Proteomic profiling of renal tissue of normo- and hypertensive rats with the renalase peptide RP220 as an affinity ligand.

Renalase (RNLS) is a recently discovered protein that plays an important role in the regulation of blood pressure by acting inside and outside cells. Intracellular RNLS is a FAD-dependent oxidoreductase that oxidizes isomeric forms of ß-NAD(P)H. Extracellular renalase lacking its N-terminal peptide and cofactor FAD exerts various protective effects via non-catalytic mechanisms. Certain experimental evidence exists in the literature that the RP220 peptide (a 20-mer peptide corresponding to the amino acid sequence RNLS 220-239) reproduces a number of non-catalytic effects of this protein, acting on receptor proteins of the plasma membrane. The possibility of interaction of this peptide with intracellular proteins has not been studied. Taking into consideration the known role of RNLS as a possible antihypertensive factor, the aim of this study was to perform proteomic profiling of the kidneys of normotensive and hypertensive rats using RP220 as an affinity ligand. Proteomic (semi-quantitative) identification revealed changes in the relative content of about 200 individual proteins in the kidneys of hypertensive rats bound to the affinity sorbent as compared to the kidneys of normotensive animals. Increased binding of SHR renal proteins to RP220 over the normotensive control was found for proteins involved in the development of cardiovascular pathology. Decreased binding of the kidney proteins from hypertensive animals to RP220 was noted for components of the ubiquitin-proteasome system, ribosomes, and cytoskeleton.

Comparative proteomic analysis of renal tissue of normotensive and hypertensive rats.

Comparative proteomic analysis of kidney tissue from normotensive (WKY) and spontaneously hypertensive (SHR) rats revealed quantitative and qualitative changes in renal proteins. The number of renal proteins specific for WKY rats (blood pressure 110-120 mm Hg) was 13-16. There were 20-24 renal proteins specific for SHR (blood pressure 180 mm Hg and more). The total number of identified renal proteins common for both rat strains included 972-975 proteins. A pairwise comparison of all possible (SHR-WKY) variants identified 8 proteins specific only for normotensive (WKY) animals, and 7 proteins specific only for hypertensive ones (SHR). Taking into consideration their biological roles, the lack of some enzyme proteins in hypertensive rats (for example, biliverdin reductase A) reduces the production of molecules exhibiting antihypertensive properties, while the appearance of others (e.g. betaine-homocysteine S-methyltransferase 2, septin 2, etc.) can be interpreted as a compensatory reaction. Renal proteins with altered relative content (with more than 2.5-fold change) accounted for no more than 5% of all identified proteins. Among the proteins with an increased relative content in hypertensive animals, the largest group consisted of proteins involved in the processes of energy generation and carbohydrate metabolism, as well as antioxidant and protective proteins. In the context of the development of hypertension, the identified relative changes can apparently be considered compensatory. Among the proteins with the most pronounced decrease in the relative content in hypertensive rats, the dramatic reduction in acyl-CoA medium-chain synthetase-3 (ACSM3) appears to make an important contribution to the development of renal pathology in these animals.

The delayed effect of rotenone on the relative content of brain isatin-binding proteins of rats with experimental parkinsonism.

Isatin (indoldione-2,3) is an endogenous biological regulator found in the brain, peripheral tissues, and biological fluids of humans and animals. Its biological activity is realized via isatin-binding proteins, many of which were identified during proteomic profiling of the brain of mice and rats. A number of these proteins are related to the development of neurodegenerative diseases. Previously, using a model of experimental Parkinsonism induced by a seven-day course of rotenone injections, we have observed behavioral disturbances, as well as changes in the profile and relative content of brain isatin-binding proteins. In this study, we have investigated behavioral responses and the relative content of brain isatin-binding proteins in rats with rotenone-induced Parkinsonism 5 days after the last administration of this neurotoxin. Despite the elimination of rotenone, animals exhibited motor and coordination impairments. Proteomic profiling of isatin-binding proteins revealed changes in the relative content of 120 proteins (the relative content of 83 proteins increased and that of 37 proteins decreased). Comparison of isatin-binding proteins characterized by the changes in the relative content observed in the brain right after the last injection of rotenone (n=16) and 5 days later (n=11) revealed only two common proteins (glyceraldehyde-3-phosphate dehydrogenase and subunit B of V-type proton ATPase). However, most of these proteins are associated with neurodegeneration, including Parkinson's and Alzheimer's diseases.

