PURIFICATION AND BIOCHEMICAL CHARACTERIZATION OF L-AMINO ACID OXIDASE FROM WESTERN REGION INDIAN COBRA (NAJA NAJA) VENOM

Objective: Purification and biochemical characterization of LAAO from western region Indian Cobra (Naja naja) venom.Methods: LAAO was purified from Indian cobra (Naja naja) venom using sequential chromatography on Sephadex G-75 gel filtration followed by Ion exchange on CM-Sephadex C-25 column. Biochemical characterization viz., pH, Temperature, Km and Vmax were determined. Molecular weight of LAAO was determined by electrophoresis. Inhibition of LAAO from cold water extracts of Curcuma zedoria, Curcuma ceasia, Curcuma aromatic, Curcuma longa, Curcuma amada, Cucumis sativus and Benincasa hispida was done.Results: Purified LAAO showed the single band on non reducing SDS-PAGE with approximate molecular weight of 65 kDa. Further biochemical characterization revealed that, LAAO from Naja naja (western region) has an optimum pH of 7.0 and is stable at room Temperature upto 37 °C and showed an optimum enzyme concentration of 4µg/ml and Km 134.1µM and Vmax is 21.87 U/min. Cold water extract of Curcuma ceasia, Curcuma aromatic and Benincasa hispida showed comparatively significant inhibition of LAAO.Conclusion: LAAO has promising therapeutic prospects because of its effects on various biological functions. Variation in snake species and their geographical distribution also contributes to the venom properties such as composition, toxicity level, pharmacological and biological activities. The significant difference in protein profiling leads to substantial lethality among different geographical regions. Thus in this study Indian Cobra (Naja naja) venom from the western region of India was subjected to purification of LAAO and biochemical characterization. Â

LATICIFEROUS PLANT PROTEASES IN WOUND CARE

Context: Since antiquity, different parts of plants such as bark, stem and leaves have been used in wound healing. Around 10% of the angiosperm plants produce a natural polymer from specialized laticiferous cells called latex. The major role played by the latex is in wound healing and defensive mechanism against infectious diseases in plants.Objective: This paper emphasizes the role of various plant latex proteases in wound healing. The review also emphasizes on the methodology to be adopted in accessing the proteases studied for procoagulant and thrombolytic activities.Methods: This review conglomerates the reports of laticiferous plants of different families viz., Altingiaceae, Amaranthaceae, Apocyanaceae, Asclepiadaceae, Asteraceae, Caricaceae, Dipterocarpaceae, Euphorbiaceae, Lamiaceae, Moraceae, Papaveraceae, Plumbaginaceae, and Solanaceae involved in wound healing. Emphasis was given on the all possible reports on laticiferous plants in wound healing with thorough literature survey.Results: A number of proteases have been studied from plant latex proteases for their role in wound healing. Some have been extensively studied with characterization while some are yet to be explored. This review enables a detailed up-to-date knowledge of laticiferous plants studied scientifically for wound care.Conclusion: In the past 20 years, with biochemical and pharmacological characterization of plant latex it has come to light that proteases are involved in wound healing. However, research on latex protease is still in budding stage. Adopting the proteases having promising applicability in wound care needs to be focussed.Â

N-[4-Cyano-3-(trifluoro­meth­yl)phen­yl]-2-meth­oxy­benzamide

In the title compound, C16H11F3N2O2, the carboxamide group connecting the two aromatic rings is in a syn-periplanar configuration; the mol­ecule is non-planar; the dihedral angle between the two aromatic rings is 13.95 (18)°. Intra­molecular N—H⋯O and C—H⋯O hydrogen bonds occur. In the crystal, mol­ecules are linked by inter­molecular C—H⋯O hydrogen bonds

Discovery of new G-quadruplex binding chemotypes.

We report here on the discovery and preliminary evaluation of a novel non-macrocyclic low molecular weight quadruplex-stabilizing chemotype. The lead compounds, based on a furan core, show high G-quadruplex stabilisation and selectivity as well as potent in vitro anti-proliferative activity

Understanding mixed sequence DNA recognition by novel designed compounds: the kinetic and thermodynamic behavior of azabenzimidazole diamidines

Sequence-specific recognition of DNA by small organic molecules offers a potentially effective approach for the external regulation of gene expression and is an important goal in cell biochemistry. Rational design of compounds from established modules can potentially yield compounds that bind strongly and selectively with specific DNA sequences. An initial approach is to start with common A·T bp recognition molecules and build in G·C recognition units. Here we report on the DNA interaction of a synthetic compound that specifically binds to a G·C bp in the minor groove of DNA by using an azabenzimidazole moiety. The detailed interactions were evaluated with biosensor-surface plasmon resonance (SPR), isothermal calorimetric (ITC), and mass spectrometry (ESI-MS) methods. The compound, DB2277, binds with single G·C bp containing sequences with subnanomolar potency and displays slow dissociation kinetics and high selectivity. A detailed thermodynamic and kinetic study at different experimental salt concentrations and temperatures shows that the binding free energy is salt concentration dependent but essentially temperature independent under our experimental conditions, and binding enthalpy is temperature dependent but salt concentration independent. The results show that in the proper compound structural context novel heterocyclic cations can be designed to strongly recognize complex DNA sequences

Design and Synthesis of Heterocyclic Cations for Specific DNA Recognition: From AT-Rich to Mixed-Base-Pair DNA Sequences

