[The Effects of the Hydrogen Sulfide Donor GYY4137 on the Proteasome Pool of Colorectal Cancer Cells].

Cancer cells are characterized by an increased level of metabolism and are highly dependent on the correct functioning of the processes that ensure homeostasis. Reactive sulfur species (RSS) are important molecular modulators of metabolic processes in both healthy and tumor cells. The effect of RSS and, in particular, H2S, on key cellular systems, including the ubiquitin-proteasome system (UPS), which provides the destruction of most intracellular proteins, has been shown. The main components of the UPS are proteasomes, multisubunit protein complexes, within which proteolysis occurs. At the same time, data on the effect of H2S directly on the pool of proteasomes in tumor cells are insufficient. Here, we studied the effect of incubation of SW620B8-mCherry colorectal adenocarcinoma cells expressing a fluorescently labeled proteasome subunit with 50, 100, and 200 µM of the hydrogen sulfide donor GYY4137. The effect of the substance on the proteasome pool was assessed 6, 24, 48, and 72 h after administration. It was shown that the chymotrypsin-like and caspase-like proteasome activity decreases in cells incubated with 200 µM of the GYY4137 for 24 h. This coincided with an increase in the expression of proteasome subunit genes. In lysates of cells incubated with 200 µM GYY4137 for 48 h an increase in the content of the constitutive ß5 subunit was observed and the activity of proteasomes leveled off. Following prolonged incubation with GYY4137 (72h), an increase in the expression levels of some proteasome genes was also observed, although this did not have a significant effect on the activity and subunit composition of proteasomes. Thus, the obtained data indicate the modulation of proteasome activity by the hydrogen sulfide donor and the effect of GYY4137 on transcription and translation of proteasome genes.

[Changes in the Activities and Contents of Individual Forms of Proteasomes in Samples of the Cerebral Cortex during Pathology Development in 5xFAD Mice].

The ubiquitin-proteasome system (UPS) provides hydrolysis of most intracellular proteins in proteasomes. There are various forms of proteasomes that differ, among other things, in the set of proteolytic subunits and the presence of activators. Alzheimer's disease (AD) is characterized by disturbances in the functional state of the UPS. At the same time, an increase in the expression of certain forms of proteasomes, in particular, proteasomes containing immune subunits (nonconstitutive proteasomes), has been shown. Here, we studied dynamic changes in the expression of catalytic proteasome subunit genes and corresponding proteins in the cerebral cortex of animals using a mouse model of AD (5xFAD transgenic mice). Increases by 4 and 6 folds in transcripts of the PSMB9 and PSMB8 genes encoding immune proteasome subunits were detected, as well as a significant increase in the content of immune ß-subunits (by 2.8 folds, ß1i; 2.2 folds, ß2i) in samples from 5xFAD mice at the age of 380 days, compared with samples from mice at 60 days of age. Moreover, the activation of both 20S and 26S proteasomes containing immune subunits were revealed in samples from 380 days old 5xFAD mice by electrophoresis in native conditions. This indicates activated synthesis of the immune subunits and assembly of nonconstitutive proteasomes at the terminal stage of pathology development. The obtained data, in combination with the available literature, indicate that the activation of nonconstitutive proteasomes is a universal phenomenon characteristic of various animal models of AD, which may reflect both the development of neuroinflammation and adaptive processes in tissues induced by the accumulation of toxic protein aggegates.

[Dynamic Changes in the Activities and Contents of Particular Proteasome Forms in the Cerebral Cortex of C57BL/6 Mice during Aging].

