In clinical and diagnostic proteomics, it is essential to develop a comprehensive and robust system for proteome analysis. Although multidimensional liquid chromatography/tandem mass spectrometry (LC/MS/MS) systems have been recently developed as powerful tools especially for identification of protein complexes, these systems still some drawbacks in their application to clinical research that requires an analysis of a large number of human samples. Therefore, in this study, we have constructed a technically simple and high throughput protein profiling system comprising a two-dimensional (2D)-LC/MS/MS system which integrates both a strong cation exchange (SCX) chromatography and a microLC/MS/MS system with micro-flowing reversed-phase chromatography. Using the microLC/MS/MS system as the second dimensional chromatography, SCX separation has been optimized as an off-line first dimensional peptide fractionation. To evaluate the performance of the constructed 2D-LC/MS/MS system, the results of detection and identification of proteins were compared using digests mixtures of 6 authentic proteins with those obtained using one-dimensional microLC/MS/MS system. The number of peptide fragments detected and the coverage of protein sequence were found to be more than double through the use of our newly built 2D-LC/MS/MS system. Furthermore, this multidimensional protein profiling system has been applied to plasma proteome in order to examine its feasibility for clinical proteomics. The experimental results revealed the identification of 174 proteins from one serum sample depleted HSA and IgG which corresponds to only 1 microL of plasma, and the total analysis run time was less than half a day, indicating a fairly high possibility of practicing clinical proteomics in a high throughput manner.
Using a prototype of the Tomo-e Gozen wide-field CMOS mosaic camera, we acquire widefield optical images at a cadence of 2 Hz and search them for transient sources of duration 1.5 to 11.5 seconds. Over the course of eight nights, our survey encompasses the equivalent of roughly two days on one square degree, to a fluence equivalent to a limiting magnitude about V = 15.6 in a 1-second exposure. After examining by eye the candidates identified by a software pipeline, we find no sources which meet all our criteria. We compute upper limits to the rate of optical transients consistent with our survey, and compare those to the rates 2 expected and observed for representative sources of ephemeral optical light.
The positive and negative ionization characteristics of 20 different α-amino acids were investigated using Direct Analysis in Real Time (DART) mass spectrometry. Almost all of the amino acids M were ionized to generate the (de)protonated analytes [M ± H](±)via proton transfer reactions with the typical background ions H3O(+)(H2O)n and O2˙(-) and resonant electron capture by M. The application of DART to amino acids also resulted in molecular ion formation, fragmentation, oxidations involving oxygen attachment and hydrogen loss, and formation of adducts [M + R](-) with negative background ions R(-) (O2˙(-), HCO2(-), NO2(-) and COO(-)(COOH)), depending on the physicochemical and/or structural properties of individual amino acids. The relationship between each amino acid and the ionization reactions observed suggested that fragmentation can be attributed to pyrolysis during analyte desorption as well as excess energy obtained via (de)protonation. Oxidation and [M + R](-) adduct formation, in contrast, most likely originate from reactions with active oxygen such as hydroxyl radicals HO˙, indicating that the typical background neutral species involved in analyte ionization in DART mass spectrometry contain HO˙.
We report the detection of an optical impact flash on Jupiter on 2021 October 15 by a dedicated telescope, Planetary ObservatioN Camera for Optical Transient Surveys, for the first time. Our temporally resolved three-band observations of the flash allowed investigations of its optical energy without the need for approximations on the impact brightness temperature. The kinetic energy of the impactor was equivalent to approximately two megatons of TNT, an order of magnitude greater than that of previously detected flashes on Jupiter and comparable with the Tunguska impact on Earth in 1908. This detection indicates that Tunguska-like impact events on Jupiter occur approximately once per year, two to three orders of magnitude more frequently than terrestrial impacts. The observed flash displayed a single-temperature blackbody spectrum with an effective temperature of approximately 8300 K without clear temporal variation, possibly representing common radiative features of terrestrial Tunguska-class superbolides.
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