Prion protein detection in serum using micromechanical resonator arrays

Prion proteins that have transformed from their normal cellular counterparts (PrP^c) into infectious form (PrP^res) are responsible for causing progressive neurodegenerative diseases in numerous species, such as bovine spongiform encephalopathy (BSE) in cattle (also known as mad cow disease), scrapie in sheep, and Creutzfeldt-Jakob disease (CJD) in humans. Due to a possible link between BSE and CJD it is highly desirable to develop non-invasive and ante mortem tests for the detection of prion proteins in bovine samples. Such ante mortem tests of all cows prior to slaughter will help to prevent the introduction of PrP^res into the human food supply. Furthermore, detection of PrP^res in donated blood will also help to prevent the transmission of CJD among humans through blood transfusion. In this study, we have continued development of a micromechanical resonator array that is capable of detecting PrP^c in bovine blood serum. The sensitivity of the resonators for the detection of PrP^c is further enhanced by the use of secondary mass labels. A pair of antibodies is used in a sandwich immunoassay format to immobilize PrP^c on the surface of resonators and attach nanoparticles as secondary mass labels to PrP^c. Secondary mass labeling is optimized in terms of incubation time to maximize the frequency shifts that correspond to the presence of PrP^c on the surface of resonators. Our results show that a minimum of 200 pg mL⁻¹ of PrP^c in blood serum can be detected using micromechanical resonator arrays.     

Synthesis of poly(butyl acrylate)/sodium silicate nanocomposite fire retardant

Poly(butyl acrylate) (PBA)/sodium silicate (SS) nanocomposites were prepared via emulsifier-free emulsion technique in presence of Cu(II)/glycine chelate complex and ammonium persulfate (APS) initiator. The strongly hydrophobic PBA was intercalated into the hydrophilic SS layer. Since the interlayers of silicate were filled with sodium cations, the hydrophilic properties were enhanced and lead to high degree of swelling. The formation of the PBA/SS nanocomposite was confirmed by infrared spectra (IR). Furthermore, as evidenced by transmission electron microscopy (TEM), the composite so obtained was found to have nanoscale structure. X-ray diffraction (XRD) was used to characterize the nanoscale dispersion of the layer silicate and useful for measurement of d-spacing in interlayer system. It was found from thermogravimetric analysis that PBA/SS nanocomposites had more thermal stability as compared to raw PBA due to intercalation. Burning test of the nanocomposites performance exhibited a flame retardant property, which was also verified from cone calorimeter analysis. For its commercialization, the ecological friendly nature was studied via biodegradation and was found to have better biodegradability than the raw PBA.     

Synthesis and electrochemical studies of charge-transfer complexes of thiazolidine-2,4-dione with σ and π acceptors

In the present work, we report the synthesis and characterization of novel charge-transfer complexes of thiazolidine-2,4-dione (TZD) with sigma acceptor (iodine) and pi acceptors (chloranil, dichlorodicyanoquinone, picric acid and duraquinone). We also evaluated their thermal and electrochemical properties and we conclude that these complexes are frequency dependent. Charge-transfer complex between thiazolidine-2,4-dione and iodine give best conductivity. In conclusion, complex with sigma acceptors are more conducting than with pi acceptors.     

Kinetics and mechanism of the oxidation of some organic sulfides by morpholinium chlorochromate

The oxidation of organic sulfides by morpholinium chlorochromate (MCC) resulted in the formation of the corresponding sulfoxides. The reaction is first order with respect to both MCC and the sulfide. The reaction is catalyzed by toluene‐p‐sulfonic acid (TsOH). The oxidation was studied in 19 different organic solvents. An analysis of the solvent effect by Swain's equation showed that both the cation‐ and anion‐solvating powers of the solvents play important roles. The correlation analyses of the rate of oxidation of 34 sulfides were performed in terms of various single and multiparametric equations. For the aryl methyl sulfides, the best correlation is obtained with Charton's localized‐delocalized‐resonance and localized‐delocalized‐resonance‐steric equations. The oxidation of alkyl phenyl sulfides exhibited a very good correlation in terms of the Pavelich–Taft equation. The polar reaction constants are negative, indicating an electron‐deficient sulfur center in the rate‐determining step. A mechanism involving formation of a sulfonium cation intermediate in the slow step has been proposed. © 2008 Wiley Periodicals, Inc. Int J Chem Kinet 41: 65–72, 2009     

A tandem mass spectrometric approach to the identification of O‐glycosylated glargine glycoforms in active pharmaceutical ingredient expressed in Pichia pastoris

Glycoforms of glargine expressed in Pichia pastoris were isolated by high‐performance liquid chromatography and analyzed by a series of chemical and mass spectrometric methods for the identification of various glycoforms, glycosylation position, nature and structure of glycans. Reduction and alkylation, peptide mapping techniques were used to decipher the amino acid site at which glycosylation had taken place. Chemical methods were coupled with mass spectrometry techniques such as electrospray ionization and matrix‐assisted laser desorption/ionization for identification of the glycosylation site. Copyright © 2009 John Wiley & Sons, Ltd.     