Proteasomes are key components of the ubiquitin-proteasome system. Various forms of proteasomes are known. During aging, disturbances in the functioning of proteasomes have been revealed, as well as increased expression of their particular forms. Considering these data, we studied the expression of genes encoding the constitutive and immune subunits of proteasomes in cerebral cortex samples from C57BL/6 mice at the ages of 60, 190, 380, and 720 days. In addition, the contents of constitutive and immune proteasome subunits, chymotrypsin-like and caspase-like activities of proteasome pools, as well as the activity of the ß5i immune subunit were studied in tissue homogenates. The chymotrypsin-like activity and the activity of the ß5i subunit of different forms of proteasomes separated by electrophoresis in native gel were characterized. Compared with samples from young animals, in the cerebral cortex of animals at an age of 720 days the following changes in the expression patterns of proteasome genes were revealed: a decreased expression of the PSMB5 gene encoding constitutive proteasome subunit ß5; increased expression of genes encoding immune proteasome subunits ß5i and ß1i. In tissue homogenates of aged mice, an increase in the content of immune subunits ß1i and ß2i was shown. In samples from old animals, chymotrypsin-like activity was decreased and a tendency to a decrease in caspase-like activity of proteasomes as well as the ß5i subunit activity was revealed. Analysis of the activity of native complexes in tissues obtained from old animals revealed decreased chymotrypsin-like activity of 26S and 20S proteasomes containing the ß5i subunit. Based on the obtained data, it can be assumed that changes in the pool of nonconstitutive proteasomes reflect aging-associated adaptive processes in the mouse brain.

[Efficiency and safety of Prostatex in patients with chronic prostatitis/chronic pelvic pain syndrome. Results of the phase IV PRESTIGE trial].

INTRODUCTION: A large number of both preclinical and clinical studies demonstrates the efficiency of bioregulatory peptides for the treatment of prostatitis and chronic pelvic pain syndrome (CPPS). A relatively new drug in this group is Prostatex, the active ingredient of which is bovine prostate extract. AIM: To evaluate the effect of taking Prostatex on the intensity of CPPS, the sexual function, and the results of microscopy of expressed prostate secretions and urinalysis. MATERIALS AND METHODS: A cohort of patients aged 25-65 years with chronic abacterial prostatitis and complaints of chronic pelvic pain was analyzed. The abacterial type of prostatitis was confirmed by bacteriological examination of expressed prostate secretions. The patients received Prostatex for 30 days according to the following scheme: 1 suppository rectally 1 time per day. The follow-up was 30 days. Before starting the drug and at the end of the 30-day course, patients completed the Chronic Prostatitis Symptom Index (NIH-CPSI) and the sexual function questionnaire. In addition, urinalysis and microscopic study of expressed prostate secretions was performed. RESULTS: A total of 1700 patients were included in the study. While taking the drug, there was a significant decrease in pain during digital rectal examination, as well as in the intensity of pain as a symptom of CPPS. The severity of symptoms after treatment was lower in all domains of NIH-CPSI. Microscopic study of the expressed prostate secretions during treatment showed a decrease in the number of patients with excessive number of leukocytes. The sexual function improved, while urinalysis and microscopy of expressed prostate secretions returned to the reference values. CONCLUSION: The use of Prostatex for the treatment of patients with CPPS reduces the severity of pain and other symptoms of chronic prostatitis, increases sexual function and normalizes the expressed prostate secretions and urinalysis. In order to obtain data of a higher level of evidence, it is necessary to carry out randomized, blind, placebo-controlled studies.

[Evaluation of long-term results of using the drug Fertiwell: impact on the likelihood of conception and pregnancy outcome].

INTRODUCTION: Current methods of treating male infertility have limited efficiency, since they are aimed to individual stages of the pathogenesis. Preparations based on testicular regulatory polypeptides are the most physiological and universal, owing to a complex effect on the self-regulation of testicular tissue. AIM: To study the delayed efficiency and safety of therapy with Fertiwell in patients with pathospermia and to assess the frequency of conception and pregnancy outcome in their partners based on the collection, analysis and interpretation of medical data. MATERIALS AND METHODS: A telephone survey of patients participating in the phase III clinical trial was carried out. The fact of conception in a couple was assessed over a period of 1 to 9 months after completion of therapy, as well as time from completion of the course to conception, pregnancy outcomes, newborn health outcomes. RESULTS: In the period from 1 to 9 months after completion of therapy, pregnancy occurred in 17 out of 34 couples (50%) in the Fertiwell group and in 13 out of 42 couples (30.95%) in the placebo group. This difference was statistically and clinically significant (p<0.05). All pregnancies resulted in a live birth. The median time from completion of the course to conception was 4 months in Fertiwell group and 6 months in the placebo group. There were no significant differences in anthropometric parameters of newborns between the two groups (p>0.05). CONCLUSION: When using the drug Fertiwell, pregnancy and live birth rate was significantly higher (2.23 times) compared to the control group. There was a trend toward earlier pregnancies in partners of men receiving Fertiwell. Thus, this drug can be recommended for the treatment of men with idiopathic infertility as monotherapy, as well as in combination with assisted reproductive technologies.

