Immunohistochemical analysis of NaPi2b protein (MX35 antigen) expression and subcellular localization in human normal and cancer tissues

Aim: To study the expression profile of the NaPi2b protein and its localization in breast, ovarian and lung cancer cells in relation to normal tissues adjacent to tumor. Methods: Immunohistochemical analysis with monoclonal antibody MX35 was applied for investigation of NaPi2b protein expression in breast, lung and ovarian carcinomas. Intensity of NaPi2b protein expression was calculated with semiquantitative scores. Results: NaPi2b (MX35) protein expression was detected in breast, lung and ovarian cancer cells and adjacent normal tissue. We have shown that in contrast to ovarian tumors in breast and lung tumors NaPi2b expression is down regulated comparing to correspondent normal tissues. Conclusion: This study provides the data on the pattern of NaPi2b expression and cellular localization in breast, lung and ovarian cancers, which might be useful for understanding the mechanism of transport and maintenance of inorganic phosphate in cancer and normal cells, as well as for developing novel immunotherapeutic approaches based on MX35 monoclonal antibody

Comparison of the effect of superplasticizing admixtures on the processes of cement hydration during mechanochemical activation

© 2016 Ruslan Ibragimov, Sergey Pimenov, Ilham Kiyamov, Ramil Mingazov, Laysan Kiyamova.The results of the study of the influence of two different super-plasticizers on the peculiarities of the process of hydration during mechanochemical activation (MCA) of a binder are given. The influence of SAS admixtures during the MCA of a binder on the kinetics of heat release and the alkalinity of the liquid phase of cement slurry is revealed. Technological properties of concrete mixture and physicomechanical properties of heavy-weight concrete, obtained by the MCA of a binder with the SAS admixtures, were studied. Dependencies of the influence of SAS on the processes and the nature of cement hydration were established, as well as on granulometric and phase composition of hydrated new formations. It was established that the application of SAS admixtures at the MCA of a binder makes it possible to substantially increase the efficiency of cement dispergation, which manifests itself in the decrease of grain sizes and the increase in specific surface area. The MCA of a binder with SAS admixtures makes it possible to obtain homogeneous cement suspension with the uniform distribution of water in the volume of cement, which positively affects physicomechanical properties and the quality of the obtained concrete. The application of SAS admixtures at the MCA of a binder results in the reduction of induction period of hydration by 2-5 hours, and also in the acceleration of crystallization period by 5-7 hours faster than in the reference compositions. MCA of cement suspension with SAS admixtures leads to the increase in the density of concrete mixture and a substantial increase in the strength limit of heavy-weight concrete at compression during all periods of hardening. With the MCA of a binder with SAS, an increase in the amount of crystalline phase by 10-12 % is observed in the first twenty-four hours of hardening, which confirms the conclusion about acceleration of cement hydration with the joint use of SAS and MCA. According to the results of a comprehensive study we received new data of the influence of super-plasticizing admixtures of different chemical bases on the peculiarities of the processes of hydration of a binder during MCA. The obtained data can be used for selection of superplasticizing admixtures in the production of the ferroconcrete products, obtained with the MCA of a binder, as well as in the technology of construction of buildings made of monolithic reinforced concrete

Quantitative analysis of SLC34A2 expression in different types of ovarian tumors

Aim: The main purpose of this study was to estimate the SLC34A2 gene expression in normal ovary and different types of ovarian tumors. Methods: We have investigated SLC34A2 gene expression level in papillary serous, endometrioid, unspecified adenocarcinomas, benign tumors, and normal ovarian tissues using real-time PCR analysis. Differences in gene expression were calculated as fold changes in gene expression in ovarian carcinomas and benign tumors compared to normal ovary. Results: We have found that SLC34A2 gene was highly expressed in well-differentiated endometrioid and papillary serous ovarian carcinomas compared to low-differentiated endometrioid carcinomas, benign serous cystoadenomas and normal ovary. Analysis of SLC34A2 gene expression according to tumor differentiation level (poor- and well-differentiated) showed that SLC34A2 is up-regulated in well differentiated tumors. Conclusion: Upregulation of SLC34A2 gene expression in well-differentiated tumors may reflect cell differentiation processes during ovarian cancerogenesis and could serve as potential marker for ovarian cancer diagnosis and prognosis

