DNA Methylation of a Group of Long Non-Coding RNA Genes at Different Stages of Ovarian Cancer Dissemination.

There are three types of metastases in ovarian cancer: lymphogenous, hematogenous, and peritoneal. Dissemination of the tumor in the peritoneum is directly related with the development of ascites and a poor prognosis. The purpose of this study is to determine changes in the methylation level of a group of long non-coding RNA (lncRNA) genes at different stages of ovarian cancer progression. The methylation level of 7 lncRNA genes (LINC00472, LINC00886, MAFG-DT, SNHG1, SNHG6, TP53TG1, and TUG1) was studied by quantitative methyl-specific PCR in 93 samples of ovarian tumors and 75 paired samples of histologically normal tissue, as well as in 29 peritoneal macroscopic metastases. Using the nonparametric Mann-Whitney test, a significant (p<0.001) increase in the level of methylation of the LINC00886, SNHG1, SNHG6, and TUG1 genes in the tumor tissue was shown. For the LINC00472, LINC00886, and SNHG6 genes, a significant relationship was found with the clinical stage (p≤0.001), as well as with the appearance of metastases for the LINC00472 (p<0.001) and SNHG6 (p=0.005) genes. There was a significant increase in the level of methylation of MAFG-DT and TP53TG1 (p<0.001) genes, as well as a decrease in LINC00886 (p=0.003) in peritoneal metastases relative to the primary focus. Methylation of the LINC00472 and SNHG6 genes can be considered as a factor in initiating ovarian cancer metastasis, and methylation of the LINC00886, MAFG-DT, and TP53TG1 genes as a colonization factor for metastases in the peritoneum. Thus, a relationship between methylation of a group of lncRNA genes at different stages of ovarian cancer dissemination was shown, which is important for understanding the mechanisms of these processes and for developing innovative approaches to ovarian cancer therapy.

The Role of Methylation of a Group of microRNA Genes in the Pathogenesis of Metastatic Renal Cell Carcinoma.

The role of methylation of 9 miRNA genes in the pathogenesis of metastatic clear cell renal cell carcinoma was determined by quantitative methylation-specific PCR (MS-PCR). For 5 genes (MIR125B-1, MIR137, MIR193A, MIR34B/C, and MIR375), a significant correlation of high methylation level with late (III-IV) stages, large size (T3+T4) of the tumor, and metastasis to lymph nodes and/or distant organs was revealed. For another group of genes (MIR125B-1, MIR1258, MIR193A, MIR34B/C, and MIR375), a statistically significant correlation of high methylation level with loss of differentiation in the tumor (G3-G4) was found, and the opposite pattern was found for MIR203A. A total of 7 microRNA genes (MIR125B-1, MIR1258, MIR137, MIR193A, MIR203A, MIR34B/C, and MIR375) were identified, the methylation of which is associated with the progression of metastatic clear cell renal cell carcinoma. For 6 of them (except MIR34B/C) these data were obtained for the first time. Thus, new factors of the development and progression of clear cell renal cell carcinoma were identified as potential biomarkers for the early diagnosis and prognosis of metastatic clear cell renal cell carcinoma.

Hypermethylation of Long Non-Coding RNA Genes Group in the Breast Cancer Development and Progression.

We analyzed changes in the level of methylation of CpG islands in four long non-coding RNA (lncRNA) genes MEG3, ZNF667-AS1, GAS5, and SEMA3B-AS1 as promising markers of breast cancer. Methylation analysis was performed by quantitative methylation-specific PCR on a set of 38 paired (tumor/normal) breast cancer samples. Significantly (p<0.001) increased methylation was shown for three of the four lncRNA genes: MEG3, ZNF667-AS1, and SEMA3B-AS1. We found significant correlations of the methylation level of all the studied lncRNA genes with the stage of cancer and with lymphogenic metastasis, and for MEG3 and ZNF667-AS1 also with the tumor size. Methylation of ZNF667-AS1, and SEMA3B-AS1 genes in breast cancer was detected for the first time. Based on these findings, new potential markers for the diagnosis and prognosis of breast cancer can be proposed.

