Coumarin constituents of Selinum tenuifolium

Three furocoumarins, bergapten, heraclenin and heraclenol have been isolated from the roots of Selinum tenuifolium (Umbelliferae). Another coumarin, m.p. 85–86°, isolated from the same source and provisionally designated as ST-1, has been proved to be mixture of imperatorin, and 8-geranyloxypsolaren by analyses of various reaction products and separation by preparative TLC on silica gel G impregnated with silver nitrate.     

Comparison among four triazole fungicides on growth and development of sheath blight of rice pathogen Rhizoctonia solani Kühn AG1-1A

The sheath blight of rice pathogen Rhizoctonia solani Kühn AG1-1A has long been known as a major crisis in global rice production. The present study aimed to compare the efficacy of four triazole fungicides at different doses on the growth and development of R. solani Kühn AG1-1A. Obtained results demonstrated inhibitory effects of all four fungicides on mycelial growth, sclerotia formation and biomass production of R. solani Kühn AG1-1A with significant variation among the treatment doses as well as fungicide molecules (p ≤ 0.05). At the respective EC₅₀ doses all four fungicides inhibited cell wall degrading enzymes viz. invertase, cellulase, polygalacturonase and pectin lyase, of the sheath blight pathogen. The extent of inhibition of the enzymes significantly varied among the fungicides. It is important to note that in spite of having common mode of action, all four triazole fungicides demonstrated significant variation in their fungicidal efficacy on R. solani Kühn AG1-1A.     

Regiospecific one-pot synthesis of pyrimido[4,5-d]pyrimidine derivatives in the solid state under microwave irradiations

Electron rich 6-[(dimethylamino)methylene]amino uracil 1, undergoes [4+2] cycloaddition reactions with various in situ generated glyoxylate imine and imine oxides 6 to provide novel pyrimido[4,5-d]pyrimidine derivatives of biological significance, after elimination of dimethylamine from the (1:1) cycloadducts and oxidative aromatisation. This procedure provides a convenient method for the direct synthesis of pyrimido[4,5-d]pyrimidines in excellent yields when carried out in the solid state and under microwave irradiations.     

Stability analysis and optimal control of an SIR epidemic model with vaccination

This paper focuses on the study of a nonlinear mathematical SIR epidemic model with a vaccination program. We have discussed the existence and the stability of both the disease free and endemic equilibrium. Vaccine induced reproduction number is determined and the impact of vaccination in reducing the vaccine induced reproduction number is discussed. Then to achieve control of the disease, a control problem is formulated and it is shown that an optimal control exists for our model. The optimality system is derived and solved numerically using the Runge-Kutta fourth order procedure.     

Molecular cloning,sequence characterization and recombinant expression of Nanog gene in goat fibroblast cells using lentiviral based expression system

Nanog is a homeodomain containing protein which plays important roles in regulation of signaling pathways for maintenance and induction of pluripotency in stem cells. Because of its unique expression in stem cells it is also regarded as pluripotency marker. In this study goat Nanog (gNanog) gene has been amplified, cloned and characterized at sequence level with successful over-expression in CHO-K1 cell line using a lentiviral based system. gNanog ORF is 903 bp long which codes for Nanog protein of size 300 amino acids (aas). Complete nucleotide sequence shows some evolutionary mutation in goat in comparision to other species. Protein sequence of goat is highly similar to other species. Overall, gNanog nucleotide sequence and predicted protein sequence showed high similarity and minimum divergence with cattle (96 % identity/4 % divergence) and buffalo (94/5 %) while low similarity and high divergence with pig (84/15 %), human (81/23 %) and mouse (69/40 %) indicating evolutionary closeness of gNanog to cattle and buffalo. gNanog lentiviral expression construct was prepared for over-expression of Nanog gene in adult goat fibroblast cells. Lentiviral expression construct of Nanog enabled continuous protein expression for induction and maintenance of pluripotency. Western blotting revealed the expression of Nanog gene at protein level which supported that the lentiviral expression system is highly promising for Nanog protein expression in differentiated goat cell.     

Optimization of Dexamethasone Mixed Nanomicellar Formulation

The purpose of this study was to develop a clear aqueous mixed nanomicellar formulation (MNF) of dexamethasone utilizing both d-α-tocopherol polyethylene glycol-1000 succinate (Vit E TPGS) and octoxynol-40 (Oc-40). In this study, Vit E TPGS and Oc-40 are independent variables. Formulations were prepared following solvent evaporation method. A three level full-factorial design was applied to optimize the formulation based on entrapment efficiency, size, and polydispersity index (PDI). A specific blend of Vit E TPGS and Oc-40 at a particular wt% ratio (4.5:2.0) produced excellent drug entrapment, loading, small mixed nanomicellar size and narrow PDI. Solubility of DEX in MNF is improved by ~6.3-fold relative to normal aqueous solubility. Critical micellar concentration (CMC) for blend of polymers (4.5:2.0) was found to be lower (0.012 wt%) than the individual polymers (Vit E TPGS (0.025 wt%) and Oc-40 (0.107 wt%)). No significant effect on mixed nanomicellar size and PDI with one-factor or multi-factor interactions was observed. Qualitative ¹H NMR studies confirmed absence of free drug in the outer aqueous MNF medium. MNF appeared to be highly stable. Cytotoxicity studies on rabbit primary corneal epithelial cells did not indicate any toxicity suggesting MNF of dexamethasone is safe and suitable for human topical ocular drops after further in vivo evaluations.KEY WORDS: aqueous mixed nanomicelles, characterization, critical micellar concentration (CMC), dexamethasone, experimental design     

Structural and Functional Insights into the Catalytic Inactivity of the Major Fraction of Buffalo Milk Xanthine Oxidoreductase

Background

Xanthine oxidoreductase (XOR) existing in two interconvertible forms, xanthine dehydrogenase (XDH) and xanthine oxidase (XO), catabolises xanthine to uric acid that is further broken down to antioxidative agent allantoin. XOR also produces free radicals serving as second messenger and microbicidal agent. Large variation in the XO activity has been observed among various species. Both hypo and hyper activity of XOR leads to pathophysiological conditions. Given the important nutritional role of buffalo milk in human health especially in south Asia, it is crucial to understand the functional properties of buffalo XOR and the underlying structural basis of variations in comparison to other species.

Methods and Findings

Buffalo XO activity of 0.75 U/mg was almost half of cattle XO activity. Enzymatic efficiency (k _cat/K _m) of 0.11 sec⁻¹ µM⁻¹ of buffalo XO was 8–10 times smaller than that of cattle XO. Buffalo XOR also showed lower antibacterial activity than cattle XOR. A CD value (Δε_{430 nm}) of 46,000 M⁻¹ cm⁻¹ suggested occupancy of 77.4% at Fe/S I centre. Buffalo XOR contained 0.31 molybdenum atom/subunit of which 48% existed in active sulfo form. The active form of XO in buffalo was only 16% in comparison to ∼30% in cattle. Sequencing revealed 97.4% similarity between buffalo and cattle XOR. FAD domain was least conserved, while metal binding domains (Fe/S and Molybdenum) were highly conserved. Homology modelling of buffalo XOR showed several variations occurring in clusters, especially close to FAD binding pocket which could affect NAD⁺ entry in the FAD centre. The difference in XO activity seems to be originating from cofactor deficiency, especially molybdenum.

Conclusion

A major fraction of buffalo milk XOR exists in a catalytically inactive form due to high content of demolybdo and desulfo forms. Lower Fe/S content and structural factors might be contributing to lower enzymatic efficiency of buffalo XOR in a minor way.