A colorimetric hydroxy naphthol blue based loop-mediated isothermal amplification detection assay targeting the ß-tubulin locus of Sarocladium oryzae infecting rice seed.

Sarocladium oryzae is a widely prevalent seed-borne pathogen of rice. The development of a rapid and on-site detection method for S. oryzae is therefore important to ensure the health of rice seeds. Loop-mediated isothermal amplification (LAMP) is ideal for field-level diagnosis since it offers quick, high-specific amplification of target template sequences at a single temperature. We designed primers based on the ß-tubulin region of S. oryzae. The LAMP technique devised was extremely sensitive, detecting the presence of the S. oryzae template at concentrations as low as 10 fg in 30 minutes at 65°C. The assay specificity was confirmed by performing the experiment with genomic DNA isolated from 22 different phytopathogens. Through the addition of hydroxy naphthol blue in the reaction process prior to amplification, a colour shift from violet to deep sky blue was seen in the vicinity of the target pathogen only. Finally, the LAMP assay was validated using live infected tissues, weeds and different varieties of seeds collected from different locations in Tamil Nadu, India. If developed into a detection kit, the LAMP assay developed in this study has potential applications in seed health laboratories, plant quarantine stations, and on-site diagnosis of S. oryzae in seeds and plants.

Response of rice (Oryza sativa L.) cultivars to elevated ozone stress.

The plant response to elevated ozone stress reveals inter-species and intra-species disparity. Ozone-induced crop yield loss is predicted to increase in the future, posing a threat to the world economy. This study aims to evaluate the cultivar specific variation in rice exposed to elevated ozone. Fifteen short-duration rice cultivars were exposed to 50 ppb ozone for 30 days at reproductive stage. The physiological, biochemical, growth and yield traits of all test cultivars were significantly affected in response to elevated ozone. On an average, ozone stress decreased the tiller number by 22.52%, number of effective tillers by 30.43%, 1000 grain weight by 0.62% and straw weight by 23.83% over control. Spikelet sterility increased by 19.26% and linear multiregression 3D model significantly fits the spikelet sterility and photosynthetic traits with the R2 of 0.74 under elevated ozone. Principal Component Analysis with total variance of 57.5% categorized 15 rice cultivars into four major groups, i.e., ozone sensitive (MDU6, TRY(R)2 and ASD16), moderately ozone sensitive (ASD18, ADT43, and MDU5), moderately ozone tolerant (ADT37, ADT(R)45, TPS5, Anna(R)4, PMK(R)3, and ADT(R)48), and ozone tolerant (CO51, CO47, and ADT36). This study indicates that the different responses of rice cultivars to elevated ozone stress through a change in plant physiology, biochemical, growth, and yield traits and the results directed to provide scientific information on plant adaptations to ozone stress and helps in efforts to search ozone tolerant gene for plant breeding.

Experimental investigation of electro-rheological properties of modeled vegetable oils.

Vegetable oil becomes polarized on oxidation and polymerization resulting in the formation of peroxide, triglycerides, etc. The quality and reusable state were investigated for sunflower, sesame, rice bran oil and model oil with the addition of oleic acid (2, 4 and 6 %) and antioxidants (citric and tert-Butyl hydroquinone-TBHQ). Excessive reclaims of cooking oil produce toxic by-products due to chemical breakdown that induce the production of polar compounds in oil. To determine the consumable fitness, variations of dielectric constant are observed at different temperatures (29 to 70 °C) and frequencies (1 to 10(7)Hz) for the cooking oil. Physical parameters, such as viscosity and density associated with the saturated and unsaturated fatty acid, are also measured at different temperatures to determine the quality of oil. Dielectric constant and viscosity are correlated and analyzed using a newly developed equation with high correlation constant (R (2) = 0.998) for oil added with citric acid. Oil added with 2-4 % of oleic acid is observed to have high determination coefficient (R (2) > 0.92). A lowest correlation (R (2) = 0.6-0.7) was observed for the oil added with TBHQ. The present study also states that addition of TBHQ to oil does not impede oxidation reaction. Besides, even the shelf life of the oil could not be enhanced and may produce adverse effects in human health.