Technological qualities of natural bitumens of Tatarstan depending on chemical and geochemical characteristics of their composition

A variety of composition of natural bitumens of the Permian sediments of Tatarstan territory is given. The necessity of differentiation of the given raw by chemical and geochemical parameters of their composition is proved. Perspective lines of use of various types of natural bitumens are considered

CRISPR/Cas9 Technique for Identification of Genes Regulating Oxaliplatin Resistance of Pancreatic Cancer Cell Line

© 2016, Springer Science+Business Media New York.Genome editing approach based on prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats) system is a simple and useful way to investigate gene functions on a genome-wide scale. It is especially important for cancer research because of genetic contribution to tumor development. We applied this technique in a high-throughput screening format to find genes that could be involved in chemotherapy resistance of pancreatic cancer. We used AsPC1 cell line expressing doxycycline-inducible Cas9 to screen two sgRNA lentiviral libraries: (1) cell cycle genes (CC, 983 genes, ∼12,000 sgRNA) and (2) genome-wide (GW, ∼90,000 sgRNA). These sets of cells with different gene knockouts were treated with oxaliplatin to identify knockouts which increase sensitivity to the drug. We have performed screening both in vitro and in vivo settings. For the in vivo arm of our experiments, peritoneal carcinomatosis model in severe combined immunodeficiency (SCID) mice was created by intraperitoneal injection of AsPC1/Cas9 cells infected with sgRNA library. Genomic DNA from cells and animal tumor material was analyzed using next generation sequencing (NGS) to obtain data about representation of sgRNA. Preliminary data allowed us to identify genes potentially modulating oxaliplatin sensitivity

Panel of SEREX-defined antigens for breast cancer autoantibodies profile detection

© 2016 Informa UK Limited, trading as Taylor & Francis Group.Content: Identification of panel of SEREX-defined antigens for breast cancer autoantibodies profile detection. Objective: To create panel of antigens that can differentiate breast cancer patients and healthy individuals. Methods: SEREX (serological analysis of cDNA expression libraries) method, ELISA (enzyme-linked immunosorbent assay), qPCR (quantitative polymerase chain reaction). Results: In large-scale screening of 16 SEREX-antigens by sera of breast cancer patients and healthy donors, a combination of six antigens (RAD50, PARD3, SPP1, SAP30BP, NY-BR-62 and NY-CO-58) was identified, which can differentiate breast cancer patients and healthy donors with 70% sensitivity and 91% specificity. Elevated mRNA expression of SPP1 gene was revealed in breast tumors (2–7-fold) that correlated with SPP1 antigen immunoreactivity in autologous patients’ sera. Conclusions: The new panel of six SEREX-antigens was proposed, which enables creation of serological assay for breast cancer diagnostics and/or prognosis

Genesis of biomarker hydrocarbons in the environment and their role in assessing an oil pollution source

Oil spills are classified according to chemical types that reflect qualitative and quantitative changes in petroleum composition during biodegradation in soil. Possible trends in composition of biomarker petroleum hydrocarbons in polluted soil, as determined by natural and climatic conditions and by activation of soil microflora, were revealed. It was shown that the use of selective bioremediation methods could lead to a noticeable redistribution of the relative amount of C27 : C28 : C29 steranes in marine crude oils owing to more intense degradation of C 27 steranes (5α, 14β, 17β, 20R and 20S, as well as 5α, 14α, 17α, 20R and 20S) as compared with their higher C29 homologues. The observed changes in the composition of steranes suggest another, namely, terrigenous origin of petroleum, and, thus, may lead to an erroneous conclusion concerning the genotype of crude oil as a pollution source. It was found that hopane hydrocarbons are highly stable under similar conditions. It was shown that the set of biomarker hydrocarbons might be useful as biological indicators in the assessment of a petroleum pollution source. © MAIK "Nauka/Interperiodica"

