Mediated glucose enzyme electrodes by cross-linking films of osmium redox complexes and glucose oxidase on electrodes

Here, we report on a novel, versatile approach for the preparation of mediated enzyme electrodes, demonstrated using cross-linked films of glucose oxidase and a range of functionalised osmium complexes on graphite electrodes. Response of enzyme electrodes are optimised by evaluation of glucose response as a function of variation in ratios of [Os(2,2′-bipyridine)₂(4-aminomethyl pyridine)Cl]⁺ redox mediator, polyallylamine support and glucose oxidase enzyme cross-linked using a di-epoxide reagent in films on graphite. Lowering of the redox potential required to mediate glucose oxidation is achieved by synthesis of complexes using (4,4′-dimethyl-2,2′-bipyridine) or (4,4′-dimethoxy-2,2′-bipyridine) as a ligand instead of (2,2′-bipyridine). Enzyme electrodes prepared using the complexes based on dimethoxy- or dimethyl-substituted bipyridines provide glucose oxidation current densities of 30 and 70 μA?cm^?2 at 0.2 and 0.35 V applied potential compared to 120 μA?cm^?2 at 0.45 V for the initial enzyme electrode, under pseudo-physiological conditions in 5 mM glucose, with stability of signals proving inadequate for long-term operation. Current output and stability may be improved by selection of alternate anchoring and cross-linking methodology, to provide enzyme electrodes capable for application to long-term glucose biosensors and anodes in enzymatic fuel cells.

A Novel Fusion Protein Diphtheria Toxin–Stem Cell Factor (DT-SCF)—Purification and Characterization

Fusion toxins are an emerging class of targeted therapeutics for the treatment of cancer. Diphtheria toxin-stem cell factor (DT-SCF) is one such novel fusion toxin designed to target malignancies expressing c-kit. Since, c-kit overexpression has been reported on many types of cancers, it appeared to be a reasonably good molecule to target. In the present study, we report construction, expression, purification, and characterization of DT-SCF. DT-SCF gene coding for 1–387 amino acids of diphtheria toxin, His-Ala linker, 2–141 amino acids of SCF was cloned into expression vector with C terminal His tag. The induced DT-SCF protein was exclusively expressed in insoluble fraction. Purification of DT-SCF was achieved by inclusion body isolation and metal affinity chromatography under denaturing and reducing conditions. Purified DT-SCF was renatured partially on-column by gradually reducing denaturant concentration followed by complete refolding through rapid dilution technique. Cell viability assay provided the evidence that DT-SCF is a potent cytotoxic agent selective to cells expressing c-kit. The novelty of this study lies in employing SCF as a ligand in construction of fusion toxin to target wide range of malignancies expressing c-kit. Efficacy of DT-SCF fusion toxin was demonstrated over a range of malignancies such as chronic myeloid leukemia (K562), acute lymphoblastic leukemia (MOLT4), pancreatic carcinoma (PANC-1), and cervical carcinoma (HeLa 229). This is the first study reporting specificity and efficacy of DT-SCF against tumor cells expressing c-kit. There was significant correlation (P = 0.007) between c-kit expression on cells and their sensitivity to DT-SCF fusion toxin.     

Acid-degradable solid-walled microcapsules for pH-responsive burst-release drug delivery

Acid-degradable microcapsules were prepared via an interfacial polymerization. Degradation of the thin wall of the capsules leads to all-or-nothing cargo release. The only byproducts of degradation are acetone, and a non-toxic triamide. Proof-of-concept experiments showed that cargo can be delivered to and released in cells.     

Synthesis of some novel annelated 1,2,3‐selena/thiadiazoles and 2h‐diazaphospholes

A number of annelated heterocyclic indene derivatives having 1,2,3‐selena/thiadiazole and 2H‐1,2,3‐diazaphosphole rings have been synthesized by exploiting the α‐ketomethylene functionality in some novel 1‐methyl‐piperidin‐4‐ones and 1‐oxo‐tetrahydrothiopyran‐4‐ones and were characterized by their physical and spectral data.     

Differential pulse adsorptive stripping voltammetric determination of dinoseb and dinoterb at a modified electrode.

A sensitive adsorptive stripping voltammetric method for the determination of dinitrophenolic herbicides, dinoseb (DSB) and dinoterb (DTB) at a bare carbon paste electrode (CPE) and a clay modified carbon paste electrode (CMCPE) was developed. A systematic study of various experimental conditions, such as the pH, accumulation variables and composition of a modifier on the adsorptive stripping response, were examined by using differential pulse voltammetry. A significant improvement was observed in the sensitivity by using the present method with CMCPE. When CMCPE was used, a linear response was obtained over the concentration range 2 x 10(-10) to 3 x 10(-7) M and 6 x 10(-10) to 6 x 10(-7) M with lower detection limits of 1 x 10(-10) M and 5.4 x 10(-10) M for dinoseb and dinoterb, respectively, at an accumulation time of 100 s. The interference from other herbicides and ions on the stripping signals of both compounds was also evaluated. The described method was applied to estimate of the dinoseb and dinoterb in environmental samples.     

