Characterization of an immobilized enzyme reactor for on-line protein digestion

Despite the developments for faster liquid chromatographic and mass spectral detection techniques, the standard in-solution protein digestion for proteomic analyses has remained relatively unchanged. The typical in-solution trypsin protein digestion is usually the slowest part of the workflow, albeit one of the most important. The development of a highly efficient immobilized enzyme reactor (IMER) with rapid performance for on-line protein digestion would greatly decrease the analysis time involved in a proteomic workflow. Presented here is the development of a silica based IMER for on-line protein digestion, which produced rapid digestions in the presence of organic mobile phase for both model proteins and a complex sample consisting of the insoluble portion of a yeast cell lysate. Protein sequence coverage and identifications evaluated between the IMER and in-solution digestions were comparable. Overall, for a yeast cell lysate with only a 10 sec volumetric residence time on-column, the IMER identified 507 proteins while the in-solution digestion identified 490. There were no significant differences observed based on identified protein’s molecular weight or isoelectric point between the two digestion methods. Implementation of the IMER into the proteomic workflow provided similar protein identification results, automation for sample analysis, and reduced the analysis time by 15 hr

Street railway equipment

Citation: Alexander, Lizzie Bea. Problems of bread making. Senior thesis, Kansas State Agricultural College, 1907.Morse Department of Special CollectionsIntroduction: Alternating current will perhaps in time supplant direct current for traction purposes as it did in lighting, with the advent of the long distance high potential transmission, but as yet the direct current plays a very important part in both power and traction. It has however the disadvantage of being difficult to transmit long distances without great loss. This is due to the inherent limitations in the production of high voltages from the direct current generator. Commutation at voltages sufficiently high to warrant economical transmission over long distances, is extremely difficult. Sparking and other difficulties at once arise which place the voltage within the limitations mentioned. The alternating current generator has not only the advantage of higher voltage, but by means of suitable transformer devices the voltage may be raised to any desired value for economical transmission. It will be readily seen, therefore, that alternating current machinery will supplant the direct current for traction purposes. In this thesis only direct current is treated. Direct current on a small scale and for local distribution fulfills more nearly the requirements demanded for traction purposes such as is needed on railways. The discussions are confined to the electrical equipment of the car only, the mechanical construction being referred to only as needed in line with the electrical connections

Extraction, Enrichment, Solubilization, and Digestion Techniques for Membrane Proteomics

The importance of membrane proteins in biological systems is indisputable; however, their amphipathic nature makes them difficult to analyze. In this study, the most popular techniques for extraction, enrichment, solubilization, and digestion are compared, resulting in an overall improved workflow for the insoluble portion of Saccharomyces cerevisiae cell lysate. Yeast cells were successfully lysed using a French press pressure cell at 20 000 psi, and resulting proteins were fractionated prior to digestion to reduce sample complexity. The proteins were best solubilized with the addition of ionic detergent sodium deoxycholate (1%) and through the application of high-frequency sonication prior to a tryptic digestion at 37 °C. Overall, the improved membrane proteomic workflow resulted in a 26% increase in membrane protein identifications for baker's yeast. In addition, more membrane protein identifications were unique to the improved protocol. When comparing membrane proteins that were identified in the improved protocol and the standard operating procedure (176 proteins), 93% of these proteins were present in greater abundance (higher intensity) when using the improved method

Airboat Use and Disturbance of Floating Mat Fen Wetlands in Interior Alaska, U.S.A.

The use of airboats is expanding in Alaska, particularly in the interior. This study describes the nature, magnitude, and distribution of disturbance caused by airboat trails over floating mat fen wetlands in the Tanana Flats near Fairbanks, Alaska. Airphoto interpretation showed over 300 km of airboat trails by 1995, with a 15% expansion of the trail system since 1989. Field sampling was done at 30 trail and adjacent control sites along this trail system to assess changes in hydrology, soils, and vegetation. Water velocities in the trails at two-thirds of the sites were at least an order of magnitude greater than velocities of less than 1.5 cm/sec in the control areas. On average, 30 cm of the 0.5-0.75 m thick floating mat has been removed or eroded by airboat traffic at the sampling locations. Nearly all emergent floating mat vegetation has been destroyed (5% cover remaining on average) so that the trails resemble a highly visible open water stream channel 2-3 m wide through the floating mats. Although the recovery and regrowth potential of floating mats in trails is high, recovery is unlikely in trails with continuing use.En Alaska, l'utilisation des hydroglisseurs est à la hausse, surtout à l'intérieur des terres. Cette étude décrit la nature, l'amplitude et la distribution des perturbations causées par le sillage des hydroglisseurs sur les zones humides de vasières à mattes flottantes situées dans les plaines marécageuses de Tanana près de Fairbanks (en Alaska). L'interprétation de photos a montré que, en 1995, les sillages d'hydroglisseurs s'étendaient sur plus de 300 km, ce qui correspond à une augmentation de 15 p. cent du réseau depuis 1989. On a procédé à un échantillonnage sur le terrain à 30 emplacements de sillages et aires témoins adjacentes le long du réseau en vue d'évaluer les changements dans l'hydrologie, les sols et la végétation. Dans deux tiers des emplacements, la vitesse de l'eau dans les sillages était d'au moins un ordre de grandeur supérieure aux vitesses de moins de 1,5 cm/sec dans les zones témoins. Aux endroits d'échantillonnage, 30 cm en moyenne de la matte flottante épaisse de 0,5 à 0,75 m a été enlevée ou érodée par la circulation de l'hydroglisseur. Presque toute la végétation de la matte flottante émergée a été détruite (il reste en moyenne 5 p. cent du couvert) de sorte que les sillages sont très visibles et forment un chenal d'eau ouverte de 2 à 3 m de large à travers les mattes flottantes. Bien que le potentiel de recouvrement et de repousse des mattes flottantes dans les sillages soit élevé, il est peu probable que ce recouvrement ait lieu dans les sillages qui sont utilisés de façon continue

Implementation of high slurry concentration and sonication to pack high-efficiency, meter-long capillary ultrahigh pressure liquid chromatography columns

Slurry packing capillary columns for ultrahigh pressure liquid chromatography is complicated by many interdependent experimental variables. Previous results have suggested that combination of high slurry concentration and sonication during packing would create homogeneous bed microstructures and yield highly efficient capillary columns. Herein, the effect of sonication while packing very high slurry concentrations is presented. A series of six, 1 m × 75 μm internal diameter columns were packed with 200 mg/mL slurries of 2.02 μm bridged-ethyl hybrid silica particles. Three of the columns underwent sonication during packing and yielded highly efficient separations with reduced plate heights as low as 1.05