An antioxidant effect of manganese (Mn) complexes due to the scavenging of the superoxide anion (O2-.) or to the decomposition of hydrogen peroxide (H2O2) has been described. We report here that Mn also can exert a prooxidant effect under certain experimental conditions. Thus Mn2+ in phosphate buffer increased the bactericidal effect of phorbol myristate acetate-stimulated polymorphonuclear leukocytes (PMNs) on extracellular Escherichia coli. This effect was inhibited by azide, catalase, and a decrease in chloride concentration and was not observed when normal PMNs were replaced by those of patients with chronic granulomatous disease or myeloperoxidase (MPO) deficiency. Mn2+ could be replaced by Mn3+ or by superoxide dismutase (SOD). These findings suggest that Mn (or SOD), by increasing the conversion of O2-. to H2O2, can increase the activity of the MPO-H2O2-chloride antimicrobial system released by stimulated PMNs.

Background Rhipicephalusmicroplus is the vector of deadly cattle pathogens, especially Babesia spp., for which a recombinant vaccine is not available. Therefore, disease control depends on tick vector control. However, R.microplus populations worldwide have developed resistance to available acaricides, prompting the search for novel acaricide targets. G protein-coupled receptors (GPCRs) are involved in the regulation of many physiological processes and have been suggested as druggable targets for the control of arthropod vectors. Arthropod-specific signaling systems of small neuropeptides are being investigated for this purpose. The pyrokinin receptor (PKR) is a GPCR previously characterized in ticks. Myotropic activity of pyrokinins in feeding-related tissues of Rhipicephalussanguineus and Ixodesscapularis was recently reported. Methods The R.microplus pyrokinin receptor (Rhimi-PKR) was silenced through RNA interference (RNAi) in female ticks. To optimize RNAi, a dual-luciferase assay was applied to determine the silencing efficiency of two Rhimi-PKR double-stranded RNAs (dsRNA) prior to injecting dsRNA in ticks to be placed on cattle. Phenotypic variables of female ticks obtained at the endpoint of the RNAi experiment were compared to those of control female ticks (non-injected and beta-lactamase dsRNA-injected). Rhimi-PKR silencing was verified by quantitative reverse-transcriptase PCR in whole females and dissected tissues. Results The Rhimi-PKR transcript was expressed in all developmental stages. Rhimi-PKR silencing was confirmed in whole ticks 4 days after injection, and in the tick carcass, ovary and synganglion 6 days after injection. Rhimi-PKR silencing was associated with an increased mortality and decreased weight of both surviving females and egg masses (P < 0.05). Delays in repletion, pre-oviposition and incubation periods were observed (P < 0.05). Conclusions Rhimi-PKR silencing negatively affected female reproductive fitness. The PKR appears to be directly or indirectly associated with the regulation of female feeding and/or reproductive output in R.microplus. Antagonists of the pyrokinin signaling system could be explored for tick control. Graphical abstract

This study reports the baculovirus expression and biochemical characterization of recombinant acetylcholinesterase from Haematobia irritans (L.) (rHiAChE) and the effect of the previously described G262A mutation on enzyme activity and sensitivity to selected organophosphates. The rHiAChE was confirmed to be an insect AChE2-type enzyme with substrate preference for acetylthiocholine (Km 31.3 microM) over butyrylthiocholine (Km 63.4 microM) and inhibition at high substrate concentration. Enzyme activity was strongly inhibited by eserine (2.3 x 10(-10) M), BW284c51 (3.4 x 10(-8) M), malaoxon (3.6 x 10(-9) M), and paraoxon (1.8 x 10(-7) M), and was less sensitive to the butyrylcholinesterase inhibitors ethopropazine (1.1 x 10(-6) M) and iso-OMPA (4.1 x 10(-4) M). rHiAChE containing the G262A substitution exhibited decreased substrate affinity for both acetylthiocholine (Km 40.9 microM) and butyrylthiocholine (Km 96.3 microM), and exhibited eight-fold decreased sensitivity to paraoxon, and approximately 1.5- to 3-fold decreased sensitivity to other inhibitors. The biochemical kinetics are consistent with previously reported bioassay analysis, suggesting that the G262A mutation contributes to, but is not solely responsible for observed phenotypic resistance to diazinon or other organophosphates.