Aberrant protein expression in plasma and kidney tissue during experimental obstructive nephropathy

Kidney failure is a major health problem worldwide. Patients with end-stage renal disease require intensive medical support by dialysis or kidney transplantation. Current methods for diagnosis of kidney disease are either invasive or insensitive, and renal function may decline by as much as 50% before it can be detected using current techniques. The goal of this study was, therefore, to identify biomarkers of kidney disease (associated with renal fibrosis) that can be used for the development of a non-invasive clinical test for early disease detection. We utilized two protein-profiling technologies (SELDI-TOF MS and 2-D) to screen the plasma and kidney proteome for aberrantly expressed proteins in an experimental mouse model of unilateral uretric obstruction, which mimics the pathology of human renal disease. Several differentially regulated proteins were detected at the plasma level of day-3-obstructed animals, which included serum amyloid A1, fibrinogen α, haptoglobin precursor protein, haptoglobin and major urinary proteins 11 and 8. Differentially expressed proteins detected at the tissue level included ras-like activator protein 2, haptoglobin precursor protein, malate dehydrogenase, α enolase and murine urinary protein (all p<0.05 versus controls). Immunohistochemistry was used to confirm the up-regulation of fibrinogen. Interestingly, these proteins are largely separated into four major classes: (i) acute-phase reactants (ii) cell-signaling molecules (iii) molecules involved in cell growth and metabolism and (iv) urinary proteins. These results provide new insights into the pathology of obstructive nephropathy and may facilitate the development of specific assay(s) to detect and monitor renal fibrosis.     

Detection of colorectal adenoma and cancer based on transthyretin and C3a-desArg serum levels

Colorectal cancer is the second leading cause of cancer death, and it develops from benign colorectal adenomas in over 95% of patients. Early detection of these cancer precursors by screening tests and their removal can potentially eradicate more than 95% of colorectal cancers before they develop. To discover sensitive and specific biomarkers for improvement of pre‐clinical diagnosis of colorectal adenoma and cancer, we analysed in two independent studies (n = 87 and n = 83 patients) serum samples from colorectal cancer (stage III), colorectal adenoma and control patients using SELDI‐TOF‐MS. Extensive statistical analysis was performed to establish homogeneous patient groups based on their clinical data. Two biomarkers that were each able to distinguish control patients from either colorectal adenoma or colorectal cancer patients (p<0.001) were identified as transthyretin (pre‐albumin) and C3a‐desArg by MS/MS and were further validated by antibody‐based assays (radial immunodiffusion, ELISA). A combination of both proteins clearly indicated the presence of colorectal adenoma or carcinoma. Using a cut‐off of <0.225 g/L for transthyretin and >1974 ng/mL for C3a‐desArg, we found a sensitivity and specificity for colorectal adenoma of 96% and 70%, respectively.     

Chemical cross-linking and mass spectrometry to map three-dimensional protein structures and protein-protein interactions

Closely related to studying the function of a protein is the analysis of its three-dimensional structure and the identification of interaction sites with its binding partners. An alternative approach to the high-resolution methods for three-dimensional protein structure analysis, such as X-ray crystallography and NMR spectroscopy, consists of covalently connecting two functional groups of the protein(s) under investigation. The location of the created cross-links imposes a distance constraint on the location of the respective side chains and allows one to draw conclusions on the three-dimensional structure of the protein or a protein complex. Recently, chemical cross-linking of proteins has been combined with a mass spectrometric analysis of the created cross-linked products. This review article describes the most popular cross-linking reagents for protein structure analysis and gives an overview of the different available strategies that employ chemical cross-linking and different mass spectrometric techniques. The challenges for mass spectrometry caused by the enormous complexity of the cross-linking reaction mixtures are emphasized. The various approaches described in the literature to facilitate the mass spectrometric detection of cross-linked products as well as computer software for data analyses are reviewed.     

有限封闭环境中基于PCA的水声测距方法

有限空间中多途效应严重影响测距的准确性,针对该问题,提出一种基于主成分分析的水声信号测距方法,对信号协方差矩阵进行降维和特征抽取,在保留协方差矩阵信息量的同时抑制多途干扰信号的影响。实验结果表明,该方法能够提高有限封闭液态场环境中水声测距的准确性和稳健性。     

TOF电子能谱仪抛物面电子反射镜参数计算

针对TOF电子能谱仪抛物面电子反射镜的参数问题，依据电磁理论和Reflect-Grid模型，通过合理的近似和简化，对抛物面电子反射镜的电场分布和电子收集效率进行了理论计算，对电子在其中的飞行轨迹进行了模拟。结果表明，抛物网有无置电压对电子的收集效率有很大影响，当偏置电压为700V时，电子收集效率比无偏置电压时高60-80倍，理论计算为100倍，二者基本相符，为装置的改造和使用提供了理论指导。     

