Journal: Biology

Article Title: Curcumin Enhances Gemcitabine Sensitivity in Breast Cancer Cells Through ROS-Associated Mitochondrial Apoptosis and Transcriptional Reprogramming

doi: 10.3390/biology15050448

Figure Lengend Snippet: Therapeutic window analysis of curcumin and gemcitabine in normal (MCF-10A) and cancer cells. ( A ) Concentration-dependent effects of curcumin and gemcitabine on MCF-10A cell viability following 48 h exposure. Curcumin exhibited moderate cytotoxicity, whereas gemcitabine showed a stronger dose–response effect. The IC 50 reference line (red dashed) and the therapeutic threshold (70%, green dotted) are indicated. The shaded green area represents the proposed therapeutic window. ( B ) Comparative analysis of cell viability in MCF-10A normal cells and cancer cells at their respective cancer cell IC 50 concentrations following 48 h treatment. MCF-10A cells maintained >70% viability across all tested conditions, whereas cancer cells exhibited approximately 50% viability at their IC 50 values, supporting the presence of a potential therapeutic window.

Article Snippet: The human normal breast epithelial cell line MCF-10A was also obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and used to evaluate therapeutic selectivity.

Techniques: Concentration Assay

Journal: Biology

Article Title: Curcumin Enhances Gemcitabine Sensitivity in Breast Cancer Cells Through ROS-Associated Mitochondrial Apoptosis and Transcriptional Reprogramming

doi: 10.3390/biology15050448

Figure Lengend Snippet: Intracellular ROS generation following curcumin (Cur), gemcitabine (Gem), and combination treatment in breast cancer and normal epithelial cells (48 h). Representative DCFH-DA flow cytometry histograms showing intracellular ROS levels in ( A ) MDA-MB-231, ( B ) MCF-7, and ( C ) MCF-10A cells after 48 h treatment with Cur (IC 50 ), Gem (IC 50 ), Cur+Gem, or Cur+Gem with NAC pre-treatment (5 mM, 2 h). DCF fluorescence was detected in the FL1 channel. Cur+Gem treatment induced a rightward shift in fluorescence intensity in both breast cancer cell lines, indicating increased ROS accumulation, whereas NAC pre-treatment reduced this shift. In MCF-10A cells, ROS elevation was limited under identical treatment conditions. Histograms are representative of three independent biological experiments. MFI values are indicated within panels.

Article Snippet: The human normal breast epithelial cell line MCF-10A was also obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and used to evaluate therapeutic selectivity.

Techniques: Flow Cytometry, Fluorescence

Journal: Biology

Article Title: Curcumin Enhances Gemcitabine Sensitivity in Breast Cancer Cells Through ROS-Associated Mitochondrial Apoptosis and Transcriptional Reprogramming

doi: 10.3390/biology15050448

Figure Lengend Snippet: (ΔΨm) alterations following Cur+Gem treatment in breast cancer and normal epithelial cells (48 h). Representative JC-1 flow cytometry dot plots showing mitochondrial membrane potential in MDA-MB-231, MCF-7, and MCF-10A cells after 48 h treatment with Cur+Gem or Cur+Gem with NAC pre-treatment (5 mM, 2 h). JC-1 monomers (FL1) indicate mitochondrial depolarization, whereas JC-1 aggregates (FL2) indicate intact membrane potential. Depo-larized cells were quantified from the lower right quadrant (FL1 + /FL2 − ). Cur+Gem treatment markedly increased the proportion of depolarized cells in MDA-MB-231 (~86%) and MCF-7 (~69%) cells compared to control conditions, where-as NAC pre-treatment reduced depolarization toward control levels. In MCF-10A cells, mitochondrial depolarization following Cur+Gem treatment was limited (~8%) and was reduced after NAC exposure. A minimum of 20,000 events were acquired per sample. Quadrant percentages are indicated within each panel. Data are representative of three inde-pendent biological experiments.

Article Snippet: The human normal breast epithelial cell line MCF-10A was also obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and used to evaluate therapeutic selectivity.

Techniques: Flow Cytometry, Membrane, Control

Journal: Biology

Article Title: Curcumin Enhances Gemcitabine Sensitivity in Breast Cancer Cells Through ROS-Associated Mitochondrial Apoptosis and Transcriptional Reprogramming

doi: 10.3390/biology15050448

Figure Lengend Snippet: Proposed integrative schematic model summarizing the molecular and cellular mechanisms associated with Cur-mediated sensitization to Gem in breast cancer cells. Combined treatment induces intracellular ROS accumulation, which is functionally linked to mitochondrial membrane depolarization (ΔΨm loss) and activation of the intrinsic apoptotic cascade via caspase-9 and caspase-3. The contribution of ROS to this response is supported by NAC-mediated attenuation of mitochondrial depolarization, apoptosis, and synergy scores. Concurrently, transcriptional reprogramming favors a pro-apoptotic shift (↑ BAX, ↑ TP53, ↑ CASP3/9; ↓ BCL2, ↓ NF-κB) and is accompanied by the suppression of angiogenic signaling through VEGFA downregulation. These interconnected redox, mitochondrial, and transcriptional alterations are more pronounced in triple-negative MDA-MB-231 cells, whereas comparatively limited ROS accumulation and mitochondrial disruption are observed in MCF-7 and especially MCF-10A cells under identical conditions, supporting subtype- and cell-selective vulnerability within this in vitro model.

Article Snippet: The human normal breast epithelial cell line MCF-10A was also obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and used to evaluate therapeutic selectivity.

