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Background: The specificity of Streptococcus pneumoniae interaction with different epithelial cells has not been fully studied. Yet, it has been demonstrated that this type of streptococcus induced bacterial inflammation in 49 % of cases of various respiratory diseases. Previous studies indicate that inhalation exposure to occupational risk factors accounts for a higher average long-term incidence rate of community-acquired pneumonia among workers. The objective of our study was to substantiate approaches to developing criteria for establishing the immunocompromised status of workers exposed to chrysotile asbestos for subsequent vaccination against pneumococcal disease. Materials and methods: We analyzed data on 304 cases of community-acquired pneumonia registered in 2005–2016 among chrysotile miners and millers, obtained from the Information System of Epidemiological Surveillance maintained by the Center for Hygiene and Epidemiology in the Sverdlovsk Region. In addition, we screened 240 employees for prevaccination immunity using immunoassay tests. Results: We established a higher incidence of community-acquired pneumonia in miners and millers (217 cases or 71.4 %) compared to the control cohort consisting of auxiliary staff (87 cases or 28.6 %). We also noted that the longer length of employment was associated with an increased secretory IgA synthesis in both miners and millers and a significant decrease in the bactericidal action of neutrophils in miners. Conclusion: Chrysotile asbestos exposure leads to adaptive stress, strain of protective and compensatory mechanisms, poor resistance causing increased susceptibility to viral and bacterial diseases, and the development of secondary immunodeficiency in experienced workers of the main occupations, all contributing to higher incidence of community-acquired pneumonia.

This article is the second communication in a series of articles devoted to the effects of a domestic preparation of macrophage-activating factor (GcMAF-RF) and assessment of its biological properties. The aim of this work was to study the effect of the GcMAF-RF upon M0 → M1 polarization of macrophages (Mph), and activation of the professional properties of ex vivo generated antigen-presenting dendritic cells (DC), as well as on ex vivo production of pro-inflammatory (TNFα, IL-1β, IL-6, IFNγ, IL-17, IL-18) and anti-inflammatory (TGF-β, IL-4, IL-10) cytokines, growth factors (IL-2, GM-CSF, G-CSF, VEGF) and chemokines (MCP, IL-8) by the whole blood cells from healthy donors. Mph and DC were generated from the monocytes (3 to 5×106 /ml) derived from adherent fraction of peripheral blood mononuclear cells (MNC) of healthy donors. Granulocyte/macrophage colony-stimulating factor (rhGM-CSF) was used to obtain Mph, whereas DC production was induced by GM-CSF and interferon-α. To provide M1 polarizing signals, bacterial lipopolysaccharide (LPS from E. coli 0114:B4) was used in controls. In experimental series, GcMAF-RF was added 48 h before the end of culture. The stimulating effect of the obtained Mph and DC upon cell proliferation was assessed in allogeneic mixed culture of leukocytes (alloMLC) using radiometric technique, by 3 H-thymidine incorporation. The influence index (IR) of Mph or DC upon allo-SCL was calculated as the ratio of the proliferative response of MNCs in the presence of Mph, or DC to the level of spontaneous MNC proliferation. To determine the cytokine production by human whole blood cells ex vivo, peripheral blood samples from 3 donors with two replicate GcMAF-RF preparations were used, at a total of 6 points. All variants of the study were carried out with mitogen-activated and non-activated blood cells. The cytokine content was determined by the ELISA assays. The effects of GcMAF-RF were quantified as a fold increase (FI), i.e., the ratio of cytokine production in the presence of GcMAF-RF to the level of their spontaneous production. It was shown that the GcMAF-RF preparation was as effective, as lipopolysaccharide (LPS), the standard Mph and DC activator which induces polarization of differentiated M0-macrophages into M1 cells and final maturation of DCs, manifesting by a significant increase in their allo-stimulatory activity in a mixed leukocyte culture (allo-MLC). Moreover, GcMAF-RF stimulates production of numerous cytokines and chemokines (TNFα, IL-1β, IL-6, IL-18, IL-4, IL-10, GM-CSF, G-CSF, VEGF, IL-8), by blood cells (granulocytes, lymphocytes, monocytes), thus indicating direct participation of the macrophage activator GcMAF-RF in various immune processes. The domestic GcMAF-RF drug induces polarization of macrophages M0 → M1, final maturation of DCs and allostimulating activity of Mf and DCs, and is also able to effectively stimulate circulating blood cells to synthesize cytokines/chemokines with pro-inflammatory and immunoregulatory activities.

Differences in the production of cytokines by tumor biopsy specimens were revealed depending on the pathological prognostic stages of The American Joint Committee on Cancer (AJCC) of invasive nonspecific breast carcinoma (INBC). The patients with a predominant absence of metastases in combination with a triple negative molecular subtype differ from the patients with other pathological prognostic stages in the cytokine-producing tumor resource of IL-18, IL-1β, IL-1Ra, TNF-α, GM-CSF, and MCP-1.

Currently, the data on the presence of adenoviral receptors in fish are very limited. We used mouse recombinant adeno-associated viral vectors (rAAV) with a calcium sensor GCaMP6m, which are typically used for the CA1 region of the mouse dorsal hippocampus but have not previously been used for gene delivery in the fish brain. The aim of this work was (1) to assess the ability of the mouse hippocampal rAAV to label brain cells in a juvenile chum salmon Oncorhynchus keta and (2) to determine the phenotype of rAAV-labeled cells. In vivo gene delivery was carried out through intracranial injection of the GFP-tagged rAAV directly into the mesencephalic tegmentum of a yearling chum salmon. The level of infectious efficacy was assessed 1 week after the injection of the vector. Using confocal scanning microscopy, rAAV-GFP expression was assessed in various tegmental areas of a juvenile O. keta, followed by the localization of the neuron-specific protein HuCD tagged with red-fluorescent Alexa Fluor 546. An immunohistochemical analysis combined with DAPI staining revealed rAAV/HuCD colocalization in various neurons of the mouse mesencephalic tegmentum. This finding provides evidence that rAAV-expressing tegmental cells in the juvenile chum salmon refer to a neuron-specific line of brain cells, indicative of the ability of mammalian hippocampal adenoviruses to infect neurons of the fish central nervous system. Thus, mammalian hippocampus-specific rAAVs can incorporate into fish brain neurons with subsequent expression of the viral proteins. It appears that tegmental neurons in a juvenile chum salmon contain a homologue of the mammalian adenoviral receptor.

The relationship between the content of supernatant cytokines and the immune histochemical (IHC) expression of epithelial-mesenchymal transition (EMT) markers was studied in biopsy samples of invasive breast carcinoma of no special type (IBC-NST) from patients with and without lymphatic spread of metastases (group II and group I, respectively). The concentrations of TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF, MCP-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, and IL-1Ra, as well as the immunohistochemical (IHC) expression of EMT markers cadherin-E (CDH1), β-1 integrin (CD29) and type II collagen (CII) were assayed. Results have shown that the patients of these groups statistically significantly differed by spontaneous production of IL-18 and G-CSF, and by the index of the polyclonal activator influence (IPAI) on G-CSF production. The study of relationships between supernatant cytokine production and the IHC expression by biopsy samples revealed correlations between the parameter of CII expression in the tumor tissue and the cytokine production by tumor biopsy specimens; these correlation links were typical for all IBC-NST patients, while correlation links (both direct and reverse ones) between CDH1, CD29, and cytokine production varied in groups of patients with and without lymphatic spread of metastases. The study revealed the features of the correlation links between the cytokine production by the tumor, its microenvironment and the IHC expression of EMT markers in IBC-NST patients with and without lymphatic spread of metastasis.

The article focuses on the influence of human leukemia differentiation factor (HLDF), carcinoembryonic antigen (CEA), and polyclonal activators (PA) on cytokine production by peripheral blood cells in breast cancer and benign breast diseases. It was found that the influence of internal factors on the production of cytokines by the peripheral blood cells is associated with lymphatic metastasis (CEA: IL-10; HLDF: IL-6, IL-1β, TNF-α, and G-CSF). One special circumstance was that there were no differences between the production of cytokines by peripheral blood cells in the patients with breast cancer compared to the patients with benign breast diseases with a high risk of malignant transformation. This is evidence of the functional similarity of peripheral blood cells in patients with these conditions. Cytokine production under the influence of PA was different only in case of TNF-α in all study groups.

The material of patients with invasive carcinoma of no special type (ICNT) and nonmalignant diseases (ND) of the mammary gland was studied. When comparing the concentrations of histidine-rich glycoprotein (HRG) and E-cadherin (CDH1), statistically significant differences between ICNT and ND by HRG in the supernatant of blood cells and its spontaneous production by biopsies and by CDH1 at its induced production, as well as by influence indices of polyclonal activators on the production of CDH1 were found. When comparing the expression of immunohistochemical markers, no statistically significant differences between ICNT and ND were obtained.

We studied the effect of the HLDF differentiation factor on production of cytokines by biopsy samples of nonmalignant breast diseases (ND) and invasive breast carcinoma of no special type (IBC-NST), in the absence and presence of lymphogenic metastasis: IBC-NST patients werw subdivided into groups on the prognostic protocol of the 8th edition of the AJCC committee. Group IA consisted of patients with T1-T2 tumor sizes, and predominantly with positive expression of estrogen and progesterone receptors (ER+/PR+/HER2-); it also included one patient with the HER2+ (ER-/PR-/HER2+) molecular subtype. The IB group was mainly composed of patients with T2 tumor size, with the presence of lymphogenic metastasis (in 8 out of 10) patients and with positive expression of estrogen and progesterone receptors (ER+/PR+/HER2-) and it also included three patients with the HER2+ (ER-/PR-/HER2+) molecular subtype. Group IIA consisted of patients with T1-T2 tumor sizes, mainly with no metastases in the lymph nodes (in 11 out of 12 patients) and with a triple negative molecular subtype. Group IIB included patients with T2 tumor size, the presence of nodal metastasis and the expression of markers of ER-/PR-/HER2 - and ER-/PR-/HER2+. Group IIIA consisted of patients with tumor size T1-T3, with the presence of nodal metastasis and the expression of markers of ER-/PR+/HER2+ and ER-/PR-/HER2+. Group IIIC consisted of patients with T3 tumor size, lymphogenic metastasis, and expression of ER-/PR-/HER2-markers (triple negative molecular subtype). Due to a limited number of patients in the groups IIB, IIIA and IIIC, as well as due to more severe clinical and pathological stages, according to the prognostic Protocol of the 8th edition of the AJCC Committee, they were pooled into group III. Concentrations of IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1Ra, TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF and MCP-1 were assayed in supernatants of biopsy specimens of breast tissue. Results have shown that with IBC-NST, a statistically significantly higher level of spontaneous production (SP) by biopsy specimens of IL-17, IL-18, IFN-γ and VEGF, and a lower level of SP IL-6 as compared with ND. Patients of all clinical and pathological groups showed a high VEGF spontaneous production as compared with ND, while statistically significant differences from patients with ND were not found in IL-17 spontaneous production in group IB patients, and IL-18 spontaneous production were absent in group IA. Only in patients with IA and IB, the IL-6 spontaneous production was lower as compared to ND, and the IL-8 spontaneous production was lower in the IA group. IFN-γ spontaneous production was higher in patients with IBC-NST group IIA as compared with ND. Under the influence of the HLDF differentiation factor, it was found that the parameters of IBC-NST patients were statistically significantly higher in the production of IL-1Ra, IL-17, IL-18 and VEGF, and statistically significantly lower in the production of IL-6 as compared to ND. HLDF had a higher impact on the content of IL-18 in IBC-NST patients than in ND. After HDLF sublimation IL-6 values were lower in patients