液相色谱-离子阱质谱和飞行时间质谱分析橄榄油中的特丁津除草剂

This application describes the use of LC/Ion Trap MS and LC/TOF for the identification of and quantization of terbutylazine in olive oil samples. The analysis by ion trap was achieved in MS/MS mode, monitoring the characteristic fragment ion at m/z 174. The identification by LC/TOF was accomplishment with accurate mass, along with the accurate mass of chlorine isotope cluster present in terbutylazine. The accuracy typically is better than 2 ppm. The method sensitivity, linearity precision accuracy matrix effect, and limit of detection were also studied.     

Development of a practical foolproof system using ultrasonic local positioning

The objective of this research is to develop a prototype for practical foolproof system which can be used in manual assembly processes. For this purpose, a high-performance and low-cost ultrasonic system is proposed to measure 3D positions of an indoor mobile object. Composed of an ultrasonic sender and a receiver, the system employs ultrasonic time-of-flight (TOF) and trilateration to estimate the positions of the object with an accuracy of few centimeters. To calculate three TOFs, ultrasonic signals are processed full-digitally with a low-cost FPGA, which provides high design flexibility keeping both high performance and low noise. Proposed system is autonomous, so there is no need of an external PC and the system development cost becomes low. As an improved thresholding method to calculate the TOFs, this paper proposes a debounce module, designed in the FPGA, to remove the pulse noises generated during the thresholding. The resulting time delay from the debounce is compensated by a microprocessor for calculating actual TOFs. Lastly, the positions of the mobile object are calculated from the TOFs values by trilateration in the microprocessor. In order to remove measurement noises, both moving average filters and Kalman filters are adopted in calculating the TOFs and positions.     

Occurrence and risk assessment of mycotoxins,acrylamide, and furan in Latvian beer

This work reports data on the occurrence of nine mycotoxins and two food processing contaminants – acrylamide and furan – in a total of 100 beers produced in Latvia. Mycotoxins were detected by high-performance liquid chromatography (HPLC) coupled with time-of-flight mass spectrometry, acrylamide by HPLC coupled with quadrupole-Orbitrap mass spectrometry, and furan by headspace gas chromatography-mass spectrometry. The most frequently occurring mycotoxins were HT-2 and deoxynivalenol (DON), which were detected in 52% and 51% of the analysed samples. The highest content was observed for DON, reaching the maximum of 248 µg kg^?1. Furan was ubiquitous, and 74% of the samples contained acrylamide. In terms of the estimated exposure, the biggest potential risk was identified for HT-2 representing more than 11% of tolerable weekly intake. The margin of exposure approach indicated the exposure to furan through beer as significant, this parameter being close to the critical limit.     

Mono‐Estolide Synthesis from trans‐8‐Hydroxy‐Fatty Acids by Lipases in Solvent‐Free Media and Their Physical Properties

Pseudomonas aeruginosa 42A2 is known to produce two hydroxy‐fatty acids, 10(S)‐hydroxy‐8(E)‐octadecenoic and 7,10(S,S)‐dihydroxy‐8(E)‐octadecenoic acids, when cultivated in a mineral medium using oleic acid as a single carbon source. These compounds were purified, 91 and 96 % respectively, to produce two new families of estolides: trans‐8‐estolides and saturated estolides from the monohydroxylated monomer. trans‐8‐estolides were produced by three different lipases (Novozym 435, Lipozyme RM IM and Lipozyme TL IM) with reaction yields between 68.4 ± 2.1 and 94.7 ± 2.4 % in a solvent‐free medium at 80 °C in 168 h under vacuum. Novozym 435 was found to be the most efficient biocatalyst for both hydroxy‐fatty acids with reaction yields of 71.7 ± 2.3 and 94.7 ± 2.4 %, respectively. Moreover, saturated estolides were also produced from a saturated 10(S)‐hydroxy‐8(E)‐octadecenoic. These estolides were chemically and enzymatically synthesized with Novozym 435, under the previous described reaction conditions with yields of 60.7 ± 2.1 and 71.2 ± 2.3 % respectively. Finally, viscosity, glass transition temperature, decomposition temperatures and enthalpies were determined to characterize both types of estolides. Thermal applications for both types of polyesters were improved since glass transition temperatures were lowered and decomposition temperatures were increased, with respect to their corresponding substrates.