Integration of mass spectrometry into early-phase discovery and development of central nervous system agents.

The early-phase discovery and development of useful central nervous system (CNS) agents present ample opportunities to exploit mass spectrometry and provide detailed compound/mixture characterization, or to make the process faster and/or more economic. Neuropeptide FF antagonists and centrally active thyrotropin-releasing hormone analogues were used as specific examples in this work. We evaluated the characterization of focused libraries of peptide derivatives by electrospray ionization, tandem mass spectrometry and liquid chromatography/tandem mass spectrometry on a quadrupole ion trap and nanoelectrospray on a Fourier transform ion cyclotron resonance mass spectrometer. Immobilized artificial-membrane chromatography was employed as a model to predict/rank new agents against lead compounds for their potential to reach the central nervous system in pharmacologically significant amounts. Measuring brain concentrations in rodents after the intravenous administration of test compounds was used as an in vivo approach, and we took advantage of microdialysis sampling that furnished samples without interfering tissue matrix and afforded the estimation of extracellular concentrations in a localized part of the brain. Overall, making atmospheric-pressure ionization mass spectrometry an integral part of the process has played a major role in increasing throughput, selectivity, specificity and detection sensitivity and thereby providing useful information about the extent or mechanism of transport and metabolic activation/inactivation in early-phase discovery and development of CNS agents.     

The population-sample decomposition approach to multivariate estimation methods

In this paper it is argued that all multivariate estimation methods, such as OLS regression, simultaneous linear equations systems and, more widely, what are known as LISREL methods, have merit as geometric approximation methods, even if the observations are not drawn from a multivariate normal parent distribution and consequently cannot be viewed as ML estimators. It is shown that for large samples the asymptotical distribution of any estimator, being a totally differentiable covariance function, may be assessed by the δ method. Finally, we stress that the design of the sample and a priori knowledge about the parent distribution may be incorporated to obtain more specific results. It turns out that some fairly traditional assumptions, such as assuming some variables to be non-random, fixed over repeated samples, or the existence of a parent normal distribution, may have dramatic effects on the assessment of standard deviations and confidence bounds, if such assumptions are not realistic. The method elaborated by us does not make use of such assumptions.     

Study of the optical properties of a reflecting coating prepared from a Kapton film with deposited aluminum

The effect of the flux density, the flux, and the energy of electrons on the change in reflection spectra and the total solar radiation absorption by the reflecting coating of a space vehicle fabricated from a Kapton film with deposited aluminum in the People's Republic of China is investigated. Tomsk Polytechnic University. Translated from Izvestiya Vysshikh Uchebnykh Zavedenii, Fizika, No. 7, pp. 29–34, July, 2000.     

Universal abelian covers of superisolated singularities

We give explicit examples of Gorenstein surface singularities with integral homology sphere link, which are not complete intersections. Their existence was shown by Luengo–Velasco, Melle–Hernández and Némethi, thereby providing counterexamples to the universal abelian covering conjecture of Neumann and Wahl (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Rheological properties of PDMS/clay nanocomposites and their sensitivity to microstructure

The effect of microstructure on the rheology of clay/polymer nanocomposites is investigated using dispersions of organically treated clay in nearly Newtonian poly(dimethylsiloxane). Degree of dispersion and floc size are altered by using two different dispersion procedures and by changing the shear history. The scaling for dynamic moduli of attractive colloids applies, except for a possible relaxation mechanism at very low frequencies. The time to reach the crossover at a given frequency is found to be extremely sensitive to the dispersion procedure used. Hydrodynamic and elastic components of the steady state stress, on the other hand, evolve in a very similar fashion for the different systems. Although the relaxation times of the elastic stress components change drastically with flow-induced changes in structure, the dispersion process hardly has an effect at all. Intermittent start-up flows in the forward and reverse directions show that anisotropy persists long after the flow has been arrested, even at shear rates where no large reversible flocs are present. The degree of dispersion only had a limited effect on the anisotropy. Finally, the effect of shear on structure recovery has been studied. Very low shear rates are found to increase the rate of recovery, even for small strains.     

Monitoring the heterogeneity in single cell responses to drugs using electrochemical impedance and electrochemical noise

Impedance spectroscopy is a widely used technique for monitoring cell–surface interactions and morphological changes, typically based on averaged signals from thousands of cells. However, acquiring impedance data at the single cell level, can potentially reveal cell-to-cell heterogeneity for example in response to chemotherapeutic agents such as doxorubicin. Here, we present a generic platform where light is used to define and localize the electroactive area, thus enabling the impedance measurements for selected single cells. We firstly tested the platform to assess phenotypic changes in breast cancer cells, at the single cell level, using the change in the cell impedance. We next show that changes in electrochemical noise reflects instantaneous responses of the cells to drugs, prior to any phenotypical changes. We used doxorubicin and monensin as model drugs and found that both drug influx and efflux events affect the impedance noise signals. Finally, we show how the electrochemical noise signal can be combined with fluorescence microscopy, to show that the noise provides information on cell susceptibility and resistance to drugs at the single cell level. Together the combination of electrochemical impedance and electrochemical noise with fluorescence microscopy provides a unique approach to understanding the heterogeneity in the response of single cells to stimuli where there is not phenotypic change.

A light addressable single-cell impedance technique for cell adhesion monitoring and measurement of a cell''s drug response based on electrochemical noise is introduced.