The aim of this study was to examine the hepatoprotective activity of multicomponent mixtures of natural origin in the BALB/C mouse model (n = 59), with acute direct toxic liver injury (DTLI) induced by the administration of streptozotocin (STZ) (100 mg/kg) in combination with a high-fat and high-fructose diet (HFFD). The hepatoprotective activity of activated hydrolytic lignin (Bp-Cx-1), methanolic fraction of Bp-Cx-1 (Bp-Cx-M) and isoflavones from kudzu Pueraria lobata roots (IFL) were evaluated on molecular level using mass spectrometry (MS)-based omics technologies. Untargeted label-free DIA quantitation resulted in 7214 protein groups identification (FDR 1%) after filtering across 40 liver tissue extracts. All treatment groups were closer to the control samples on the liver proteomic landscape compared to the untreated DTLI group, with the best results shown for the Bp-Cx-M and IFL groups. In order to identify differences between specific groups, we applied the post hoc Dunn's test and used Hedges' g as the effect size metric, revealing 64 proteins that tended to return to their normal level after treatment. In-depth proteomic liver tissue analysis enabled us not only to reveal the main pathways such as inflammation and oxidative stress, which are in a good agreement with DTLI and non-alcoholic liver disease pathophysiology, but also to evaluate hepatoprotective activity of multicomponent mixtures of natural origin containing polyphenols and mostly associated with protein metabolism (e.g., PSMD7, HCFC1) and deubiquitination pathways (e.g., UCHL3). It is worth noting that the Bp-Cx-1 isolated methanol fraction (Bp-Cx-M) demonstrated a pronounced increased hepatoprotective activity compared to the parent material due to the enrichment with active components such as polyphenols. Consistent with the proteomic findings of restored ubiquitin-proteasome function, assessment by comet assay revealed that treatments with Bp-Cx-M and IFL significantly reduced DNA damage by 50% compared to the untreated DTLI group. The developed MS-based multi-omics approach may be implemented for the robust and high-throughput screening method during assessment of new hepatoprotective agents of synthetic or natural origin.

The Corinth Rift in Greece is very active, with high rates of extension, sedimentation and environmental change. International Ocean Discovery Program (IODP) Expedition 381 drilled three sites sampling syn-rift sediments, complementing previous fault and stratigraphic interpretations and providing the longest high-resolution stratigraphic record for an early phase rift. Sedimentation in the Gulf of Corinth started-2.0-2.5 Ma as an isolated basin, with the most recent rift phase starting at-0.75 Ma, marking a shift to increasingly orbitallycontrolled marine incursions. A wide range of paleoenvironmental conditions have been generated, reflected in the diverse microfossil assemblages and sedimentary lithologies. Drilling results highlight a recent acceleration of strain rate and fault activity connected to rapid strain localization, with linkage of the border fault system over 10s-100s kyr timescales. Over long timescales (100s kyr), these variations in fault slip rate control sediment accumulation. On shorter time scales (10s kyr), changes in accumulation rate and type of sediment are primarily controlled by glacial-interglacial climate change, with accumulation rates in glacial periods at least double that of interglacial periods, accompanied by enhanced basin stratification and dominance of non-marine faunal assemblages. The mud-dominated sediments have three stratal package types (bioturbated, bedded, laminated) that record distinct hydrological conditions linked to climate and sea level which influence the landscape and basin conditions. Results from the Corinth Rift are compared with other active basins and rift systems for a better understanding of the tectonic and climatic processes shaping these environments.

Background: Schizophrenia (SZ) is associated with chronic oxidative stress in the patient’s body. Previous studies revealed an increased copy number of genes for 47S pre-ribosomal RNA (pre-rRNA) in SZ patients. In this study, levels of oxidative stress and factors involved in the adaptive response to chronic stress (rDNA transcription) were, for the first time, compared in blood cells of patients with catatonic SZ(C) and paranoid SZ(P), chronic forms of schizophrenia, as well as healthy controls (HC). Methods: Ribosomal DNA (rDNA) and telomere repeat (TR) were quantified in leukocyte DNA using non-radioactive quantitative hybridization. Fragments of 5′ external transcribed spacer (5′ ETS) and 18S rRNA were assayed in leukocyte RNA using quantitative reverse transcription PCR (RT-qPCR). Proteins γ-histone H2AX (γH2AX), NADPH-oxidase 4 (NOX4), nuclear factor erythroid 2-related factor 2 (NRF2), BCL2-like protein 4 (BAX), BCL2, and oxidation marker 8-oxo-2′-deoxyguanosine (8-oxodG) were quantified in blood lymphocytes using flow cytometry. Results: SZ(C) cells exhibited higher levels of the oxidative stress markers than SZ(P) and HC cells. The rDNA copy numbers in SZ(C) genomes negatively correlated with the amounts of the oxidative stress markers levels. Thus, genomes of blood cells isolated from catatonic patients harbor more copies of ribosomal genes than those from paranoid schizophrenia patients, correlating with higher levels of rRNA in catatonic patients. Conclusions: The upregulated ribosome biogenesis appears to be required for adaptive response to the elevated levels of oxidative stress in catatonic compared to paranoid patients.

The development of novel polymers for biomedical applications remains a highly demanded field of polymer science. In this study, the radical polymerization of N-vinylsuccinimide (VSI) using S,S-dibenzyl trithiocarbonate (DBTTC) as a reversible chain-transfer agent was investigated both in bulk and in 1,4-dioxane solution. The resulting polymers were characterized by SEC, 1H, 13 & Scy; and HSQC NMR spectroscopy to determine their molecular weight distribution, conversion, and microstructure. The polymerization followed a controlled mechanism, as evidenced by linear molecular weight increase with conversion, suppression of the gel effect, and living chain behavior. The position of the trithiocarbonate group within the polymer chain was elucidated via nucleophilic and reduction reactions, and radical initiator substitution. The trithiocarbonate group was found to shift from a central to asymmetric position depending on synthesis conditions and chain length. PVSI-DBTTC was subsequently employed as a macro-RAFT agent for the synthesis of triblock copolymers with O-cholesteryl acrylate (ChA). These copolymers were analyzed by SEC, 1H NMR spectroscopy and DSC. Hydrolysis of the VSI units to N-vinylsuccinamic acid (VSAA) units yielded the amphiphilic triblock copolymer - PVSAA-b-PChA-bPVSAA, which self-assembled into micelles in aqueous media. The resulting dispersions were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM), which revealed the formation of spherical nanoparticles with a hydrodynamic diameter of up to 200 nm and a polydispersity index (PDI) below 0.3 in phosphate-buffered saline solution (pH 7.4). The developed micelles successfully encapsulated irinotecan and exhibited pH-responsive release. Furthermore, the nanoparticles demonstrated low cytotoxicity in various cells and low uptake by macrophages, as well as preserved the potent anticancer activity of the loaded drug, making them as promising candidate for effective drug delivery.

Today, biocompatible and bioinspired well-defined copolymers with the ability to form nanoparticles are of great interest as potential drug delivery systems. In this study, we report the synthesis of novel biocompatible copolymers from monomers with different activity and capable of forming gradient copolymers by RAFT polymerization. In particular, the copolymerization of N-vinylsuccinimide (VSI) and O-cholesteryl (meth)acrylate (Ch (M)A) mediated by S,S'-dibenzyl trithiocarbonate (DBTTC) has been thoroughly studied by varying the monomer ratio and the ratio of monomer to RAFT agent. Important dependencies such as molecular weight and monomer conversion versus time, and molar fraction of monomer units as a function of monomer conversion were investigated. The obtained copolymers were thoroughly characterized using a number of physicochemical methods such as 1H NMR, 1H-13C HSQC and ATR-IR-spectroscopy, size-exclusion chromatography, static and dynamic light-scattering, as well as thermogravimetric analysis. In addition, the reactivity ratios of VSI and ChMA were determined and the dyad and triad compositions of the copolymers were calculated from the obtained values. The synthesized P(VSI-co-Ch(M)A) were subjected to selective hydrolysis of succinimide ring to convert it into succinamic acid. This approach yields a set of bioinspired amphiphilic copolymers based on Nvinylsuccinamic acid (VSAA) and Ch(M)A. The synthesized series of P(VSI-co-Ch(M)A) and P(VSAA-co-Ch(M)A) were used to obtain nanoparticles by nanoprecipitation or self-assembly via direct dissolution in aqueous medium. In addition, the method of surface hydrolysis of VSI units in pre-formed P(VSI-co-Ch(M)A) nanoparticles was applied to produce nanoparticles with hydrophilic negatively charged surface and enhanced stability. All techniques were optimized to prepare nanoparticles with characteristics suitable for systems considered for drug delivery. Successful loading of the antitumor drug irinotecan into nanoparticles was achieved with high encapsulation efficacies. The storage stability of empty and irinotecan loaded nanoparticles were studied in various media (water, saline solution, serum containing cell culture medium) under room and refrigerator conditions. The developed empty nanoparticles exhibited low rate of uptake by macrophages and low cytotoxicity to irinotecan-sensitive colon cancer cells (Caco-2). In turn, the irinotecan-loaded nanoparticles demonstrated inhibitory activity against Caco-2 cells comparable to the free drug.

The aim of this study was to examine the hepatoprotective activity of multicomponent mixtures of natural origin in the BALB/C mouse model (59 female mice), with non-alcoholic fatty liver disease (NAFLD) induced by the administration of streptozotocin (STZ) in combination with a high-fat, high-fructose diet. The hepatoprotective activity of activated hydrolytic lignin (BP-Cx-1), metanolic fraction of BP-Cx-1 (BP-Cx-M) and isoflavones from kudzu Pueraria lobata roots (IFL) was evaluated using mass spectrometry (MS)-based omics technologies. Untargeted label-free DIA quantitation resulted in 8088 protein groups identification (FDR 1%) in 40 liver tissue extracts. All treatment NAFLD groups were closer to the control samples on the liver proteomic landscape with the best results shown for BP-Cx-1-m and ISF groups. This corroborated well with metabolomic fingerprinting by FTICR MS. In order to identify differences between specific groups, we applied the post-hoc Dunn’s Test and used Hedges' g as the Effect Size metric and 64 proteins that tended to return to their normal level after treatment were revealed. Pathophysiologically specific proteomics changes occurring during NAFLD were mostly associated with metabolism of the proteins (PSMD7, HCFC1) and deubiquitination pathways (UCHL3). It is worth noting that BP-Cx-1 isolated methanol fraction (Bp-Cx-m) demonstrated the pronounced increased hepatoprotective activity due to the enrichment with active components such as polyphenols. Additionally, it was shown that treatments with Bp-Cx-M and IFL significantly reduced DNA damage by 50% compared to the NAFLD untreated group. Finally, MS-based multiomics approach demonstrated its power as an advanced screening method for the assessment of the hepatoprotective activity and molecular mechanisms of action of complex multicomponent mixtures of natural origin.

Early recognition of a risk of Alzheimer's disease (AD) remains a global challenge, and blood proteomic markers are of particular interest for wide-scale diagnostic use. Quantitative multiple reaction monitoring (MRM) approach demonstrates good reproducibility in the characteristic changes in the levels of reported candidate biomarkers (CBs) in different cohorts in AD. Following up on our previous study, we performed a joint analysis of 331 blood plasma samples from two different clinical cohorts of participants, comprising a total of 95 samples from patients with AD, 136 samples from patients with mild cognitive impairment (MCI), and 100 samples from controls. The obtained results confirm the significance of 37 CBs. A logistic regression-based algorithm was used to build protein classifiers, and a total of 21 important proteins were selected, 13 of which (ORM1, APOA4, LBP, HP, FN1, BCHE, APOE, PZP, A1BG, TF, SERPINA7, TTR, and F12) formed a universal panel that demonstrated strong classification performance in distinguishing AD patients from controls (ROC-AUC = 0.90) and in separating stable and progressing patients with MCI (ROC-AUC = 0.81). Overall, the analysis confirms the high potential of the MRM method for validating CBs in independent cohorts.

INTRODUCTION:Identifying early risks of developing Alzheimer's disease (AD) is a major challenge as the number of patients with AD steadily increases and requires innovative solutions. Current molecular diagnostic modalities, such as cerebrospinal fluid (CSF) testing and positron emission tomography (PET) imaging, exhibit limitations in their applicability for large-scale screening. In recent years, there has been a marked shift toward the development of blood plasma-based diagnostic tests, which offer a more accessible and clinically viable alternative for widespread use. Furthermore, advances in large-scale proteomics technologies have boosted an interest in identifying novel biomarkers and developing panels of AD-associated proteins. AREAS COVERED:This review mainly examines the results of recent searches for proteomic markers of AD in blood plasma (from 2022-2024 PubMed), focuses on some aspects for special attention in further studies, and discusses the prospects for their further application. EXPERT OPINION:Recent advances in AD plasma/serum proteomic studies are largely driven using novel Olink/PEA and SomaScan/aptamer technologies, which complement the 'gold standard' of MS-based quantitative proteomics (MRM/SRM), and particularly expand the capabilities for studying low-abundant proteins.

Structural changes of the brain in schizophrenia progress over time and their extent differs depending on the patient, yet their mechanisms remain unclear. Polymorphisms in the genes regulating immune response and inflammation have associations with the risk of schizophrenia. Studying their impact on the structural parameters of the brain in the patients is relevant for prediction of cognitive impairment. The goal of this work was to investigate the possible associations of polymorphisms of IL10 immonoregulatory gene with neuroanatomic changes in schizophrenia to gain new insights on the role of immunogenetic factors in the development of neurodegeneration in the patients. 89 patients and 99 healthy volunteers were enrolled. SNPs of IL10 gene (G-1082A and C-592A) were determined by polymerase chain reaction. MRI scans were performed on a Siemens Magnetom Verio 3T MRI scanner. It was found that the A allele for G-1082A SNP and CC genotype for C-592A SNP are more frequent in the patients than in the control group, and are significantly (p < 0.01) associated with decreased mean thickness in a number of areas of the frontal cortex in the patients. These findings are important for revealing the role of immunogenetic factors regulating IL-10 production in the pathogenesis of neurodegeneration in schizophrenia. Further research directed at finding and confirming new immunogenetic markers of structural brain changes in schizophrenia is important for identifying the various biological variants of the disease that can differ in pathogenesis, clinical presentation and therapeutic opportunities.

The study of urinary peptidome is an important area of research, which concerns the characterization of endogenous peptides, as well as the identification of biomarkers for a wide range of socially significant diseases. First of all, this relates to renal and genitourinary pathologies and/or pathologies associated with proteinuria, such as kidney diseases, bladder, prostate and ovarian cancers, diabetic nephropathy, and pre-eclampsia. Unlike proteins, peptides do not require proteolytic hydrolysis, can be analyzed in their native form and can provide certain information about occurring (patho)physiological processes. Mass spectrometry (MS)-based approaches are the most unbiased and sensitive instruments with high multiplexing capacity and provided most of the current information about endogenous urine peptides. However, despite the large number of urine peptidomic studies, there are certain issues related to the insufficient comparability of their results due to the lack of consistent approaches to their interpretation. Also the development of a custom project-specific protein library for endogenous peptides search and identification is another important point that should be noted in the context of high-throughput peptidomic analysis. Here we propose the custom-specific urinary protein database and the grouping of endogenous urinary peptides with overlapping sequences as useful tools, which can facilitate the acquisition and analysis of LC-MS peptidomic data, as well as the comparison of results of different studies, which should facilitate their more efficient further application.

Accumulation of neurotoxic aggregates of beta-amyloid peptides (Aβ) is a hallmark of Alzheimer's disease (AD) progression. Post-translational modifications (PTMs) increase Aβ aggregation and cytotoxicity, and the content of specific Aβ proteoforms is elevated in senile plaques of AD patients. The pathophysiological mechanisms of aggregate formation and the role of Aβ proteoforms need thorough study both to understand the role played by specific processes in the initiation of neuronal degradation and to find effective preventive means of therapeutic action. The present work investigates the dynamics of accumulation of phosphorylated serine-8 proteoform Aβ (pSer8-Aβ) using the 5xFAD mouse amyloid model. Aβ samples from human cerebrospinal fluid (CSF) and brain were also investigated. Western blot studies using 1E4E11 and 4G8 antibodies showed that accumulation of pSer8-Aβ in mouse brain starts as early as at the age of 3 months and reaches a maximum by the age of 14-17 months, which is generally similar to the dynamics of accumulation of the total pool of Aβ peptides. The pSer8-Aβ level in human CSF in AD patients can reach ~ 1-10% of the total amount of Aβ. Mass spectrometric analysis showed that Aβ phosphorylation by the Ser8, Tyr10, and Ser26 residues in brain tissues, as well as phosphorylation of the APP by Thr719 residue, is possible. These findings support the assumption that pSer8-Aβ proteoforms are involved in amyloidosis in AD. KEYWORDS Beta-amyloid, mass spectrometry, Alzheimer's disease, phosphorylation.

As a young (<5 Myr old) active rift with high resolution spatial and temporal constraints, the Corinth Rift is a natural laboratory for testing models of rift and fault network development in the early stages of continental rifting. New analyses of the rift fault network in the offshore syn-rift sequence are combined with ocean drilling borehole data from IODP Expedition 381. The expedition drilled and sampled syn-rift sediments from the last few Myr and provides the first absolute age framework for the offshore rift, allowing determination of robust fault slip rates and temporal patterns in fault network activity. Spatial variations in activity and rates throughout the rift fault network, for four time intervals over the past similar to 2 Myr, illustrate changes in strain distribution and highlight three dominant processes controlling the development of the fault network: 1) progressive strain localisation and transfer of strain from major S-dipping to major N-dipping faults from similar to 2 Ma - 130 ka; 2) linkage of a southern border fault system and subsequent acceleration of fault slip rates on major N-dipping faults at similar to 335 ka; 3) increased rift margin flexure and subsequent deformation since similar to 130 ka, a response to rapid subsidence in the hanging wall of an established crustal scale border fault system. Since similar to 130 ka the rift fault network has experienced a two-fold increase in average cumulative slip rates, with the highest slip rates (>7 mm/yr) occurring on major segments of the border fault system in the central rift. A comparison of seismic moment rates from historical earthquakes (last 320 years) is consistent with the geological timescale of fault slip rates (highest rates in the western and central rift), but not with the distribution of very recent activity (from 50-year earthquake records). As a result, a moment deficit is present along the central rift, which could be accommodated by a large (Mw 6.5) earthquake, potentially even rupturing multiple linked fault segments. The details of rift fault network activity from this study reveal how quickly strain can migrate and become localised during early continental rifting, and how rapidly fault slip accelerates in response to the establishment of major rift border fault systems. Identifying the nature and timescales of these important rift processes furthers our models of early rift evolution and has implications for assessing seismic hazard in regions of active continental rifting.