Application of serex-analysis for identification of human colon cancer antigens

Copyright © Experimental Oncology, 2015. Background: Colorectal, lung and breast tumors are the most devastating and frequent malignances in clinical oncology. SEREX-analysis of colon cancer leads to identification of more than hundred antigens which are potential tumor markers. With idea that immunoscreening with pool of allogeneic sera is more productive for antigen isolation, SEREX-analysis was applied to four cases of stages II-IV primary colon tumor and 22 new antigens were isolated. Objective: To characterize 22 primary colon cancer antigens isolated by SEREXtechnique. Materials and Methods: Allogenic screening, real-time PCR analysis. Results: After allogeneic immunoscreening, for 5 of 22 (22%) isolated antigens were confirmed colon cancer restricted serological profile solely positive for 14% of tested colon cancer sera. Through these five antigens, KY-CC-17/β-actin has cytoskeleton function; KY-CC-14/ACTR1A and KY-CC-19/TSGA2 participate in chromosome segregation; KY-CC-12/FKBP4 regulates steroid receptor function and KY-CC-15/PLRG1 is a component of spliceosome complex. For the last four antigens tested were found aberrant mRNA expression in some cases of colon tumor. Conclusion: The exploration of identified antigens may define suitable targets for immunotherapy or diagnostic of colon cancer

Autoantibody Response to ZRF1 and KRR1 SEREX Antigens in Patients with Breast Tumors of Different Histological Types and Grades

© 2016 Lada Dyachenko et al.Purpose. To investigate a frequency of antibody response to SEREX-identified medullary breast carcinoma autoantigens ZRF1 and KRR1 in sera of breast cancer patients taking into account clinical and molecular characteristics of tumors for opening of new perspectives in creation of minimally invasive immunological tests for cancer diagnostics. Methods. Enzyme-linked immunosorbent assay and bioinformatics analysis. Results. Increased frequency of antibody response was found in sera of breast cancer patients to ZRF and KRR1 antigens. The antibody response to these antigens was higher in sera of patients with invasive ductal carcinoma than in sera of patients with other histological types of breast tumors. Moreover, more frequent antibody response to ZRF antigen was found in sera of patients with less aggressive tumors. The sequence analysis of ZRF1 antigen SEREX clones obtained from cDNA libraries of different tumors demonstrates that they encode different protein isoforms. Conclusion. Tumor-associated antigens KRR1 and ZRF1 and their cognate autoantibodies could be considered as potential molecular markers of breast cancer which need to be further investigated

The study of phosphate transporter NaPi2b expression in different histological types of epithelial ovarian cancer

The identification of markers that are specifically expressed by different histological types of epithelial ovarian cancer (EOC) may lead to the development of novel and more specific diagnostic and therapeutic strategies. Sodium-dependent phosphate transporter NaPi2b (or MX35 ovarian cancer antigen) is a novel perspective marker of EOC. To date, the studies on NaPi2b/MX35 expression in different histological types of EOC are limited. Aim: To examine NaPi2b/MX35 expression in different histological types of epithelial ovarian tumors. Methods: Here, we describe the analysis of NaPi2b expression in serous (n = 17), endometrioid (n = 8), and mucinous ovarian tumors (n = 3) by Western-blotting (WB), immunohistochemistry and RT-PCR. Results: The results of immunohistochemical and WB analysis showed that benign and well-differentiated malignant papillary serous tumors as well as well-differentiated malignant endometriod tumors overexpress NaPi2b protein. However, no overexpression of NaPi2b was detected in benign and malignant mucinous tumors as well as in poorly differentiated endometriod tumors. Notably, the expression NaPi2b mRNA was detected in all investigated histological types of EOC. Conclusion: We have shown the differential expression profile of NaPi2b phosphate transporter at protein level in various histological types of epithelial ovarian cancer. This finding might facilitate the development of more effective approaches for diagnosis and treatment of this disease