All-fiber high-power erbium-doped laser system generating optical pulses with a duration of 200 µs to 5 ms for fractional photo-rejuvenation.

An all-fiber high-power erbium-doped fiber laser (EDFL) source generating optical pulses from 200 µs to 5 ms with a stable rectangular envelope for fractional photo-rejuvenation is proposed and experimentally demonstrated. A master oscillator power amplifier (MOPA) configuration composed of a master oscillator, an acousto-optic modulator (AOM), and a one-stage amplifier is designed and employed in the EDFL to serve as an efficient laser system with excellent output performance. To avoid multistage amplifiers, the master oscillator generates 1.5 W, and a Yb-free Er-doped large-mode-area (LMA) active fiber is used for a one-stage power amplifier. There are two benefits to this approach: first, modulation of both pump and seed pulses is used to achieve clear rectangular shaped pulses without amplified spontaneous emission (ASE) growth; and second, there are no power limitations in the amplifier and undesirable 1 µm ASE compared to Er/Yb systems. We have reached 28.6 W of peak power with 26% slope efficiency limited only by available pump power, so the system can be easily scaled for achieving a higher peak power.

[The role of expression of monocarboxylates of the first and fourth types (MCT1, MCT4) by tumor and stromal cells of prostate cancer in determining the prognosis and the efficiency of definitive treatment].

AIM: A search for new methods for diagnosing clinically significant prostate cancer is of importance due to the insufficient accuracy of modern methods in detecting aggressive tumors. One of the promising opportunities for the early diagnosis of clinically significant prostate cancer is the assessment of the glycolytic profile of the tumor by determining the expression of monocarboxylates (MCT) types 1 and 4 in tumor cells, as well as in adjacent stromal cells. MATERIALS AND METHODS: An analysis of patients of who underwent radical prostatectomy at the Institute of Urology and Reproductive Health of Sechenov University from 2015 to 2017 was carried out. The patients with histologically confirmed prostate adenocarcinoma were included in the study. Among them, the presence or absence of biochemical recurrence during the first year was studied. An immunohistochemical (IHC) study of postoperative specimen was performed to determine the expression of MCT1 and MCT4 by tumor and stromal cells. The correlation between the intensity of their expression and the risk of biochemical recurrence and the tumor characteristics was evaluated. RESULTS: High membrane expression of MCT1 directly correlated with high stromal expression of MCT4 (r=0.314, p<0.003). A significant direct correlation was found between the predominance of stromal expression of MCT4 over membrane expression and biochemical recurrence (r=0.403, p<0.001), as well as a high ISUP group (4 and 5) (r=0.294, p=0.005). CONCLUSIONS: Determination of the level of expression of type 1 and 4 monocarboxylate transporters in adenocarcinoma cells and tumor stromal cells can become an effective tool for risk stratification, and may also predict the biological behaviors of the prostate cancer and the efficiency of definitive treatment.

[Prospective comparison of cognitive and mpMR/US fusion biopsy for prostate cancer detection].