Objectives The COVID-19 pandemic has exposed a number of key challenges that need to be urgently addressed. Mass spectrometric studies of blood plasma proteomics provide a deep understanding of the relationship between the severe course of infection and activation of specific pathophysiological pathways. Analysis of plasma proteins in whole blood may also be relevant for the pandemic as it requires minimal sample preparation.Methods The frozen whole blood samples were used to analyze 203 plasma proteins using multiple reaction monitoring (MRM) mass spectrometry and stable isotope-labeled peptide standards (SIS). A total of 131 samples (FRCC, Russia) from patients with mild (n=41), moderate (n=39) and severe (n=19) COVID-19 infection and healthy controls (n=32) were analyzed.Results Levels of 94 proteins were quantified and compared. Significant differences between all of the groups were revealed for 44 proteins. Changes in the levels of 61 reproducible COVID-19 markers (SERPINA3, SERPING1, ORM1, HRG, LBP, APOA1, AHSG, AFM, ITIH2, etc.) were consistent with studies performed with serum/plasma samples. The best-performing classifier built with 10 proteins achieved the best combination of ROC-AUC (0.97-0.98) and accuracy (0.90-0.93) metrics and distinguished patients from controls, as well as patients by severity.Conclusions Here, for the first time, frozen whole blood samples were used for proteomic analysis and assessment of the status of patients with COVID-19. The results obtained with frozen whole blood samples are consistent with those from plasma and serum.

Background/Objectives: The development of blood tests for the early detection of individual predisposition to socially significant diseases remains a pressing issue. Methods: In this pilot study, multiple reaction monitoring mass spectrometry (MRM-MS) with a BAK-270 assay was applied for protein concentrations analysis in blood plasma from 21 healthy volunteers of the European cohort. Results: The levels of 138 plasma proteins were reliably and precisely quantified in no less than 50% of samples. The quantified proteins included 66 FDA-approved markers of cardiovascular diseases (CVD), and other potential biomarkers of pathologies such as cancer, diabetes mellitus, and Alzheimer’s disease. The analysis of individual variations of the plasma proteins revealed significant differences between the male (11) and female (10) groups. In total, fifteen proteins had a significantly different concentration in plasma; this included four proteins that exhibited changes greater than ±1.5-fold, three proteins (RBP4, APCS, and TTR) with higher levels in males, and one (SHBG) elevated in females. The obtained results demonstrated considerable agreement with the data collected from 20 samples of a North American cohort, which were analyzed with the similar MRM assay. The most significant differences between the cohorts of the two continents were observed in the level of 42 plasma proteins (including 24 FDA markers), of which 17 proteins showed a ≥1.5-fold change, and included proteins increased in North Americans (APOB, CRTAC1, C1QB, C1QC, C9, CRP, HP, IGHG1, IGKV4-1, SERPING1, RBP4, and AZGP1), as well as those elevated in Europeans (APOF, CD5L, HBG2, SELPLG, and TNA). Conclusions: The results suggest a different contribution of specific (patho)physiological pathways (e.g., immune system and blood coagulation) to the development of socially significant diseases in Europeans and North Americans, and they should be taken into account when refining diagnostic panels.

The development of effective anti-cancer therapeutics remains one of the current pharmaceutical challenges. The joint delivery of chemotherapeutic agents and biopharmaceuticals is a cutting-edge approach to creating therapeutic agents of enhanced efficacy. In this study, amphiphilic polypeptide delivery systems capable of loading both hydrophobic drug and small interfering RNA (siRNA) were developed. The synthesis of amphiphilic polypeptides included two steps: (i) synthesis of poly-αl-lysine by ring-opening polymerization and (ii) its post-polymerization modification with hydrophobic l-amino acid and l-arginine/l-histidine. The obtained polymers were used for the preparation of single and dual delivery systems of PTX and short double-stranded nucleic acid. The obtained double component systems were quite compact and had a hydrodynamic diameter in the range of 90–200 nm depending on the polypeptide. The release of PTX from the formulations was studied, and the release profiles were approximated using a number of mathematical dissolution models to establish the most probable release mechanism. A determination of the cytotoxicity in normal (HEK 293T) and cancer (HeLa and A549) cells revealed the higher toxicity of the polypeptide particles to cancer cells. The separate evaluation of the biological activity of PTX and anti-GFP siRNA formulations testified the inhibitory efficiency of PTX formulations based on all polypeptides (IC50 4.5–6.2 ng/mL), while gene silencing was effective only for the Tyr-Arg-containing polypeptide (56–70% GFP knockdown).

Despite of multiple systematic studies of schizophrenia based on proteomics, metabolomics, and genome-wide significant loci, reconstruction of underlying mechanism is still a challenging task. Combination of the advanced data for quantitative proteomics, metabolomics, and genome-wide association study (GWAS) can enhance the current fundamental knowledge about molecular pathogenesis of schizophrenia. In this study, we utilized quantitative proteomic and metabolomic assay, and high throughput genotyping for the GWAS study. We identified 20 differently expressed proteins that were validated on an independent cohort of patients with schizophrenia, including ALS, A1AG1, PEDF, VTDB, CERU, APOB, APOH, FASN, GPX3, etc. and almost half of them are new for schizophrenia. The metabolomic survey revealed 18 group-specific compounds, most of which were the part of transformation of tyrosine and steroids with the prevalence to androgens (androsterone sulfate, thyroliberin, thyroxine, dihydrotestosterone, androstenedione, cholesterol sulfate, metanephrine, dopaquinone, etc.). The GWAS assay mostly failed to reveal significantly associated loci therefore 52 loci with the smoothened p < 10−5 were fractionally integrated into proteome-metabolome data. We integrated three omics layers and powered them by the quantitative analysis to propose a map of molecular events associated with schizophrenia psychopathology. The resulting interplay between different molecular layers emphasizes a strict implication of lipids transport, oxidative stress, imbalance in steroidogenesis and associated impartments of thyroid hormones as key interconnected nodes essential for understanding of how the regulation of distinct metabolic axis is achieved and what happens in the conditioned proteome and metabolome to produce a schizophrenia-specific pattern.

Glomerulopathies with nephrotic syndrome that are resistant to therapy often progress to end-stage chronic kidney disease (CKD) and require timely and accurate diagnosis. Targeted quantitative urine proteome analysis by mass spectrometry (MS) with multiple-reaction monitoring (MRM) is a promising tool for early CKD diagnostics that could replace the invasive biopsy procedure. However, there are few studies regarding the development of highly multiplexed MRM assays for urine proteome analysis, and the two MRM assays for urine proteomics described so far demonstrate very low consistency. Thus, the further development of targeted urine proteome assays for CKD is actual task. Herein, a BAK270 MRM assay previously validated for blood plasma protein analysis was adapted for urine-targeted proteomics. Because proteinuria associated with renal impairment is usually associated with an increased diversity of plasma proteins being present in urine, the use of this panel was appropriate. Another advantage of the BAK270 MRM assay is that it includes 35 potential CKD markers described previously. Targeted LC-MRM MS analysis was performed for 69 urine samples from 46 CKD patients and 23 healthy controls, revealing 138 proteins that were found in ≥2/3 of the samples from at least one of the groups. The results obtained confirm 31 previously proposed CKD markers. Combination of MRM analysis with machine learning for data processing was performed. As a result, a highly accurate classifier was developed (AUC = 0.99) that enables distinguishing between mild and severe glomerulopathies based on the assessment of only three urine proteins (GPX3, PLMN, and A1AT or SHBG).

