Lzfeld, Germany) peritoneal cells. Animals, killed by cervical dislocation, have been i.p. injected with 8 mL of sterile saline. Pooled peritoneal cells collected from mice (n = four in person experiments) had been washed, re-suspended in culture medium, and seeded into 96-well round-bottom microplates (Costar, Corning, NY, USA) in 100- volumes, 2 105 cells/well. All experimental variants have been run in duplicate. Complete RPMI-1640 culture medium (Sigma-Aldrich, St. Louis, MO, USA) contained 10 heat-inactivated foetal bovine serum, two mM L-glutamine, 50 /mL gentamicin, and 5 10-5 M 2-mercaptoethanol (all Sigma-Aldrich). Cultures had been maintained at 37 C, 5 CO2 in humidified incubator (Sanyo Electric Biomedical, Osaka, Japan). The Institution Animal Ethics Committee (No. 13/2006) approved the animal welfare and all experimental procedures. three.six.1. Nitric Oxide (NO) Production Higher output NO production was induced by mixture of lipopolysaccharide (LPS from E. coli 0111:B4, 0.1 ng/mL; Sigma) and murine recombinant interferon- (IFN-, five ng/mL; R D Systems, Minneapolis, MN, USA) in mouse peritoneal cells. Tested compounds have been applied concomitantly with these priming stimuli. The concentration of nitrites in supernatants of cells was assayed in the interval of 24 h. It was detected in individual, cell-free samples (50 ) incubated five min at ambient temperature with an aliquot of a Griess reagent (1 sulphanilamide/0.1 naphtylendiamine/2.5 H3 PO4 ). The FAUC 365 Purity identified by MS and NMR evaluation. Yayoisaponin A (3) is often a new compound identified in Allium porrum, although it was already recognized in a further species on the genus Allium. Leek flower saponins 1 have been tested collectively with other structurally connected spirostanol Compounds 4 for in vitro cytotoxicity and for effects on NO production. The obtained toxicity information closely correlated together with the suppression of NO production. The highest inhibitory effects on viability (LDH assay) have been exhibited by 6-deoxyaginoside (2),Molecules 2021, 26,12 ofwhich is comparable to the well-known digitonin (five). However, the activity variations in between all tested saponins are negligible. Immune cells play an essential function in overall health and diseases. Our final results showed for t.