INTRODUCTION: According to the recommendations of the European Association of Urology the presence of a suspicious lesion on MRI is an indication for both primary and secondary MR-targeted biopsies. At the same time, the Russian Society of Urologists recommends to perform mpMR/US fusion biopsy only in patients with a prior negative biopsy. In clinical practice, mpMR/US fusion and cognitive biopsies are the most frequently performed. However, when comparing them, contradictory data on detection of clinically significant prostate cancer is obtained. OBJECTIVE: to compare the detection rate of clinically significant prostate cancer performing cognitive and mpMR/US fusion biopsies. MATERIALS AND METHODS: Inclusion criteria: PSA >2 ng/mL and/or a positive DRE, and/or a suspicious lesion on TRUS, and PI-RADSv2.1 lesion more or equal 3. At first, "unblinded" urologist performed a transperineal mpMR/ultrasound fusion and saturation biopsy. Then "blinded" urologist obtained transrectal cognitive biopsy Clinically significant cancer was defined as ISUP more or equal 2. RESULTS: We enrolled 96 patients. Median age was 63 years, prostate volume - 47 cm3 and PSA - 6.82 ng/mL. MpMR/US fusion and cognitive biopsies were comparable in regard to the detection rate of clinically significant (32.3% vs 25.0%; p=0.264), clinically insignificant cancer (25.0% and 26.0%; p=0.869) and overall detection rate (57.3% and 51%;p=0.385). Both biopsies missed clinically significant cancer with equal frequency (5.2%; p=0.839). Histological efficacy also was comparable. The number of positive cores between mpMR/US fusion and cognitive biopsy was equal (34.1% and 31.1% respectively; p= 0.415). At the same time, no statistically significant difference was found with respect to maximum cancer core length (53.1% vs 47.7%, respectively; p=0.293). CONCLUSION: The results suggest that both cognitive and mpMR/US fusion biopsies are equally accurate diagnostic methods for clinically significant prostate cancer detection, thus their wider introduction into clinical practice is necessary.

Synchronization of quasiperiodic oscillations in nearly Hamiltonian systems: The degenerate case.

Quasiperiodic perturbations of two-dimensional nearly Hamiltonian systems with a limit cycle are considered. The behavior of solutions in a small neighborhood of a degenerate resonance is studied. Special attention is paid to the synchronization problem. Bifurcations of quasiperiodic solutions that arise when the limit cycle passes through the neighborhood of a resonance phase curve are investigated. The study is based on an analysis of an autonomous pendulum-type system, which is obtained by the method of averaging and determines the dynamics in the resonance zone. Two possible topological structures of the unperturbed averaged system are distinguished. For each case, the intervals of a control parameter that correspond to oscillatory synchronization are found. The results are applied to a Duffing-Van der Pol-type equation.

Combinatorial invariant for Morse-Smale diffeomorphisms on surfaces with orientable heteroclinic.

In this paper, we consider a class of orientation-preserving Morse-Smale diffeomorphisms defined on an orientable surface. The papers by Bezdenezhnykh and Grines showed that such diffeomorphisms have a finite number of heteroclinic orbits. In addition, the classification problem for such diffeomorphisms is reduced to the problem of distinguishing orientable graphs with substitutions describing the geometry of a heteroclinic intersection. However, such graphs generally do not admit polynomial discriminating algorithms. This article proposes a new approach to the classification of these cascades. For this, each diffeomorphism under consideration is associated with a graph that allows the construction of an effective algorithm for determining whether graphs are isomorphic. We also identified a class of admissible graphs, each isomorphism class of which can be realized by a diffeomorphism of a surface with an orientable heteroclinic. The results obtained are directly related to the realization problem of homotopy classes of homeomorphisms on closed orientable surfaces. In particular, they give an approach to constructing a representative in each homotopy class of homeomorphisms of algebraically finite type according to the Nielsen classification, which is an open problem today.

[Structures of the Mouse Central Nervous System Contain Different Quantities of Proteasome Gene Transcripts].

Proteasomes are multisubunit complexes that degrade most intracellular proteins. Three of the 14 subunits of the 20S proteasome, specifically ß1, ß2, and ß5, demonstrate catalytic activity and hydrolyze peptide bonds after acidic, basic, and hydrophobic amino acids, respectively. Within proteasome, the constitutive catalytic subunits ß1, ß2, and ß5 can be substituted by the immune ßli, ß2i, and ß5i subunits, respectively. However, proteasomes do not always contain all the immune subunits at once; some proteasomes contain both immune and constitutive catalytic subunits simultaneously. Incorporation of immune subunits modifies the pattern of peptides produced by proteasomes. This is essential for antigen presentation and cellular response to stress as well as for a number of intracellular signaling pathways. We have developed a quantitative PCR-based system for the determination of the absolute levels of murine constitutive and immune proteasome subunits gene expression. Using the obtained system, we have estimated the expression levels of genes encoding proteasome subunits in the mouse central nervous system (CNS) tissues. We have shown that the quantity of transcripts of proteasome catalytic subunits in different CNS structures differed significantly. These data allow us to assume that the studied brain regions can be divided into two groups, with relatively "high" (cerebral cortex and spinal cord) and "low" (hippocampus and cerebellum) levels of proteasome subunit genes expression. Moreover, it was possible to distinguish structures with similar and significantly different gene expression profiles of proteasome catalytic subunits. Thus, the gene expression profiles in the cortex, spinal cord, and cerebellum were similar, but different from the expression profile in the hippocampus. Based on the obtained data, we suggest that there are differences in the proteasome pool, as well as in the functional load on the ubiquitin-proteasome system in different parts of the CNS.

H2S counteracts proinflammatory effects of LPS through modulation of multiple pathways in human cells.

BACKGROUND: Hydrogen sulfide donors reduce inflammatory signaling in vitro and in vivo. The biological effect mediated by H2S donors depends on the kinetics of the gas release from the donor molecule. However, the molecular mechanisms of H2S-induced immunomodulation were poorly addressed. Here, we studied the effect of two different hydrogen sulfide (H2S)-producing agents on the generation of the LPS-induced inflammatory mediators. Importantly, we investigated the transcriptomic changes that take place in human cells after the LPS challenge, combined with the pretreatment with a slow-releasing H2S donor-GYY4137. METHODS: We investigated the effects of GYY4137 and sodium hydrosulfide on the release of proinflammatory molecules such as ROS, NO and TNF-α from LPS-treated human SH-SY5Y neuroblastoma and the THP-1 promonocytic cell lines. Transcriptomic and RT-qPCR studies using THP-1 cells were performed to monitor the effects of the GYY4137 on multiple signaling pathways, including various immune-related and proinflammatory genes after combined action of LPS and GYY4137. RESULTS: The GYY4137 and sodium hydrosulfide differed in the ability to reduce the production of the LPS-evoked proinflammatory mediators. The pre-treatment with GYY4137 resulted in a drastic down-regulation of many TNF-α effectors that are induced by LPS treatment in THP-1 cells. Furthermore, GYY4137 pretreatment of LPS-exposed cells ameliorates the LPS-mediated induction of multiple pro-inflammatory genes and decreases expression of immunoproteasome genes. Besides, in these experiments we detected the up-regulation of several important pathways that are inhibited by LPS. CONCLUSION: Based on the obtained results we believe that our transcriptomic analysis significantly contributes to the understanding of the molecular mechanisms of anti-inflammatory and cytoprotective activity of hydrogen sulfide donors, and highlights their potential against LPS challenges and other forms of inflammation.

Peculiarities of Histological Structure of Pulmonary Veins in Patients with Total Anomalous Pulmonary Venous Drainage as Morphological Substrates Relating to Formation of Postoperative Pulmonary Venous Obstruction.

The histological and immunohistochemical methods were employed to examine the peculiarities of histological structure of pulmonary veins and left atrium of the heart in norm and in various types of total anomalous drainage of pulmonary veins. In contrast to normal pulmonary vein covered with external multiple muscle layers (myocardial sleeve), such sleeve is absent in veins that have no connection with the left atrium irrespective of the type of the defect. In patients with total anomalous pulmonary venous drainage, the structure of left atrium was heterogeneous featuring either the presence or absence of inner angiomural lining in this atrium. The structural peculiarities are important for insight into etiology of the development of postoperative pulmonary venous obstruction in patients with total anomalous pulmonary venous drainage.

