Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Repurposed clindamycin suppresses pyroptosis in tumor-associated macrophages through Inhibition of caspase-1

doi: 10.1186/s13046-025-03478-5

Figure Lengend Snippet: Computational screening of chemical compounds against TAM-expressed protein targets identifies the antibiotic clindamycin as a TAM immunomodulator. (A) Pharmacophore modeling and screening of FDA-approved drugs. Except for ZFP36, protein structures of the selected targets were processed to derive pharmacophore models representing potential binding sites. FDA-approved drugs were then subjected to virtual screening, followed by refinement of promising binders using flexible molecular docking. This process led to the identification of the clindamycin- CASP1 interaction. (B) Predicted interactions of clindamycin and CASP1 residues in the binding pocket

Article Snippet: Cells were either kept untreated or incubated with 25 μg/ml clindamycin (Selleckchem, USA, #S2830) for 24 h. Subsequently, we measured the caspase-1 activity with the FLICA ® 660 caspase-1 assay kit from ImmunoChemistry Technologies (Bloomington, USA) according to the manufacturer’s instructions.

Techniques: Binding Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Repurposed clindamycin suppresses pyroptosis in tumor-associated macrophages through Inhibition of caspase-1

doi: 10.1186/s13046-025-03478-5

Figure Lengend Snippet: Clindamycin suppresses the pyroptotic phenotype in monocyte-derived macrophages (MDMs). MDMs were incubated with (white circle) or without (red circle) clindamycin (2 h, 10 µg/ml) before being exposed to LPS (100 ng/ml) alone or in combination with nigericin (10 µM) for 3 h. Supernatants were assayed for IL-1β ( A ) and IL-18 ( B ) by ELISA. (C) MDMs were treated as indicated and caspase-1 activation was estimated as Mean Fluorescence Intensity (MFI) using FLICA by flow cytometry. ( D ) MDMs were treated as indicated and caspase-1 activation was measured by Western blotting. The band at a height of 20 kDa is the cleaved active caspase-1, while ASC and GAPDH served as controls in the same lysates. ( E ) MDM were treated with LPS (100 ng/ml) and nigericin (10 µM) in the presence or absence of clindamycin, and supernatants were assayed for LDH release after 24 h. ( F ) MDM were treated as indicated and cleavage of gasdermin D was measured by Western blotting. The band at a height of 30 kDa is the cleaved, active form of gasdermin D, and ß-actin serves as a loading control in the same lysates. ( G ) MDM were incubated with or without clindamycin (25 µg/ml, 18 h) and then challenged with nigericin (10 µM, 3 h). To investigate the membrane structure, macrophages were stained with Wheat Germ Agglutinin (WGA, green). DAPI (blue) was used as a counterstain for nuclei. The arrows indicate cell membrane damage. Scale bar: 10 µM. ( H ) MDM were incubated with or without clindamycin (25 µg/ml, 18 h) and then challenged with nigericin (10 µM) for 15 h. Cell membrane integrity loss was measured as uptake of Cytotox Green dye under a live imaging microscope (Incucyte). The graph shows the increase in green fluorescent area (µm 2 /image) over the challenge period

Article Snippet: Cells were either kept untreated or incubated with 25 μg/ml clindamycin (Selleckchem, USA, #S2830) for 24 h. Subsequently, we measured the caspase-1 activity with the FLICA ® 660 caspase-1 assay kit from ImmunoChemistry Technologies (Bloomington, USA) according to the manufacturer’s instructions.

Techniques: Derivative Assay, Incubation, Enzyme-linked Immunosorbent Assay, Activation Assay, Fluorescence, Flow Cytometry, Western Blot, Control, Membrane, Staining, Imaging, Microscopy

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Repurposed clindamycin suppresses pyroptosis in tumor-associated macrophages through Inhibition of caspase-1

doi: 10.1186/s13046-025-03478-5

Figure Lengend Snippet: Inflammasome hyperactivity in multiple myeloma TAMs is attenuated by clindamycin. Bone marrow (BM) from MM patients with initial diagnosis was incubated in the presence (25 µg/ml, red) or absence (grey) of clindamycin for 18 h. Subsequently, the activity of caspase-1 in TAMs (CD163+/CD15-) ( A , left) was analyzed using FLICA by flow cytometry ( A , right). ( B ) Quantification of FLICA signal from (A) in material from six patients. Caspase-1 activity was estimated as Mean Fluorescence Intensity (MFI). Two-tailed paired Student’s t-test was performed. Error bars show SEM

Article Snippet: Cells were either kept untreated or incubated with 25 μg/ml clindamycin (Selleckchem, USA, #S2830) for 24 h. Subsequently, we measured the caspase-1 activity with the FLICA ® 660 caspase-1 assay kit from ImmunoChemistry Technologies (Bloomington, USA) according to the manufacturer’s instructions.

Techniques: Biomarker Discovery, Incubation, Activity Assay, Flow Cytometry, Fluorescence, Two Tailed Test

Journal: Molecules

Article Title: Determination of Pharmaceuticals, Heavy Metals, and Oxysterols in Fish Muscle

doi: 10.3390/molecules26051229

Figure Lengend Snippet: Pharmaceuticals and selected metabolites which were not detected in fish muscle with their limit of detection (ng/g).

Article Snippet: Acetaminophen D4, caffeine D9, salbutamol D7, trimethoprim D9, fluoxetine D5, bisoprolol D5, ciprofloxacin D8, mycophenolic acid D3, clindamycin D3, valsartan D3, erythromycin C13D3, dulfamethoxazole D4 were used as internal standards and were purchased from Toronto Research Chemicals (Toronto, Ontario, ON, Canada).

Techniques: