Journal: Química Nova

Article Title: Cat's claw oxindole alkaloid isomerization induced by common extraction methods

doi: 10.1590/s0100-40422013000600012

Figure Lengend Snippet: Figure 2. HPLC-PDA monitoring of alkaloid isomerization after heating under reflux of pentacyclic oxindole alkaloid (POA) (a) and tetracyclic oxindole alka loids (TOA) (b) in dynamic maceration (DM) extract; Comparison of interconversion between mitraphylline (1) and isomitraphylline (2) in DM extract (c) and reference solution (d); Black line and gray line for POA with cis and trans D/E ring junction, respectively. Mitraphylline (1), isomitraphylline (2), speciophylline (3), uncarine F (4), pteropodine (5), isopteropodine (6) rhyncophylline (7), and isorhyncophylline (8)

Article Snippet: The mitraphylline (1) (Phytolab, batch 2946, Germany) and isomitraphylline (2) (Chromadex, batch 09417-101, USA) were used as external standards.

Techniques: Reflux, Comparison

Journal: Química Nova

Article Title: Cat's claw oxindole alkaloid isomerization induced by common extraction methods

doi: 10.1590/s0100-40422013000600012

Figure Lengend Snippet: Figure 3. The first derivative (dx/dy) from the data fitted by Weibull equation in function of reaction time of pentacyclic oxindole alkaloid (POA) having cis D/E (a) and trans D/E ring junctions (b), and tetracyclic oxindole alkaloids (TOA) (c). Mitraphylline (1), isomitraphylline (2), speciophylline (3), ptero podine (5), isopteropodine (6), rhyncophylline (7), and isorhyncophylline (8)

Article Snippet: The mitraphylline (1) (Phytolab, batch 2946, Germany) and isomitraphylline (2) (Chromadex, batch 09417-101, USA) were used as external standards.

Techniques:

Journal: Química Nova

Article Title: Cat's claw oxindole alkaloid isomerization induced by common extraction methods

doi: 10.1590/s0100-40422013000600012

Figure Lengend Snippet: Figure 1S. HPLC-PDA profile of oxindole alkaloids in DM extract, at 245 nm (a); HPLC-MS/MS profiles monitoring the pseudomolecular ions [M+H]+ at m/z 369.3 for pentacyclic oxindole alkaloid (POA), and m/z 385.4 for tetracyclic oxindole alkaloids (TOA) (b); Distinctive transitions of POA at m/z 369.3 → 337.1 (c), and TOA at m/z 385.4 → 353.4 (d); MS/MS spectra of POA (e), and TOA (f). Mitraphylline (1), isomitraphylline (2), speciophylline (3), uncarine F (4), pteropodine (5), isopteropodine (6), rhyncophylline (7), and isorhyncophylline (8)

Article Snippet: The mitraphylline (1) (Phytolab, batch 2946, Germany) and isomitraphylline (2) (Chromadex, batch 09417-101, USA) were used as external standards.

Techniques: Tandem Mass Spectroscopy

Journal: Pharmaceutics

Article Title: Modulation of Multispecific Transporters by Uncaria tomentosa Extract and Its Major Phytoconstituents

doi: 10.3390/pharmaceutics16111363

Figure Lengend Snippet: Relative transporter-specific transport of the validated SLC transporter substrates in the presence of 100 µM uncarine D, mitraphylline, or isopteropodine. Data are relative to the vehicle control. All data are presented as the mean ± SEM.

Article Snippet: Mitraphylline (purity 99.66%) and uncarine D (purity 97%) were from PhytoLab GmbH & Co. KG (Vestenbergsgreuth, Germany), and isopteropodine (USP reference standard) was obtained from Merck KGaA.

Techniques: Control

Journal: Pharmaceutics

Article Title: Modulation of Multispecific Transporters by Uncaria tomentosa Extract and Its Major Phytoconstituents

doi: 10.3390/pharmaceutics16111363

Figure Lengend Snippet: Relative transporter-specific transport of the validated ABC transporter substrates in the presence of 100 µM uncarine D, mitraphylline, or isopteropodine. Data are relative to the vehicle control. All data are presented as the mean ± SEM.

Article Snippet: Mitraphylline (purity 99.66%) and uncarine D (purity 97%) were from PhytoLab GmbH & Co. KG (Vestenbergsgreuth, Germany), and isopteropodine (USP reference standard) was obtained from Merck KGaA.

Techniques: Control

Journal: Pharmaceutics

Article Title: Modulation of Multispecific Transporters by Uncaria tomentosa Extract and Its Major Phytoconstituents

doi: 10.3390/pharmaceutics16111363

Figure Lengend Snippet: IC 50 values of mitraphylline and isopteropodine determined in transporter-overexpressing cell lines.

Article Snippet: Mitraphylline (purity 99.66%) and uncarine D (purity 97%) were from PhytoLab GmbH & Co. KG (Vestenbergsgreuth, Germany), and isopteropodine (USP reference standard) was obtained from Merck KGaA.