Aim . To study the effect of inhalation therapy with an active hydrogen (AH) on the protein composition of exhaled breath condensate (EBC) in patients with post-COVID syndrome (PCS). Material and methods . This randomized controlled parallel prospective study included 60 patients after coronavirus disease 2019 (COVID-19) with PCS during the recovery period and clinical manifestations of chronic fatigue syndrome who received standard therapy according to the protocol for managing patients with chronic fatigue syndrome (CFS). The patients were divided into 2 groups: group 1 (main) — 30 people who received standard therapy and AH inhalations (SUISONIA, Japan) for 10 days, and group 2 (control) — 30 medical workers who received only standard therapy. Patients in both groups were comparable in sex and mean age. All participants in the study were sampled with EBC on days 1 and 10. Samples were subjected to tryptic digestion and high-performance liquid chromatography combined with tandem mass spectrometry analysis using a nanoflow chromatograph (Dionex 3000) in tandem with a high-resolution time-of-flight mass spectrometer (timsTOF Pro). Results . A total of 478 proteins and 1350 peptides were identified using high resolution mass spectrometry. The number of proteins in samples after AH therapy, on average, is 12% more than before treatment. An analysis of the distribution of proteins in different groups of patients showed that only half of these proteins (112) are common for all groups of samples and are detected in EBC before, after, and regardless of hydrogen therapy. In addition to the qualitative difference in the EBC protein compositions in different groups, quantitative changes in the concentration of 36 proteins (mainly structural and protective) were also revealed, which together made it possible to reliably distinguish between subgroups before and after treatment. It is worth noting that among these proteins there are participants of blood coagulation (а-1-antitrypsin), chemokine- and cytokine-mediated inflammation, and a number of signaling pathways (cytoplasmic actin 2), response to oxidative stress (thioredoxin), glycolysis (glyceraldehyde-3- phosphate dehydrogenase), etc. Conclusion . The use of hydrogen therapy can contribute to the switching of a number of physiological processes, which may affect the success of recovery in PCS patients. In particular, the obtained results indicate the activation of aerobic synthesis of adenosine triphosphate in mitochondria by hydrogen therapy, which correlates well with the decrease in the blood lactate level detected by laboratory studies. At the same time, this therapy can inhibit pro-inflammatory activity, negatively affecting the coagulation and signaling pathways of integrins and apoptosis, and, in addition, activate protective pathways, tricarboxylic acid cycle, FAS signaling, and purine metabolism, which may be essential for effective recovery after COVID-19.

Schizophrenia is a multifactorial and clinically polymorphic disease with a significant hereditary component. The study of the influence of functionally significant polymorphisms of the genes regulating immune response and inflammation on the immune parameters in patients with schizophrenia is relevant for the development of new methods for predicting the course of the disease and finding new therapy targets. The aim of the study:To study the association of polymorphisms in cytokine and Toll-like receptor genes with the serum levels of immune mediators in patients with schizophrenia. Materials and methods: 95 patients diagnosed with schizophrenia and 99 healthy volunteers were recruited into the study. Polymorphisms of genes regulating the immune response (IL1B rs1143627, IL4 rs2243250, IL2 rs2069762, IL10 (rs1800896, rs1800872, rs1800871), IFNL rs8099917, IFNL rs12979860, TLR9 rs5743836, TLR9 rs352140, TLR3 rs3775291), the serum level of major cytokines and markers of systemic inflammation were determined. Results: The patients were found to have elevated levels of key pro-inflammatory cytokines IL-8 and IL-17A and other markers of systemic inflammation, as well as an increase in the content of antigen-antibody complexes in the bloodstream. Increased frequency of IL10 gene polymorphisms (rs1800896, rs1800872, rs1800871) was revealed in the patients with schizophrenia. A relationship between IL-6 and IL-8 levels and the carriage of IL1B T-511C (rs16944), IL10 1082A (rs1800896), IL10 592 C>A (rs1800872) and TLR3 Leu412Phe (G/A) (rs3775291) single nucleotide polymorphisms was found. Conclusion: The revealed associations may indicate a role of SNPs in cytokine and Toll-like receptor genes in modulating the network genetic interactions underlying certain immunological endophenotypes in the patients. Based on the results of this work and literature data, it is important to further study the relationship of immunogenetic markers in schizophrenia with the immune profiles of the disease and its clinical manifestations, including in larger samples of patients.