The search for potential hypotensive peptides in the amino acid sequence of human renalase and their identification in proteolytic fragments of this protein.

Renalase (RNLS) is a secretory protein discovered in 2005. It plays an important role in the regulation of blood pressure. Studies by two independent laboratories have shown that administration of purified recombinant RNLS reduced blood pressure in experimental animals. However, the mechanisms of the antihypertensive effect of RNLS still remain unclear, especially in the context of the shift in the catalytic paradigm of this protein. In addition, there is growing evidence that endogenous plasma/serum RNLS, detected by enzyme immunoassay, is not an intact protein secreted into the extracellular space, and exogenous recombinant RNLS is effectively cleaved during short-term incubation with human plasma samples. This suggests that the antihypertensive effect of RNLS may be due to peptides formed during proteolytic processing. Based on the results of a bioinformatics analysis of potential RNLS cleavage sites (Fedchenko et al., Medical Hypotheses, 2022; DOI: 10.1016/j.mehy.2022.110895), a number of short peptides have been identified in the RNLS sequence that show similarity to fragments of known peptide inhibitors of angiotensin-converting enzyme. Some of them were found as a part of larger RNLS peptides, formed during RNLS cleavage by chymotrypsin and, and to a lesser extent, by trypsin.

Neuroprotective effects of isatin and afobazole in rats with rotenone-induced Parkinsonism are accompanied by increased brain levels of Triton X-100 soluble alpha-synuclein.

Effects of the endogenous neuroprotector isatin and the pharmacological drug afobazole (exhibiting neuroprotective properties) on behavioral reactions and quantitative changes in the brain proteomic profile have been investigated in rats with experimental rotenone Parkinsonism. A single dose of isatin (100 mg/kg subcutaneously on the last day of a 7-day course of rotenone administration) improved the motor activity of rats with rotenone-induced Parkinsonism in the open field test (horizontal movements) and the rotating rod test. Afobazole (10 mg/kg intraperitoneally, daily during the 7-day course of rotenone administration) reduced the manifestations of rigidity and postural instability. Proteomic analysis, performed using brain samples obtained the day after the last administration of rotenone and neuroprotectors, revealed similar quantitative changes in the brain of rats with rotenone Parkinsonism. An increase in the relative content of 65 proteins and a decrease in the relative content of 21 proteins were detected. The most pronounced changes - an almost ninety-fold increase in the alpha-synuclein content - were found in the brains of rats treated with isatin. In animals of the experimental groups treated with "Rotenone + Isatin", as well as "Rotenone + Afobazole", the increase in the relative content of this protein in the brain was almost 60 and 50 times higher than the control values. Taking into consideration the known data on the physiological role of alpha-synuclein, an increase in the content of this protein in the brain upon administration of neuroprotectors to animals with rotenone Parkinsonism may represent a compensatory reaction, at least in the early stages of this disease and the beginning of its treatment.

The effect of renalase-derived peptides on viability of HepG2 and PC3 cells.