Discovery of a latent calcineurin inhibitory peptide from its autoinhibitory domain by docking, dynamic simulation, and in vitro methods.

Autoinhibitory domain (AID) of calcineurin (CN) was discovered two decades ago. Fewer investigations are reported to find out shortest possible peptide from the AID for CN inhibition. Hence, this study has focused on screening of nearly 150 peptide fragments derived from the AID using in silico method. Therefore, we have employed docking studies, aiming to analyze the best pose of AID-derived peptides on CN active site. We also analyzed binding free energy (ΔG) of docked complex using molecular mechanics/generalized Born surface area (MM/GBSA). MM/GBSA predicts two short peptides P1 and P2 found to be lowest binding free energy. Two peptides exhibit better binding affinity with CN, suggests that the possible candidates for potential CN inhibition. Further, the stability of the docked complex was analyzed using molecular dynamic (MD) simulation. MD study shows that CNA:P2 is the most stable complex than CN A:P1 and CN A:AID. Besides, we have synthesized and purified P1 and P2 peptides over high performance liquid chromatography (HPLC) found to be 90.31% and 98.93% of purity, respectively. In addition, AID peptides were characterized over mass spectral analysis. Peptides were subjected to CN inhibitory assay using malachite green method. Where, P1 and P2 exhibit CN inhibition better than AID. In particular, shortest peptide P2 shows highest inhibitory activity than AID. Enzyme assay reveals CN inhibitory activity of P2 peptide is consistent within silico results. In silico and in vitro, results corroborated each other to confirm short peptide P2 can be used as a potential CN inhibitor.

A novel assay method for calcium calmodulin dependent phosphatase from bovine brain extract.

Calcium calmodulin dependent protein ser/thr phosphatase, also referred to as protein phosphatase 2B (PP2B), is rich in neural tissue, and plays an important role in the overall function of the nervous system. Routinely phosphatase assay employs, para-Nitrophenlylphosphate (p-NPP), as a substrate, is also extended to assay PP2B. However, in the present study, the differential spectral characterstic property of tyrosine and phopshotyrosine has been exploited to employ the latter as a candidate substrate for the PP2B assay. The specific activity of PP2B using phosphortyrosine in bovine Bos Taurus indicus brain extract (Bos Taurus indicus), was measured in presence of different metal ions like Ca(2+), Mn(2+) and Mg(2+). Further modulators like dithiothreitol (DTT), calmodulin (CaM) and metal chelators such as EGTA and EDTA were applied to confirm the role of divalent cations and to determine calcium calmodulin dependent phoshphatase activity. PP2B activity was higher with phosphotyrosine in presence of Ca(2+) than with p-NPP. Further experiments, involving calmodulin as a modulator, confirmed phosphotyrosine as a better substrate over p-NPP. Calmodulin further enhanced the effect of phosphotyrosine as a potential substrate confirming calcium calmodulin dependent phosphatase activity. Phosphotyrosine is proposed as a better substrate in assaying calcium dependent phosphatase activity when compared to para-nitrophenylphosphate.

Development and characterization of lecithin stabilized glibenclamide nanocrystals for enhanced solubility and drug delivery.

Novel LNCs (lipid nanocrystals) were developed with an aim to improve the solubility, stability and targeting efficiency of the model drug glibenclamide (GLB). PEG 20000, Tween 80 and soybean lecithin were used as polymer, surfactant and complexing agent, respectively. GLB nanocrystals (NCs) were prepared by precipitation process and complexed using hot and cold melt technique. The LNCs were evaluated by drug loading, saturation solubility (SL), optical clarity, in vitro dissolution, solid state characterization, in vivo and stability analysis. LNCs exhibited a threefold increase in SL and a higher dissolution rate than GLB. The percentage dissolution efficiency was found to decrease with increase in PEG 20000. The average particle size was in the range of 155-842 nm and zeta potential values tend to increase after complexation. X-ray powder diffractometry and differential scanning calorimetry results proved the crystallinity prevailed in the samples. Spherical shaped particles (<1000 nm) with a lipid coat on the surface were observed in scanning electron microscopy analysis. Fourier transform infrared results proved the absence of interaction between drug and polymer and stability study findings proved that LNCs were stable. In vivo study findings showed a decrease in drug concentration to pancreas in male Wistar rats. It can be concluded that LNCs are could offer enhanced solubility, dissolution rate and stability for poorly water soluble drugs. The targeting efficiency of LNCs was decreased and further membrane permeability studies ought to be carried out.

