[Production and internalization of extracellular vesicules in normal and under conditions of hyperglycemia and insulin resistance]. / Produktsiia i internalizatsiia vnekletochnykh vezikul v norme i v usloviiakh giperglikemii i insulinorezistentnosti.

Extracellular vesicles (EVs) are spherical structures of cell membrane origin, ranging in the size from 40 nm to 5000 nm. They are involved in the horizontal transfer of many proteins and microRNAs. The mechanisms EV internalization include clathrin-dependent endocytosis, caveolin-dependent endocytosis, raft-mediated endocytosis, and macropinocytosis. Type 2 diabetes mellitus (T2DM) is a common group of metabolic disorders in adults; the incidence and prevalence increase in parallel with the obesity epidemic. Since adipose tissue plays a crucial role in the development of insulin resistance, EVs secreted by adipose tissue can be a kind of information transmitter in this process. EVs of adipocytic origin are predominantly absorbed by tissue macrophages, adipocytes themselves, hepatocytes, and skeletal muscles. This contributes to the M1 polarization of macrophages, a decrease in glucose uptake by hepatocytes and myocytes due to the transfer of functionally active microRNAs by these EVs, which affect carbohydrate and lipid metabolism. Patients with T2DM and impaired glucose tolerance have significantly higher levels of CD235a-positive (erythrocyte) EVs, as well as a tendency to increase CD68-positive (leukocyte) and CD62p-positive (platelets/endothelial cells) EVs. The levels of CD31+/CD146-positive BB (endothelial cells) were comparable between diabetic and euglycemic patients. EVs from diabetic patients were preferably internalized by monocytes (mainly classical and intermediate monocyte fractions and to a lesser extent by non-classical monocyte fractions) and B cells compared to euglycemic patients. Internalization of EVs from patients with T2DM by monocytes leads to decreased apoptosis, changes in differentiation, and suppression of reactions controlling oxidative stress in monocytes. Thus, insulin resistance increases secretion of EVs, which are preferentially internalized by monocytes and influence their function. EVs are considered as sources of promising clinical markers of insulin resistance, complications of diabetes mellitus (endothelial dysfunction, retinopathy, nephropathy, neuropathy), and markers of EVs can also be used to monitor the effectiveness of therapy for these complications.

Serum levels of cytoskeleton remodeling proteins and their mRNA expression in tumor tissue of metastatic laryngeal and hypopharyngeal cancers.

Actin-binding proteins (ABPs) and various signaling systems are involved in the process of squamous cell carcinoma of the larynx and hypopharynx (SCCLH) metastasis. The clinical significance of these proteins has not yet been determined. We analyzed the relationship between the mRNA levels of cofilin 1 (CFL1), profilin 1 (PFN1), adenylyl cyclase-associated protein 1 (CAP1), SNAI1 and RND3 and SCCLH metastasis. The serum levels of the above ABPs were estimated and the relationship between them and their mRNA expressions was analyzed. The expression levels of ABP mRNAs were measured by real-time RT-PCR in paired tissue samples taken from 54 patients with SCCLH (T1-4N0-1M0). Expression analysis was performed using the 2-ΔΔCT method. The levels of ABPs in the blood serum were measured by ELISA. Statistical analysis was carried out using the SPSS Statistica 20.0 software package. No significant difference in the mRNA gene expression in tumor tissue of patients with T1-3N0M0 SCCLH and patients with T2-4N1-2M0 SCCLH was found. High expression of RND3 mRNA was accompanied by an increase in mRNA expression of all studied ABPs. In the blood serum of T2-4N1-2M0 patients, the level of PFN1 was lower by 21% and the level of CAP1 was higher by 75% than those observed in T1-4N0M0 patients. The data obtained showed that RND3 is involved in the regulation of molecular cascades of SCCLH metastasis. PFN1 and CAP1 serum levels can be good classifiers of metastases in patients with SCCLH.

Circulating Actin-Binding Proteins in Laryngeal Cancer: Its Relationship with Circulating Tumor Cells and Cells of the Immune System.

We previously exposed the role of actin-binding proteins (ABPs) in cancer development and progression. In this paper, we studied the relationship between circulating ABPs and the number of ABP-expressing leukocytes and circulating tumor cells (CTCs) in patients with highly aggressive laryngeal squamous cell carcinoma (LSCC). The levels of cofilin (CFL1), profilin (PFN1), ezrin (EZR), fascin (FSCN1), and adenylate cyclase-associated protein 1 (CAP1) were determined using enzyme immunoassay. The ABP expression by the cellular pools was analyzed by flow cytometry. The highest levels of FSCN1 and EZR were found in the blood serum of LSCC patients. There was a difference in ABP expression between the pools of leukocytes and CTCs. Leukocytes were mainly represented by CAP1+ and FSCN1+ pools, and CTCs contained CAP1+, FSCN1+, and EZR+ cells. The serum FSCN1 level correlated with the number of FSCN1-containing and CFL1-containing leukocytes. Thus, the level of circulating EZR is likely related to its expression in CTCs. The levels of CFL1 and PFN1 are likely to be supported by the expression of these proteins by leukocytes. Both CTCs and leukocytes can be a source of FSCN1 and CAP1 in blood serum. The results suggest that serum proteins can be produced by various cells, thus indicating both cancer development and the response of the immune system to this process.

Relationship between the Levels of mRNA Expression for Protein Phosphatase 1B and Proteins Involved in Cytoskeleton Remodeling in Squamous Cell Carcinoma of the Larynx and Hypopharynx.

We analyzed the expression of genes encoding proteins involved in cytoskeleton remodeling (RND3, SNAI1, vimentin, cofilin, adenylate cyclase-associated protein 1, ezrin, and profilin) depending on the level of expression of protein phosphatase 1B (PPM1B) mRNA on the example of squamous cell carcinoma of the larynx and hypopharynx. Against the background of a high level of PPM1B expression, a significantly high level of profilin expression was noted. Metastasis correlated with the level of snai1 expression, while relapse after combination treatment was negatively associated with the level of vimentin expression. The obtained new data can reflect molecular peculiarities of the tumor growth in squamous cell carcinoma of the larynx and hypopharynx.

Relationship between the mRNA Expression Levels of Calpains 1/2 and Proteins Involved in Cytoskeleton Remodeling.

Remodeling of the cytoskeleton underlies various cellular processes, including those associated with metastasis. The role of the proteases and proteins involved in cytoskeletal reorganization is being actively studied. However, there are no published data on the relationship between the mRNA expression levels of calpains 1/2 (CAPN 1/2) and the proteins associated with cytoskeleton remodeling. Therefore, the purpose of our study was to establish the relationship between the mRNA expression levels of CAPN 1/2 and the proteins involved in cytoskeletal reorganization, such as cell motility markers (SNAI1, VIM, and RND3) and actin-binding proteins (CFN1, PFN1, EZR, FSCN1, and CAP1) using the model of laryngeal/laryngopharyngeal squamous cell carcinoma (LC). The gene expression level was determined by reverse transcriptase real-time PCR and calculated using the 2-ΔΔCt method in paired tissue samples of 44 patients with LC (T1-4N0-2M0). The patients were divided into two groups: those with low and those with high CAPN 1/2 expression levels. It was found that metastasis in LC patients was associated with decreased expression levels of VIM and CAP1, and increased levels of CAPN1. A high level of CAPN2 was accompanied by a high expression level of EZR, indicating the activation of invasion processes. The results obtained need to be confirmed in further studies using a larger sample of patients and target genes. Our study is important in elucidating the mechanisms that underlie cancer progression and metastasis, a development that could subsequently open the way to a search for new prognostic and predictive markers of laryngeal/laryngopharyngeal cancer progression.

Expression of Genes Encoding Cell Motility Proteins during Progression of Head and Neck Squamous Cell Carcinoma.