Renalase (RNLS) is a recently discovered protein, which plays different roles inside and outside cells. Intracellular RNLS is a FAD-dependent oxidoreductase (EC 1.6.3.5), while extracellular RNLS lacks its N-terminal peptide, FAD cofactor, and exhibits various protective effects in a non-catalytic manner. Certain evidence exists, that plasma/serum RNLS is not an intact protein secreted into the extracellular space, and exogenous recombinant RNLS is effectively degraded during short-term incubation with human plasma samples. Some synthetic analogues of the RNLS sequence (e.g. the Desir's peptide RP-220, a 20-mer peptide corresponding to the RNLS sequence 220-239) have effects on cell survival. This suggests that RNLS-derived peptides, formed during proteolytic processing, may have own biological activity. Based on results of a recent bioinformatics analysis of potential cleavage sites of RNLS (Fedchenko et al., Medical Hypotheses, 2022) we have investigated the effect of four RNLS-derived peptides as well as RP-220 and its fragment (RP-224) on the viability of two cancer cell lines: HepG2 (human hepatoma) and PC3 (prostate cancer). Two RNLS-derived peptides (RP-207 and RP-220) decreased the viability of HepG2 cells in a concentration dependent manner. The most pronounced and statistically significant effect (30-40% inhibition of cell growth) was observed at 50 µM concentration of each peptide. In the experiments with PC3 cells five of six RNLS-derived peptides had a significant impact on the cell viability. RP-220 and RP-224 decreased cell viability; however, no concentration dependence of this effect was observed in the range of concentrations studied (1-50 µM). Three other RNLS-derived peptides (RP-207, RP-233, and RP-265) increased viability of PC3 cells by 20-30%, but no concentration-dependence of this effect was found. Data obtained suggest that some RNLS-derived peptides may influence the viability of various cells and manifestation and direction of the effect (increase of decrease of the cell viability) is cell-type-specific.

[Serum creatine phosphokinase as a predictor of upper limb amputation in electrical trauma]. / Soderzhanie kreatinfosfokinazy v plazme krovi kak prediktor amputatsii verkhnei konechnosti pri elektrotravme.

OBJECTIVE: To study the relationship between serum creatine phosphokinase and outcomes of injury in victims with electrical burns. MATERIAL AND METHODS: Among 40 patients with electrical injury, 7 (18%) ones underwent upper limb amputation. There were 37 (92.5%) men and 3 (7.5%) women aged 37 (28; 47) years. We analyzed total serum creatine phosphokinase and MB fraction on the first day in patients with and without amputations. RESULTS: Total serum creatine phosphokinase exceeded the upper reference value in 11 out of 33 patients without amputation and in all 7 patients with limb amputation (p=0.001). Patients with limb amputation had significantly higher total serum creatine phosphokinase and MB fraction (p<0.001 and p=0.030, respectively). Logistic regression equation showed that high total serum creatine phosphokinase significantly influenced amputation rate (p<0.001), as evidenced by odds ratio (42.7, 95% CI 3.5-514.8). ROC analysis revealed the cut-off value of total serum creatine phosphokinase (950 IU/L). Sensitivity was 100% (63; 100), specificity - 94% (86; 94), positive predictive value - 78% (49; 78), negative predictive value - 100% (92; 100). CONCLUSION: Total serum creatine phosphokinase depends only on severity of electrical and flame burns. Serum creatine phosphokinase is a predictor of upper limb amputation in patients with electrical injury. Total serum creatine phosphokinase ≥ 950 IU/L is significant for upper limb amputation (in CK-MB fraction within the reference values).

[Characteristics of behavioral reactions and the profile of brain isatin-binding proteins of rats with the rotenone-induced experimental parkinsonism]. / Osobennosti povedencheskikh reaktsii i profilia izatin-sviazyvaiushchikh belkov mozga u krys s indutsirovannym rotenonom éksperimental'nym parkinsonizmom.

