Mass Spectrogram Research Articles

The Enhanced Anti-anxiety Potentiality and Relevant Material Basis for the Extracts of Bile Processed Coptidis Rhizoma

Objective Bile-processed Coptidis Rhizoma (CR) exhibits stronger cold properties and central effect. However, the commonly used bile includes pig/cattle/sheep bile. It is currently unclear which kind of bile performs the best synergistic effect, and the enhanced efficacy is mainly contributed by what components. In this work, a comparative study on pig/cattle/sheep bile processed CR (pCR/cCR/sCR) was carried out. Materials and Methods Firstly, the anxiety model of sthenic heat type was established. Then, pharmacological research involving general state observation, biochemical examination (succinate dehydrogenase, glucokinase, corticosterone, adrenocor ticotropic hormore), pathological evaluation (liver and hippocampus tissue), and behavioral tests (elevated plus maze, open field test) was implemented. Finally, chemical assessment involving high-performance liquid chromatography (HPLC) fingerprints of pCR/cCR/sCR extracts and mass spectrograms of pig/cattle/sheep bile were performed in combination. Results cCR performed the most prominent enhancement in improving hyperactive energy metabolism in liver and abnormal hyperactivity of hypothalamic–pituitary–adrenal axis axis. As for chemical assessment, HPLC fingerprints of pCR/cCR/sCR extracts indicated that alkaloid components detected in cCR extracts were relatively low, mass spectrograms of pig/cattle/sheep bile suggested that bile acids in cattle bile were relatively abundant, and the contents of taurocholic acid, glycocholic acid, and glycodeoxycholic acid were particularly high. Conclusion Cattle bile could be specified as the processing excipient for bile-processed CR, and the high levels of taurocholic acid, glycocholic acid, and glycodeoxycholic acid mainly contributed to the enhanced efficacy of cCR.

Preparation and Characterization of Guaiacol-Furfuramine Benzoxazine and Its Modification of Bisphenol A-Aniline Oxazine Resin.

A new type of benzoxazine resin has been synthesized using a natural phenol source, guaiacol, and a biomass amines, furfuramine. The synthesis conditions were optimized; when the reaction molar ratio of guaiacol, furfuramine, and polyformaldehyde was 1:1:4, the highest synthetic yield was reached. The product was characterized via testing using transform infrared spectroscopy (FT-IR), gel permeation chromatography (GPC), mass spectrogram (MS), and nuclear magnetic resonance (1H-NMR) to confirm its molecular structure. A differential scanning calorimetry (DSC) test was conducted to analyze the thermodynamic properties of the product, and the results showed that the product decomposed and evaporated at around 180 °C, making it impossible to achieve self-curing. However, the prepared guaiacol-furfuramine benzoxazine resin (GFZ) can be blended and cured in certain proportions with bisphenol A-aniline oxazine resin (BAZ) as a GFZ/BAZ binary system (5:95, 10:90, 20:80, and 40:60). Dynamic mechanical analysis (DMA) test results showed that when the content of GFZ was 10%, the storage modulus of the copolymer resin was greatly improved. After conducting impact strength tests on the copolymer resin, it was found that the toughness of the copolymer resin had improved, and the maximum impact strength had increased by nearly three times. This indicates that the flexible long-chain structure in GFZ can effectively improve the toughness of the cured copolymer system. The reaction of active groups on benzoxazine molecules with other resins can not only improve the mechanical properties of their cured products, but also has important significance in the preparation of low-cost and environmentally friendly sustainable composite materials with excellent comprehensive performance.

Two flavors in adulterated sesame oil: discovery, confirmation, and content regularity study.

Exploring and accurately detecting new adulteration markers in sesame oil is an important measure for sesame oil adulteration monitoring. In this study, two endogenous flavors sulfurol and γ-nonalactone which can be used as potential adulteration markers were first discovered in sesame oil and accurately quantified. First, the two endogenous flavors were discovered using gas chromatography-mass spectrometry (GC-MS), and their structures were confirmed by comparing the mass spectrograms with the NIST spectral library. Then the liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using direct methanol extraction pretreatment and vanillin-D3 as an internal standard was developed for rapid quantitation and application. The method was successfully validated with recoveries ranging from 88.5% to 102.2% and relative standard deviations between 2.6% and 10.5% (n = 6). The combined method of GC-MS and LC-MS/MS was indicated to be efficient and highly sensitive for detection of sulfurol and γ-nonalactone in edible oil. Subsequently, 31 sesame oils from the market were detected, revealing that 31 samples contained the identified flavors within a relatively consistent range. However, the concentration of these flavor substances in one sample was abnormally high, indicating that there was a potential risk of adulteration. Therefore, the developed method shows good potential for quality evaluation and adulteration screening of sesame oil.