The model of head and neck squamous cell carcinoma (HNSCC) was used to study the expression of genes encoding actin-binding proteins depending on the type of cell motility. The expression of SNAIL1 and CAPN2 mRNA in HNSCC tissue was higher than in specimens of dysplastic epithelium of the larynx and hypopharynx, which can be explained by activation of mesenchymal and amoeboid types of cell motility. In biopsy material of HNSCC patients with T1-2N0M0, expression of genes responsible for actin-binding proteins differed from that of patients with pretumor pathology of the larynx and hypopharynx: expression of FSCN was lower, while expressions of EZR and CAP1 were higher. The data attest that progression of HNSCC is associated with activation of both types of cell motility and with the changes in the expression of mRNA encoding cell motility proteins.

Circulating Proteasomes in the Pathogenesis of Head and Neck Squamous Cell Carcinoma.

Increased proteasome activity was revealed in blood serum of patients with stage T1N0M0 head and neck squamous cell carcinoma in comparison with patients with chronic diseases of the larynx and laryngopharynx. This opens prospects of using chymotrypsin-like activity measurement for differential diagnosis of squamous cell carcinoma, screening for high-risk groups, and evaluation of the degree of tumor differentiation.

Adenylyl Cyclase-Associated Protein 1 in the Development of Head and Neck Squamous Cell Carcinomas.

We compared the content of adenylyl cyclase-associated protein 1 (CAP1) in the blood and tissues of patients with head and neck squamous cell carcinomas (with and without regional metastases), patients with chronic inflammatory diseases aggravated by laryngeal and laryngopharyngeal dysplasia, and healthy individuals. The data suggest that serum CAP1 concentration correlated with the depth of primary tumor invasion and the presence of regional metastases. In cancer patients, the serum level of CAP1 was lower than in patients with laryngeal and laryngopharyngeal dysplasia, which can be of importance for differential and timely diagnostics of malignant tumors.

[Proteasome and calpain activities in colon cancer: the relation with metastasis and prognosis].

The chymotrypsin-like total proteasome activity and activity of 26S and 20S pools of proteasomes as well as calpain activitiy were found to be significantly increased in colon cancer tissue as compared to normal colon tissue. Patients with stage T2-4N1-3M0-lcolon cancer had higher activity level of the 26/20S proteasome and total calpain activities than patients with stage T2-4NOMO-1 colon cancer. The chymotrypsin-like total activity of proteasomes and the total activity of caplains in patients with stage T2-4N0-3M1 colon cancer were re- spectively 1.6 and 2.0-fold higher compared to those in patients with stage T2-4N0-3M0 colon cancer. Survival analysis of patients with colon cancer showed that the high level of chymotrypsin-like protesome activity in colon cancer and normal colon tissues (more than 72-1000 units/mg protein and 35-1000 units/mg protein, respectively) and the high level of total calpain activity in colon cancer and normal colon tissues (more than 100.5-1000 Units/mg protein and 52-1000 Units/ mg protein, respectively) were unfavorable factors in terms of 2-year metastasis-free survival.

[Papillomatosis as a criteria for the formation of the group at risk of laryngeal cancer].

The objective of the present study was to determine the incidence of dysplastic pre-neoplastic alterations in the patients presenting with papillomatosis. The results of the clinical, endoscopic, and morphological examination of 42 patients presenting with laryngeal papillomas made it possible to diagnose grade II-III laryngeal cancer in 7 (16.7%) patients. Half of them exhibited pre-neoplastic changes in laryngeal mucosa. The following criteria for the formation of laryngeal cancer risk groups, besides grade II-III epithelial dysplasia, were identified: long (over 12 months) history of papillomatosis, human papilloma virus infection, the history of smoking over 20 years, professional contacts with petroleum, oil, and lubrication materials, and the male gender. An algorithm of observation and treatment for each group of patients differing in the degree of dysplastic changes in laryngeal mucosa was proposed. It is concluded that adequate clinical and endoscopic monitoring of the patients at risk of laryngeal cancer makes it possible to increase the frequency of detection of this condition at the early stages of its development.

Chymotrypsin-like activity of proteasomes and total calpain activity in gastric and colorectal cancer.

