V-FITC/PI double staining. The apoptotic rate was measured by flow cytometric analysis. Data are presented as the mean ± SD. ∗∗ P < 0.01 vs. control; # P < 0.05 vs. DEX. (c) MC3T3-E1 cells were stained by Hoechst to show the extent of apoptosis after interventions of DEX and paeoniflorin. (d) The expressions of osteogenic protein-Runx2 (runt-related transcription factor2) and apoptotic proteins such as Bcl-2 (B cell leukemia 2) and Bax (BCL2-associated X protein) were measured by western blot. ∗ P < 0.05, ∗∗ P < 0.01 vs. control; # P < 0.05 vs. DEX; ## P < 0.01 vs. DEX. (e) Protein levels of caspase-3 and cleaved caspase-3 were determined by western blot analysis, shown in quantitative analysis. Data are presented as the mean ± SD. ∗∗ P < 0.01 vs. control; # P < 0.05 vs. DEX.

Article Snippet: For Hoechst staining, 10 μ L Hoechst live cell staining solution (Beyotime Institute of Biotechnology) was added to the culture medium and mixed gently.

Techniques: Flow Cytometry, Double Staining, Control, Staining, Western Blot