The neurotoxins rotenone and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (МPTP) are used for modeling Parkinson's disease in animals (PD). They induce the mitochondrial respiratory chain dysfunction, which leads to the dopaminergic (DA) neuron degeneration. The advantage of the rotenone model consists in ability of rotenone to cause neurodegeneration showing symptoms and molecular biological characteristics similar to those of PD. Isatin (indoldione-2,3) is an endogenous regulator found in tissues and biological fluids of humans and animals. It exhibits a broad range of biological activity mediated by numerous isatin-binding proteins. In this work we have investigated behavioral reactions and profiles of brain isatin-binding proteins of rats with Parkinson's syndrome (PS) in comparison with the corresponding parameters of MPTP-induced Parkinsonism in mice. Systemic injection of rotenone caused severe PS comparable with the effect of MPTP injection. It was accompanied by significant body weight loss, death, oligokinesia, muscular rigidity, and postural instability of animals. In spite of the same pathogenic basis of PS caused by rotenone and MPTP, the molecular mechanisms of their action differ. In the case of rotenone-induced PS, the pool of isatin-binding proteins common of the control rats and the rats with PS (146) significantly exceeded the pool of the common proteins of control mice and mice with PS induced by MPTP, whether right after neurotoxin injection (27), or (all the more) in a week after the MPTP injection (14). The comparison of isatin-binding proteins specific of the animals with MPTP-induced PS and with the rotenone-induced PS (as compared with the control animals) revealed total absence of proteins common of these two models of PD. It is to be noted that both neurotoxins particularly affected the proteins participating in the signal transmission and enzyme activity regulation. The changes of the profile of isatin-binding proteins in response to the injection of rotenone suggest that the neuroprotector isatin could also influence positively in the case of the rotenone model of PD.

Dynamics of Metabolic and Oxidative Parameters of Erythrocytes during Treatment of Chronic Heart Failure with Molecular Hydrogen.

We studied the effect of molecular hydrogen (H2) on the content of 2,3-diphosphoglyceric acid (2,3-DPG), ATP, malondialdehyde, and catalase activity in erythrocytes in chronic heart failure. Inhalation of 2% molecular hydrogen H2 was carried out for 40 min repeatedly (5 days) or once. Inhalation of H2 caused an increase in ATP concentration in both research groups, but was more pronounced after repeated inhalation. The content of 2,3-DPG increased after repeated exposure to H2. The increase in metabolic activity under the effect of H2 was accompanied by a decrease in malondialdehyde concentration and an increase in catalase activity. Thus, the application of H2 in chronic heart failure reduced oxidative stress and improved metabolism of erythrocytes, which contributes to improvement of microcirculation. This allows us to recommend H2 for protection against ischemic and reperfusion damage to the myocardium.

[The key role of the regulatory 19S subunit in changes in the brain proteasome subproteome induced by the neuroprotector isatin]. / Kliuchevaia rol' reguliatornoi 19S subchastitsy v izmenenii subproteoma proteasom mozga, indutsirovannom neiroprotektorom izatinom.

Isatin (indole-2,3-dione) is an endogenous regulator exhibiting various effects mediated by numerous isatin-binding proteins localized in different compartments of cells of the brain and peripheral tissues. It attenuates manifestations of experimental parkinsonism induced by administration of the MPTP neurotoxin and reduces the movement disorders characteristic of this disease. The molecular mechanisms of the neuroprotective action of isatin include its direct interaction with proteasomes, intracellular supramolecular complexes responsible for the targeted elimination of proteins. Incubation of fractions of 26S and 20S rabbit brain proteasomes, containing the whole spectrum of proteasomal subunits, as well as a number of proteasome-associated proteins, with isatin (100 µM) had a significant impact on the profile of released proteins. In the case of 26S proteasomes containing, in addition to the core part (20S proteasome), 19S regulatory subparticles, incubation with isatin resulted in a more than threefold increase in the number of dissociated proteins. In the case of 20S proteasomes (containing only the 20S core particle), incubation with isatin resulted in a significant decrease in the number of dissociated proteins compared to the control. Our results indicate an important role of the regulatory 19S subunit components in the formation of the proteasome subproteome and the sensitivity of these supramolecular complexes to isatin.

[Comparative analysis of proteins associated with 26S and 20S proteasomes isolated from rabbit brain and liver]. / Sravnitel'nyi analiz belkov, assotsiirovannykh s 26S i 20S proteasomami mozga i pecheni krolika.

