INTRATUMORAL AMPLIFICATION HETEROGENEITY IN HER2/neu-POSITIVE BREAST CANCER MOLECULAR-GENETIC SUBTYPES

The defining feature of HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtype breast cancer is HER2/neu gene amplification and protein overexpression on cancer cell membrane. The HER2-targeted therapy is nowadays available for patients with HER2-positive breast cancer However, a significant fraction of HER2+ tumors acquire or possess intrinsic mechanisms of resistance, based on multiple factors, and genetic heterogeneity among them. The aim of our study was to quantify the heterogeneity of HER2/neu amplification in HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes of breast cancer. Material and methods. A retrospective analysis of 210 cases referred for dual probe fluorescence in situ hybridization (FISH) confirmation of an immunohistochemical equivocal 2+ result was performed. Results. Our results demonstrated a heterogeneous amplification pattern of HER2/neu gene, whose expression is a substantial cause of HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes of breast cancer, in 31 % of invasive breast cancer cases. As heterogeneous, we interpreted tumors containing cells with HER2/CEP17 ratio < 2 and gene copies 4 ≤ HER2/neu < 6, that is, those without HER2/neu amplification. The amount of heterogeneous tumors between HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes was not statistically significant. ROC analyses identified optimal cutoff point for HER2/CEP17 ratio as 2.6 for distinguishing heterogeneous tumors. Conclusion. The heterogeneity of HER2/neu amplification is determined by FISH in 31 % of cases and is independent of molecular breast cancer subtype. If a HER2/neu-positive breast cancer has HER2/CEP17 ratio ≤ 2,6, it contains minor subclones without HER2/neu amplification with a probability of 95 %. Our results demonstrated that HER2/neu amplification heterogeneity may be important for prognosis of survival and treatment decisions

Immunohistochemical evaluation of Ki-67, Cyclin D1 and β-catenin expression in the subtypes of triple negative breast cancer

Aim. To evaluate the expression levels of Ki-67 and cyclin D1 and β-catenin in the subtypes of triple negative breast cancer. Methods. The study was conducted on the surgical material from 60 patients of clinical stage 2A (T1N1M0 or T2N0M0) who were treated at the Rostov Research Institute of Oncology from 2012 to 2015. For immunohistochemistry, antibodies to estrogen and progesterone receptors, cytokeratins 5/6, Ki-67, cyclin D1, β-catenin, HER2/neu and EGFR proteins were used. Results. Triple negative breast cancer with the signs of basal epithelium was found to have a significantly higher expression level of Ki-67 compared to non-basal-like one. In some part of triple negative breast cancer samples overexpression of cyclin D1 was observed. The high level of cyclin D1 in the basal-like subtype was less common than in the subtypes without the signs of basal epithelium, but its average value was significantly higher. In triple negative cancer with cyclin D1 overexpression, the loss of β-catenin on the cell membrane and its abnormal accumulation in the cytoplasm was significantly more frequent. β-catenin translocation into the cell nucleus was observed only in basal-like triple negative cancer, and 2 times more often in case of cyclin D1 overexpression. Conclusion. In triple negative breast cancer tumors with overexpression of cyclin D1 and abnormal expression of β-catenin are observed in some cases; these biomarkers can be considered as potential therapeutic targets for this group of tumors

Achievements and prospects of cellular technologies based on the activated lymphocytes in the treatment of malignant tumors

