Site-specific N-linked glycosylation analysis of human carcinoembryonic antigen by sheathless capillary electrophoresis-tandem mass spectrometry

With 28 potential N-glycosylation sites, human carcinoembryonic antigen (CEA) bears an extreme amount of N-linked glycosylation, and approximately 60% of its molecular mass can be attributed to its carbohydrates. CEA is often overexpressed and released by many solid tumors, including colorectal carcinomas. CEA displays an impressive heterogeneity and variability in sugar content, however site-specific distribution of carbohydrate structures has not been reported so far. The present study investigated CEA samples purified from human colon carcinoma and human liver metastases and enabled the characterization of 21 out of 28 potential N-glycosylation sites with respect to their occupancy. The coverage was achieved by a multi-enzymatic digestion approach with specific enzymes, such as trypsin, endoproteinase Glu-C, and the non-specific enzyme, pronase, followed by analysis using sheathless CE-MS/MS. In total, 893 different N-glycopeptides and 128 unique N-glycan compositions were identified. Overall, a great heterogeneity was found both within (micro) and in between (macro) individual N-glycosylation sites. Moreover, notable differences were found on certain N-glycosylation sites between primary adenocarcinoma and metastatic tumor in regard to branching, bisection, sialylation and fucosylation. Those features, if further investigated in a targeted manner, may pave the way towards improved diagnostics and monitoring of colorectal cancer progression and recurrence. Raw mass spectrometric data and Skyline processed data files that support the findings of this study are available in the MassIVE repository with the identifier MSV000086774 [https://doi.org/doi:10.25345/C5Z50X]

Інтелектуальна система керування електропривода камери телевізійного спостереження

Інтелектуальна система керування електропривода камери телевізійного спостереження = The intelligent control system of the television camera's electric drive / Д. Ю. Шарейко, І. С. Білюк, А. М. Фоменко, О. В. Савченко, О. В. Майборода // Вісн. аграр. науки Причорномор'я. – Миколаїв : МДАУ, 2018. – № 4. – C. 128-133.Розглянуто тиристорний електропривод постійного струму, система керування в якому побудована на основі самонавчальної нейронної сітки. Використовуючи властивості нелінійності здійснено налаштування з покращенням швидкодійності та зменшенням помилки керування. Проведено дослідження та порівняльний аналіз інтелектуальної системи керування та лінійної системи керування електропривода постійного струму. Доведено перевагу інтелектуальної системи керування в електроприводі камери спостереження.The thyristor electric drive of a direct current, the control system in which is built on the basis of a self-learning neural network, is considered. Using the properties of nonlinearity, tuning is performed with improved performance and reduced control error. A study and comparative analysis of the intelligent control system and the linear control system of the electric drive of the direct current was carried out. The advantage of the intelligent control system in the electric drive of the surveillance camera has been proved.Рассмотрен тиристорный электропривод постоянного тока, система управления в котором построена на основе самообучающейся нейронной сети. Осуществлена настройка с использованием свойств нелинейности улучшением быстродействия и уменьшением ошибки управления. Проведено исследование и сравнительный анализ интеллектуальной системы управления и линейной системы управления электропривода постоянного тока. Доказано преимущество интеллектуальной системы управления в электроприводе камеры наблюдения

O-glycomic and proteomic signatures of spontaneous and butyrate-stimulated colorectal cancer cell line differentiation

Gut microbiota of the gastrointestinal tract provide health benefits to the human host via bacterial metabolites. Bacterial butyrate has beneficial effects on intestinal homeostasis and is the preferred energy source of intestinal epithelial cells, capable of inducing differentiation. It was previously observed that changes in the expression of specific proteins as well as protein glycosylation occur with differentiation. In this study, specific mucin O-glycans were identified that mark butyrate-induced epithelial differentiation of the intestinal cell line CaCo-2 (Cancer Coli-2), by applying porous graphitized carbon nano-liquid chromatography with electrospray ionization tandem mass spectrometry. Moreover, a quantitative proteomic approach was used to decipher changes in the cell proteome. It was found that the fully differentiated butyrate-stimulated cells are characterized by a higher expression of sialylated O-glycan structures, whereas fucosylation is downregulated with differentiation. By performing an integrative approach, we generated hypotheses about the origin of the observed O-glycome changes. These insights pave the way for future endeavors to study the dynamic O-glycosylation patterns in the gut, either produced via cellular biosynthesis or through the action of bacterial glycosidases as well as the functional role of these patterns in homeostasis and dysbiosis at the gut-microbiota interface