Synergy between the Levels of Methylation of microRNA Gene Sets in Primary Tumors and Metastases of Ovarian Cancer Patients.

We studied the correlations between the levels of methylation of a group of 21 microRNA genes in 99 primary tumors and 29 macroscopic peritoneal metastases of ovarian cancer. Analysis of the level of methylation by quantitative methylation-specific PCR showed that co-methylation was detected for 13 pairs of microRNA genes in primary tumors and for 22 pairs in metastases. Pairs of microRNA genes that have shown significant co-methylation can be involved in common processes and pathways of gene regulation and interaction and can have common target genes. The results are highly significant and pairs of microRNA genes can be proposed as new potential markers for the diagnosis and prognosis of ovarian cancer metastasis.

Optimized Marker System for Early Diagnosis of Breast Cancer.

Changes in the methylation levels of 21 microRNA genes in 91 breast cancer samples in comparison with paired samples of histologically unchanged tissue were studied by quantitative methylation-specific PCR. For 19 microRNA genes, a significant increase in the methylation level in tumors in comparison with normal tissues was shown (Mann-Whitney test). When considering the data for breast cancer samples only from patients with clinical stages I and II (59samples), 17 genes with a significantly increased level of methylation were identified. Increased methylation level for 11 genes (MIR124-1, MIR124-3, MIR125B-1, MIR127, MIR129-2, MIR132, MIR137, MIR193a, MIR34B/C, MIR375, and MIR9-1) compared to the paired norm was highly significant (p<0.001, FDR=0.01). The ROC analysis was used to optimize a set of markers for diagnosing breast cancer at the early stages consisting of 4 microRNA genes: MIR125B1, MIR127, MIR1258, and MIR132; the system is characterized by 100% specificity, 85% sensitivity, and AUC=0.924. Importantly, 100% specificity eliminates false positive results. Detection of methylation of at least one of the 4 genes of this set is sufficient to classify the patient's sample as breast cancer.

Hypermethylation of Genes in New Long Noncoding RNA in Ovarian Tumors and Metastases: A Dual Effect.

The role of methylation in the regulation of genes of long noncoding RNA (lncRNA) is still poorly understood. We revealed new hypermethylated lncRNA genes in ovarian tumors and their effect on metastasis of ovarian cancer. A multiple and significant (p<0.001) increase in methylation of a group of lncRNA genes (MEG3, SEMA3B-AS1, ZNF667-AS1, and TINCR) was shown by quantitative methylation-specific PCR using the non-parametric Mann-Whitney test. Moreover, methylation of SEMA3B-AS1, ZNF667-AS1, and TINCR genes in ovarian cancer tumors was detected for the first time. Comparative analysis of 19 samples of peritoneal metastases and paired primary tumors showed a significant decrease in the methylation level of the same 4 genes: MEG3 (p=0.004), SEMA3B-AS1 (p=0.002), TINCR (p=0.002), and ZNF667-AS1 (p<0.001). Reduced methylation of suppressor lncRNA genes in peritoneal metastases is probably associated with the involvement of these lncRNA in the regulation of plastic reversion of the epithelial-mesenchymal transition to the mesenchymal-epithelial transition. Thus, the effect of lncRNA and their methylation on the development of tumors and metastases of ovarian cancer was demonstrated, which is important for understanding of the pathogenesis and mechanisms of metastasis of ovarian cancer. New properties of lncRNA can find application in the development of new approaches in the therapy of ovarian cancer.

Droplet Shape and Wetting Behavior under the Influence of Cyclically Changing Humidity.

Relative humidity (RH) plays a crucial role in wetting and spreading phenomena by affecting the evaporation rate, evaporation modes, and spreading dynamics via precursor film formation, surface modification, and surface tension alteration. We examined the effect of the periodically varied relative humidity (RH) between low (20%) and high (85%) levels on the wetting of the droplet of nonhygroscopic (pure surfactants) and hygroscopic (ethylene glycol, glycerol) liquids on a hydrophobic surface. It was revealed that the changing RH induces two modes of transition between the wetting states of the droplet: with hysteresis and without hysteresis. Droplets of both nonhygroscopic and hygroscopic liquids exhibit shape hysteresis during the first cycle: (i) droplets of surfactants irreversibly spread saving an initial volume; and (ii) ethylene glycol and glycerol droplets irreversibly absorb the moisture, increasing the volume and the base diameter. Further, cyclically changing the RH results in the droplet breathing effect, i.e., the nonhysteresis transition of the droplet shape between two wetting states corresponding to the minimum and maximum RH levels. In the case of the glycerol droplet for three cycles of the RH variation, the volume hysteresis (the droplet volume increases in each cycle) was observed. This is determined by the moisture absorption due to high hygroscopicity of glycerol. We also revealed that for all liquids studied, the droplet spreading at each increase in RH started at reaching the RH threshold level.