[Long-term results of combined treatment of patient with multiple primary cancers, including locally-advanced renal cell cancer, bladder cancer and transitional-cell cancer of the contralateral kidney].

A clinical case of the successful surgical treatment of patient with multiple primary cancers, including locally-advanced right renal cell cancer, transitional-cell cancer bladder cancer and metachronous transitional-cell cancer of the left kidney with one of the longest follow-up and survival time described in the literature.

[Risk factors for complications of ureterolithotripsy].

AIM: to determine main risk factors for complications of ureterolithotripsy. MATERIALS AND METHODS: a retrospective analysis of the results of 545 ureteroscopies performed in 506 patients with ureteral stones over the past 7 years at the urological clinic named after M.F. Vladimirsky was carried out. The relationship between preoperative and intraoperative factors and complications of ureterolithotripsy was analyzed. RESULTS: The overall complication rate was 22.4%. The risk of intraoperative complications was proved to increase (p<0.05) along with the stone size, location in proximal ureter, stone impaction for more than 3 weeks and the degree of hydronephrosis. No preoperative stenting or nephrostomy tube prior to ureteroscopy was associated to an increased risk of intraoperative complications (RR=2.88; p=0.03). Patients with preoperative drainage of upper urinary tract has lower probability of intraoperative complications (OR=0.35; p=0.03). The risk of developing stricture and ureteral obliteration in uncomplicated ureteroscopy was minimal (RR=0.008, p=0.0001). Small ureteral perforation and pronounced mucosal inflammation around the stone have the similar influence on the risk of complications, which was more than 7.5 times (p=0.0001) higher than in uncomplicated ureteroscopy. Large ureteral perforation was associated with the highest risk of stricture formation and ureteral obliteration, which was 64 times (p=0.0001) higher than in uncomplicated ureteroscopy. CONCLUSION: The rate of complications of ureterolithotripsy directly depends on the following factors: size and location of the stone, the stone impaction, pre- and intraoperative (nephrostomy tube) drainage of the upper urinary tract, the degree of hydronephrosis, level of bacteriuria and intraoperative trauma complications.

[The contribution of M. N. Akhutin in field surgery (to the 75th anniversary of Victory in the Great Patriotic war)].

The article is devoted to M. N. Akhutin (1899-1848), one of the founders of the Russian military field surgery, lieutenant general of the medical service, professor, corresponding member of the USSR Academy of medical Sciences, chief of the Military medical Academy (1940-1941). M. N. Akhutin, disciple of V. A. Oppel, was active participant in the battles at the lake Hasan and the Khalkhin-Gol river (1838-1939). During the Great Patriotic War, he was the chief surgeon of the Bryansk, 2nd Baltic and 1st Ukrainian fronts. Since 1945 he was deputy chief surgeon of the Soviet Army. He is author of works on military field surgery. He was the first director of the A. V. Vishnevsky institute of surgery and the head of the department of faculty surgery of the I. M. Sechenov 1st Moscow medical Institute.

Thermodynamic properties and lattice dynamics investigation of LuB2C: experiment and ab initio calculations.

A sample of lutetium carboboride LuB2C was synthesized from a mixture of lutetium hydride, boron and carbon by annealing in argon. The temperature dependence of the heat capacity Cp(T) (2-300 K) and lattice parameters a(T), b(T), and c(T) (5-300 K) of the carboboride was experimentally determined. The experimental values of the heat capacity were fitted with the approximation Cp(T) = aT + ΣCD + CE + CTLS(T). Here the first term is the electronic contribution, the second is the sum of the Debye components, the third is the Einstein contribution, and the fourth is the contribution to the heat capacity due to the vibrations of the two-level systems which are formed in the Lu-subsystem due to the asymmetry of the B-C atomic arrangement around the Lu3+-ions and, as a consequence, the possible transition of the lutetium atoms between spatially close, but energetically non-equivalent positions. A strong anisotropy of the thermal expansion of the carboboride was revealed. Along the c axis the coefficient of thermal expansion monotonically increases; in the basal plane, the expansion is practically not observed. The temperature dependence of the unit cell volume Vu(T) has been analyzed in the Debye-Einstein approximation taking into account the electronic contribution and effect of two-level systems. The values of the Gruneisen parameters corresponding to different modes of the phonon spectrum of the carboboride have been determined. The frequencies of the lattice vibrations, determined in a Raman scattering experiment, are in satisfactory agreement with the parameters obtained from Cp(T) using the Debye-Einstein approximation. Using ab initio band theory methods and an exchange-correlation functional in the PBE form in the VASP package, it was established that the total energies of these two crystal structures differ by no more than 0.01 eV f.u.-1. Calculations of the thermodynamic properties of LuB2C yielded similar results for orthorhombic and tetragonal phases of the carboboride.