Bioprotein Spectrum Availability Analysis Based on ANN

The most common way to analysis protein components is using mass spectrum, however there are errors cause in the operating process and made by equipment, some mass spectrometry images cannot be used for research as the image loses its original features or the features are covered by noise. To exclude unavailable image the equipment can use the method of ANN. ANN uses neural networks to find features of images, and automatically classify images. It successively adjusts parameters in the neural network and produces an output increasingly similar to the target output. This paper analyses the pre-train mass spectrometry of proteins from mouse organs and uses machine learning methods to judge the availability of these mass spectrometry. By using different optimizers and network structures and trying various hyper parameters, this study trained a model that has 85% precision when judging the availability of the mass spectrogram. In particular, it is very effective in excluding noisy images. This greatly reduces the time cost of manually judging the availability of images and it has a good classification effect.

Ultra-Fast Portable and Wearable Sensing Design for Continuous and Wide-Spectrum Molecular Analysis and Diagnostics.

The design and characterization of spatiotemporal nano-/micro-structural arrangement that enable real-time and wide-spectrum molecular analysis is reported and demonestrated in new horizons of biomedical applications, such as wearable-spectrometry, ultra-fast and onsite biopsy-decision-making for intraoperative surgical oncology, chiral-drug identification, etc. The spatiotemporal sesning arrangement is achieved by scalable, binder-free, functionalized hybrid spin-sensitive (<↑| or <↓|) graphene-ink printed sensing layers on free-standing films made of porous, fibrous, and naturally helical cellulose networks in hierarchically stacked geometrical configuration (HSGC). The HSGC operates according toa time-space-resolved architecture that modulate the mass-transfer rate for separation, eluation and detection of each individual compound within a mixture of the like, hereby providing a mass spectrogram. The HSGC could be used for a wide range of applictions, including fast and real-time spectrogram generator of volatile organic compounds during liquid-biopsy, without the need of any immunochemistry-staining and complex power-hungry cryogenic machines; and wearable spectrometry that provide spectral signature of molecular profiles emiited from skin in the course of various dietry conditions.

Environmental Audio Tagging Using Deep Convolution Neural Network and Digital Signal Processing

Machine learning has experienced a strong growth in recent years, due to increased dataset sizes and computational power, and to advances in deep learning methods that can learn to make predictions in extremely non-linear problem settings. The intense problem of automatic environmental sound classification has received alarming attention from the research community in recent years. In this paper the audio dataset is converted into mass spectrogram using Digital Signal Processing (DSP). The spectrogram thus obtained is fed to the Convolutional Neural Network (CNN) for the classification of the audio signal. In this we present a deep convolutional neural network architecture with localized kernels for environmental sound. By training the network on another additional deformed data, the hope is that the network becomes invariant to all deformations and generalizes better to all unseen data. We show that the proposed DSP in combination with CNN architecture, yields state-of-the-art performance for environmental sound classification.

Synthesis, Characterization and Biological Evaluation of Novel Triazolyl - Acridine Derivatives as Cytotoxic Agents

Acridine and Triazols both are biologically active heterocyclic rings with cytotoxic potential. Triazolyl- acridine adduct attract the attention in the field of medicinal chemistry. Here we synthesized a series of triazolyl- acridine compounds by the appropriate procedures. Structure of all synthesized compounds was confirmed by the various spectroscopic methods e.g. FT-IR, proton NMR and Mass spectrograms. All synthesized triazolyl-acirdines compounds were assayed in-vitro for cytotoxic activity against MCF-7 (human breast adenocarcinoma cell line) and HT-29 (human colon adenocarcinoma cell line) cells by MTT- assay. All target compounds shows increasing activity in dose dependent manner. However MPSP-9 was sensitive against MCF-7 but compound MPSP-1 was most sensitive with minimum inhibitory concentration (IC50 value) against MCF-7 and HT-29 both cell lines.