Chymotrypsin-like activity of proteasomes and total calpain activity were studied in patients with stage T2-4N0-3M0 gastric cancer and stage T2-4N0-2M0-1 colorectal cancer. Activities of proteasomes and calpains in gastric and colorectal cancer tissues were higher than in the corresponding normal tissues. Changes in activities of proteasomes and calpains were mutually related. The appearance of lymphogenic metastases in gastric cancer was associated with the increase in calpain activity. The progress of colorectal cancer and development of lymphogenic and hematogenic metastases were associated with elevated chymotrypsin-like activity of proteasomes.

[Effect of tissue inhibitors of metalloproteinases (TIMP) on course and prognosis of head and neck squamous cell carcinoma].

The paper compares the levels of TIMP-1 and TIMP-2 in blood serum from patients with head and neck squamous cell carcinoma (HNSCC) to establish correlations with clinico-morphological parameters of the disease and prognosis. Our study involved 140 patients with HNSCC (T1-3N0-3M0) and 38 healthy donors. The level of serum TIMP-1 in patients was significantly higher (p < or = 0.02) while that of TIMP-2 was in inverse correlation with histological grade of tumor (p < or = 0.04) and age (p < or = 0.05). High level of TIMP-1 appeared an unfavorable prognosticator for 2-year relapse-free survival in HNSCC patients (p < or = 0.05). Those of both TIMP-1 and TIMP-2 identified in blood serum prior to therapy involved high risk of metastasis development (p < or = 0.05) and (p < or = 0.05), respectively.

[A relationship of squamous-cell metaplasia with the pattern of the course of a respiratory epithelial dysplastic process].

The morphological signs of squamous-cell metaplasia were revealed in 65 out of 193 patients with varying bronchial mucosal changes. A follow-up of the patients indicated that in the group of patients with epithelial dysplasia without squamous-cell metaplasia, the prognosis was good in 65.6% of cases. In the group of patients with epithelial dysplasia and squamous-cell metaplasia, the prognosis was poor in 86.2% of cases.

[Association of the degree of bronchial epithelial dysplasia to the serum level of non-enzymatic antioxidants].

The serum levels of non-enzymatic antioxidants were studied in patients with chronic nonspecific lung diseases with first-to-third-degree dysplasia of the bronchial epithelium (BE) before and after therapeutic correction. The development of BE dysplastic changes was ascertained to cause a considerable reduction in the content of antioxidant vitamin A. During the therapy contributing to reversal of BE dysplastic alterations, there was an increase in the serum levels of vitamin A and uric acid in patients with simple chronic bronchitis with both first- and second-degree dysplasia. If no therapeutic effect occurred, the systemic level of the antioxidants remained unchanged.

[Genetic polymorphism of glutathione-s-transferases theta1 and mu1 in lung cancer].

Polymorphism of GSTT1 and GSTM1 variant genes of phase II biotransformation of xenobiotics was studied in 181 patients with lung cancer. Null genotype frequency in cancer patients was higher than in healthy subjects. It was still higher in metastatic cancer disseminated to the regional lymph nodes as compared with localized tumor.

[Intravenous blood laser irradiation in patients with chronic nonspecific lung diseases and bronchial epithelial dysplasia]. / Vnutrivennaia lazernaia terapiia u bol'nykh HNZL s displaziei bronkhial'nogo épiteliia.

The study included patients with chronic nonspecific lung diseases (CNLD) and a morphologically verified diagnosis of first-second degree bronchial epithelial dysplasia (BED). Forty-six patients underwent intravenous blood laser irradiation (IBLI) (a study group); 45 patients received routine anti-inflammatory therapy (a control group). After therapy in the study group, the clinical and endoscopic signs of an inflammatory process were eliminated in 80.4 and 78.2% of cases, respectively. With morphological monitoring, a full and partial regression of BED occurred in 63.1 and 17.4%, respectively. In the control group, clinical and endoscopic improvements were achieved in 53.3 and 52.0% of cases. Dysplastic changes could be arrested only in 8.9% of cases. The differences were statistically significant (p < 0.05). Thus, IBLI is a highly effective treatment of dysplastic bronchial epithelial changes in patients with CNLD as compared with the controls.