[Dynamics of the Functional Activity and Expression of Proteasome Subunits during Cellular Adaptation to Heat Shock].

The ubiquitin-proteasome system (UPS) performs proteolysis of most intracellular proteins. The key components of the UPS are the proteasomes, multi-subunit protein complexes, playing an important role in cellular adaptation to various types of stress. We analyzed the dynamics of the proteasome activity, the content of proteasome subunits, and the expression levels of genes encoding catalytic subunits of proteasomes in the human histiocytic lymphoma U937 cell line immediately, 2, 4, 6, 9, 24, and 48 h after a heat shock (HS). The initial decrease (up to 62%) in the proteasome activity in cellular lysates was revealed, then 10 h after HS the activity began to recover. The amount of proteasomal α-subunits in the cells decreased 2 h after HS, and was restored to 24-48 h after HS. Fluctuations in the levels of mRNAs encoding proteasome catalytic subunits with the maximum expression 2 h after HS and a gradual decrease to 48 h after HS were observed. The average estimated number of mRNA copies per cell ranged from 10 for weakly to 150 for highly expressed proteasome genes. Thus, the recovery efficiency of UPS functionality after HS, which reflects the important role of proteasomes in maintaining cell homeostasis, was evaluated.

Soluble Ligand of the Immune Checkpoint Receptor (sPD-L1) in Blood Serum of Patients with Renal Cell Carcinoma.

The content of the soluble ligand of the immune checkpoint receptor (sPD-L1) was determined in the blood serum of 106 patients with renal cell carcinoma and 11 patients with benign kidney tumors by direct ELISA (Human sPD-L1 Platinum ELISA; Affimetrix, eBioscience). The control group included 19 healthy men and 18 women. Serum level of sPD-L1 significantly surpassed the control values in both patients with primary renal cancer (p<0.0001) and in patients examined during disease progression (p<0.05). In patients with benign kidney tumors, the level of this marker was significantly higher than in the control (p<0.05), but lower than in patients with renal cell carcinoma. The sPD-L1 level significantly increased with disease stage (p<0.001); it was higher in the presence of metastases in regional lymph nodes irrespective of their number (N1 or N2) than in the absence of metastases (N0); it was also increased in patients with distant metastases (M1) and patients with grade III-IV tumors in comparison with grade III-IV tumors (p<0.05). The highest sPD-L1 levels were recorded in patients with tumor size corresponding to T2 and T3 and decreased in patients with T4 tumors. Thus, sPD-L1 level in patients with renal cell carcinoma correlated with tumor grade and metastasizing and can be considered as a promising marker in monitoring of the effect of anti-PD1/PD-L1 therapy.

Comparative Analysis of Blood Plasma Proteome in Patients with Renal Cell Carcinoma.

Comparative mass spectrometric analysis of protein composition was carried out in 36 blood plasma specimens from patients with renal cell carcinoma and 20 specimens from donors. Analysis of protein composition of plasma specimens devoid of the major protein fractions showed a 20-50% higher level of protein identifications in patient' specimens. Specimens of the control and experimental series were similar by protein composition, 70-80% identifications in experimental and control series coinciding. High similarity of biological processes with participation of the proteins identified in both series was observed. The greater part of proteins in both series were located extracellularly and were exosomal (specimens from renal cancer patients) or vesicular (specimens from healthy volunteers).