Evaluation of an in-capillary approach for performing quantitative cytochrome P450 activity studies

An automated in-capillary assay requiring very small quantities of reagents was developed for performing in vitro cytochrome P450 (CYP450) drug metabolism studies. The approach is based on the following: (i) hydrodynamic introduction of nanoliter volumes of substrate and enzyme solutions in the sandwich mode, within a capillary; (ii) mixing the reagents by diffusion across the interfaces between the injected solutions; (iii) collection of the capillary content at the end of the in-capillary assay; and (iv) off-line analysis of the incubation mixture by ultrahigh pressure liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). After optimizing the injection sequence of the reagents, the in-capillary approach was applied to the quantitative determination of the kinetics of drug metabolism reactions catalyzed by three CYP450 isozymes involved in human drug metabolism: CYP1A2, CYP2D6, and CYP3A4. It was demonstrated that this in-capillary method was able to provide similar kinetic parameters for CYP450 activity (e.g., Michaelis constants and turnover values) as the classical in vitro method, with a drastic reduction of reagent consumption. Injection setups used for in-capillary CYP450 assay

Chiral Anomaly and γ3π\gamma 3\pi

Measurement of the $\gamma 3\pi$ process has revealed a possible conflict with what should be a solid prediction generated by the chiral anomaly. We show that inclusion of appropirate energy-momentum dependence in the matrix element reduces the discrepancy.Comment: 8 page standard Latex fil

Partial Derivation of Transformation Properties of Quarks and Leptons

Under the assumptions that $SU(3)_c\times U(1)_Y \times G^{\prime}$ with $G^{\prime}$ simple is a local symmetry group at high energies, that color is parity-conserving, and the Y-charges are irreducible, we show that anomaly constraints imply the minimal set of fermions is fifteen in number. Given this minimal set, we further show that $G^{\prime}$ must be $SU(2)$ and the unbroken gauge symmetry is {\it either} color {\it or} the product of color with electric charge.Comment: 9 pages, UMDHEP 94-72 and IFP-487-UN

Innovative methodology to transfer conventional GC-MS heroin profiling to UHPLC-MS/MS

Nowadays, in forensic laboratories, heroin profiling is frequently carried out by gas chromatography coupled with mass spectrometry (GC-MS). This analytical technique is well established, provides good sensitivity and reproducibility, and allows the use of large databases. Despite those benefits, recently introduced analytical techniques, such as ultra-high-pressure liquid chromatography (UHPLC), could offer better chromatographic performance, which needs to be considered to increase the analysis throughput for heroin profiling. With the latter, chromatographic conditions were optimized through commercial modeling software and two atmospheric pressure ionization sources were evaluated. Data obtained from UHPLC-MS/MS were thus transferred, thanks to mathematical models to mimic GC-MS data. A calibration and a validation set of representative heroin samples were selected among the database to establish a transfer methodology and assess the models' abilities to transfer using principal component analysis and hierarchical classification analysis. These abilities were evaluated by computing the frequency of successful classification of UHPLC-MS/MS data among GC-MS database. Seven mathematical models were tested to adjust UHPLC-MS/MS data to GC-MS data. A simplified mathematical model was finally selected and offered a frequency of successful transfer equal to 95%. Figur

Single-run separation of closely related cationic and anionic compounds by CE-ESI-MS: application to the simultaneous analysis of melamine and its analogs in milk

In recent years, two adulteration incidents concerning the addition of melamine, a nitrogen-rich industrial small polar compound, to pet food and infant formula products have occurred in China. These issues prompted laboratories to develop methods for the analysis of melamine and related compounds in a wide variety of food products and ingredients. In this context, a CE-ESI-MS method was developed to simultaneously analyze melamine and its related products (ammeline, ammelide and cyanuric acid) that possess close physico-chemical properties. This method allows the simultaneous analysis of both cations and anions in a single run, using CE to divide the run into two time segments in normal polarity mode. For this purpose, ESI polarity was switched once during the run, increasing sensitivity and data quality. The method was applied to spiked powdered milk and melamine-contaminated powdered milk, with two sample preparation procedures