“On water” ultrasound-assisted one pot efficient synthesis of functionalized 2-oxo-benzo[1,4]oxazines: First application to the synthesis of anticancer indole alkaloid,Cephalandole A

For the first time, an efficient, simple, synthetic green protocol for the one-pot synthesis of functionalized 2-oxo-benzo[1,4]oxazines 24–29 in water under ultrasound irradiation is presented. As compared to conventional methods, the present protocol avoids traditional chromatography and purification steps and furnished the target molecules in excellent yields (upto 98%) with no side products. The methodology was also demonstrated on gram scale synthesis. Moreover, functionalized 2-oxo-quinoxaline analogues 31–33, another class of bio-active heterocyclic scaffolds, were also prepared using this method. For the first time, this protocol was successfully applied in the synthesis of the anticancer indole alkaloid, Cephalandole A 35.     

Orientation of tie-lines in the phase diagram of DOPC/DPPC/cholesterol model biomembranes

We report the direction of tie-lines of coexisting phases in a ternary diagram of DOPC/DPPC/cholesterol lipid bilayers, which has been a system of interest in the discussion of biological rafts. For coexisting Ld and Lo phases, we find that the orientation angle α of the tie-lines increases as the cholesterol concentration increases and it also increases as temperature increases from T = 15 °C to T = 30 °C. Results at lower cholesterol concentrations support the existence of a different two-phase coexistence region of Ld and So phases and the existence of a three-phase region separating the two two-phase regions. Our method uses the X-ray lamellar D-spacings observed in oriented bilayers as a function of varying hydration. Although this method does not obtain the ends of the tie-lines, it gives precise values (±1°) of their angles α in the ternary phase diagram.     

Pt3Te4 nanoparticles from tellurium nanowires

We report the synthesis of platinum telluride nanoparticles (Pt(3)Te(4) NPs) in the solution phase at room temperature using a template-assisted method. The dendrimeric aggregates formed are composed of several small units of Pt(3)Te(4) NPs of ～4 nm diameter. Tellurium nanowires (Te NWs) are used as the template and the reducing agent in the growth of NPs which occurs due to the galvanic replacement reaction between Te NWs and PtCl(6)(2-). Surface-enhanced Raman scattering (SERS) of the dispersed Pt(3)Te(4) NPs was studied using crystal violet (CV) as the analyte. SERS sensitivity up to 10(-8) M of CV was observed. The Raman enhancement factor (EF) of adsorbed CV on NP aggregates was calculated to be 1.74 × 10(5). The catalytic ability of the as-synthesized Pt(3)Te(4) NPs for the reduction of 4-nitrophenol (4-NP) was studied.     

Ag(9) quantum cluster through a solid-state route

A silver cluster having the composition Ag(9)(H(2)MSA)(7) (H(2)MSA = mercaptosuccinic acid) was synthesized in macroscopic quantities using a solid-state route. The clusters were purified by PAGE and characterized by UV-vis, FTIR, luminescence, and NMR spectroscopy, TEM, XPS, XRD, TG, SEM/EDAX, elemental analysis, and ESI MS. The solid-state route provides nearly pure Ag(9) clusters, and nanoparticle contamination was insignificant for routine studies. Formation of various clusters was observed by modifying the conditions. The effect of ligands on the synthesis was checked. The cluster decomposed slowly in water, and the decomposition followed first-order kinetics. However, it could be stabilized in solvent mixtures and in the solid state. Such materials may be important in cluster research because of their characteristic absorption profiles, which are similar to those of Au(25) and Au(38). The cluster showed luminescence with a quantum yield of 8 × 10(-3) at 5 °C.     

Binding of the bioflavonoid robinetin with model membranes and hemoglobin: Inhibition of lipid peroxidation and protein glycosylation

Recent years have witnessed burgeoning interest in plant flavonoids as novel therapeutic drugs targeting cellular membranes and proteins. Motivated by this scenario, we explored the binding of robinetin (3,7,3′,4′,5′-pentahydroxyflavone, a bioflavonoid with remarkable ‘two color’ intrinsic fluorescence properties), with egg yolk phosphatidylcholine (EYPC) liposomes and normal human hemoglobin (HbA), using steady state and time resolved fluorescence spectroscopy. Distinctive fluorescence signatures obtained for robinetin indicate its partitioning (K_p = 8.65 × 10⁴) into the hydrophobic core of the membrane lipid bilayer. HbA–robinetin interaction was examined using both robinetin fluorescence and flavonoid-induced quenching of the protein tryptophan fluorescence. Specific interaction with HbA was confirmed from three lines of evidence: (a) bimolecular quenching constant K_q ? diffusion controlled limit; (b) closely matched values of Stern–Volmer quenching constant and binding constant; (c) τ₀/τ = 1 (where τ₀ and τ are the unquenched and quenched tryptophan fluorescence lifetimes, respectively). Absorption spectrophotometric assays reveal that robinetin inhibits EYPC membrane lipid peroxidation and HbA glycosylation with high efficiency.