The objective of this study is to induce experimental diabetes mellitus by Streptozotocin in normal adult

A Dyp-type peroxidase enzyme from thermophilic cellulose degrader Thermobifida fusca (TfuDyP) was investigated for catalytic ability towards lignin oxidation. TfuDyP was characterised kinetically against a range of phenolic substrates, and a compound I reaction intermediate was observed via pre-steady state kinetic analysis at max 404 nm. TfuDyP showed reactivity towards Kraft lignin, and was found to oxidise a -aryl ether lignin model compound, forming an oxidised dimer. A crystal structure of TfuDyP was determined, to 1.8Å resolution, which was found to contain a diatomic oxygen ligand bound to the heme centre, positioned close to active site residues Asp-203 and Arg-315. The structure contains two channels providing access to the heme cofactor for organic substrates and hydrogen peroxide. Site-directed mutant D203A showed no activity towards phenolic substrates, but reduced activity towards ABTS, while mutant R315Q showed no activity towards phenolic substrates, nor ABTS.

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Platelet P2Y receptor signalling via RhoGTPases is necessary for platelet-dependent leukocyte recruitment, where no platelet aggregation is observed. We investigated signalling cascades involved in distinct P2Y-dependent platelet activities in vitro, using specific inhibitors for phospholipase C (PLC) (U73122, to inhibit the canonical pathway), and RhoGTPases: Rac1 (NSC23766) and RhoA (ROCK inhibitor GSK429286). Human platelet rich plasma (for platelet aggregation) or isolated washed platelets (for chemotaxis assays) was treated with U73122, GSK429286 or NSC23766 prior to stimulation with adenosine diphosphate (ADP) or the P2Y specific agonist MRS2365. Aggregation, chemotaxis (towards f-MLP), or platelet-induced human neutrophil chemotaxis (PINC) towards macrophage derived chemokine (MDC) was assessed. Molecular docking of ADP and MRS2365 to P2Y was analysed using AutoDock Smina followed by GOLD molecular docking in the Accelrys Discovery Studio software. Inhibition of PLC, but not Rac1 or RhoA, suppressed platelet aggregation induced by ADP and MRS2365. In contrast, platelet chemotaxis and PINC, were significantly attenuated by inhibition of platelet Rac1 or RhoA, but not PLC. MRS2365, compared to ADP had a less pronounced effect on P2Y-induced aggregation, but a similar efficacy to stimulate platelet chemotaxis and PINC, which might be explained by differences in molecular interaction of ADP compared to MRS2365 with the P2Y receptor. Platelet P2Y receptor activation during inflammation signals through alternate pathways involving Rho GTPases in contrast to canonical P2Y receptor induced PLC signalling. This might be explained by selective molecular interactions of ligands within the orthosteric site of the P2Y receptor.

Antimicrobial resistance represents a significant challenge to future healthcare provision. An acronym ESKAPEE has been derived from the names of the organisms recognised as the major threats although there are a number of other organisms, notably Neisseria gonorrhoeae, that have become equally challenging to treat in the clinic. These pathogens are characterised by the ability to rapidly develop and/or acquire resistance mechanisms in response to exposure to different antimicrobial agents. A key part of the armoury of these pathogens is a series of efflux pumps, which effectively exclude or reduce the intracellular concentration of a large number of antibiotics, making the pathogens significantly more resistant. These efflux pumps are the topic of considerable interest, both from the perspective of basic understanding of efflux pump function, and its role in drug resistance but also as targets for the development of novel adjunct therapies. The necessity to overcome antimicrobial resistance has encouraged investigations into the characterisation of resistance-modifying efflux pump inhibitors to block the mechanisms of drug extrusion, thereby restoring antibacterial susceptibility and returning existing antibiotics into the clinic. A greater understanding of drug recognition and transport by multidrug efflux pumps is needed to develop clinically useful inhibitors, given the breadth of molecules that can be effluxed by these systems. This review discusses different bacterial EPIs originating from both natural source and chemical synthesis and examines the challenges to designing successful EPIs that can be useful against multidrug resistant bacteria.

