Targeting Vancomycin-Resistant Enterococci (VRE) Infections and Van Operon-Mediated Drug Resistance Using Dimeric Cholic Acid-Peptide Conjugates.

Emergence of vancomycin resistance in Gram-positive bacteria and the prevalence of vancomycin-resistant Enterococci (VRE) infections are highly alarming as very limited antibiotic options are available against VRE infections. Here, we present the synthesis of cholic acid-derived dimeric amphiphiles where two cholic acid moieties are tethered through carboxyl terminals using different alkylene spacers. Our investigations revealed that dimer 5 possessing a propylene spacer and glycine-valine peptides tethered on hydroxyl groups is the most effective antimicrobial against VRE. Dimer 5 can permeabilize bacterial membranes, generate reactive oxygen species, and clear preformed biofilms. We further demonstrate that dimer 5 downregulates vancomycin-mediated transcriptional activation of the vanHAX gene cluster and does not allow VSE to develop vancomycin resistance until 100 generations. Therefore, this study, for the first time, presents a bacterial membrane-targeting amphiphile that can mitigate VRE infections and inhibit the emergence of vancomycin resistance.

Priming alleviates high temperature induced oxidative DNA damage and repair using Apurinic/apyrimidinic endonuclease (Ape1L) homologue in wheat (Triticum aestivum L.).

Crop plants require an optimum range of temperature for normal growth and development however high temperature can adversely affect the plants, induce oxidative stress and disintegrate biomolecules especially DNA and proteins. In wheat, high temperature stress (35-40 °C) during ripening stage hampers the yield tremendously. In this study, we assessed high temperature (HT) induced oxidative stress, subsequent DNA damage and role of priming in stress tolerance by analyzing DNA repair enzyme Triticum aestivum AP endonuclease (TaApe1L). Sixteen days old seedlings of wheat varieties PBW 550 and PBW 343 were primed with mild drought and exposed to HT (38 °C) for 2, 4, and 6 h. Hydrogen peroxide (H2O2) was used as oxidative stress marker and quantified on regular time intervals. DNA damage was analyzed by DNA laddering and TaApe1L gene expression was analyzed using RT PCR and western blotting. Phylogenetic analysis of Ape1 revealed presence of some key amino acids that are evolutionary conserved. A significant increase in H2O2 content was observed after 6 h of exposure especially in PBW 343. Similarly, the DNA damage was also increased with HT exposure especially in PBW 343. The TaApe1L mRNA expression increased after priming in both the varieties after 4 h. But APE1 protein expression was higher in PBW 343, which can be correlated with DNA damage and repair. Lastly, it can be concluded that there is varietal difference in the HT sensitivity but 6 h exposure was detrimental to both the varieties. Also, drought priming improved HT tolerance by over expressing APE1.