The compounds synthesized in this research were designed with the goal of establishing a new paradigm for mixed-base-pair DNA sequence-specific recognition. The design scheme starts with a cell-permeable heterocyclic cation that binds to AT base pair sites in the DNA minor groove. Modifications were introduced in the original compound to include an Hbond accepting group to specifically recognize the G-NH that projects into the minor groove. Therefore, a series of heterocyclic cations substituted with an azabenzimidazole ring has been designed and synthesized for mixed-base-pair DNA recognition. The most successful compound, 12a, had an azabenzimidazole to recognize G and additional modifications for general minor groove interactions. It binds to the DNA site −AAAGTTT− more strongly than the −AAATTT− site without GC and indicates the design success. Structural modifications of 12a generally weakened binding. The interactions of the new compound with a variety of DNA sequences with and without GC base pairs were evaluated by thermal melting analysis, circular dichroism, fluorescence emission spectroscopy, surface plasmon resonance, and molecular modeling

Robust encryption for secure image transmission over wireless channels

The security of multimedia data transmitted over wireless networks is of increased interest. Encryption mechanisms securely transmit multimedia data over insecure networks. A major issue that has received very little attention so far is that the very same properties that gives ciphers (encryption mechanisms) their cryptographic strength make them sensitive to channel errors as well. In addition, this would enhance the error propagation inherent in compressed data. Therefore provision of security for multimedia transimission over wireless channel results in throughput loss. Nevertheless this lost throughput is traded for increased security. To our knowledge there has been no substantial effort to optimize this tradeoff. Opportunistic encryption proposed in this work is a way to optimize the tradeoff between security offered and the throughput lost due to a cipher. We show that opportunistic encryption methods that adapt to channel variations will lead to an overall increase in the system performance. Two broad scenarios are considered. (a) exact channel state information upto a finite time horizon is known and (b) only the average Signal-to-Noise Ratio (SNR) is known. Proposed opportunistic encryption framework is found to achive significant gains in throughput compared to fixed block length encryption methods for a wide range of average SNR values. We have shown that applying opportunistic encryption on JPEG compressed image results in a better quality of received image and improved security compared to fixed block length encryption

Effects of bacterial inoculants and an enzyme on the fermentation quality and aerobic stability of ensiled whole-crop sweet sorghum

A study was conducted to evaluate the effects of bacterial inoculation and cellulase on the fermentation quality of ensiled whole-crop sweet sorghum (WCSS, Sorghum bicolor L. Moench). The WCSS (323 g dry matter (DM)/kg, 251 g water soluble carbohydrates (WSC)/kg DM, 43 g crude protein (CP)/kg DM and 439 g neutral detergent fibre (NDF)/kg DM) was ensiled with i) no additive (control); ii) Lactobacillus buchneri (LB); iii) Lactobacillus plantarum (LP); and iv) LB+E, a combination of LB and enzyme. These treatments were ensiled in 1 L anaerobic jars for 25 days. The jars were opened on days 3, 7 and 15 to determine pH, while those of day 25 were sampled to determine nutrient composition, fermentation characteristics and aerobic stability. Inoculation reduced pH, butyric acid and ammonia-N and increased lactic acid content in sweet sorghum silage compared with the control. The aerobic stability of WCSS was improved with LB, while it was reduced with the homofermentative LP treatment compared with the control. The LB+E reduced the fibre, but increased residual WSC of silage. The aerobic stability of LB+E silage was lower than LB treated silage. Using enzymes to increase the WSC content of crops that already have high levels of WSC may result in reduced aerobic stability of silage. Further work is needed to evaluate these effects on silage produced on farm scale and on animal production performance.Keywords: Aerobic stability, enzyme, fermentation, inoculants, silag

Synthesis, characterization and bioactivity studies of novel 1,3,4-oxadiazole small molecule that targets basic phospholipase A2 from Vipera russelli

Secretory phospholipase A2 (sPLA2) is a key enzyme participating in the inflammatory cascade followed by the action of cyclooxygenase-2 and lipoxygenases. Therefore, inhibitors of sPLA2 could be used as potent anti-inflammatory agents to treat the early phase of inflammation. In this study, we have prepared the fenoprofen and ibuprofen analogs containing 1,3,4-oxadiazole nucleus and tested against Vipera russelli venom's basic sPLA2 (VRV-PL-VIIIa). Among the tested ligands 5(a--t),2-(2-chlorophenyl)-5-(1-(4-phenoxyphenyl) ethyl)-1,3,4-oxadiazole (5m) inhibited the catalytic activity of VRV-PL-VIIIa with an IC50 value of 11.52 µM. Biophysical studies revealed that the 5m quenches the intrinsic fluorescence of VRV-PL-VIIIa, in a concentration dependent manner. Also, the compound 5m affected VRV-PL-VIIIa conformation, which was observed by circular dichroism spectra that recorded the prominent shift in the α-helix peak and the random coil formation of VRV-PL-VIIIa. Further, molecular docking analysis revealed that the compound 5m possess strong hydrophobic interactions at catalytic triad region of the VRV-PL-VIIIa. Evident to in vitro and in silico studies, 5m strongly inhibited the hemolysis of red blood cells. Our in vivo pharmacological studies revealed that the compound 5m inhibited the edematogenic activity of VRV-PL-VIIIa in mouse foot pad. Additionally, the 5m inhibited VRV-PL-VIIIa-induced myotoxicity and lung hemorrhage in mice. Overall, our ADMET results depicted that 5m possess better druggable property. Thus, this study explored the new fenoprofen and ibuprofen analog 5m as the lead-structure that serves as an anti-inflammatory agent