High-resolution atmospheric pressure infrared laser desorption/ionization mass spectrometry imaging of biological tissue

An atmospheric pressure laser desorption/ionization mass spectrometry imaging ion source has been developed that combines high spatial resolution and high mass resolution for the in situ analysis of biological tissue. The system is based on an infrared laser system working at 2.94 to 3.10 μm wavelength, employing a Nd:YAG laser-pumped optical parametrical oscillator. A Raman-shifted Nd:YAG laser system was also tested as an alternative irradiation source. A dedicated optical setup was used to focus the laser beam, coaxially with the ion optical axis and normal to the sample surface, to a spot size of 30 μm in diameter. No additional matrix was needed for laser desorption/ionization. A cooling stage was developed to reduce evaporation of physiological cell water. Ions were formed under atmospheric pressure and transferred by an extended heated capillary into the atmospheric pressure inlet of an orbital trapping mass spectrometer. Various phospholipid compounds were detected, identified, and imaged at a pixel resolution of up to 25 μm from mouse brain tissue sections. Mass accuracies of better than 2 ppm and a mass resolution of 30,000 at m/z?=?400 were achieved for these measurements.

On a Conjecture Concerning a Theorem of Cramér and Wold

A conjecture concerning the Cramér–Wold device is answered in the negative by giving a Fourier-free, probabilistic proof using only elementary techniques. It is also shown how a geometric idea allows one to interpret the Cramér–Wold device as a special case of a more general concept.     

Determining temperature in a magic-angle spinning probe using the temperature dependence of the isotropic chemical shift of lead nitrate

The calibration of temperature in a magic-angle spinning probe with lead nitrate is discussed. The effects of rotation frequency on temperature are demonstrated.     

Ions and atoms in superfluid helium (4He)

New experimental results for mobilities in superfluid helium of the alkali earth ions Be⁺, Mg⁺, Ca⁺, Sr⁺ and Ba⁺ in the temperature region from 1.27 up to 1.66 K are reported. Surprisingly, the temperature dependence of the Be⁺ ion mobility, measured here for the first time, is more similar to that of the He⁺ ion than to the heavier alkali earth ions. This behavior may suggest a snowball like structure for the defect around Be⁺ in contrast to the bubble like defects around the heavier alkali earth ions.     

Monoalkylidene Derivatives of Meldrum's Acid

We were in need of several aliphatic monoalkylidene derivatives of Meldrum's acid (1) for use in conjunction with a current synthetic problem. Implementation of base^1,2 or Lewis acid³ initiated Knoevenagel conditions were found to be unsatisfactory, in our hands, due to the intercedence of subsequent Michael addition of Meldrum's acid to the desired product. We reasoned, therefore, that a method should be employed which generates the alkylidene derivative in the absence of Meldrum's acid.     

Remarks on parameter identification. I

Summary In this paper a method for constructing a spatially varying diffusion coefficient for a parabolic, partial differential equation is given. This function is obtained as the limit of a sequence of functions which are obtained by solving a sequence of finite dimensional optimization problems.Dedicated to Professor Lothar Collatz on the occasion of his 75th birthdaySupported in part by a grant from NORCUS with funds provided by the Department of Energy as part of the Basalt Waste Isolation Project     

Remote polar substituent effects on 13C NMR spectra of olefinic and aromatic carbons

Chemical shifts of the olefinic carbons of ω-substituted 1-pentenes and 1-butenes and the aromatic carbons of phenylethanes and phenylpropanes have been measured. Correlations with various substitutent constants, electronegativities, NMR data from related chemical systems and calculated electrical fields were evaluated. Although the pattern of results is quite consistent with expectations for a field-induced shift of π-electron density, simple correlations of high statistical precision are not found. The general problem of correlating substitutent effects with reactivity and NMR shifts is discussed.     

Online coupling of thin layer chromatography with matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry: synthesis and application of a new material for the identification of carbohydrates by thin layer chromatography/matrix free material enhanced laser desorption/ionization mass spectrometry

This article describes the online hyphenation of thin layer chromatography with matrix free material enhanced laser desorption/ionization mass spectrometry (mf‐MELDI‐MS), the preparation of new material for MELDI and application of this newly synthesized material using TLC/MELDI‐MS for the analysis of carbohydrate reference standards and plant extracts. Samples included within these analyses are standard solutions of glucose, sucrose, raffinose and a plant extract of Quercus robur, which is used for its anti‐inflammatory, anti‐viral and anthelminitc properties in phytomedicine. A new material for mf‐MELDI‐MS is prepared by immobilizing bradykinin – a peptide, on silica gel coupled to 4‐(3‐triethoxysilylpropylureido)azobenzene. This modification enables the absorption of laser energy sufficient for desorption and ionization of low molecular weight molecules like carbohydrates and amino acids. The newly synthesized material delivered excellent results in respect to signal‐to‐noise (S/N) ratio (S/N ratio: >9/1) and sensitivity (limit of detection (LOD): lower to ng/µL). Hyphenation of TLC to MELDI‐MS employing the novel developed material simultaneously as chromatographic and mass spectrometric sorbent was shown for the first time for the analysis of low molecular weight molecules like mono‐ and oligosaccharides. Copyright © 2010 John Wiley & Sons, Ltd.     

Optimization of Streptomyces bacteriophage ��C31 integrase system to prevent post integrative gene silencing in pulmonary type II cells

φC31 integrase has emerged as a potent tool for achieving long-term gene expression in different tissues. The present study aimed at optimizing elements of φC31 integrase system for alveolar type II cells. Luciferase and β-galactosidase activities were measured at different time points post transfection. 5-Aza-2''deoxycytidine (AZA) and trichostatin A (TSA) were used to inhibit DNA methyltransferase and histone deacetylase complex (HDAC) respectively. In A549 cells, expression of the integrase using a CMV promoter resulted in highest integrase activity, whereas in MLE12 cells, both CAG and CMV promoter were equally effective. Effect of polyA site was observed only in A549 cells, where replacement of SV40 polyA by bovine growth hormone (BGH) polyA site resulted in an enhancement of integrase activity. Addition of a C-terminal SV40 nuclear localization signal (NLS) did not result in any significant increase in integrase activity. Long-term expression studies with AZA and TSA, provided evidence for post-integrative gene silencing. In MLE12 cells, both DNA methylases and HDACs played a significant role in silencing, whereas in A549 cells, it could be attributed majorly to HDAC activity. Donor plasmids comprising cellular promoters ubiquitin B (UBB), ubiquitin C (UCC) and elongation factor 1α (EF1α) in an improved backbone prevented post-integrative gene silencing. In contrast to A549 and MLE12 cells, no silencing could be observed in human bronchial epithelial cells, BEAS-2B. Donor plasmid coding for murine erythropoietin under the EF1α promoter when combined with φC31 integrase resulted in higher long-term erythropoietin expression and subsequently higher hematocrit levels in mice after intravenous delivery to the lungs. These results provide evidence for cell specific post integrative gene silencing with φC31 integrase and demonstrate the pivotal role of donor plasmid in long-term expression attained with this system.