As proteome-wide C-terminal sequence analysis has been largely intractable, we developed a polymer-based enrichment approach to profile protein C-terminal peptides by mass spectrometry and identified ...hundreds of C-terminal peptides in the Escherichia coli proteome. We isotopically labeled GluC protease-digested and undigested samples and identified GluC substrates and their cleavage sites by quantification of neo-C-terminal peptides. Our method thus enables global annotation of protein C-terminal posttranslational modifications, including proteolytic truncations.
The windings insulation of electrical machines will remain a topic that is updated frequently. The criteria severity requested by the electrical machine applications increases continuously. ...Manufacturers and designers are always confronted with new requirements or new criteria with enhanced performances. The most problematic requirements that will be investigated here are the extremely long lifespan coupled to critical operating conditions (overload, supply grid instabilities, and critical operating environments). Increasing lifespan does not have a considerable benefit because the purchasing price of usual machines has to be compared to the purchasing price and maintenance price of long lifespan machines. A machine having a 40-year lifespan will cost more than twice the usual price of a 20-year lifetime machine. Systems which need a long lifetime are systems which are crucial for a country, and those for which outage costs are exorbitant. Nuclear power stations are such systems. It is certain that the used technologies have evolved since the first nuclear power plant, but they cannot evolve as quickly as in other sectors of activities. No-one wants to use an immature technology in such power plants. Even if the electrical machines have exceeded 100 years of age, their improvements are linked to a patient and continuous work. Nowadays, the windings insulation systems have a well-established structure, especially high voltage windings. Unfortunately, a high life span is not only linked to this result. Several manufacturers’ improvements induced by many years of experiment have led to the writing of standards that help the customers and the manufacturers to regularly enhance the insulation specifications or qualifications. Hence, in this publication, the authors will give a step by step exhaustive review of one insulation layout and will take time to give a detailed report on the standards that are linked to insulation systems. No standard can provide insurance about lifespan, nor do any insulation tests incorporate all of the operating conditions: thermal, mechanical, moisture and chemical. Even if one manufacturer uses the standards compliance to demonstrate the quality of its realization; in the end, the successful use in operation remains an objective test. Thereafter, both customer and manufacturers will use the standards while knowing that such documents cannot fully satisfy their wishes. In one 20-year historical review, the authors will highlight the duration in insulation improvements and small breakthroughs in standards writing. High lifespan machines are not the main interest of standards. A large part of this publication is dedicated to the improvements of the insulation wall to achieve the lifespan. Even if the choice of raw materials is fundamental, the understanding of ageing phenomena also leads to improvements.
In electric power generation, customers want generators with high efficiency. Nowadays, modern turbo-generators have efficiencies greater than 98%. Although this amount should not be obtained for all ...kind of machines, efficiency will remain one of the main parameters for customer choice. Efficiency is also linked to the life of the machine: the higher the efficiency is, the longer the machine's lifetime. During the past decade, new forms of energy production have appeared and generators have been developed to fit well into this market. For example, wind generators evolved towards permanent magnet generators having high polarity and running at low speed. Nevertheless, their structure is not fixed. An industrial company has built a prototype of such a generator which uses fractional-slot concentrated-windings (FSCW). This kind of winding is not the structure used by default in such electrical machines. Another field of interest is in autonomous generators which can be used on boats. Even if everyone has in mind large merchant ships, we must not forget smaller ships, such as fishing boats and short-range cruise ships, which spend the most of their time near the coast. This kind of ship does nothave large areas for installing the electric generation or the electric propulsion. It is the reason why, in this article, we focus on the efficiency of machines using fractional-slot concentrated-windings. In many publications which compare performances between distributed and concentrated windings, the result is almost the same. The efficiency of FSCW is not as high as the efficiency associated to the machines which are using distributed windings. Design methods have to be redrawn to integrate, as soon as possible, the loss mitigation in order to provide the best efficiency in power conversion. The following discussion, step by step, introduces the loss mitigation in every part of a machine using FSCW. To close the discussion, a design is produced and it appears that efficiency can be enhanced with suitable design methods.
To evaluate the accuracy of dual-section helical computed tomography (CT) in acute pulmonary embolism (PE) diagnosis.
Of 204 consecutive patients with clinically suspected acute PE (mean age, 58 ...years +/- 14 SD), 158 were enrolled. All patients underwent dual-section helical CT (2.7-mm effective section thickness) and selective pulmonary arteriography within 12 hours of each other. Each image was analyzed independently by two observers, who determined image quality and presence of PE among arterial segments, including at the subsegmental level. The final diagnosis was made with consensus.