Rapid Identification of the Chemical Components of Ilex rotunda Thunb Using UPLC-Q-TOF-MS/MS

Ilicis Rotundae Cortex (IRC) consists of the bark of Ilex rotunda Thunb, and its chemical constituents mainly include flavonoid glycosides, phenols, and triterpenoid saponins. In this study, a preliminary analysis was performed to identify and obtain the chemical components from IRC to better control the quality of the medicinal materials and provide a chemical basis for the study of the efficacy of the active components. Simple and efficient sample pretreatment and ultrasonic-assisted extraction methods were used to analyze the mass spectrum fragments and fracture modes in the anion mode by UPLC-Q-TOF-MS/MS. Using a two-step strategy, the neutral loss, diagnostic ions, and characteristic fragments were studied to screen diverse skeletons and substitutions, and the possible compounds were identified by comparison with databases. The representative compounds were compared with the standard, and the mass spectrogram was found to match perfectly. Thus, our findings reveal that this method is feasible and reliable and can be used to analyze the chemical components of IRC. We identified 105 compounds, including 22 triterpenoid saponins, 15 chlorogenic acids, 33 phenylpropanoids and phenylpropanosides, 3 iridoids, 1 flavonoid, 10 lignans, 12 glycosides, and 9 other compounds. This method lays the foundation for further elucidating the pharmacodynamics of IRC and provides a practical method for the identification of IRC.

Isoeucommin A attenuates kidney injury in diabetic nephropathy through the Nrf2/HO-1 pathway.

Diabetic nephropathy (DN) is a common complication in patients with diabetes and a leading cause of mortality. The management of DN in the clinic still remains a challenge. Therefore, the identification of novel compounds for DN treatment and their characterization in preclinical DN models are crucial. Isoeucommin A is a lignan compound isolated from Eucommia ulmoides Oliv, which has not been studied in detail. Our aim was to investigate the effect of Isoeucommin A in DN and to elucidate the molecular mechanisms though which Isoeucommin A acts in vitro and in vivo. We first isolated and purified Isoeucommin A by microporous resin column chromatography and studied the mass spectrogram, as well as the structure of Isoeucommin A, by high‐resolution electrospray ionization mass spectroscopy and NMR, respectively. We further established an in vivo rat DN model and measured the changes of blood glucose, body weight, kidney index (KI), blood urea nitrogen, creatinine (CRE), glutathione, malondialdehyde (MDA), SOD, albumin (ALB) and urinary ALB to CRE ratios on treatment with Isoeucommin A. In addition, we measured SOD, MDA, glycogen synthase kinase‐3β (GSK‐3β), phosphorylated (p)‐GSK‐3β, nuclear factor erythroid‐derived 2‐related factor 2 (Nrf2) and heme oxygenase‐1 (HO‐1) levels by quantitative real‐time PCR and western blot, and estimated cell viability by a 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐tetrazolium bromide assay. After Isoeucommin A treatment, body weight, as well as SOD, glutathione, HO‐1 and Nrf2 expression levels, in DN rats increased in a dose‐dependent manner. In contrast, the levels of blood glucose, KI, blood urea nitrogen, CRE, urinary ALB to CRE ratio, tumor necrosis factor‐α, interleukin‐1β, interleukin‐6 and MDA decreased significantly. In addition, Isoeucommin A protected H2O2‐stimulated renal tubular epithelial cells from oxidative stress and activated the Nrf2/HO‐1 signaling pathway in high‐glucose‐stimulated human renal mesangial cells. In conclusion, Isoeucommin A could alleviate inflammation and oxidative stress in in vitro and in vivo DN models and thus attenuate kidney injury by activating the Nrf2/HO‐1 signaling pathway. Isoeucommin A could have the potential to be used as an effective drug for the treatment of DN.

Proteomic Imaging of Vestibular Schwannomas and Normal Nerves. Histopathologic Correlations.

Proteomic analysis of vestibular schwannoma (VS), non-vestibular schwannoma (NVS), and normal nerve (NN) using mass spectrometry and imaging of matrix assisted laser desorption ionization-time of flight (MALDI-TOF). Retrospective, qualitative, and descriptive study on VS, NVS, and NN. Samples were provided by our Tumor Bank. They were analyzed histologically then sprayed by acid matrix. The laser beam of MALDI performed desorption-ionization of the sample. A mass spectrogram (MS) was drawn depending on time of flight of ionized peptides, and MALDI-imaging was obtained which is a summation color spectrum depending on sample's peptide content. The slice was reexamined histologically and results compared with MALDI-imaging. Fifty schwannomas were sampled, of which 27 exploitable: 22 VS (17 Antoni type A and five type B) and five NVS (all Antoni type B). Eleven NN were analyzed. Among the 22 VS, near-total correlation between MALDI-imaging and pathology was found in two cases (9.1%), partial correlation in four (18.2%), and no correlation in 16 (72.7%); correlations were more frequent in VS of the Antoni type B. MS showed a peptide spike at 2,000 m/z in 7 (31.8%) and 5,000 m/z in 21 (95.5%). Among the five NVS, near-total correlation was found in three cases (60%), partial correlation in one (20%), and no correlation in one (20%). MS showed a peptide spike at 2,000 m/z in two (40%) and 5,000 m/z in all (100%). Among the 11 NN, near-total correlation was found in nine cases (81.8%), partial correlation in one (9.1%), and no correlation in one (9.1%). MS showed no peptide spike at 2,000 or 5,000 m/z. Behind homogeneous areas on histology, there was great heterogeneity on MALDI-imaging and MS, regarding VS and NVS, but not NN. There was a lack of correlation between MALDI-imaging and pathology in VS (except Antoni type B) as compared with NVS and NN. The lack of correlation in VS of the type A as compared with type B VS and NVS could be attributed to the overexpression of degeneration-associated proteins/peptides in VS of the type B as well as NVS that are better correlated with histologic findings. The two peptide spikes detected in schwannoma and not in NN opens up the prospect of tumor biomarkers identifiable by sequencing. The proteomic polymorphism found in VS and NVS was absent on histology which is a new morphologic characteristic of schwannoma. Further studies should be performed in the future to confirm the benefit and usefulness of the MALDI in the analysis of VS and NVS.

Peak Identification and Quantification by Proteomic Mass Spectrogram Decomposition.

Recent advances in the liquid chromatography/mass spectrometry (LC/MS) technology have improved the sensitivity, resolution, and speed of proteome analysis, resulting in increasing demand for more sophisticated algorithms to interpret complex mass spectrograms. Here, we propose a novel statistical method, proteomic mass spectrogram decomposition (ProtMSD), for joint identification and quantification of peptides and proteins. Given the proteomic mass spectrogram and the reference mass spectra of all possible peptide ions associated with proteins as a dictionary, ProtMSD estimates the chromatograms of those peptide ions under a group sparsity constraint without using the conventional careful preprocessing (e.g., thresholding and peak picking). We show that the method was significantly improved using protein-peptide hierarchical relationships, isotopic distribution profiles, reference retention times of peptide ions, and prelearned mass spectra of noise. We examined the concept of database search, library search, and match-between-runs. Our ProtMSD showed excellent agreements of 3277 peptide ions (94.79%) and 493 proteins (98.21%) with Mascot/Skyline for an Escherichia coli proteome sample and of 4460 peptide ions (103%) and 588 proteins (101%) with match-between-runs by MaxQuant for a yeast proteome sample. This is the first attempt to use a matrix decomposition technique as a tool for LC/MS-based proteome identification and quantification.

Composition and high performance liquid phase analysis of paphia undulate in Marine shellfish

Study of Marine shellfish ripple and the composition of the clams and HPLC analysis of the ripple, the clam, by high performance liquid technical analysis corrugated and the composition of the clam, the experimental results show that the high performance liquid chromatographic separation of small molecular peptides in the mass spectrogram corresponding molecular weight range of 120-896, mixed small peptide molecular weight below mainly concentrated in the 1000 u, meet the needs of the preparation of small molecular peptides.

SILIA-Based 4C Quantitative PTM Proteomics.

To absolutely and relatively quantitate the alteration of a posttranslationally modified (PTM) proteome in response to a specific internal or external signal, a 15N-stable isotope labeling in Arabidopsis (SILIA) protocol has been integrated into the 4C quantitative PTM proteomics, named as SILIA-based 4C quantitative PTM proteomics (S4Quap). The isotope metabolic labeling produces both forward (F) and reciprocal (R) mixings of either 14N/15N-coded tissues or the 14N/15N-coded total cellular proteins. Plant protein is isolated using a urea-based extraction buffer (UEB). The presence of 8M urea, 2% polyvinylpolypyrrolidone (PVPP), and 5mM ascorbic acid allows to instantly denature protein, remove the phenolic compounds, and curb the oxidation by free radicals once plant cells are broken. The total cellular proteins are routinely processed into peptides by trypsin. The PTM peptide yield of affinity enrichment and preparation is 0.1-0.2% in general. Ion exchange chromatographic fractionation prepares the PTM peptides for LC-MS/MS analysis. The collected mass spectrograms are subjected to a target-decoy sequence analysis using various search engines. The computational programs are subsequently applied to analyze the ratios of the extracted ion chromatogram (XIC) of the 14N/15N isotope-coded PTM peptide ions and to perform the statistical evaluation of the quantitation results. The Student t-test values of ratios of quantifiable 14N/15N-coded PTM peptides are normally corrected using a Benjamini-Hochberg (BH) multiple hypothesis test to select the significantly regulated PTM peptide groups (BH-FDR<5%). Consequently, the highly selected prospect candidate(s) of PTM proteins are confirmed and validated using biochemical, molecular, cellular, and transgenic plant analysis.

МАСС-СПЕКТРОМЕТРИЧЕСКИЕ ИССЛЕДОВАНИЯ ДИНАМИКИ СОСТАВА ВЫДЫХАЕМОГО ВОЗДУХА В ПРОЦЕССЕ ДИНАМИЧЕСКИХ ПОСТУРАЛЬНЫХ ВОЗДЕЙСТВИЙ

The paper studies the possibility of using a quadrupole gas mass spectrometer as part of a diagnostic complex based on a rotary (mechanorgical) table to assess the effectiveness of the dynamic postural effects on patients. Analysis of the obtained mass spectrograms indicates that the response of patients is individual and can vary significantly. Taking into account the mass spectrometric parameters of exhaled air allows the patient to be assigned to a specific group. Information on the concentration of oxygen and carbon dioxide in the patient's exhalation, as well as the analysis of the envelope shape of the measured dynamic mass spectrograms can be used for comprehensive diagnostics of the patient's condition in addition to the data of the main sensors in the complex.

Non-target screening method for VOCs analysis based on GC-QTOF

Volatile organic compounds (VOCs) are an important family of pollutants in the atmosphere, which will produce a series of ecological and environmental effects as well as human health effects. China has issued a series of documents to strengthen the control of VOCs. The monitoring and analysis of VOCs is the premise and foundation of its management and control. Although target analysis has the advantage of clear compound list, it also has the problem of pollutant missing because VOCs are a great variety in structure. To date, non-target screening has become a research hotspot in the field of environmental analysis. In this study, a non-target VOCs screening method was established based on qualitative screening of environmental air samples by GC-QTOF. To optimize mass spectrum signal, the effects of ion source temperature and electron ionization energy on molecular ion peaks were investigated. The results show that the ion source temperature in the range from 200 to 270°C has little influence on the abundance of molecular ion peaks. However, the response of molecular ion peaks for different VOCs varies significantly with the electron ionization energies from 15 to 70 eV, which results from the difference in its reaction ability with electrons because of diverse structure. According to the variation trend of molecular ion peaks, the tested VOCs can be divided into three categories: first, the molecular ion peaks show little change with the increase of electron ionization energy, such as benzene, chlorobenzene and phenol. Second, the molecular ion peak shows an upward trend with the increase of electron ionization energy, as dichloromethane, dibromodichloromethane, etc. Third, molecular ion peak decreases with the increase of electron ionization energy, and even molecular ion peak loss occurs such as phthalates, straight chain alkanes, and some haloalkanes under 70 eV. Therefore, ionization at 70 eV was convenient to match with the NIST mass spectrum library, and Ionization at 15 eV was helpful to obtain the molecular ion signal. Based on the boiling point, a prediction model of VOCs retention time was established. The final non-target screening process includes: (1) Firstly analyze VOCs in the sample under 70 eV, compare the obtained mass spectrogram with the spectrogram library of the instrument to screen out possible candidates; (2) the molecular ion peak was obtained under the condition of 15 eV, and the compound was identified according to the accuracy error of the candidate molecular ion 5 ppm as the mass threshold value; (3) isotopic pattern analysis if it involves; (4) further identification of compounds using the retention time prediction model; (5) standard samples can be used to confirm the identified substances finally. Preliminary testing of 108 standard substances as unknown VOCs in the non-target screening process demonstrated that 94 species could be correctly identified with 87% accuracy. The constructed non-target screening method was used to analyze VOCs in the environmental air samples collected from the workshop and boundary of one factory in Dalian, and 30 VOCs were screened out, among which 23 compounds were confirmed by standard samples. The sample analysis indicates that the non-target screening method constructed in this study can be used for the qualitative identification of VOCs in environment air samples.