Simultaneous optimization of multiple performance characteristics in coagulation-flocculation process for Indian paper industry wastewater.

The goal of this study was to optimize the coagulation-flocculation process in wastewater generated from the paper and pulp industry using a grey relational analysis (GRA)-based Taguchi method. Process parameters included types and doses of natural coagulants and coagulant aid, and pH. To track the efficiency of the treatment process, the following responses were chosen for optimization: chemical oxygen demand (COD), total dissolved solids (TDS) and turbidity of wastewater, alone or in combination or all together. Analysis of variance showed that the type and dose of the coagulant aid were the most significant parameters, followed by pH and the dose of the coagulant; the type of coagulant used was found to be insignificant in the coagulation-flocculation process. Optimization of process parameters to achieve lower turbidity and greater removal of COD and TDS was verified in a separate confirmatory experiment, which showed improvements in COD and TDS removal and a decrease in turbidity of 8.2, 6.35 and 26.17%, respectively, with the application of the Taguchi method and GRA.

Size-selective voltammetry: modification of the interface between two immiscible electrolyte solutions by zeolite Y.

Size- and charge-selective ion transfer across the zeolite-Y-modified interface between two immiscible electrolyte solutions (ZM-ITIES) is described. The zeolite-Y membrane is prepared from pressed disks by healing with tetraethyl orthosilicate (TEOS). Size- and charge-selective transfer of the tetraethylammonium cation, size-selective exclusion of tetrabutylammonium cation, and charge-selective exclusion of the tetrafluoroborate and perchlorate anions are demonstrated at the ZM-ITIES. The exclusion studies suggest that the membrane is coherent and contains a low density of pinholes, after healing with TEOS. Various factors affecting the ion transfer such as analyte concentration, supporting electrolyte concentration, and scan rate are investigated. The diffusion coefficient of tetraethylammonium ions within the zeolite-Y pores is found to be on the order of 10(-8) cm2 s(-1).

Effect of sodium molybdate on the status of lipids, lipid peroxidation and antioxidant systems in alloxan-induced diabetic rats.

BACKGROUND: Diabetes mellitus manifests itself in a wide variety of complications and the symptoms of the disease are multi-factorial. METHODS: The lipid peroxidation (LPO) and antioxidant status were investigated in hemolysate, liver and kidney in alloxan-induced diabetic rats and the effect of molybdate supplementation on antioxidant defense systems. RESULTS: Diabetic rats exhibited an increase in the levels of lipids, lipid peroxidation and decrease in antioxidant status when compared to control rats. Oral administration of molybdate to diabetic rats for 30 days significantly reduced the levels of lipids like cholesterol, triglycerides, phospholipids and lipid peroxidation whereas increased the activities of antioxidants like superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and reduced glutathione (GSH). CONCLUSION: Molybdate prevents oxidation of lipids and protects antioxidant systems in experimental diabetic rats, and thus, may be useful for the prevention or early treatment of diabetic mellitus.

Effect of DL-alpha-lipoic acid on the status of lipid peroxidation and lipids in aged rats.

The effect of dextro and levo (DL)-alpha-lipoic acid on lipid peroxidation and lipids has been evaluated in plasma, liver, and kidney of young and aged rats. The levels of thiobarbituric acid reactive substances (TBARS) and lipids were considerably higher in aged rats compared with younger controls. DL-alpha-lipoic acid (100 mg/kg body wt/day) was administered intraperitoneally for 7 and 14 days. Supplementation of lipoic acid in aged rats prevents the elevated levels of TBARS and lipids. From our observations, we conclude that lipoic acid is very effective in normalizing age-related alterations in lipids, and it can be implemented in the aged to minimize age-associated disorders where free radicals are the major cause.