[Comparative analysis of side-by-side antegrade stenting and Y-biliary stenting for malignant biliary stricture]. / Sravnitel'nyi analiz antegradnogo stentirovaniia side-by-side i Y-stentirovaniia pri zlokachestvennoi zhelchnoi strikture vorot pecheni.

In the time period from 2009 to 2017 in Botkin Hospital (Moscow) bilateral biliary stenting was performed in 43 patients with malignant common hepatic duct stricture. Patients were divided into 2 statistically homogeneous groups: 'side-by-side' stenting in 28 patients and percutaneous 'Y'-biliary stent placement in 15 patients. The causes of malignant obstruction were as follows: in the 1st group 13 (46.4%) patients with extrahepatic cholangiocarcinoma (46.4%), 8 (28.6%) patients with intrahepatic cholangiocarcinoma, 4 (14.3%) patients with gallbladder cancer, 3 (10.7%) patients with metastatic cancer. In the 2nd group 6 (40%) patients with extrahepatic cholangiocarcinoma, 4 (26.6%) patients with intrahepatic cholangiocarcinoma, 2 (13.3%) patients with gallbladder cancer, 3 (20%) patients with metastatic cancer. Characteristics of patients: age - 1st group 71.2±5.1 years, 2nd group 74.3±5.5 years; sex - (m/f) 1st group 18/10, 2nd group 9/6; location of stricture - 1st group Bismuth IIIa/IIIb 17/11, 2nd group Bismuth IIIa/IIIb 11/4; mean level of bilirubin - 1st group 284±8.2 µmol/l; 2nd group 311±7.4 µmol/l. Technically all procedures were successful (100%). No complications and mortality associated with the procedure was recorded. Clinically significant results were achieved in 26 (92.8%) patients in Group 1 and in 13 (86.7%) patients in Group 2 (p=0.043). Following stenting procedures, 23 (82.1%) patients in Group 1 and 11 (68.8%) patients in the Group 2 had chemotherapy (p=0.047). 19 patients from the 1st group and 10 patients from the 2nd group died due to tumor progression of the underlying disease, other patients are under care of a physician. Median survival rate: 1st group (12 patients - 50 days, 7 patients - 100 days, 9 patients are alive at the time of writing); 2nd group (7 patients - 50 days, 4 patients - 100 days, 4 patients are alive at the time of writing). The results of this study showed that 6 (21.4%) patients from the 1st group and 4 (26.7%) patients from the 2nd group had biliary stent occlusion (p=0.041). The average period of stent function in the 1st group was 78±4.5 days, and 63±4.8 days in the 2nd group (p=0.036). Based on the obtained results, it is recommended to use the 'side-by-side' method of bilobar biliary stenting in patients with malignant common hepatic duct strictures.

Removal of micrometer size particles from surfaces using laser-induced thermocapillary flow: Experimental results.

HYPOTHESIS: Reducing particle contaminations on solid and delicate surfaces is of great importance in a number of industries. A new non-destructive method is proposed, which is based on the laser-induced thermocapillary effect for the removal of micron size particles from surfaces. The cleaning mechanism is related to the surface-tension-driven flows produced by the laser heating of thin layer of a cleaning liquid deposited onto a surface contaminated with particles. EXPERIMENTS: Focusing the laser irradiation into the line laser beam allowed using this method for a large-scale cleaning of surfaces. Hexadecane was used as a cleaning liquid to remove micron-sized polyethylene, Teflon, talc and Al2O3 particles from surfaces of welding glass, carbolite and soft magnetic disc using the line beam of the IR laser. FINDINGS: A good cleaning efficiency was achieved for cases of polyethylene and Teflon particles on both the complete wettable welding glass and the low-wettable soft magnetic disc, while in case of oleophilic talc and Al2O3 particles the effectiveness of the cleaning method was lower on all three substrates investigated. The thermal influence of the laser irradiation on substrates used was measured with infrared camera. It was shown that temperature in the irradiated area during the long-time heating increases insignificantly and cannot cause any damage of the substrate.