We have isolated fractions of 26S and 20S proteasomes were from the rabbit liver and the brain. According to mass spectrometric (MS) analysis, the 26S proteasome fractions from these organs contained catalytic and regulatory subunits characteristic of the proteasome core and regulatory subunits. The 20S fractions of brain and liver proteasomes contained only catalytic proteasome subunits. In addition to proteasome subunits, the isolated fractions contained components of the ubiquitin-proteasome system, ubiquitinated proteins, enzymes that play an important role in metabolic processes, cytoskeletal components, signaling, regulatory, and protective proteins, as well as proteins regulating gene expression, cell division, and differentiation. The abundance of a number of proteasome-associated proteins was comparable or exceeded the abundance of intrinsic proteasome components. About a third of the proteins common to all studied fractions (26S and 20S of brain and liver proteasomes) belong to the group of multifunctional proteins. Selective biosensor validation confirmed the affinity binding of proteins (aldolase, phosphoglycerate kinase) identified during MS analysis to the brain 20S proteasome. Comparison of the subproteomes of the 26S and 20S brain proteasomes showed that removal of components of the regulatory (19S) subparticles caused almost two-fold increase in the total number of individual proteins associated with the core part of the proteasome (20S). In the liver, the number of proteins associated with the core part of the proteasome remained basically unchanged after the removal of the components of the regulatory (19S) subparticles. This indicates that in the brain and, possibly, in other organs, proteins of the regulatory (19S) subunit play an important role in the formation of the proteasome interactome.

Infrared Spectroscopy in Differential Diagnosis of Pulmonary Embolism.

The aim of the study was to assess the effectiveness of infrared spectroscopy for verification of pulmonary embolism (PE) and a number of similar diseases. MATERIALS AND METHODS: Infrared spectroscopy was used to investigate blood serum of 19 healthy volunteers and 30 patients with intraoperatively confirmed PE as well as with chronic obstructive pulmonary disease (COPD) (n=10), pneumonia (n=10), tuberculosis (n=10), lung abscess (n=10) and lung cancer (n=10), acute disorder of cerebral circulation (ADCC) (n=10), ischemic heart disease (IHD) (n=10). Peak height ratios of absorption band were taken as diagnostic parameters (cm-1/сm-1): P1 - 1160/1165; P2 - 1165/1070; P3 - 1165/1150; P4 - 1165/1050; P5 - 1100/1050; P6 - 1025/1165. These parameters of IR spectrum are significant for the given nosology. RESULTS: The calculated indicators have demonstrated statistically significant difference of IR spectra parameters for the studied nosologies (p<0.001) even on the small samples supplementing each other and enabling step-by-step exclusion of lung abscess and pulmonary tuberculosis, COPD and pneumonia, cancer, IHD, ADCC, and PE.The presented radar charts, built with consideration of the values of all peak height ratios of the absorption bands with diagnostically significant maxima, provided the possibility to visualize the IR profiles making the differentiation of PE and its clinical analogs not only more objective and reliable but also more explicit and compelling. CONCLUSION: Infrared spectroscopy is a potentially effective method of PE differential diagnosis. Sample expansion will allow researchers to evaluate the sensitivity and specificity of this technique compared to the existing standard schemes of PE verification.

New Technology for the Use of Inhaled Nitric Oxide to Protect the Heart and Lungs during Operations with Cardiopulmonary Bypass.

The aim of the study was to evaluate the effectiveness of a new technology for the use of inhaled nitric oxide (NO) for the heart and lung protection during operations with cardiopulmonary bypass (СРВ). MATERIALS AND METHODS: The study included 90 patients who underwent heart valve surgery and combined procedures under CPB and pharmacological cardioplegia. Three groups were created: group 1 (control, n=30); group 2 (n=30) - NO inhalation (20 ppm) was conducted traditionally, that is, before and after CPB; group 3 (n=30) - NO inhalation was performed using a new technology - during the entire operation, with pulmonary artery perfusion and lung ventilation performed during CPB. Troponin I (cTn I) level, changes in the pulmonary function parameters, and clinical indicators were studied. RESULTS: Statistically significant lower levels of postoperative cTn I were registered in the patients of groups 2 and 3, at the same time, the levels were significantly lower in group 3 compared to group 2. The patients in group 1 (standardized anesthesia protocol) demonstrated an increase in the alveolar-arterial oxygen difference, an increase in intrapulmonary shunting, a decrease in blood oxygenation, and static lung compliance after СРВ. In both cases, NO inhalation retained the values of lung compliance and pulmonary oxygenating function after CPB, and in the patients of group 3, it also significantly reduced intrapulmonary shunting and alveolar-arterial difference after CPB. NO inhalation allowed a statistically significant decrease in the incidence of pulmonary dysfunction, acute respiratory failure, as well as the time of respiratory support in the ICU. CONCLUSION: The developed technology for the use of inhaled NO in surgery with CPB provides a clinically marked protective effect on the heart and lungs. The effectiveness of the protective action of NO depends on the duration of its administration and is most pronounced when used during the entire operation, including CPB time.