Fate of deoxynivalenol and degradation products degraded by aqueous ozone in contaminated wheat

Deoxynivalenol (DON) is a secondary metabolite produced by Fusarium species in cereals that can be harmful to the health of humans and animals. In this work, a treatment system of aqueous ozone hybrid hydroxyl free radical (·OH) was establishedto degrade DON. An ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF) method was applied to detect and follow the fate of DON throughout the degradation process. The electrospray ionization mass spectrometry (ESI/MS) analysis confirmed the presence of DON and two degradation products (at 313.0215 and 325.0101 m/z) with the secondary mass spectrogram in the ESI positive mode. Based on the ESI/MS data, a multiple reaction monitoring (MRM) method was established to screen and monitor DON and the two degradation products in contaminated wheat. The MRM method was applied to analyze the degree of DON degradation in the samples. Overall, this work investigated the characteristics of DON and products in the ozone degradation process, and provided new insights into the application of an aqueous ozone system for the detoxification of mycotoxin-contaminated cereals.

Thermal maturity, source characteristics, and migration directions for the Ordovician oil in the Central Tabei Uplift, Tarim Basin: Insight from biomarker geochemistry

The central of Tabei Uplift, Tarim Basin in western China, including Rewapu, Yueman, and Fuyuan (RYF) blocks, can offer biomarker geochemistry of Ordovician oils and their valuable information for deep petroleum system. Biomarker analysis was performed on the Ordovician oils from the RYF using gas chromatography–mass spectrometry to constrain thermal maturity, oil source, and possible oil migration directions. The homogenization temperature of fluid inclusions and the burial history corrected by vitrinite reflectivity were used to reflect the oil charging periods. The oil samples were characterized by the complete occurrence of n-alkanes, dominated C20–C21–C23 terpanes, low abundance of homohopanes in the m/z 191 mass spectrogram, and a dominance of C29 among C27–29 regular steranes in the m/z 217 mass spectrogram. A weak and negligible biodegradation indicated by the biomarker signatures suggested favorable preservative conditions for the oils. Biomarker maturity parameters indicated that the oils were mainly in the late “oil window.” The oils were suggested to be obtained from reducing marine sedimentary environments and belong to the same crude oil family. The ratios related to the triaromatic steroids indicated that the Cambrian–Lower Ordovician and Middle–Upper Ordovician source rocks contributed to the Ordovician petroleum system, and the Middle–Upper Ordovician source rocks were likely the major contributors for the current oil samples. The filling tracers suggested that the main oil migration was from southwest to northeast along the Shunbei No. 1 fault rather than from the Manjiaer Depression in the southeast.