This article reviews the immune system and its role in the relationship between the tumor and the body of a patient with tumor diseases. It is about controlling homeostasis by recognizing and eliminating genetically alien substances (antigens). Antitumor treatment is now not only considered as an “instrument” for eliminating and destroying tumor cells, but also its ability to change/restore impaired functions of the immune system attracts attention. The used antitumor treatment is widely known to be immunosuppressive, stress and radiation effects also cause and/or enhance immunosuppression. In this work, the authors provide literature data demonstrating current status and problems of cellular immunotherapy of malignant tumors with the use of activated lymphocytes, and the role of antigen-specific T-lymphocytes as one of its most important agents is reviewed. Currently, among the immunotherapeutic methods, a special place is occupied by approaches involving the use of autologous or allogenic ex vivo stimulated immunocompetent cells (adoptive immunotherapy). The importance of complex influence on various links (T-, B-, NK-cell) and stages (presentation, recognition, proliferation, differentiation, migration, activation, effector functions) of the immune response is considered. The emergence of targeted drugs based on antibodies, as well as vaccines, especially dendritic cells, has provoked the emergence of a new wave of interest in the formation of specific antitumoral immune response mediated by T lymphocytes, so the introduction of the latter can be classified as a kind of targeted therapy. The value of antigen-specific T-lymphocytes in the formation of antitumor immunity is shown, which emphasizes the importance not only of CD8+, but also of CD4+ T-lymphocytes. In addition, there are suggestions of the possible significance of both T- and B-cells for developing a strategy of cellular immunotherapy. The literature data suggest that not only cytotoxic lymphocytes, but also T-helpers and even B-lymphocytes can be effective as antigen-specific lymphocytes as a component of antitumor treatment. The authors consider the possibility of obtaining antigen-specific T cells, as well as their further storage. The possibility of elimination or selective inhibition of regulatory T-cells during adoptive immunotherapy aimed at removing the suppressor effect on cytotoxic lymphocytes is studied. Various strategies for the use of cell therapy are also discussed

Intermolecular interactions of decamethoxinum and acetylsalicylic acid in systems of various complexity levels

Intermolecular interactions between decamethoxinum (DEC) and acetylsalicylic acid (ASА) have been studied in the phospholipid-containing systems of escalating complexity levels. The host media for these substances were solvents, L-α-dipalmitoylphosphatidylcholine (DPPC) membranes, and samples of human erythrocytes. Peculiar effects caused by DEC-ASА interaction have been observed in each system using appropriate techniques: (a) DEC-ASА non-covalent complexes formation in DPPC-containing systems were revealed by mass spectrometry with electrospray ionization; (b) joint DEC-ASА action on DPPC model membranes led to increasing of membrane melting temperature Tm, whereas individual drugs caused pronounced Tm decreasing, which was demonstrated by differential scanning calorimetry; (c) deceleration of DEC-induced haemolysis of erythrocytes under joint DEC-ASА application was observed by optical microscopy

Comparative DNA Cytometry of Primary and Recurrent Soft Tissue Sarcomas

The goal of comparative investigation was to reveal the distinctive features of the DNA content and cell distribution in the phases of the cell cycle of recurrent STS. DNA cytometry in the tumor tissue of 30 primary soft tissue sarcomas (t2a-2bn0M0) and 30 STS recurrences (t2-3n0M1) was carried out using the method of flow cytofluorometry. the tumor ploidy and cell distribution in the cell cycle phases were analyzed. Results. A number of differences in the DNA cytometric parameters of primary and recurrent STS have been revealed, they include: an increase in the proportion of aneuploid tumors in case of recurrence, the number of tumors with DNA index within the mitotic cycle, an increase in the proportion of cells in G2+M- phase of diploid and aneuploidy tumors and a decrease in S- phase of aneuploid ones. It has been shown that with a G2 differentiation degree, the proportion of cells in G2+M, S- and IP of recurrent STS is significantly lower than the primary parameters. An increase in the proportion of cells in G2+M- phase and a decrease in the rate of proliferation of recurrent STS, depending on the stage, are shown only in case of stage III. Conclusion. The revealed features of DNA content and cell cycle of tumor cells of soft tissue sarcomas will allow to approach to understanding of biological bases of recurrence of this malignant disease.Целью исследования было выявить в сравнительном аспекте отличительные особенности содержания ДНК и распределения клеток по фазам клеточного цикла рецидивных СМТ. Методом проточной цитофлуориметрии проводили ДНК-цитометрию в опухолевой ткани 30 первичных сарком мягких тканей (T2a-2bN0M0) и 30 – рецидивов СМТ (t2-3n0M1). анализировали плоидность опухоли и распределение клеток по фазам клеточного цикла. результаты. Выявлен ряд различий ДНК-цитометрических показателей первичных и рецидивных СМТ, которые заключаются: в увеличении доли анеуплоидных опухолей при рецидивах, числа опухолей с ИДНК в пределах митотического цикла, увеличение доли клеток в G2+M- фазе диплоидных и анеуплоидных опухолей и снижение в S-фазе анеуплоидных. Показано, что при степени дифференцировки G2 доля клеток в фазах G2+M, S- и ИП рецидивных СМТ значимо ниже параметров первичных. Увеличение доли клеток в G2+M-фазе и снижение темпов пролиферации рецидивных СМТ в зависимости от стадии показаны только при III стадии. Выводы. Выявленные особенности содержания ДНК и параметров клеточного цикла опухолевых клеток сарком мягких тканей позволят приблизиться к пониманию биологических основ рецидивирования этого злокачественного заболевания