Techniques: Membrane, Activation Assay, Disruption, In Vitro

Journal: Advanced Science

Article Title: A Versatile Biomimic Nanotemplating Fluidic Assay for Multiplex Quantitative Monitoring of Viral Respiratory Infections and Immune Responses in Saliva and Blood

doi: 10.1002/advs.202204246

Figure Lengend Snippet: Summary of Imprinting Template Proteins

Article Snippet: Heat‐inactivated SARS‐CoV‐2 (ATCC VR‐1986HK; Cedarlane), heat‐inactivated Influenza A (0810248CFHI; Cedarlane), human coronavirus spike protein (RDC3141; Cedarlane), MERS‐CoV spike protein (40069‐V08B1; SinoBiological), recombinant SARS‐CoV spike protein (10683‐CV; Novus Biological), anti‐H1N1 Influenza A virus nucleocapsid protein antibody (ab104870; Abcam), anti‐influenza (Ab01106‐15.0 & Ab01103‐15.0), and anti‐MERS‐CoV spike protein antibody (Ab01675‐21.0) (Absolute Antibody), MERS‐CoV/NCov spike protein antibody (NBP3‐06391) and Influenza A Haemagglutinin H1N1 antibody (NBP3‐06578) (Novus Biological), human coronavirus antibody (40640‐T62), and MERS‐CoV nucleocapsid protein antibody (100213‐RP02) (SinoBiological).

Techniques: Recombinant, Variant Assay

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: Neurobiology of disease

Article Title: Inhibition of CXCR2 enhances CNS remyelination via modulating PDE10A/cAMP signaling pathway.

doi: 10.1016/j.nbd.2023.105988

Figure Lengend Snippet: Fig. 3. PDE10A regulated differentiation and maturation in primary cultured OPCs. Primary OPCs were maintained for 5 days in differentiation medium containing 0.1 μM and 1 μM TAK-063 or DMSO as control. (A) Confocal results of A2B5 expression in primary cultured OPCs. (B) Confocal results of MBP expression in primary cultured OPCs. Cells were stained with A2B5 (green), MBP (red) and DAPI (blue). (Scale bar = 50 μm). (C, D) Western blot bands and statistical analysis of PDGFRα, MBP, PLP1, MOG and MOBP expression. Primary cultured OPCs were transfected with PDE10A or control cDNA for 6 h, and then incubated with SB225002 at 1 μM for another 24 h. (E) Western blot bands of PDGFRα and MBP in PDE10A overexpressed cells. (F, G) Relative value of PDGFRα and MBP intensity normalized by β-actin. Results are shown as the mean ± SD form at least five experiments, and the confocal assay was performed three independent times with duplication. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. Control cells, &&&P < 0.01 vs. OPC overexpressed PDE10A. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The blots were incubated with primary antibodies overnight at 4 ◦C, including CXCR2 (A3301), PLP1 (A14251), MAG (A16914), MOG (A5353), Caspr (A3721), SOX10 (A8655) (1:1000), β-actin (AC026, 1:100000) (Abclonal, China), MOBP (12472–1-AP) (1:500, Proteintech, China), PDE10A (sc-515,023), PDGFRα (sc-398,206) (1:500, Santa Cruze, USA), MBP (Ab40390), OLIG2 (Ab109186) (1:1000, Abcam, USA), pERK1/2 (4695 s), ERK1/2 (4370 s), Lamin B1 (13435 s) (1:1000, Cell signaling technology, USA).

Techniques: Cell Culture, Control, Expressing, Staining, Western Blot, Transfection, Incubation, Confocal Assay

Journal: Neurobiology of disease

Article Title: Inhibition of CXCR2 enhances CNS remyelination via modulating PDE10A/cAMP signaling pathway.

doi: 10.1016/j.nbd.2023.105988

Figure Lengend Snippet: Fig. 4. Inhibition of PDE10A promoted remyelination in EB intoxicated rats. MS model rats were induced with EB injection into right lateral ventricle and then treated with TAK-063 (2 mg/kg) for 14 days. (A) Rotarod test of rats treated with TAK-063 (n = 10). (B, D) LFB staining and statistical analysis of demyelinated area in the rat corpus callosum. (C, E) Expression and calculation of the mean IOD of MBP in the rat corpus callosum. (F) Western blot assay of PDGFRα, MBP, MOG, MOBP and Caspr expression in the rat corpus callosum. (G) Relative value of PDGFRα, MBP, MOG, MOBP and Caspr intensity normalized by β-actin. Results are presented as the mean ± SD from at least four rats. **P < 0.01 and ***P < 0.001 vs. Control rats, ##P < 0.01 and ###P < 0.001 vs. EB-injected rats.

Article Snippet: The blots were incubated with primary antibodies overnight at 4 ◦C, including CXCR2 (A3301), PLP1 (A14251), MAG (A16914), MOG (A5353), Caspr (A3721), SOX10 (A8655) (1:1000), β-actin (AC026, 1:100000) (Abclonal, China), MOBP (12472–1-AP) (1:500, Proteintech, China), PDE10A (sc-515,023), PDGFRα (sc-398,206) (1:500, Santa Cruze, USA), MBP (Ab40390), OLIG2 (Ab109186) (1:1000, Abcam, USA), pERK1/2 (4695 s), ERK1/2 (4370 s), Lamin B1 (13435 s) (1:1000, Cell signaling technology, USA).

Techniques: Inhibition, Injection, Staining, Expressing, Western Blot, Control