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Aim. Breast cancer (BC) is a heterogeneous disease which develops under the selective “pressure” of the tumor microenvironment. Therefore, it is promising and important to study tumor-associated macrophages as the innate immunity cells. The innate immunity system supposedly is able to regulate tumor immune surveillance and does not necessarily require the innate link. Object and methods. Materials of the study were intraoperative tissue of tumors and ipsilateral lymph nodes in radical mastectomy. Immunohistochemical characteristics of removed tumors (ER, PR, HER2, Ki67) were used for organizing two balanced groups of patients with primary BC. In the first group, there were no metastases into the regional lymph nodes, N0, whereas in the second one there was primary metastatic BC, N1. The groups included three patients with non-luminal HER2+, luminal A, luminal HER2-negative, luminal HER2+, and triple-negative BC, with 15 patients in each group, with a total of 30 patients. Results. The main idea of the work was to find the differences between the quantities of CD68+ and CD163+ macrophages, localized in tumor biopsy samples between the groups of patients with non-metastatic BC (N0) and with metastases (N1), balanced by the clinical values of ER, PR, HER2 (non-luminal HER2+, luminal A, luminal B HER2+, luminal B HER2-, and triple-negative BC). The results of statistical processing of average quantitative indicators of CD68+TAM, CD163+M2-like macrophages and CD68/CD163 (macrophage index M1) between groups N0 and N1 did not reveal a significant difference. However, when comparing the calculated immune-positive macrophages from all fields of view (40×lens), which were 5-17 for each specimen, a significant increase in CD68+TAM in the metastasis group was established. Comparison of average quantitative indicators of CD68+, CD163+ and CD68/CD163 between groups BC N0 and N1 showed no significant differences (the Mann-Whitney test). We determined the IHC characteristics of BC for each biopsy sample, including Ki67 and G-grade. Therefore, a statistical check of their correlation relationships with average quantitative indicators of CD68+, CD163+ and CD68/CD163 macrophages was conducted, which revealed a reliable relationship between CD68+ and Ki67, expressed as a percentage, and between ER/PR (in the clinical interpretation: +/-) and CD68+, CD163+, but not CD68/CD163.Conclusions. 1. The number of CD68+TAM was significantly higher in the focus of the primary BC of patients with metastases in the regional lymph nodes. 2. We detected a reverse correlation between CD68+TAM, CD163+M2-like macrophages and the clinical value of ER/PR. Consequently, an increase in the number of TAMs (in particular, M2) in the primary ER+/ PR+BC with metastases may not have a predictive value for metastasis, or ER+/PR+ BC possibly metastasizes without a significant increase in TAM. 3. The revealed differences are confirmed by reliable statistical calculations and relate to the effect of an exceptionally pathological process, since patients did not receive treatment before radical mastectomy. 4. Interpretation of personal data on the number of CD68+TAM and CD163+M2- like macrophages still needs to be developed.

The problem of diagnosing scars of the head and neck begins important in modern society. This is due to the high incidence of different types of scar tissue, their ineffective treatment, and possible recurrences. One of the causes of pathological scars is the disruption of the collagen gene polymorphism that encodes it. However, data on the value of the polymorphic variant of the collagen type I gene in the formation of pathological conditions of the skin are rather limited. Today according to literature there are disparate data on the differential diagnosis of pathological scars, all of them are not systematized and need the optimization. Object and methods. To study the polymorphism of the COL1A2 gene (rs42524), 60 patients were examined. All patients have got cicatricial tissue localized in different areas of the head and neck and were treated at the Department of Maxillofacial Surgery of Poltava State Clinical Hospital. Additional examination of scar tissue was performed by analyzing digital images in Adobe Photoshop CC using a color system RGB. Results and Discussion. We have studied the differences in the external state of scar tissue, depending on the genotype in a group of patients with pathological scars in different topographoanatomical areas of the head and neck. The scar tissues were evaluated using features such as scar consistency, color, sensitivity, and scar area, without considering the scar tissue type by the author’s standardized system for the external evaluation of cicatricial tissue. Patients with pathological scars in different topographic and anatomical regions of the head and neck, which are carriers of the allele C (genotypes GC and CC), have more pronounced signs of low-grade chronic inflammation of the skin. The obtained data about association of the polymorphism gene COL1A2 (rs42524) with signs of low-grade chronic inflammation of the skin in patients with pathological scars in different areas of the head and neck, probably indicate a possible effect of single-nucleus replacement on the physiological functions of collagen, namely the collagen specific collaboration with different molecules and cells. Using the RGB system, it was established that using the RGB color systems in Adobe Photoshop makes it possible to differentiate the types of pathological scars of the head and neck depending on the intensity of color spectrum. It was noted the relationship between the presence of the polymorphic variant of the COL1A2 gene (rs42524) in the genotype and data of RGB examination among patients with scars in different areas of the head and neck that were separated in previous studies, by the types of scars in 4 groups according to the type of scar. At present, there is no optimized method of differential diagnosis of different types of scars of the head and neck, which is why there is a necessity to develop an informative and economically feasible method of diagnosis. Conclusions. It is possible to determine precisely the type of scar and, depending on the type, to choose the optimal method of surgery or conservative therapy by using the RGB system and determining the type of scar, taking into account the effect gene polymorphism of collagen 1 alpha-2 (COL1A2) (rs42524) on cicatricial tissue formation.

The triple-negative subgroup of breast cancer (TNBC) (estrogen, progesterone-, and HER2-receptor negatives) often affects young women and usually has a worse prognosis as compared to other types of BC. Recently, a complex role of the immune system in the growth, elimination and metastasis became an object of increased attention, promising to provide clinical prognostic markers of progression, as well as targets for the treatment of a wide range of tumors, including TNBC. In BC in general and in TNBC in particular, tumor-associated macrophages (TAM) are mainly associated with a worse prognosis, but their composition, apparently, varies between individual cases of TNBC. Thus, the research of TAM in TNBC is relevant, and therefore the aim of this study was to examine the quantitative parameters and features of localization of CD68+ and CD163+M2-like TAMs, which infiltrate TNBC in the primary focus without metastases and in paired samples with metastases in the lymph nodes, as well as the pathomorphological characteristics of this type of BC to determine their contribution to metastasis. Biopsy samples and clinical data were obtained from patients undergoing treatment at Poltava Regional Clinical Dispensary. The study was approved by the Ethics Commission of Ukrainian Medical Stomatological Academy. Materials of the study were intraoperative tumor tissues and ipsilateral lymph nodes in radically removed mammary glands without neoadjuvant treatment. Immunohistochemical (IHC) characteristics of the removed tumors (HER2, ER, PR, Ki67) were used to distinguish the TNBC group. According to the meta-analysis, the use of CD68 as a biomarker for TAM for IHC evaluation has its priorities, as compared with the individual definition of CD163 or CD206. We analyzed two subgroups of TNBC, balanced by the N0/1 status, 3 samples in each, and 3 samples of metastases in the lymph nodes, respectively. The average age of patients was 64 years, from 52 to 76. Primary-metastatic BC was observed in all patients in the 2nd, N1 group. Morphological and IHC findings were obtained according to the criteria for determining the molecular and biological subtype. IHC studies for determining TAM and M2-like macrophages were performed using the streptavidin-peroxidase method. The evaluation of immunohistochemical coloring was performed by counting CD68+TAM and CD163+M2-like TAMs under the light microscope (Biolam, LOMO, Russia: lens ?40, eyepiece ?7) in 7-10 fields of view of the intensive ICH-reaction of each section, calculating the arithmetic mean, within the tumor nests and tumor stroma. The count included immunopositive cells with macrophage morphology. Microphotographs were obtained using the Leica DM500 microscope, Leica, Germany (lens ? 4, 40). The statistical analysis was carried out using the GraphPad Prism 5 software, using nonparametric, parametric and simulation methods. The study is organized as a descriptive pathomorphological comparison between the two groups: N0 and N1, triple-negative BC, comparison of quantitative characteristics of CD68+ and CD163+M2-like fraction of TAM, as well as for verification of possible correlation relations between the quantitative characteristics of macrophages in the primary tumor and in the corresponding metastasis in the ipsilateral axillary lymph nodes. Increased level of infiltration of the primary focus of TNBC with CD68+TAM and CD163+M2-like macrophages is reliably associated with metastasis. It is assumed that the M1-like subpopulation of TAM most closely contacts with tumor nests in TNBC, which may be a prerequisite for the spread of the tumor, or metastasis. Primary premetastatic effects of TNBC may be related to the increase of not only M2, but also M1 macrophages. In metastatic lymph nodes, the amount of CD68+TAM was significantly lower than that of primary TNBC, whereas the quantitative indices of CD163+M2 correlated. Along with the localization and co-localization of CD68+ and CD163+M2-like TAMs, the results conform with the generally accepted paradigm that M1 TAM has an anti-tumoral but lower potential and M2 the protumoral one, but this is shown for metastases in the regional lymph nodes in primary-metastatic TNBC. Precise quantitative TAM indices for the use in clinical practice have not been developed, although their pathogenetic significance and quantitative variations are reliable according to metastasis and other clinical data of TNBC.

The effects of fullerene C60 (FC60) on the level of free radical and destruction processes were studied in rats with experimental adjuvant arthritis (AA). It was shown the protective effect of FC60 during AA. The effect was accompanied by an increase of the antioxidant enzymes activity, superoxide dismutase in the liver (15.96 ± 0.38 μmol/kg x s) and in the kidneys (5.36 ± 0.27 μmol/kg x s) and catalase in the kidneys (9.56 ± 0.78 μmol/kg x s) and in the heart (2.26 ± 0.41 μmol/kg x s) in comparison to control group (43.83± 5.69%; 54.55 ± 6.18%; 11.68 ± 0.52 μmol/kg x s; 3.43 ± 0.47 μmol/kg x s; 4.77 ± 0.5 μmol/kg x s; 0.98 ± 0.12 μmol/kg x s accordingly). It was shown a protective effect of FC60 during AA directed on the depression of the destructive processes in connective tissue that was expressed through the reduction of the total collagenolitic activity level in cartilage (10.05 ± 0.06 μmol/g/min) and bone (11.21 ± 0.04 μmol/g/min) tissues, free hydroxyproline contents (1.54 ± 0.04 μg/ml) and alkaline phasphatase activity (1.24 ± 0,14 μmol/l x sec) in comparison to control group (11.91 ± 0.49 μmol/g/min; 13.19 ± 0.15 μmol/g/min; 2.25 ± 0.07 μg/ ml; 2.19 ± 0.24 μmol/l x sec accordingly). Taken together, these results accentuate the perspective of future investigations of action FC60 during rheumatoid arthritis as a feasible therapeutic agent.