Evaluating the role of a trypsin inhibitor from soap nut <Emphasis Type="Italic">(Sapindus trifoliatus L. Var. Emarginatus)</Emphasis> seeds against larval gut proteases,its purification and characterization

Background

The defensive capacities of plant protease Inhibitors (PI) rely on inhibition of proteases in insect guts or those secreted by microorganisms; and also prevent uncontrolled proteolysis and offer protection against proteolytic enzymes of pathogens.

Methods

An array of chromatographic techniques were employed for purification, homogeneity was assessed by electrophoresis. Specificity, Ki value, nature of inhibition, complex formation was carried out by standard protocols. Action of SNTI on insect gut proteases was computationally evaluated by modeling the proteins by threading and docking studies by piper using Schrodinger tools.

Results

We have isolated and purified Soap Nut Trypsin Inhibitor (SNTI) by acetone fractionation, ammonium sulphate precipitation, ion exchange and gel permeation chromatography. The purified inhibitor was homogeneous by both gel filtration and polyacrylamide gel electrophoresis (PAGE). SNTI exhibited a molecular weight of 29 kDa on SDS-PAGE, gel filtration and was negative to Periodic Acid Schiff’s stain. SNTI inhibited trypsin and pronase of serine class. SNTI demonstrated non-competitive inhibition with a Ki value of 0.75?±?0.05×10-10 M. The monoheaded inhibitor formed a stable complex in 1:1 molar ratio. Action of SNTI was computationally evaluated on larval gut proteases from Helicoverpa armigera and Spodoptera frugiperda. SNTI and larval gut proteases were modeled and docked using Schrodinger software. Docking studies revealed strong hydrogen bond interactions between Lys10 and Pro71, Lys299 and Met80 and Van Der Waals interactions between Leu11 and Cys76amino acid residues of SNTI and protease from H. Armigera. Strong hydrogen bonds were observed between SNTI and protease of S. frugiperda at positions Thr79 and Arg80, Asp90 and Gly73, Asp2 and Gly160 respectively.

Conclusion

We conclude that SNTI potentially inhibits larval gut proteases of insects and the kinetics exhibited by the protease inhibitor further substantiates its efficacy against serine proteases.

     

QSAR studies of 2-substituted 2,3-dihydro-1h-naphtho[1,8-de]-1,3,2-diazaphosphorine 2-oxides and sulphides

A quantitative structure activity relationship (QSAR) study of 2-substituted 2,3-dihydro-1H-naphtho[1,8,de]-1,3,2-diazaphosphorine 2-oxides and sulphides (DND), examines the extent of the contribution by various physicochemical parameters with respect to their antimicrobial activity. Simple bivariant regression analysis, based on the least squares method, is applied in order to predict models. The predicted models reveal that the steric factor, MR, is the major contributor influencing antimicrobial activity. Bulky groups at the C-19 (C=0 group) position positively influence the potency of the compounds     

Catalyst-free synthesis of pyrazole-aniline linked coumarin derivatives and their antimicrobial evaluation

Catalyst-free one-pot CN and CC bond formation is described as a simple and ecofriendly method for the synthesis of pyrazole-aniline linked coumarin derivatives. Employing this protocol, a series of derivatives were synthesized in good to excellent yields and tested against different bacterial strains as well as fungal strains. Most of the compounds exhibited potential antimicrobial activity against both Gram-positive and Gram-negative bacterial strains. Among them, the compounds 4b, 4e, 4?h, 4i and 4?k exhibited promising activity on all the tested bacterial strains with values ranging between 1.9 and 7.8?µg/mL. In addition, these compounds were tested against various fungal strains and were found to exhibit potential antifungal activity. Fascinatingly, among the tested derivatives, the compounds 4e, 4?h and 4i were found to be equipotent to miconazole (positive control) against some of the tested fungal strains. Moreover, these compounds showed promising bactericidal, Candida-cidal and biofilm inhibition activities. Further, mechanistic study was carried out with the most active derivative 4i indicated that these compounds inhibit the ergosterol biosynthesis pathway.     

Drug repositioning for anti-tuberculosis drugs: an in silico polypharmacology approach

Molecular Diversity - Development of potential antitubercular molecules is a challenging task due to the rapidly emerging drug-resistant strains of Mycobacterium tuberculosis (M.tb)....