P081 The structure of the proteomic profile and gut microbiota in patients with primary immunodeficiency, common variable immunodeficiency and inflammatory bowel disease

Abstract Background Infringement of intestinal microbiota in patients with primary immunodeficiency (PID), common variable immunodeficiency (CVID) and ulcerative colitis (UC) continues to be studied. The aim of the research is a comprehensive study of the structure of gut microbiota and mucosal protein profile (MPP) in patients with PID, CVID and UC in different phases of the disease course. Methods The research included 15 patients with PID, 22 CVID and 46 UC (28 with relapse and 18 with persistent remission of the disease), control group—20 healthy volunteers. Estimation of gut microbiota was performed by bacteriological seeding, hydrogen breath test (HBT) with lactulose. Content of short-chain fatty acids (SCFAs) and microbial lipid markers (MLM) in faeces and colon mucosa was determined by gas–liquid chromatographic and gas chromatography–mass spectrometry (GC–MS). MPP was based on isoelectric focusing techniques (SDS–PAGE, 2DGE). Mass spectrograms were obtained using MALDI-TOF-MS/MS (Bruker, USA). Results HBT showed a 4.5-, 8- and 11-fold increase in hydrogen production on a 150th-minute study. SCFA showed a 6-, 9- and 11-fold decrease in propionic and butyric acids, mainly in patients with PID and UC: 0.2 + 0.1, 0.14 + 0.03 and 0.04 + 0.02 mg/g, respectively. Microbiological analysis showed a decrease in titers of E. coli, bifido- and lactobacilli on average 4.6 + 0.8 Lg. The conditionally pathogenic microflora was represented by E. coli, lactose-negative strains (n = 51,105/g) and haemolytic strains (n = 34,104/g, Proteus spp. (n = 48,105/g), Staphylococcus spp. (n = 29,105–6/g), Candida (n = 51,104–6/g) and Clostridium spp. (n = 38,104/g) in stool culture. MLN GS-MS results showed a 5.5-, 3- and 6-fold increase in total bacterial load in patients with PID, CVID and UC, respectively, which was represented by resident anaerobic microflora: Streptococcus mutants, Clostridium difficile, Candida albicans. Results of MPP (detection rate of 75% or more) of the colon mucosa in patients with PID were detected: 1, 2, 4 okkludin, kininogen 1, interleukin-1B, interleukin 8, B2-glycoprotein, heat shock protein 27, in patients with CVID—translational elongation factor, apolipoprotein C-III. In patients with UC—NF-kB, alipoprotein C-III, TNF-α, interleukin-2 and 8 were presented. Conclusion In patients with PID, CVID and UC markers of colon excess growth and significant decrease in SCFA are recorded during the relapse period. MPP in patients with PID, CVID and UC was characterised by proteins characteristic of inflammation, apoptosis, proliferation and proteins reflecting the activity of the autoimmune process. Patients with UC in the remission have recovery trend of gut microbiota, production of SCFA, disappearance of specific components of altered MPP.

P016 Efficiency of evaluating the expression of Toll-like receptors 2, 4, 6 and proteins proteomic profiling as integral indicators for predicting the risk of early relapse of ulcerative colitis

