pH-Dependent peroxidase activity of yeast cytochrome c and its triple mutant adsorbed on kaolinite

The peroxidase activity of wild-type yeast cytochrome c and its triple mutant K72AK73AK79A adsorbed onto kaolinite was investigated as a function of pH and temperature. Both adsorbed proteins displayed an appreciable catalytic activity, which remained constant from pH 7 to pH 10, decreased below pH 7, and showed a remarkable increase at pH values lower than 4. In the whole pH range investigated the catalytic activity of the adsorbed wild-type cytochrome c was higher than that of the mutant. Both diffuse-reflectance UV-vis and resonance Raman spectroscopies applied on solid samples were used to probe the structural features responsible for the catalytic activity of the immobilized proteins. At neutral and alkaline pH values a six-coordinate low-spin form of cytochrome c was observed, while at pH < 7 the formation of a high-spin species occurred whose population increased at decreasing pH. The orientation and exposure of the heme to the substrate-strictly dependent on adsorption-was found to affect the peroxidase activity.     

Recent advances in the applications of CE to forensic sciences (2001-2004)

The present article reviews the applications of CE in forensic science covering the period from 2001 until the first part of 2005. The overview includes the most relevant examples of analytical applications of capillary electrophoretic and electrokinetic techniques in the following fields: (i) Forensic drugs and poisons, (ii) explosive analysis and gunshot residues, (iii) small ions of forensic interest, (iv) forensic DNA and RNA analysis, (v) proteins of forensic interest, and (vi) ink analysis.     

Capillary electrochromatographic separation of illicit drugs employing a cyano stationary phase

In this study, we present a capillary electrochromatographic method for separation of basic compounds of interest in forensic science (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, 3,4-methylenedioxyethylamphetamine, cocaine, codeine, heroin, morphine, and 6-monoacethylmorphine). Several analytical conditions were taken into account to completely separate in the same run the 10 drugs of abuse analyzed. Chromatographic retention, selectivity and efficiency were evaluated in dependence of the type of stationary phase (CN and RP-C₁₈ derivatized silica particles), mobile phase composition, buffer type and pH, sample injection. The optimum separation parameters were set up using a mixture of aqueous sodium phosphate buffer (pH 2.5)/acetonitrile (80/20, v/v) as the mobile phase, 10 kV and 20 °C as applied voltage and capillary temperature, respectively. Under these conditions all the studied analytes were baseline resolved within 20 min. The method performance was investigated in terms of precision, linearity, sensitivity and accuracy to demonstrate the applicability of the developed capillary electrochromatographic system to forensic analysis. Calibration curves provided a good linearity over a working range of 100–1200 ng/mL for all analytes. Limits of detection and quantification were in the range 5–12 ng/mL and 10–30 ng/mL, respectively. Then the method was applied to the analysis of a human urine sample spiked with a basic compounds’ mixture. Urine samples’ pre-treatment was carried out through a solid phase extraction (SPE) procedure on strong cation exchange (SCX) cartridges.     

Scattering length instability in dipolar Bose-Einstein condensates

We predict a new kind of instability in a Bose-Einstein condensate composed of dipolar particles. Namely, a comparatively weak dipole moment can produce a large, negative two-body scattering length that can collapse the Bose-Einstein condensate. To verify this effect, we validate mean-field solutions to this problem using exact, diffusion Monte Carlo methods. We show that the diffusion Monte Carlo energies are reproduced accurately within a mean-field framework if the variation of the s-wave scattering length with the dipole strength is accounted for properly.     

Reflection properties of meteor trails in a forward-scatter radar experiment

Summary The distribution of the energy reflected by a meteor trail is strongly connected to the ionization density distribution across and along a meteor trail. Recent forward-scatter observations over the Bologna-Lecce baseline in Italy carried out in 1991–92, show that the number of underdense meteor trails (with electron line densityq≤2.10¹⁴ electrons/mvs. peak signal amplitudes decreases more rapidly with the increasing signal level than what predicted. An exponential (simple and/or log-normal) model is proposed, by taking into account different signal-to-noise ratio thresholds and the Kolmogorov-Smirnov (K-S) procedure is used to test the overall goodness of fit between the set of observations and specified theoretical distributions. It is shown that this model provides a better fit to data mainly for middle-high levels of received power and for values ofq≥10¹³ electrons/m.     

Grabbing yeast iso-1-cytochrome c by Cys102: an effective approach for the assembly of functionally active metalloprotein carpets

We report an approach for immobilizing iso-1-cytochrome c from Saccharomyces cerevisiae on oxygen exposing surfaces derivatized with SH-terminated silanes. The SH moieties from silanes have been brought to react with the partially buried Cys102, forming an intermolecular disulfide bond which anchored covalently cytochrome c to the surface. The presence of a single cysteine residue on the protein surface imparted a well-defined orientation to the molecular edifice. Molecular constructs obtained with native cytochrome c and with a cysteine-depleted mutant (C102T) have been investigated by means of scanning force microscopy under liquid, which was performed to assay the quality of the molecular carpet, showing that the native protein formed a robust monolayer at the surface, whereas only a negligible amount of physisorbed molecules were detected in the case of a mutant. UV-vis absorption spectroscopy was performed to confirm that immobilization takes place via the Cys102 residue. Linear sweep voltammetric measurements showed retention of the redox activity of the covalently immobilized cytochrome c, confirming the viability of the proposed immobilization method for obtaining monolayers of redox active molecules.