Neuronal Profilin Isoforms Are Addressed by Different Signalling Pathways

Profilins are prominent regulators of actin dynamics. While most mammalian cells express only one profilin, two isoforms, PFN1 and PFN2a are present in the CNS. To challenge the hypothesis that the expression of two profilin isoforms is linked to the complex shape of neurons and to the activity-dependent structural plasticity, we analysed how PFN1 and PFN2a respond to changes of neuronal activity. Simultaneous labelling of rodent embryonic neurons with isoform-specific monoclonal antibodies revealed both isoforms in the same synapse. Immunoelectron microscopy on brain sections demonstrated both profilins in synapses of the mature rodent cortex, hippocampus and cerebellum. Both isoforms were significantly more abundant in postsynaptic than in presynaptic structures. Immunofluorescence showed PFN2a associated with gephyrin clusters of the postsynaptic active zone in inhibitory synapses of embryonic neurons. When cultures were stimulated in order to change their activity level, active synapses that were identified by the uptake of synaptotagmin antibodies, displayed significantly higher amounts of both isoforms than non-stimulated controls. Specific inhibition of NMDA receptors by the antagonist APV in cultured rat hippocampal neurons resulted in a decrease of PFN2a but left PFN1 unaffected. Stimulation by the brain derived neurotrophic factor (BDNF), on the other hand, led to a significant increase in both synaptic PFN1 and PFN2a. Analogous results were obtained for neuronal nuclei: both isoforms were localized in the same nucleus, and their levels rose significantly in response to KCl stimulation, whereas BDNF caused here a higher increase in PFN1 than in PFN2a. Our results strongly support the notion of an isoform specific role for profilins as regulators of actin dynamics in different signalling pathways, in excitatory as well as in inhibitory synapses. Furthermore, they suggest a functional role for both profilins in neuronal nuclei

USE OF MIVAR EXPERT SYSTEMS FOR SOLVING PROBLEMS OF TEXT UNDERSTANDING AND IMAGE RECOGNITION

Показан практический пример применения миварных экспертных систем для решения основных задач искусственного интеллекта (ИИ): понимания текста и распознавания изображений. В «НИИ МИВАР» ведется создание систем логического искусственного интеллекта на основе российских миварных технологий многомерного накопления информации и линейной сложности логической обработки. Программный комплекс «Конструктор экспертных систем КЭСМИ 1.1» позволяет обрабатывать 5 млн продукционных правил в секунду на персональных компьютерах. Для решения задач понимания текстов на русском языке в 15-мерной миварной базе данных и правил реализована информационная модель «Картина мира» с несколькими контекстами и представлением миварной сети, содержащей 160 000 объектов (вершин графа) и более 600 000 правил (ребер графа). Для отработки подходов по семантическому распознаванию изображений к этой же многомерной миварной базе данных и правил добавлены несколько тысяч картинок.In this article the example of practical application of mivar expert systems for solving the main problems of artificial intelligence is presented: text understanding and image recognition. At «SRI MIVAR» we develop systems of logical inference which are based on Russian mivar technologies characterized by multidimensional accumulation of information and logical processing with linear complexity. “Constructor of expert systems CES 1.1” (IDE) was developed which allows to process 5 million production rules per second on personal computers. To solve problems of Russian text understanding the informational model “world picture version 3” in fifteen dimensional mivar database with multiple contexts and presentation of mivar network containing 160,000 objects (vertices of the graph), and more than 600,000 rules (edges) was developed. For testing approaches of semantic image recognition to the same mivar multidimensional database several thousand pictures were added. The usage examples of textual semantic information in recognizing image content for removal of context uncertainty are presented

Correction for out-of-focus emission in fluorescence fluctuation spectroscopy; generalization of the algorithms

One of the challenges of FFS is an adequate approximation of brightness profile. The key feature of the Fluorescence Intensity Distribution Analysis (FIDA) [1] is, for example, a polynomial model used to approximate broad spectrum of brightness profile functions. On the other hand Perroud [2] and Huang [3] have suggested an approach for brightness profile correction, as an extension of Photon Counting Histogram (PCH) analysis. In this case the additional fitting parameters are defined as a relative difference between integrals of the actual brightness profile and its Gaussian approximation. In current work we show that this approach can be derived straightforward from the FFS theory based on generation functions (GF). One of the advantages of GF is better performance of the algorithm; mainly because the applications of successive convolutions for total distribution calculation are not necessary anymore. As a logical consequence of the developed theory we apply corrections for out-of-focus emission in Fluorescence Cumulants Analysis (FCA) and show that applied corrections improve results of analysis that makes FCA an appropriate method for the analysis of FFS data alongside with PCH and FIDA. [1] Kask, P., K. Palo, D. Ullmann, and K. Gall. Fluorescence-intensity distribution analysis and its application in biomolecular detection technology. Proc. Natl. Acad. Set USA. 96:13756-13761 (1999). [2] Perroud T.D., B. Huang, M.I. Wallace, and R.N. Zare. Photon Counting Histogram for One-Photon Excitation. ChemPhysChem. 4:1121-1123 (2003). [3] Huang B., T.D. Perroud, and R.N. Zare. Photon Counting Histogram: One-Photon Excitation. ChemPhysChem. 5:1523-1531 (2004)

Comparative effectiveness of hypertensive crise management in the post-surgery period after carotid endarterectomy: first results

