COMPREHENSIVE OVERVIEW OF ADAPTOR PROTEIN RUK/CIN85 ROLES IN CANCER

Aim. This study is focused on a comprehensive overview of mechanisms and processes involved in the acquisition of cancer cell plasticity in a manner dependent on the adapter protein Ruk/CIN85 (in rodents, Ruk — regulator of ubiquitous kinase; in human CIN85 — Cbl-interacting protein of 85 kDa, encoded by SH3KBP1 gene).. Methods. Gene expression was evaluated using RT2-PCR and Western blotting, cell proliferation and survival were analyzed using MTT and/or dye exclusion assays, motility was assessed by scratch test and Transwell assay, enzyme activities were measured using spectrophotometric assays. In vivo metastasis were studies using experimental metastasis model. Conclusion. This study discloses various aspects of cancer cells plasticity, such as EMT, stemness, metabolic changes, ECM components, and drug resistance in dependence on adaptor protein Ruk/CIN85 expression level

ПОЛУЧЕНИЕ НИКЕЛЬ-КОБАЛЬТОВОГО КОНЦЕНТРАТА ПРИ ПЕРЕРАБОТКЕ РЕНИЙСОДЕРЖАЩЕГО ЖАРОПРОЧНОГО СПЛАВА

The article describes the results of HAS32-VI superalloy electrochemical processing in nitric acid solutions in galvanostatic mode. Experiments were conducted for electrochemical dissolution of HAS32-VI superalloy in galvanostatic mode using nitric acid with a concentration of 100 g/l at different values of anode current density. It is found that this leads to quantitative separation of superalloy HAS32-VI components. The anode slurry is the concentrate of refractory metals – niobium, tantalum, molybdenum and tungsten. Cobalt, rhenium and the principal amount of aluminum, chromium and nickel partially pass into electrolyte. The flow diagram of HAS32-VI superalloy processing was suggested that provides for generation and separation of the principal nickel and cobalt amount at the first stage with the formation of Ni–Co-containing metal sediment.Изложены результаты электрохимической переработки жаропрочного сплава марки ЖС32-ВИ, реализованной в гальваностатическом режиме в азотнокислом электролите. Проведены эксперименты по электрохимическому растворению в гальваностатическом режиме этого сплава с использованием раствора азотной кислоты с концентрацией 100 г/л при различных значениях плотности тока. Показано, что происходит количественное разделение составляющих его компонентов: в анодном шламе концентрируются тугоплавкие металлы – ниобий, тантал, молибден и вольфрам, а в электролит переходят частично кобальт и рений и основное количество алюминия, хрома и никеля. Предложена принципиальная технологическая схема переработки сплава ЖС32-ВИ, в которой получение и отделение основной массы никеля и кобальта осуществляются на первой стадии, с формированием металлического Ni–Co-содержащего осадка

МУЛЬТИСПИРАЛЬНАЯ КОМПЬЮТЕРНАЯ ТОМОГРАФИЯ В ДИАГНОСТИКЕ ПАТОЛОГИИ ДУГИ АОРТЫ И АНОМАЛИЙ БРАХИОЦЕФАЛЬНЫХ АРТЕРИЙ У ДЕТЕЙ ПЕРВОГО ГОДА ЖИЗНИ

Aortic malformations often cause critical states in the neonatal period. MSCT method allows you to get a detailed understanding of the anatomy of vice and identify concomitant bronchopulmonary disease. This paper presents a comparative analysis of 27 patients of the first year of life with the pathology of the aortic arch, which were performed echocardiography and MSCT. After analysis of both research methods in 4 patients revealed a discrepancy echocardiography and MSCT data which led to a change in surgical approach. Just according to SITC 2 patients revealed tracheal stenosis, in one case, isolated, in another due to compression of the vascular ring. MSCT is a highly informative method for diagnosing pathology of the aortic arch and associated bronchopulmonary disease in children in the first year of life.Пороки развития аорты зачастую являются причиной критических состояний в неонатальном периоде. Метод МСКТ позволяет получить детальное представление об анатомии порока и выявить сопутствующую бронхолегочную патологию. В работе представлен сравнительный анализ данных 27 пациентов первого года жизни с патологией дуги аорты, которым была выполнена ЭхоКГ и МСКТ. После анализа обоих методов исследования у 4 пациентов выявлено несоответствие данных ЭхоКГ и МСКТ, что привело к изменению хирургической тактики. Также по данным МСТК, у 2 пациентов выявлен стеноз трахеи, в одном случае - изолированный, в другом - за счет сдавления сосудистым кольцом. МСКТ является высокоинформативным методом диагностики патологии дуги аорты и сопутствующей бронхолегочной патологии у детей первого года жизни

Expression of adaptor protein Ruk/CIN85 isoforms in cell lines of various tissue origins and human melanoma