Techniques:

Journal: Pharmaceutics

Article Title: Modulation of Multispecific Transporters by Uncaria tomentosa Extract and Its Major Phytoconstituents

doi: 10.3390/pharmaceutics16111363

Figure Lengend Snippet: IC 50 values of mitraphylline and isopteropodine determined for BCRP and MDR1.

Article Snippet: Mitraphylline (purity 99.66%) and uncarine D (purity 97%) were from PhytoLab GmbH & Co. KG (Vestenbergsgreuth, Germany), and isopteropodine (USP reference standard) was obtained from Merck KGaA.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Targeting the Achilles’ Heel of Multidrug-Resistant Staphylococcus aureus by the Endocannabinoid Anandamide

doi: 10.3390/ijms23147798

Figure Lengend Snippet: AEA increases the intracellular concentration of Bocillin-FL with enhanced binding of Bocillin-FL to membrane proteins. Protein extracts were prepared from MDRSA CI-M and MSSA 25923 that were incubated in the absence or presence of 50 µg/mL AEA for 1 h followed by a 1 h incubation with 10 µg/mL Bocillin-FL. ( A ). The soluble fraction was run on a 10% SDS-PAGE gel under reducing conditions. Upper panel shows the green fluorescence of Bocillin-FL and the lower panel the gel after silver staining. The free Bocillin-FL ran at the front of the gel. ( B ). The membrane fraction was run on a 10% SDS-PAGE gel under non-reducing conditions. Upper panel shows the green fluorescence of Bocillin-FL bound to membrane proteins and the lower panel the gel after silver staining.

Article Snippet: Thirty μL was loaded on a 16 cm-long non-reduced SDS-PAGE composed of 5%, 7.5%, and 15% gel at a ratio of 1:1:1 (Hoefer apparatus), or 10 μL was loaded on a 7 cm-long non-reduced SDS-PAGE composed of 7.5% and 15% gel at a ratio of 1:1 using the Bio-Rad Mini-Protean system.

Techniques: Concentration Assay, Binding Assay, Incubation, SDS Page, Fluorescence, Silver Staining

Journal: International Journal of Molecular Sciences

Article Title: Targeting the Achilles’ Heel of Multidrug-Resistant Staphylococcus aureus by the Endocannabinoid Anandamide

doi: 10.3390/ijms23147798

Figure Lengend Snippet: AEA reduces the cell wall content of wall teichoic acid (WTA). ( A ). MDRSA CI-M was incubated in the absence or presence of 50 µg/mL AEA for 2 h, and the WTA content analyzed on a 20% PAGE followed by Alcian blue–silver staining. Duplicate samples are presented. ( B ). Silver staining of SDS-extracted proteins run on a 10% SDS-PAGE from the same samples as in ( A ), but from a processing step prior to WTA isolation.

Article Snippet: Thirty μL was loaded on a 16 cm-long non-reduced SDS-PAGE composed of 5%, 7.5%, and 15% gel at a ratio of 1:1:1 (Hoefer apparatus), or 10 μL was loaded on a 7 cm-long non-reduced SDS-PAGE composed of 7.5% and 15% gel at a ratio of 1:1 using the Bio-Rad Mini-Protean system.

Techniques: Incubation, Silver Staining, SDS Page, Isolation

Journal: International Journal of Molecular Sciences

Article Title: Targeting the Achilles’ Heel of Multidrug-Resistant Staphylococcus aureus by the Endocannabinoid Anandamide

doi: 10.3390/ijms23147798

Figure Lengend Snippet: AEA treatment leads to increased spontaneous lysis, but reduced cell wall autolysin Sle-1 and mature Atl1 activities. ( A ). Kinetic autolysis assay of MDRSA CI-M that has been incubated in the absence or presence of 50 μg/mL AEA for 2 h, followed by two washes in sterile water, and incubation in 50 mM Tris HCl pH 7.6 in the absence or presence of 0.05% Triton X-100 at 37 °C. ( B ). Zymogram of autolysin extracts of MDRSA CI-M that has been incubated in the absence or presence of 50 μg/mL AEA and/or 50 μg/mL MET for 2 h using S. aureus cell wall as substrate in a 10% SDS-PAGE. Duplicate samples are presented (Series 1 and 2). C = Control; A = AEA; M = Met; AM = AEA + MET. The upper panel is the Coomassie blue staining as captured by a Samsung smartphone camera, while the lower panel is the inverted gel image as captured by the Bio-Rad ChemiDoc XRS+ imaging system. The identities of the autolysin bands were determined according to Vaz and Filipe .

Article Snippet: Thirty μL was loaded on a 16 cm-long non-reduced SDS-PAGE composed of 5%, 7.5%, and 15% gel at a ratio of 1:1:1 (Hoefer apparatus), or 10 μL was loaded on a 7 cm-long non-reduced SDS-PAGE composed of 7.5% and 15% gel at a ratio of 1:1 using the Bio-Rad Mini-Protean system.

Techniques: Lysis, Incubation, SDS Page, Staining, Imaging