Wetting films of aqueous solutions of Silwet L-77 on a hydrophobic surface.

The formation of wetting films of aqueous solutions of Silwet L-77 on hydrophobic substrates takes place only at concentrations above the critical aggregation concentration (CAC). At concentrations above the critical wetting concentration (CWC) a new phenomenon was found: the formation of multilayered spots of thicker films in the wetting film of aqueous solutions of Silwet L-77 on hydrophobic surfaces. An expansion of the thicker spots within the film and the formation of "channels" between the spots and the edge of the film led to a continuous shrinkage of the wetting film and its disappearance in the end. We suggested that the multiple thicker films originate from the multilayer structuring of trisiloxane bilayers within the wetting film.

[Intracellular immunoglobulins in Namalwa and U266 cells co-cultivated with mesenchymal stromal cells].

There are contradictory data concerning the influence of mesenchymal stromal cells (MSC) on immunoglobulin (Ig) production. Most of them were obtained using MSC from bone marrow. Properties of MSC from other tissues are elusive. In the present work MSC cultures were derived from umbilical cord, adipose tissue, and bone marrow of healthy donors, as well as from bone marrow of patients with autoimmune diseases. MSC from all these sources had similar surface markers phenotype. The influence of co-cultivation with MSC at exponential or stationary phase on IgM and IgE content in Namalva and U266 cells was evaluated. MSC from bone marrow of healthy donors had no effect on IgM and IgE production. Proliferating MSC obtained from patients with Crohn's disease and multiple sclerosis stimulated Ig production. Exponentially growing MSC derived from umbilical cord and adipose tissue also stimulated Ig synthesis. MSC at stationary cultures amplified IgM production in Namalva cells and suppressed IgE synthesis in U266. Thus, MSC with similar phenotype but derived from different sources differ in their capacity to modulate Ig production in B-lymphoid cells. The effect of MSC depends on their growth stage and may differ for lymphoblastoid and myeloma cells.

[The detection of clozapine in the tissues of an exhumed corpse].

The authors report the case of finding clozapine in the tissues of an exhumed corpse. The circumstances surrounding the death and the results of primary examination of the corpse are described; in addition, the results of repeated expertise are presented. The data obtained in this study are compared with the relevant literature publications. Special attention is given to the peculiarities of interpretation of the results of forensic chemical expertise essential for the formulation of the scientifically-sound conclusion.

[Comparative studies on the genotoxic activity of a new palladium (II) acidocomplex vs cisplatin in human blood lymphocytes in vitro].

A comparative study on the genotoxic activity of cisplatin versus morfozol, the first representative of a new class of cation-anion complexes of palladium [AH]2[PdCl4] (where A-methylmorpholine) has been performed using human lymphocytes in vitro. The results of the DNA-DNA cross-linking activity investigations showed that both compounds studied exhibited biphasic dose-effect relationship: a linear decrease in the DNA percent in the comet tail and the region of the "plateau". However, in the "plateau" region, morfozol reduced the DNA percent in the comet tail up to 6 times while cisplatin caused a 2-fold decrease only. Morfozol, like cisplatin, inducing DNA-protein cross-linking and generating reactive oxygen species, was more effective than cisplatin.

[Clozapine intoxication: theoretical aspects and forensic-medical examination ].