Selective pulmonary arteriography was considered optimal in 147 (93%), suboptimal in 10 (6%), and inconclusive in one (0.6%) of 158 patients. Dual-section helical CT findings were considered technically optimal in 140 (89%), suboptimal in 11 (7%), and inconclusive in six (4%). Selective pulmonary arteriography demonstrated PE in 62 patients. Four (6%) of 62 patients had isolated subsegmental PE. The sensitivity of dual-section helical CT was 90%, and the specificity was 94%. The positive and negative predictive values were 90% and 94%, respectively.
Dual-section helical CT is an improvement in helical CT that offers a high sensitivity and specificity for the depiction of PE, including at the subsegmental level. Dual-section helical CT can replace pulmonary arteriography for the direct demonstration of PE in a majority of patients.
This study was designed to define and evaluate a specific index to quantify arterial obstruction with helical CT in acute pulmonary embolism.
Fifty-four patients (mean age, 56 years) with proven ...pulmonary emboli among 158 consecutive patients, who had undergone both CT and pulmonary angiography for clinically suspected pulmonary embolism, were eligible for the study. The CT obstruction index was defined as (n. d) (n, value of the proximal clot site, equal to the number of segmental branches arising distally; d, degree of obstruction scored as partial obstruction value of 1 or total obstruction value of 2). We compared the CT obstruction index with pulmonary arterial obstruction on angiography (assessed by the Miller index), using linear regression, and correlated it with findings on echocardiography. Interobserver variability was determined for both CT and pulmonary angiography indexes.
The CT obstruction index (29% +/- 17%) and the Miller index (43% +/- 25%) were well correlated (r = 0.867, p < 0.0001) with an excellent concordance between investigators for both the CT index (r = 0.944, p < 0.0001) and the Miller index (r = 0.904, p < 0.0001). A CT obstruction index greater than 40% identified more than 90% of patients with right ventricular dilatation.
The degree of arterial obstruction in pulmonary embolism may be quantified by a specific CT index that appears reproducible and highly correlated to the previously described index with pulmonary angiography. Further evaluations are needed to investigate the usefulness of the CT obstruction index for stratification of patient risk and determining therapeutic options.
Beckman‐Coulter has recently introduced the ProteomeLab PF2D™ for 2‐D liquid separation of protein samples. The system features separation in the first dimension by chromatofocusing, followed by RP ...chromatography in the second dimension, allowing the analysis of complex proteomics samples. When used by the standard protocol, reproducibility and column life times are limited, making the use of the instrument very costly. We here present an improved protocol for chromatofocusing, which enhances column life by at least fivefold.
Type II transmembrane serine proteases (TTSPs) are a family of cell membrane tethered serine proteases with unclear roles as their cleavage site specificities and substrate degradomes have not been ...fully elucidated. Indeed just 52 cleavage sites are annotated in MEROPS, the database of proteases, their substrates and inhibitors.
To profile the active site specificities of the TTSPs, we applied Proteomic Identification of protease Cleavage Sites (PICS). Human proteome-derived database searchable peptide libraries were assayed with six human TTSPs (matriptase, matriptase-2, matriptase-3, HAT, DESC and hepsin) to simultaneously determine sequence preferences on the N-terminal non-prime (P) and C-terminal prime (P') sides of the scissile bond. Prime-side cleavage products were isolated following biotinylation and identified by tandem mass spectrometry. The corresponding non-prime side sequences were derived from human proteome databases using bioinformatics. Sequencing of 2,405 individual cleaved peptides allowed for the development of the family consensus protease cleavage site specificity revealing a strong specificity for arginine in the P1 position and surprisingly a lysine in P1' position. TTSP cleavage between R↓K was confirmed using synthetic peptides. By parsing through known substrates and known structures of TTSP catalytic domains, and by modeling the remainder, structural explanations for this strong specificity were derived.
Degradomics analysis of 2,405 cleavage sites revealed a similar and characteristic TTSP family specificity at the P1 and P1' positions for arginine and lysine in unfolded peptides. The prime side is important for cleavage specificity, thus making these proteases unusual within the tryptic-enzyme class that generally has overriding non-prime side specificity.
To link cleaved substrates in complex systems with a specific protease, the protease active site specificity is required. Proteomic identification of cleavage sites (PICS) simultaneously determines ...both the prime- and non-prime-side specificities of individual proteases through identification of hundreds of individual cleavage sequences from biologically relevant, proteome-derived peptide libraries. PICS also identifies subsite cooperativity. To generate PICS peptide libraries, cellular proteomes are digested with a specific protease such as trypsin. Following protease inactivation, primary amines are protected. After incubation with a test protease, each prime-side cleavage fragment has a free newly formed N-terminus, which is biotinylated for affinity isolation and identification by liquid chromatography-tandem mass spectrometry. The corresponding non-prime sequences are derived bioinformatically. The step-by-step protocol also presents a web service for PICS data analysis, as well as introducing and validating PICS peptide libraries made from Escherichia coli.
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