Application of Molecular Hydrogen in Heart Surgery under Cardiopulmonary Bypass.

The aim of this work was to study the effect of molecular hydrogen on oxidative processes in cardiac surgery patients with acquired valve heart disease applied during surgery under cardiopulmonary bypass (CPB). Materials and Methods: The study involved 20 patients (16 men and 4 women) with acquired heart valve disease who were operated on under CPB. Two groups of patients were formed. In group 1 (n=11), anesthesia included inhalations of molecular hydrogen, which was supplied to the breathing circuit of the ventilator at a concentration of 1.5-2.0% immediately after tracheal intubation and throughout the operation. In group 2 (n=9), inhalation of molecular hydrogen was not performed. Blood sampling was taken at 4 stages: immediately after anesthesia induction, before CPB and after its termination, and also one day after the operation. The intensity of the processes of lipid peroxidation was evaluated by the level of diene (DC) and triene (TC) conjugates, Schiff bases (SB). Results: In the patients of group 1, the arterial blood samples showed a decrease in the level of TC and SB, as compared to the first stage of the study, before the initiation of CPB and one day after the operation. An increase in the level of DC and TC was detected after the termination of CPB (p<0.05). In the venous blood samples, an increase in the level of DC was noted before the initiation of CPB, which was restored by the third stage of the study (p<0.05). At the same time, after the termination of CPB, a tendency towards a decrease in TC and SB was observed, which persisted one day after the operation.In the patients of group 2, an increase in the concentration of SB in the arterial blood samples was recorded during the study as compared to the first stage. The level of TC and SB in the venous blood samples increased one day after the operation. Conclusion: Intraoperative inhalation of molecular hydrogen leads to a decrease in the oxidative stress manifestation, it being most pronounced one day after the operation. This suggests that molecular hydrogen can be used in cardiac surgery as an effective and safe antioxidant.

Modified Mitral Valve Repair with Its Insufficiency of Ischemic Genesis.

The aim of the study was to assess the effectiveness of modified mitral valve repair in comparison with traditional methods of correcting ischemic mitral regurgitation. Materials and Methods: The results of surgical treatment of 80 patients with coronary artery disease complicated by ischemic mitral regurgitation were analyzed. The mean age of the patients was 58.95±8.36 years; the ratio of men and women was 67:13. Heart failure of FC II (according to the NYHA classification) was detected in 6 patients (7.50%), FC III - in 69 (86.25%) patients, FC IV - in 5 (6.25%) patients.Echocardiographic examination was used to determine the significance and genesis of mitral regurgitation in the preoperative period. 57 patients (71.25%) were detected with grade II mitral regurgitation, 23 (28.75%) had grade III.Annuloplasty was chosen as the operation for the correction of the valve apparatus. The patients of group 1 (n=23) underwent reconstructive surgery on the mitral valve using an autopericardial strip according to the technique, which we have developed, in combination with coronary artery bypass grafting (CABG), the patients of group 2 (n=26) underwent plastic surgery using a support ring in combination with CABG, patients of group 3 (n=31) had myocardial revascularization without correction of the valve apparatus. Results: The patients of group 2 underwent restrictive mitral annuloplasty performed with rigid support rings, group 1 - with an autopericardial strip as a soft support ring, the patients of group 3 underwent CABG alone.One patient from group 2 died in the early postoperative period due to acute perioperative myocardial infarction.The most common complications were pleurisy, acute cardiovascular failure, acute respiratory failure, and cardiac arrhythmias. The smallest number of complications was noted in the group 3, where patients underwent CABG alone. After surgery, all the patients showed a decrease in mitral regurgitation, which was most pronounced in the groups with annuloplasty.When analyzing the immediate results of the operations, it was revealed that the patients of groups 1 and 2, who underwent combined interventions, had a higher percentage of complications, and the length of their stay in the ICU increased. However, these groups showed a significant improvement in mitral valve functioning. Plasty of the mitral valve with an autopericardial strip according to the technique, which we have developed, demonstrated a good hemodynamic effect, the absence of significant regurgitation in the postoperative period.