[Cytometric criteria of evaluation of bronchial epithelium dysplasia with various types of therapy]. / Tsitometricheskie kriterii otsenki displazii bronkhial'nogo épiteliia pri razlichnykh vidakh lecheniia.

Examinations of 111 patients with chronic non-specific pulmonary lesion revealed the dysplastic changes in the bronchial wall in 90 of them. 47 patients received the routine anti-inflammatory therapy course and another 43 patients underwent the endoscopic laser therapy. Cytometry, made before and after therapy, provided for an objective differentiation of signs in different-severity bronchial dysplasia. The application of endoscopic laser irradiation in the correction of pretumor changers in the bronchial tree was shown to be effective in 95.3% of cases, whereas the routine treatment turned out to be ineffective.

[Complications of intraoperative radiotherapy and adjuvant chemotherapy in patients with lung cancer]. / Oslozhneniia pri sochetanii intraoperatsionnogo oblucheniia i ad''iuvantnoi khimioluchevoi terpii u bol'nykh rakom lëgkogo.

A high single dose of radiation can be delivered precisely to tumor or regional metastasis area by means of intraoperative radiation therapy (IORT). A program of combined treatment including radical IORT, postoperative distant gamma-therapy and adjuvant polychemotherapy with cisplatin and etoposide has been developed at the Center. Treatment was given to 43 patients with stage III lung cancer. Complications were reported in 23.3 +/- 6.0%. Prophylactic methods have been worked out to minimize IORT--related postoperative complications. The postoperative lethality rate was 2.3 +/- 1.4%. No program-related hazards have been observed. Complications from adjuvant polychemotherapy were moderate.

[Detection of herpesvirus markers in patients with precancerous lesions of the lungs and stomach]. / Rezul'taty vyiavleniia markerov gerpesvirusov u patsientov s predrakovymi zabolevaniiami lëgkikh i zheludka.

Blood-serum concentrations of antibodies to such viruses as herpes simplex virus, cytomegalovirus and Epstein--Barr virus have been assayed versus frequency of viral DNA detection in blood and saliva suspensions from patients with lung and stomach pre-cancers. Condition of local immunity of oral mucosa was assessed on the basis of saliva levels of secretory IgA and lysozyme. It was found that as local immunity of oral mucosa deteriorated, irrespective of pre-cancer localization, high titers of antiviral antibodies (class G) were established. The frequency of herpes DNA was much higher in lymphocytes and cellular sediment from saliva of patients with gastric pre-cancers. The role of herpes viruses in the pathogenesis of precancerous diseases and malignances is discussed.

[Morphologic changes of bronchial epithelium after intraoperative radiotherapy of lung cancer]. / Morfologicheskie izmeneniia bronkhial'nogo épiteliia posle provedeniia intraoperatsionnoi luchevoi terapii.

Integration of intraoperative radiotherapy (IORT) with combined treatment of non-small cell carcinoma of the lung seems to be promising. Of particular interest are studies of morphological changes in the bronchial epithelium under the influence of large single dose of radiation (15 Gy). Our clinical investigation included patients with non-small cell carcinoma stage III who had received different schedules of treatment at the Center's clinics. Examination was carried out of biopsy material sampled from the bronchus stump as well as from the contralateral segment of the bronchial tree following combined therapy. Exacerbation of the chronic process in the bronchus stump after IORT was reported. One month after bronchial resection, 80% of patients suffered from catarrhal sclerosing bronchitis. Its progression triggered on bronchial sclerosis in 20%. Catarrhal sclerosing bronchitis in the contralateral segment of the bronchial tree occurred in 60%; there was no significant correlation between catarrhal sclerosis, on the one hand, and the initial condition of the bronchus, on the other. In the control group, there were also signs of chronic bronchitis exacerbation on the site of operation. On the whole, the distribution of patients by bronchitis patterns remained unchanged.