Purification,Characterisation and Cloning of a 2S Albumin with DNase,RNase and Antifungal Activities from Putranjiva Roxburghii

The present study reports the characterisation of a novel ~12-kDa heterodimeric protein, designated as putrin, from the seeds of Putranjiva roxburghii. The purification of putrin to homogeneity was accomplished using DEAE-sepharose where protein was unbound, CM-sepharose and Cibacron blue 3GA where it was bound and appeared as single peak on a size-exclusion chromatography column. A 15 % sodium dodecyl sulphate polyacrylamide electrophoresis gel, under reducing condition, demonstrated that putrin is made of two polypeptide chains of approximately 4.5 and 7.5 kDa. Circular dichroism studies demonstrated the helical nature and conformational stability of protein at increasing temperatures. Putrin exhibited both RNase and DNase activities and exerted antifungal activity but possessed relatively weak translation–inhibitory activity in cell-free system. The cloning and sequence analysis revealed a 414 bp open reading frame encoding a preproprotein of 137 amino acid residues. The amino acid sequence comparisons and phylogenetic analysis of putrin showed significant homology to 2S seed storage family proteins. The results demonstrated that putrin belongs to 2S albumin family and exhibits a spectrum of biotechnologically exploitable functions.     

Similarity solutions in free convection boundary-layer flows adjacent to vertical permeable surfaces in porous media: II prescribed surface heat flux

The equation which governs the similarity solution for free convection boundary-layer flow along a vertical permeable surface with prescribed surface heating and mass transfer rate is discussed. The solution is seen to depend on two non-dimensional parameters;m, the power-law exponent, and γ, the mass transfer parameter. It is shown that solutions exist for allm>?1 for γ>0 (fluid injection) whereas for γ<0 (fluid withdrawal), solution exist form>m ₀(γ), wherem ₀ is determined as a function of γ. Solutions for large mass transfer rates are obtained, for both γ>0 and γ<0. For γ>0 the form of the asymptotic solution for γ large is seen to depend on the value ofm. Solutions form large are derived, these are seen to be different depending on whether γ is positive or negative.     

Natural convection from a horizontal permeable surface in a porous medium - numerical and asymptotic solutions

The equations which govern the similarity solution for free convection boundary-layer flow above a permeable, horizontal surface in a fluid-saturated porous medium are considered. These are seen to depend on the dimensionless parameters and m measuring mass transport rate and the wall temperature variation, respectively. Numerical solutions are presented for a wide range of values of and m. Asymptotic solutions are obtained for ¦¦ large (for both fluid injection, > 0, and fluid withdrawal, < 0) and for m large. These are compared with the numerical solutions.     

热幅射对抽吸或喷射三维Couette流温度分布的影响

在固定的底板上有横向正弦射流,而匀速运动的多孔介质顶板以常速率完全抽出的情况下,理论分析了热幅射对三维Couette流动温度分布的影响.在这种射流速度下,流动呈现三维流动.利用图形分析了Prandtl数、幅射参数和射流参数对传热速率的影响.Prandtl数对温度分布的影响比射流参数或幅射参数大得多.     

Domino addition of allylzinc bromide to nitrile oxides: synthesis of 5-butenylisoxazolines

Nitrile oxides undergo addition to allylzinc bromide to generate 5-butenylisoxazolines in good yields.     

Exploring the Transition of Human α-Synuclein from Native to the Fibrillar State: Insights into the Pathogenesis of Parkinson’s Disease

The etiology of Parkinson’s disease involves the interplay between the environmental and genetic factors. Here in this study human α-synuclein upon exposure to 100 μM pendimethalin for 12 h in vitro passes through a partially folded state which proceeds to the aggregated state and terminally ends in the fibrillar phase. Variations in the ANS fluorescence intensities led to the detection of intermediate and aggregated states at 6 and 10 h respectively. Far-UV CD analysis depicted significant α-helical content for intermediate state at 6 h in presence of 100 μM pendimethalin. Further increasing the incubation time to 12 h resulted in a predominant β-sheet content which was confirmed to be fibrillar by TEM. Turbidity, Rayleigh scattering analysis, Congo red assay and ThT measurements supported the TEM data i.e. the formation of fibrillar structure of human α-synuclein upon 12 h incubation. Thus, our observation could suggest a possible underlying molecular basis for Parkinson’s disease.

