(2012) have showed that in nontypeable H. influenzae, the two-component signaling system QseB/C was involved in biofilm formation. Daines et al. (2005) and Selleck Sotrastaurin Armbruster et al. (2009) have observed the role of LuxS and AI-2 luxS-dependent factors which control biofilm formation in non-typable H. influenzae, but they considered as controversial its importance as virulence factor in pathogenesis of the biofilm-associated infections. The change in the structure of the substituent has a significant impact on the physicochemical

properties of the compound (Hulzebos et al., 2001; Martin et al., 2002). In our study, we synthesized and tested derivatives differing from each other by the type of substituents in the thioamide group. The best results were obtained for the compound having the ethyl substituent. From the microbiological point of view, the key factor is the presence of ethyl group which only slightly increases the mass and the volume of the compound compared to derivatives with cyclohexyl or 4-metoxyphenyl substituents. Additionally, lower molecular weight of ethyl derivative can have a significant effect on the antimicrobial properties of this compound. In our opinion, replacement of ethyl group on cyclohexyl or 4-metoxyphenyl in the tested pyrazole derivatives causes a significant decrease of their activity against Haemophilus spp. Besides, ethyl substituent has a limited conformational freedom which may affect selleck chemical selectivity

(Graham, 2001). This is very important information from the point of view of the further modifications of these derivatives and their activity against either biofilm-forming cells or against mature biofilm of Haemophilus spp. In addition, further work is needed to

evaluate the role of pyrazole derivatives during biofilm formation and their influence either on adhesive capabilities of haemophili rods or on quorum-sensing phenomenon. Conclusions N-ethyl-3-amino-5-oxo-4-phenyl-2,5-dihydro-1H-pyrazole-1-carbothioamide, N-(4-metoxyphenyl)-3-amino-5-oxo-4-phenyl-2,5-dihydro-1H-pyrazole-1-carbothioamide, and N-cyclohexyl-3-amino-5-oxo-4-phenyl-2,5-dihydro-1H-pyrazole-1-carbothioamide Bortezomib were tested against H. influenzae and H. parainfluenzae in form of planktonic or biofilm-forming cells. Our study shows that the Adriamycin molecular weight pyrazoles can be inhibitors acting on planktonic or biofilm-forming cells of Haemophilus spp. Additionally, these results allow to expect that this compound will be the starting substance in the search of antimicrobials with low toxicity, showing inhibitory effect against Gram-negative haemophili rods and including anti-biofilm activity. Further investigations should clarify the mechanism of pyrazoles against biofilm formed by haemophili rods. Materials and methods N-substituted-3-amino-5-oxo-4-phenyl-2,5-dihydro-1H-pyrazole-1-carbothioamide derivatives Three N-substituted-3-amino-5-oxo-4-phenyl-2,5-dihydro-1H-pyrazole-1-carbothioamide derivatives have been screened for the antibacterial investigations.