Aim: Development of monoclonal and polyclonal antibodies against recombinant GST-fused proteins including correspondingly N- and C-terminal parts of Ruk/CIN85 adaptor protein. Analysis of Ruk/CIN85 expression patterns in cell lines of various tissue origins and human melanoma. Methods: Recombinant GST-fused fragments of Ruk/CIN85 were expressed in bacterial system and affinity purified. Monoclonal antibodies against SH3A domain of Ruk/CIN85 were produced using hybridoma technique. The specificity of generated antibodies was examined by ELISA. Polyclonal antibodies against C-terminal coiled-coil region of Ruk/CIN85 were affinity purified from serum of immunized rabbit. Expression patterns of Ruk/CIN85 isoforms and their subcellular localization in cell lines of various tissue origins and human melanoma samples were analyzed by immunoblotting, immunoprecipitation and immunofluorescence microcopy. Results: Ruk/CIN85 is ubiquitously expressed SH3-containing adaptor/scaffold protein which plays important roles in signalling processes. N-terminal half of Ruk/CIN85 molecule, including three SH3 domains, and its C-terminal coiled-coil region were used as antigens to produce monoclonal and polyclonal antibodies, respectively. Hybridoma cell lines secreting monoclonal antibodies (mAbs) to SH3 fragment of Ruk/CIN85 were established. One of the mAbs was extensively characterized and designated as MISh-A1. It was shown that this mAb recognizes an epitope, which resides within first SH3A domain. Polyclonal anti-Ruks Abs affinity purified from serum of immunized rabbit specifically recognized main Ruk/CIN85 isoforms, both endogenous and recombinant, in lysates of HEK293 cells. Notably, produced Abs did not cross-react with CD2AP, the member of the same family of adaptor/scaffold proteins. Multiple molecular forms of Ruk/CIN85 with apparent molecular weights of 130, 80–85, 70–75, 50–56, 34–40 and 29 kD were detected in cell lyzates of NIH3T3, Cos1, L1210, HEK293, Ramos, HeLa S3, MDCK, C6, A549 and U937 using anti-Ruk antibodies. Oligomerization between p85 and p50–56 forms of Ruk/CIN85 was revealed in C6 and NIH3T3 cells, but not in HeLa S3 and HEK293 cells by immunoprecipitation using MISh-A1 antibody following anti-Ruk Western-blot analysis. Using immunofluorescent microscopy and anti-Ruk antibodies, endogenous Ruk-variates were found mostly in cytoplasm of C6, NIH3T3, HEK293 cells and at lower level — in nuclei. Conclusion: Patterns of Ruk/CIN85 molecular forms expression are cell-specific and determined by cellular context. Assembly of oligomeric complexes between p85 and p50–56 Ruk/CIN85 isoforms in C6 and NIH3T3 cells but not in HeLa S3 and HEK293 cells may reflect their specific biological roles in different cell lines. High level of full-length Rukl/CIN85 form expression was revealed in extracts of human melanoma samples. Abs described in this paper may prove useful in future studies of Ruk/CIN85 expression and function in normal and transformed cells

MULTISPIRAL COMPUTER TOMOGRAPHY IN DIAGNOSIS OF THE CONGENITAL DISEASE OF THE AORTA AND BRACHIOCEPHALIC ARTERIES IN CHILDREN FIRST YEAR OF LIFE

Aortic malformations often cause critical states in the neonatal period. MSCT method allows you to get a detailed understanding of the anatomy of vice and identify concomitant bronchopulmonary disease. This paper presents a comparative analysis of 27 patients of the first year of life with the pathology of the aortic arch, which were performed echocardiography and MSCT. After analysis of both research methods in 4 patients revealed a discrepancy echocardiography and MSCT data which led to a change in surgical approach. Just according to SITC 2 patients revealed tracheal stenosis, in one case, isolated, in another due to compression of the vascular ring. MSCT is a highly informative method for diagnosing pathology of the aortic arch and associated bronchopulmonary disease in children in the first year of life

Increased levels of the HER1 adaptor protein Ruk(l)/CIN85 contribute to breast cancer malignancy

The adaptor protein regulator for ubiquitous kinase/c-Cbl-interacting protein of 85kDa (Ruk/CIN85) was found to modulate HER1/EGFR signaling and processes like cell adhesion and apoptosis. Although these features imply a role in carcinogenesis, it is so far unknown how and by which molecular mechanisms Ruk/CIN85 could affect a certain tumor phenotype. By analyzing samples from breast cancer patients, we found high levels of Rukl/CIN85 especially in lymph node metastases from patients with invasive breast adenocarcinomas, suggesting that Rukl/CIN85 contributes to malignancy. Expression of Rukl/CIN85 in weakly invasive breast adenocarcinoma cells deficient of Rukl/CIN85 indeed converted them into more malignant cells. In particular, Rukl/CIN85 reduced the growth rate, decreased cell adhesion, enhanced anchorage-independent growth, increased motility in both transwell migration and wound healing assays as well as affected the response to epidermal growth factor. Thereby, Rukl/CIN85 led to a more rapid and prolonged epidermal growth factor-dependent activation of Src, Akt and ERK1/2 and treatment with the Src inhibitor PP2 and the PI3K inhibitor LY294002 abolished the Rukl/CIN85-dependent changes in cell motility. Together, this study indicates that high levels of Rukl/CIN85 contribute to the conversion of breast adenocarcinoma cells into a more malignant phenotype via modulation of the Src/Akt pathway