[Changes in the mitochondrial subproteome of mouse brain Rpn13-binding proteins induced by the neurotoxin MPTP and the neuroprotector isatin]. / Izmenenie mitokhondrial'nogo subproteoma Rpn13-sviazyvaiushchikh belkov mozga myshi pod deistviem neirotoksina MFTP i neiroprotektora izatina.

Mitochondrial dysfunction and ubiquitin-proteasome system (UPS) failure contribute significantly to the development of Parkinson's disease (PD). The proteasome subunit Rpn13 located on the regulatory (19S) subparticle play an important role in the delivery of proteins, subjected to degradation, to the proteolytic (20S) part of proteasome. We have previously found several brain mitochondrial proteins specifically bound to Rpn13 (Buneeva et al. (2020) Biochemistry (Moscow), Supplement Series B: Biomedical Chemistry, 14, 297-305). In this study we have investigated the effect of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and the neuroprotector isatin on the mitochondrial subproteome of Rpn13-binding proteins of the mouse brain. Administration of MPTP (30 mg/kg) to animals caused movement disorders typical of PD, while pretreatment with isatin (100 mg/kg, 30 min before MPTP) reduced their severity. At the same time, the injection of MPTP, isatin, or their combination (isatin + MPTP) had a significant impact on the total number and the composition of Rpn13-binding proteins. The injection of MPTP decreased the total number of Rpn13-binding proteins in comparison with control, and the injection of isatin prior to MPTP or without MPTP caused an essential increase in the number of Rpn13-binding proteins, mainly of the functional group of proteins participating in the protein metabolism regulation, gene expression, and differentiation. Selected biosensor validation confirmed the interaction of Rpn13 subunit of proteasome with some proteins (glyceraldehyde-3-phosphate dehydrogenase, pyruvate kinase, histones H2A and H2B) revealed while proteomic profiling. The results obtained testify that under the conditions of experimental MPTP-induced parkinsonism the neuroprotective effect of isatin may be aimed at the interaction of mitochondria with the components of UPS.

[Effectiveness of the Cox Maze-IV Procedure in Cardiac Surgery Patients With Atrial Fibrillation].

Aim Analyzing a 5-year experience of surgical treatment of cardiosurgical patients with atrial fibrillation (AF).Material and methods The study analyzed results of surgical treatment with extracorporeal circulation in 132 patients with AF who underwent the Maze-IV procedure using a radiofrequency ablator with transmurality feedback from 2013 through 2018.Results Two fatal outcomes were observed in the study group. These outcomes took place in the early postoperative period and were associated with progressive acute heart failure in patients with repeated surgery for mitral valve restenosis. 61.2% of the patients had no AF. Recurrent AF was observed during the first three years after surgery in association with withdrawal of the antiarrhythmic medication, which confirmed a need for long-term antiarrhythmic therapy. Analysis of risk factors for AF relapse identified significant predictors, including left ventricular dilatation larger than 5.5 cm at baseline and more than two-year duration of a history of arrhythmias.Conclusion The Maze-IV procedure proved an effective and safe method of surgical treatment in AF patients with acquired heart defects and ischemic heart disease, which allowed maintaining sinus rhythm in 61.2% of patients for 5 years. Preventive amiodarone saturation reduced the risk of AF relapse by 24.2 % (p=0.038) and incidence of postoperative arrhythmic complications by 34.9 % (p=0.008) in cardiosurgical patients.