This literature review is focused on diagnostics of acute clozapine intoxication with the fatal outcome. According to the Russian authors, clozapine intoxication ranks first in the structure of criminal poisoning and accounted for 99.7% of all the cases that occurred in Moscow during the period from 2003 to 2006. Toximetric investigations of clinical manifestations of clozapine intoxication revealed that the threshold clozapine concentration in blood is 0.12 ± 0.06 mg/I, the critical and lethal concentrations are 1.01 ± 0.2 mg/I and 3.5 ± 1.5 mg/I respectively. Autopsy on corpses of the victims of clozapine intoxication showed that most clozapine-induced pathological changes have a non-specific character (including largely circulatory disorders and dystrophic changes in parenchymatous organs). Clozapine poisoning is associated with the lengthening of QT-interval on ECG; at the values in excess of 500 ms, the risk of severe arrhythmia and sudden death significantly increases. Clozapine intake may lead to the development of potentially fatal myocarditis (the so-called clozapine-associated eosinophilic myocardium) in somatically healthy subjects. Foreign researchers report the possibility of a post-mortem increase of blood clozapine content compared with its antemortem level. They also showed that simultaneous use of substances stimulating activity of cytochrome P-450 enzymes (ethyl alcohol, finlepsin, fenitrin, nicotine) and clozapine accelerates metabolism and thereby reduces clozapine concentration in blood. It is concluded that comprehensive investigations of clozapine intoxication are needed taking into consideration pathomorphological changes induced by this agent, its potential interaction with other factors influencing human body, and the results of forensic chemical expertise of the fatal cases.

[Abortifacient properties and effect of an misoprostol analogue 11-deoxy-misoprostol on the contractile activity of the rat uterus].

Ethyl ether of 11-deoxy-16-hydroxy-16-metylprostaglandin E1 (11-deoxymisoprostol) increases the contractile activity of uterine horn segments isolated from nonpregnant rats and produces abortive effect when given in a period of time within 1 - 16 days of pregnancy. The drug action is related to a decrease of the progesterone level in ovarian incubates of pregnant rats.

Choleretic activity of 2-demethoxycarbonyl-2-ethoxycarbonyl-11-deoxymisoprostol on the model of CCl4-induced hepatitis.

Therapeutic administration of 11-deoxymisoprostol had a hepatoprotective effect, which manifested in a decrease in the content of alanine transaminase and aspartate transaminase in blood plasma, and produced a choleretic effect in rats with CCI4-induced toxic hepatitis.

[Hepatoprotective effect of 2-demethoxycarbonyl-2-ethoxycarbonyl-11-deoxymisoprostol].

2-Demethoxycarbonyl-2-ethoxycarbonyl-11-deoxymisoprostol (11-DMP) produces antioxidant effect on the models of toxic hepatitis induced by paracetamol and carbon tetrachloride. The drug normalizes the lipid peroxidation (LPO) prosess in rat liver of the rat and the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the rat blood, thus demonstrating hepatoprotective action.

[Optimal therapy of children with bronchial asthma at Pyatigorsk spa].

The study of efficacy of bronchial asthma children's rehabilitation at Pyatigorsk spa with different schemes of treatment has shown that combined treatment with dry air-radon baths and halotherapy is more effective than each of these modalities alone.

Gastroprotective properties of 11-deoxymisoprostol (prostaglandin E1 analog) and its effect on the level of sialic acids in gastric tissue of rats with peptic ulcer.

11-Deoxymisoprostol (prostaglandin E1 analog) exhibited a pronounced gastroprotective effect on various models of experimental ulcers induced by nonsteroid antiinflammatory drugs. A relationship between high resistance of the gastroduodenal mucosa under the effect of 11-deoxymisoprostol and changes in the level of sialic acid was detected.

[Efficacy of transcutaneous endobiliary procedures in patients with obstructive jaundice].

Transcutaneous endobiliary procedures (TEP) were performed in 378 patients with obstructive jaundice (OJ). External decompression of the biliary ducts was carried out as the first stage of treatment in all the patients with OJ of various etiology. This procedure led to reduction of cholangitis symptoms and hepatic failure, improvement of general condition of the patients before radical surgery. Endoprosthesis of the bile ducts made in 121 patients allowed physiological passage of bile into the intestine and improved quality of life especially in inoperable patients with hepatic and pancreatoduodenal tumors. During external drainage in 27 (7.1%) patients dislocation of drainage of the common bile duct occurred. Postoperative lethality was 8.5%. After open surgeries 2 (1.2%) patients died.