Aim. To compare the effectiveness of different antihypertensive medications in the management of hypertensive crise (HC) in the early post-surgery period after carotid endarterectomy (CEE). Material and methods. The study included 39 patients (14 women, 25 men; mean age 65,6±6,6 years) hospitalised to the Cardiovascular Surgery Department of a major Moscow City hospital. All patients underwent CEE, with a HC developed in the early post-surgery period. All participants were divided into three groups: in Group I, HC was treated with nimodipine, while Groups II and III received enalaprilat and uradipil, respectively. Results. Intravenous administration of all three medications resulted in an antihypertensive effect. A significant reduction in blood pressure (BP) levels was observed after the administration of nimodipine and uradipil. However, nimodipine caused reflectory tachycardia, which was not observed in patients treated with uradipil. Conclusion. The results obtained are consistent that out of the three studied medications, uradipil appeared the most appropriate for the HC management in the early post-CEE period. It reduced BP levels without causing reflectory tachycardia or clinical hypotension, which confirms clinical effectiveness and safety of uradipil as a medication of choice for the urgent HC management

P158

Opisthorchiasis is a form of foodborne trematodiasis which is caused by liver flukes. It has been shown that a chronic Opisthorchiasis infection increases a risk of cholagiocarcinoma of liver. It is commonly believed that a gradual change of homeostasis in a parasite microenvironment (bile) leads to liver fluke-induced cancer. Nevertheless, no systematic, analytically driven studies confirming this hypothesis have been published yet. The restricted access to clinical material and extreme complexity of the biological matrix (bile) both are the important “rate limiting factors” for a progress in the field. Here we present for the first time a cross-platform mass spectrometric analysis of bile juice collected from the patients with cholangiocarcinoma-associated diseases. We show that an effective analysis of such complex biological matrix as bile juice requires a combination of orthogonal analytical platforms (e.g. RPLC–MS and HILIC–MS) maximizing coverage of the metabolic space. Materials and methods: 28 patients with O. felineus infection and 30 negative controls were included in the study. The infection status was confirmed using microscopy analysis of the bile. Bile samples were collected from the gallbladder using sterile puncture, directly frozen and stored at −80 °C until analysis. The samples were randomized and organized into the acquisition blocks consisting of the samples and quality controls (QC). Experiments were carried out with a Dionex Ultimate 3000 LC system (Thermo Scientific/Dionex, The Netherlands) equipped with a Dual Gradient Separation pump allowing for parallel LC analysis, and hyphenated to an Impact UHR-qTOF mass analyzer (Bruker Daltonics, Germany). Reversed-phase experiments (RPLC) were performed with an UHPLC BEH Shield RP18 column 100 × 2.1 mm, 1.7 μm (Waters) and HILIC experiments with a Luna HILIC column (Phenomenex, The Netherlands) of 100 × 2.00 mm, 3 μ m. RPLC data were acquired in ESI positive mode and HILIC in negative mode, respectively. The data acquisition rate was set to 1 Hz over a mass range of m/z 50–1000. The LC–MS data files were aligned by using the in-house developed alignment algorithm MS-Align 2 tool (www.ms-utils.org/msalign2). Results: After the data prepressing, which includes alignment, noise filtering and peak picking two data matrixes costing of 412 features (metabolites) for RPLC and 428 ones for HILIC were generated. To evaluate a degree of similarity between the two data matrixes the RV coefficient (a multivariate extension of correlation coefficient) was used. The coefficient has flattened at 0.58 showing that despite a strong overlap between the datasets there is a substantial number of the “platform specific” metabolites. Those structures will certainly be missed if a single platform strategy is applied. Conclusion: Here we present for the first time a cross-platform mass spectrometric analysis of bile juice collected from the patients cholangiocarcinoma-associated diseases. We show that a combination of the two platforms greatly improves the coverage of the metabolome and as such should be a firstchoice for exploratory studies of the complex biological matrixes. Work was conducted with the application of the Tomsk regional common use center technical equipment acquired thanks to a grant of the Russian Ministry of the Agreement No. 14.594.21.0001 (RFMEFI59414X0001). This project is supported by “The Tomsk State University Academic D.I. Mendeleev Fund Program” under Grant (No. 18.1.52.2015)

Initial Guesses Generation for Fluorescence Intensity Distribution Analysis

The growing number of applications of Fluorescence Intensity Distribution Analysis (FIDA) demands for new approaches in data processing, which should increase speed and robustness of the method, iterative algorithms of parameter estimation, although proven to be universal and accurate, require some initial guesses (IG) for the unknown parameters. To develop a general algorithm for IG generation is not a trivial task, but in case of FIDA, we are in an advantageous position since available ranges of parameters values are restricted due to numerical limitation of the used model. In the present work we introduce an approach for IG generation in FIDA experiments (sample parameters - brightness and concentrations, and an experimental set-up related parameters - background, observation volume profile) based on the method of moments. A number of analytical simplifications have been introduced in order to increase the accuracy and robustness of the numerical algorithms. The performance of the developed method has been tested on simulated and experimental data. Iterative fitting with calculated IG proved to be at least five times (and often more) faster than with the arbitrarily chosen IG. Using simulated data we have also shown that at least three sets of parameters a and b, characterizing the observation volume, ensure a comparable quality of fit. A rule for selection the best set is suggested. An outline of further investigations to explore comprehensively statistical robustness of the parameters is given