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Graphical Abstract Schematic elucidation of the factors involved in the fibrillation of α-Synuclein during Parkinson’s pathogenesis.

     

Photodetachment Spectrum of Hydrogen Negative Ion Near a Partially Reflecting Spherical Surface

Photodetachment of negative ions near surfaces is of great interest in view of its fundamental significance and technological applications. We reinvestigate the dynamics of photoelectrons in H^? photodetachment near a partially reflecting spherical surface by the semiclassical closed-orbit theory. Reflection parameter R and curvature K is used to observe inelastic and spherical effects of the surface, respectively. The classical action is evaluated from the photodetached electron trajectories incident normally at the surface, arising simultaneously from the source and its image. The derived analytical formula of photodetachment cross section correctly recovers the results of reflective spherical surface published recently based on theoretical imaging method.     

Assessing the issue of instability due to Michael adduct formation in novel chemical entities possessing a carbon-carbon double bond during early drug development--applicability of common laboratory analytical protocols

The discovery of small-molecule novel chemical entities (NCEs) is often a complex play between appropriate structural requirements and optimization of the desired efficacy, safety and pharmacokinetic properties. One of the typical structural variants such as having an active carbon-carbon double bond (alpha, beta-unsaturated carbonyl group) in xenobiotics may lead to stability issues. Such functionalities are extremely reactive, paving way to nucleophilic attack by endogenously occurring and ubiquitous nucleophiles like thiols. While it is easy to make a unilateral decision to not pursue the development of xenobiotics with such functionalities, we question the wisdom of such a decision. In this report, we present in vitro methodologies with appropriate examples to illustrate the ease of assessing the reactivity of the xenobiotics containing double bonds with a known nucleophile. The protocols involve simple reaction procedures followed by measurements using standard laboratory equipments (UV spectrophotometer, HPLC and LC-MS). Our data suggests that not all xenobiotics with carbon-carbon double bonds readily form a Michael's adduct product with glutathione. Hence, the criterion for dropping discovery compounds because of alpha,beta-unsaturated double bonds needs to be reconsidered. Copyright (c) 2008 John Wiley & Sons, Ltd.     

Two Step Purification of <Emphasis Type="Italic">Acinetobacter</Emphasis> sp. Lipase and Its Evaluation as a Detergent Additive at Low Temperatures

Acinetobacter sp. lipase was purified to homogeneity by a two-step process. The crude enzyme (along with biomass) was subjected to partial purification by aqueous two phase system (ATPS), avoiding centrifugation and filtration steps. Conditions for lipase partitioning by ATPS were optimized by response surface methodology (RSM) and a combination of 29.45% polyethylene glycol 8000, 15.5% phosphate, and a pH of 7.0 resulted in an optimal partition coefficient. Partially pure lipase was further purified by a modified batch process using Octyl Sepharose CL-4B in a vacuum filtration apparatus. This two-step process resulted in a purified lipase with a yield of 74.6% having a specific activity of 88.8 U/mg of protein and a purification fold of 14.92. The homogeneity of the lipase preparation obtained by the purification process was confirmed by reversed phase high performance liquid chromatography profile. The molecular weight of the purified lipase was found to be around 32 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified lipase exhibited pH and temperature optima of 8.5 and 37 degrees C, respectively. The lipase was active at low temperatures and it retained 86.8% activity at 10 degrees C. It also displayed other features such as stability over a broad range of pH (3.0-9.0) as well as stability in the presence of hydrogen peroxide and commercial detergents. Based on these characteristics, the potential of this lipase as an additive in laundry detergent formulation was evaluated under low temperature wash conditions. The results indicated that Acinetobacter sp. lipase increased the washing efficiency of the detergent Nirma by 21-24% at 15 degrees C-20 degrees C, respectively.