The multidrug resistant (MDR) opportunistic pathogen Klebsiella pneumoniae has previously been shown to adapt to chlorhexidine by increasing expression of the MFS efflux pump smvA. Here we show that loss of the regulator SmvR, through adaptation to chlorhexidine, results in increased resistance to a number of cationic biocides in K. pneumoniae and other members of the Enterobacteriaceae. Clinical Enterobacteriaceae isolates which lack smvA and smvR also have an increased susceptibility to chlorhexidine. When smvA from Salmonella and K. pneumoniae are expressed in Escherichia coli, which lacks a homologue to SmvAR, resistance to chlorhexidine increased (4-fold) but plasmid carriage of smvA alone was detrimental to the cell. Challenge of K. pneumoniae with chlorhexidine and another cationic biocide, octenidine, resulted in increased expression of smvA (approx. 70 fold). Adaptation to octenidine was achieved through mutating key residues in SmvA (A363V; Y391N) rather than abolishing the function of SmvR, as with chlorhexidine adaptation. Molecular modelling was able to predict that octenidine interacted more strongly with these mutated SmvA forms. These results show that SmvA is a major efflux pump for cationic biocides in several bacterial species and that increased efflux through SmvA can lead to increased chlorhexidine and octenidine tolerance.

Bioactivity-guided fractionation of the ethyl acetate extract obtained from the culture of the endophytic fungus Fusarium solani resulted in the isolation of one new naphthoquinone, 9-desmethylherbarine (1), and two azaanthraquinone derivatives, 7-desmethylscorpinone (2) and 7-desmethyl-6-methylbostrycoidin (3), along with four known compounds. Their structures were elucidated by spectral analysis, as well as a direct comparison of spectral data with those of known compounds. Azaanthraquinones 2 and 3 showed cytotoxic activity against four human tumor cell lines, MDA MB 231, MIA PaCa2, HeLa, and NCI H1975. A molecular docking study suggested DNA interactions as the mode of action of these naphthoquinones and azaanthraquinones.

Increased oxidative stress is a widely accepted factor in the development and progression of diabetes and its complications. In this study, we evaluated the antioxidative potential of Teucrium polium (Family Lamiaceae) aqueous extract for protecting rat pancreatic tissue against streptozotocin (STZ)-induced oxidative stress. Diabetes was induced in rats by intraperitoneal injections of at a single dose of STZ at 40 mg/kg. The crude extract (equivalent to 0.5 g of plant powder/kg of body weight) was administered orally (intragastrically) to a group of STZ diabetic rats for 30 consecutive days. Changes in antioxidant status were evaluated by determining catalase (CAT) and superoxide dismutase (SOD) activities and the level of reduced glutathione (GSH) in pancreatic tissue. In addition, serum nitric oxide (NO) concentration, pancreatic tissue malondialdehyde (MDA) (an index of lipid peroxidation) level, and reliable markers of protein oxidation such as protein carbonyl content (PCO) and advanced oxidation protein products (AOPP) were also determined. Under diabetic conditions, blood glucose level, serum NO concentration, and pancreatic MDA, PCO, and AOPP levels were all increased. The diabetic rats also exhibited pancreatic GSH depletion along with significant reductions in activities of CAT and SOD. Rats treated with T. polium extract had significantly higher GSH levels along with enhanced CAT and SOD activities in pancreatic tissue. In addition to suppressed blood glucose levels, serum NO, pancreatic MDA, PCO, and AOPP levels were all lower than in the diabetic group. Our results strongly support the proposal that antioxidative activity of T. polium occurs by quenching the extent of lipid and protein oxidation. Based on these observations, it is concluded that T. polium may have protective effect(s) on pancreatic tissue in STZ-induced oxidative stress due to its high antioxidative potential.