Аdvanced endodontic developments in pulpectomy of primary teeth

Objective. To appraise studies about pulpectomy in primary teeth with nickel-titanium rotary files and smear layer removal and to find out may these developments enhance pulpectomy outcomes in primary teeth. Material and methods. A systematic search was implemented for PubMed, Google and Google Scholar between the years 1995-January 2016 to identify eligible studies. Studies design was established according to the CEBM recommendations. Evidence quality of studies was appraised by risk of bias. Results. Six studies about pulpectomy met the inclusion criteria, of which five were randomized controlled trials. Only one research demonstrates the enhanced outcome of pulpectomy in primary teeth with smear layer removal. Chosen studies have low overall evidence quality. Conclusions. Given the paucity, high heterogeneity of high-quality articles and their level of bias, recommendation for the use of nickel-titanium rotary files and smear layer removal in pulpectomy in primary teeth can yet not be formulated

Membranothropic properties of the urocanic acid

The effects of urocanic acid (UA) on thermodynamic parameters of model dipalmitoylphosphatidylcholine (DPPC) lipid membrane have been studied by means of differential scanning calorimet­ry (DSC). The observed ordering effect of UA on the lipid bilayer is reflected in the increase in both the main phase transition temperature and cooperative unit size of the lipid membrane. Analysis of FTIR spectra suggests localization of UA molecules in the vicinity of the polar heads and carbonyl groups of DPPC due to electrostatic interactions and H-bonds. On the basis of experimental data obtained and geometry parameters of UA and DPPC molecules, some variants of the UA localization in DPPC bilayer were discussed

Photo- and electroluminescent properties of zinc(II) complexes with tetradentate Schiff bases, derivatives of salicylic aldehyde

International audienceIt is studied how the introduction of various substituents into the composition of organic ligands affects the photoluminescence spectra of new zinc(II) complexes with tetradentate Schiff bases H2L (derivatives of salicylic aldehyde (H2SAL1, H2SAL2) and o-vanillin (H2MO1, H2MO2) with ethylenediamine and o-phenylenediamine) in the form of bulk solids and thin films. It is demonstrated that the emission spectra of bulk solid complexes without o-phenylenediamine bridges (ZnSAL1 and ZnMO1) contain additional long-wavelength bands compared to the spectra of corresponding thin films. In the case of films obtained from [ZnSAL1]2 dimer complexes, the long-wavelength band is dominant. At the same time, the photoluminescence spectra of ZnSAL2 and ZnMO2 complexes with o-phenylenediamine bridges are similar in the case of solid samples and thin films. The electroluminescent properties of organic light-emitting diodes (OLEDs) with the ITO/α-NPD/ZnL/Ca:Al structure are studied. The bathochromic shift of the electroluminescence peaks of OLEDs with respect to the photoluminescence spectra of bulk solid samples and thin films is probably related to the formation of exciplexes at the α-NPD/ZnL interface. The electroluminescence spectra of OLEDs based on [ZnSAL1]2 show a hypsochromic shift of the emission maximum, which can be caused by a shift of the recombination region into the α-NPD layer