[Qualitative difference of mitochondrial subproteoms of brain RPN10- and RPN13-binding proteins]. / Kachestvennoe razlichie mitokhondrial'nykh subproteomov RPN10- i RPN13-sviazyvaiushchikh belkov mozga.

Good evidence exists that the ubiquitin-proteasome system (UPS) plays an important role in degradation of mitochondrial proteins and membrane proteins associated with mitochondria (MAM proteins). Mitochondria contain all components of the ubiquitin-conjugating system, which are necessary for the attachment of ubiquitin molecules to target proteins, subjected to subsequent degradation in proteasomes. An important stage in the delivery of proteins for proteolytic degradation in proteasomes is their interaction with ubiquitin receptors located on the regulatory subunit (19S) of the proteasome: the Rpn10 or Rpn13 subunit. These subunits make basically the same contribution to the subsequent translocation of target proteins to the core part of the proteasome. A comparative study of mouse brain mitochondrial subproteomes bound to Rpn10 and Rpn13 subunits revealed a high specificity of the repertoire of Rpn10 and Rpn13-binding proteins. Moreover, proteins, for which mitochondrial localization or association with mitochondrial membranes was previously shown, prevailed in the case of using the Rpn13 subunit as an affinity ligand (Rpn13-binding proteins). This suggests that Rpn10 and Rpn13 play different roles in the degradation of mitochondrial proteins and MAM.

[Circular Sympathetic Pulmonary Artery Denervation in Cardiac Surgery Patients With Mitral Valve Defect, Atrial Fibrillation and High Pulmonary Hypertension].

Objective Investigate the influence of the sympathetic denervation of the pulmonary trunk and the orifices of the pulmonary arteries on the degree of pulmonary hypertension (PH) and outcomes of the surgical treatment of atrial fibrillation (AF) in patients with mitral valve defects, complicated AF, and high PH.Material and methods We analyzed the surgical treatment of 140 patients with mitral valve defect, concomitant AF, and high PH - pulmonary artery systolic pressure (PASP) gradient more than 40 mm Hg. The group of interest included 51 patients (46 patients with severe mitral stenosis and five patients with grade 4 mitral valve regurgitation). All patients underwent mitral valve correction (47 valve replacement surgeries and 4 valve-sparing interventions), biatrial Maze IV procedure, and additionally, denervation of the pulmonary trunk and the orifices of the pulmonary arteries. The control group included 89 patients diagnosed with mitral valve defect, AF, and PH with PASP > 40 mm Hg. However, unlike in patients of interest, denervation of the pulmonary arteries was not performed.Results Circular radiofrequency denervation of the pulmonary trunk and the orifices of the pulmonary arteries using a clamp-destructor is an effective and safe method, significantly reduces secondary PH (p=0.018), promotes reverse remodeling of the heart chambers, left atrium in particular (p=0.01), and improves outcomes of the Maze IV procedure (p=0.022) by restoring sinus rhythm in patients with mitral valve defects, complicated AF, and high PH.Conclusion This technique must be studied further involving a more significant number of patients, analyzing long-term results, and using this technique in patients with non-valvular causes of secondary PH.

A first principles study of the spin-orbit coupling effect in LiM (M = Na, K, Rb, Cs) molecules.

The spin-orbit (SO) interactions in low-lying electronic states of the LiM (M = Na, K, Rb, Cs) molecular series are studied through ab initio calculations of potential energy curves and SO coupling matrix elements as functions of the interatomic distance, R. Two different approaches are employed: (a) the Fock-space relativistic coupled-cluster calculations (FS-RCC) which directly yield full relativistic energies, Urel(R); the SO coupling functions, ξso(R), are extracted a posteriori through projecting scalar-relativistic wave functions onto the subspaces spanned by their full-relativistic counterparts; (b) the evaluation of the scalar-relativistic electronic energies, Usr(R), and relevant ξso(R) functions using the configuration interaction method with core-valence correlation accounted for using core polarization potentials (CI-CPP). The SO-free potentials and SO coupling functions obtained within the framework of both approaches are in good agreement with each other and their prior theoretical and empirical counterparts.