Study of γπ→ππ\gamma\pi \to \pi\pi below 1 GeV using Integral Equation Approach

The scattering of $\gamma \pi \to \pi \pi$ is studied using the axial anomaly, elastic unitarity, analyticity and crossing symmetry. Using the technique to derive the Roy's equation, an integral equation for the P-wave amplitude is obtained in terms of the strong P-wave pion pion phase shifts. Its solution is obtained numerically by an iteration procedure using the starting point as the solution of the integral equation of the Muskelshsvilli-Omnes type. It is, however, ambiguous and depends sensitively on the second derivative of the P-wave amplitude at $s=m_\pi^2$ which cannot directly be measured.Comment: 26 pages, 10 figure

Single-run separation of closely related cationic and anionic compounds by CE-ESI-MS: application to the simultaneous analysis of melamine and its analogs in milk

In recent years, two adulteration incidents concerning the addition of melamine, a nitrogen-rich industrial small polar compound, to pet food and infant formula products have occurred in China. These issues prompted laboratories to develop methods for the analysis of melamine and related compounds in a wide variety of food products and ingredients. In this context, a CE-ESI-MS method was developed to simultaneously analyze melamine and its related products (ammeline, ammelide and cyanuric acid) that possess close physico-chemical properties. This method allows the simultaneous analysis of both cations and anions in a single run, using CE to divide the run into two time segments in normal polarity mode. For this purpose, ESI polarity was switched once during the run, increasing sensitivity and data quality. The method was applied to spiked powdered milk and melamine-contaminated powdered milk, with two sample preparation procedures

Kinetic decoupling of neutralino dark matter

After neutralinos cease annihilating in the early Universe, they may still scatter elastically from other particles in the primordial plasma. At some point in time, however, they will eventually stop scattering. We calculate the cross sections for neutralino elastic scattering from standard-model particles to determine the time at which this kinetic decoupling occurs. We show that kinetic decoupling occurs above a temperature $T\sim$ MeV. Thereafter, neutralinos act as collisionless cold dark matter.Comment: Replaced with revised version, new references adde

Enhanced roughness of lipid membranes caused by external electric fields

The behavior of lipid membranes in the presence of an external electric field is studied and used to examine the influence of such fields on membrane parameters such as roughness and show that for a micro sized membrane, roughness grows as the field increases. The dependence of bending rigidity on the electric field is also studied and an estimation of thickness of the accumulated charges around lipid membranes in a free-salt solution is presented.Comment: 9 pages, 6 figures, to appear in Computational Materials Scienc

Positrons from particle dark-matter annihilation in the Galactic halo: propagation Green's functions

We have made a calculation of the propagation of positrons from dark-matter particle annihilation in the Galactic halo in different models of the dark matter halo distribution using our 3D code, and present fits to our numerical propagation Green's functions. We show that the Green's functions are not very sensitive to the dark matter distribution for the same local dark matter energy density. We compare our predictions with computed cosmic ray positron spectra (``background'') for the ``conventional'' CR nucleon spectrum which matches the local measurements, and a modified spectrum which respects the limits imposed by measurements of diffuse Galactic gamma-rays, antiprotons, and positrons. We conclude that significant detection of a dark matter signal requires favourable conditions and precise measurements unless the dark matter is clumpy which would produce a stronger signal. Although our conclusion qualitatively agrees with that of previous authors, it is based on a more realistic model of particle propagation and thus reduces the scope for future speculations. Reliable background evaluation requires new accurate positron measurements and further developments in modelling production and propagation of cosmic ray species in the Galaxy.Comment: 8 pages, 6 ps-figures, 3 tables, uses revtex. Accepted for publication in Physical Review D. More details can be found at http://www.gamma.mpe-garching.mpg.de/~aws/aws.htm