Abstract Background Success was made in the study of TLR in congenital and adaptive immunity, which determined a new look at immune processes at ulcerative colitis (UC). Modern achievements of proteomic methods of analysis research allow to define molecular characteristics of the inflammation in colon mucosa of patients with UC. Methods The study included 86 patients with UC, an average age of 39.0 ± 1.4 years. Groups: 1–15 (17.4%) patients with distal form of UC, 2–42 (48%) left-sided form, 3–29 (33.7%) patients with total UC. The expression of TLR on peripheral blood monocytes was determined in the immunofluorescence test. Two-colour analysis was performed on a flowthrough laser cytofluorimeter (Cytomics FC500, Beckman Coulte). The percentage of monocytes (CD14 + cells) carrying TLR2, TLR4, and TLR6 on their surface was assessed. The separation of proteins of colon mucosa was based on technologies of IEF, SDS–PAGE, 2DPAGE, by standard sets (MB-HIC C8 Kit, MB-IMAC Cu, MB-Wax Kit, Bruker, USA). The getting of mass spectrogram was determined by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF-MS/MS, Ultraflex II, Bruker, USA). Statistical analysis was performed using the software Statistica 10.0 (Statsoft). Results The direct average relationship was established between the number of monocytes expressing CD14 + CD282 +, CD14 + CD284 +, CD14 + CD286 + and the area of inflammation (r = 0.49, r = 0.55, r = 0.42, p &amp;lt; 0.05). The nonlinear regression equation was used. Calculation example: the risk of recurrence UC = exp (−26.1 + (0.4) × TLR 2)/(1 + exp (−26.1 + (0.4) × TLR 2)), χ2 = 130, 59, p &amp;lt; 0.0001. Thus, when the number of monocytes expressing TLR2 is not more than 60%, the risk of recurrence of the UC is not more than 11%, with values above 70%, the probability of recurrence exceeds 80%. We identified potentially new molecular markers of the early relapse of ulcerative colitis: SMAD2 activates the transcription of TFG1β and leads to development of fibrosis in colon submucosa in patients with UC; significant decrease of the expression of PPARγ promotes the activation of STAT and AP-1 signalling pathways that promotes the increase of the synthesis of IL-2, 6, 8, 12, TNFα, the activity of immune and inflammation processes in colon mucosa; the reduction of expression of β-defensin-1 in cells of colon mucosa is accompanied with increased expression of CCR6 that promotes the formation of inflammatory infiltrates in colonic submucosa in UC. Conclusion Expression of TLR 2, 4, 6 in blood monocytes (the risk of recurrence UC ratio) and new protein molecular markers of colon mucosa (SMAD2, PPARγ, and apoС-III) can be used as a tool for prediction of early relapse UC.

P078 Efficiency of evaluating the expression of Toll-like receptors 2, 4, 6 and proteins proteomic profiling as integral indicators for predicting the risk of early relapse of ulcerative colitis

Abstract Background Success was made in the study of TLR in congenital and adaptive immunity, which determined a new look at immune processes at ulcerative colitis (UC). Modern achievements of proteomic methods of analysis research allow to define molecular characteristics of the inflammation in colon mucosa of patients with UC. Methods The study included 86 patients with UC, an average age of 39.0 ± 1.4 years. Groups: 1–15 (17.4%) patients with distal form of UC, 2–42 (48%) left-sided form, 3–29 (33.7%) patients with total UC. The expression of TLR on peripheral blood monocytes was determined in the immunofluorescence test. Two-colour analysis was performed on a flow-through laser cytofluorimeter (Cytomics FC500, Beckman Coulte). The percentage of monocytes (CD14 + cells) carrying TLR2, TLR4, TLR6 on their surface was assessed. The separation of proteins of colon mucosa was based on technologies of IEF, SDS–PAGE, 2DPAGE (Bruker, USA). The getting of mass spectrogram was determined by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry (MALDI-TOF-MS/MS, Ultraflex II, Bruker, USA). Statistical analysis was performed using the software Statistica 10.0 (Statsoft). Results The direct average relationship was established between the number of monocytes expressing CD14 + CD282 +, CD14 + CD284 +, CD14 + CD286 + and the area of inflammation (r = 0.49, r = 0.55, r = 0.42, p &amp;lt; 0.05). The nonlinear regression equation was used. Calculation example: the risk of recurrence UC = exp (−26.1 + (0.4) × TLR 2)/(1 + exp (−26.1 + (0.4) × TLR 2)), χ2 = 130, 59, p &amp;lt; 0.0001. Thus, when the number of monocytes expressing TLR2 is not more than 60%, the risk of recurrence of the UC is not more than 11%, with values above 70%, the probability of recurrence exceeds 80%. We identified potentially new molecular markers of the early relapse of ulcerative colitis: SMAD2 activates the transcription of TFG1β and leads to development of fibrosis in colon submucosa in patients with UC; significant decrease of the expression of PPARγ promotes the activation of STAT and AP-1 signalling pathways that promotes the increase of the synthesis of IL-2,6,8,12,TNFα, the activity of immune and inflammation processes in colon mucosa; the reduction of expression of β-defensin-1 in cells of colon mucosa are accompanied with increased expression of CCR6, that promotes the formation of inflammatory infiltrates in colonic submucosa in UC. Conclusion Expression of TLR 2,4,6 in blood monocytes (the risk of recurrence UC ratio) and new protein molecular markers of colon mucosa (SMAD2, PPARγ and apoС-III) can be used as a tool for prediction of early relapse UC.