of groups IA and IB, and HLDF-induced IL-17 production was higher only in patients of group IA. Statistically significant differences in the index of influence of HLDF (IVHLDF), representing ratio of the cytokine concentration in the supernatants of a biopsy specimen stimulated by HLDF to spontaneous cytokine production, were found between ND and IBC-NST in the case of on IFN-γ production, and also in the case of IL-4 production (between patients in the absence and presence of lymphogenic metastasis). IVHLDF for production of IL-6, IL-8 and TNF-α was lower in group IIA patients compared to group IA, and IVHLDF for production of GM-CSF and MCP-1 was lower in group IIA as compared to group III, in addition IVHLDF for MCP-1 products was lower in group IIA as compared to ND. The HLDF effect on the cytokine production by the tumor and its microenvironment was different in ND patients and IBC-NST patients. HDLF suppressed IFN-γ production in the pooled group of IBC-NST patients; HLDF mainly had a suppressive effect on the production of IL-6, IL-8, TNF-α, GM-CSF and MCP-1 in IBC-NST patients of group IIA.Issledovali vliianie faktora differentsirovki HLDF na produktsiiu tsitokinov bioptatami pri nezlokachestvennykh zabolevaniiakh molochnoĭ zhelezy (NZ) i pri invazivnoĭ kartsinome molochnoĭ zhelezy nespetsificheskogo tipa (IKNT), pri otsutstvii i nalichii limfogennogo metastazirovaniia, a takzhe u patsientov s IKNT, raspredelennykh po podtipam na osnove prognosticheskogo protokola 8 izdaniia komiteta AJCC. Gruppu IA sostavili patsienty s razmerami opukholi T1-T2 i preimushchestvenno s polozhitel'noĭ ékspressieĭ retseptorov éstrogena i progesterona (ER+/PR+/HER2-) i odin patsient s HER2+ (ER-/PR-/HER2+) molekuliarnym podtipom. V gruppu IB byli vkliucheny patsienty s razmerami opukholi T2, preimushchestvenno s nalichiem limfogennogo metastazirovaniia (8 iz 10) i s polozhitel'noĭ ékspressieĭ retseptorov éstrogena i progesterona (ER+/PR+/HER2-), a takzhe tri patsienta s molekuliarnym podtipom HER2+ (ER-/PR-/HER2+). Gruppu IIA sostavili patsienty s razmerami opukholi T1-T2, preimushchestvenno s otsutstviem metastazov v limfouzlakh (11 iz 12 bol'nykh) i s troĭnym negativnym molekuliarnym podtipom. Gruppu IIB sostavili patsienty s razmerami opukholi T2, nalichiem limfogennogo metastazirovaniia i ékspressieĭ markerov: ER-/PR-/HER2- i ER-/PR-/HER2+. Gruppu IIIA sostavili patsienty s razmerami opukholi T1-T3, nalichiem limfogennogo metastazirovaniia i ékspressieĭ markerov: ER-/PR+/HER2+ i ER-/PR-/HER2+. Gruppu IIIC sostavili patsienty s razmerami opukholi T3, s nalichiem limfogennogo metastazirovaniia i s ékspressieĭ markerov ER-/PR-/HER2- (troĭnoĭ negativnyĭ molekuliarnyĭ podtip). Vvidu malogo kolichestva patsientov v gruppakh IIB, IIIA i IIIC, a takzhe v sviazi s bolee tiazhelymi kliniko-patologicheskimi stadiiami soglasno prognosticheskomu protokolu 8 izdaniia komiteta AJCC oni byli ob"edineny v odnu gruppu III. Kontsentratsii IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IL- 1β, IL-1Ra TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF i MCP-1 opredeliali v supernatantakh bioptatov tkani molochnoĭ zhelezy. Issledovaniia pokazali, chto pri IKNT otmechalsia statisticheski znachimo bolee vysokiĭ uroven' spontannoĭ produktsii (SP) IL-17, IL-18, IFN-γ i VEGF i bolee nizkiĭ uroven' SP IL-6 po sravneniiu s NZ molochnoĭ zhelezy. U patsientov vsekh kliniko-patologicheskikh grupp IKNT otmechena vysokaia SP VEGF po sravneniiu s NZ, togda kak statisticheski znachimye razlichiia ot patsientov s NZ po SP IL-17 otsutstvovali u patsientov gruppy IB, a po SP IL-18 otsutstvovali u patsientov IA. Tol'ko u patsientov grupp IA i IB SP IL-6 byla nizhe po sravneniiu s NZ, a SP IL-8 byla nizhe u patsientov gruppy IA. SP IFN-γ byla vyshe u patsientov s IKNT gruppy IIA po sravneniiu s NZ. Pri vozdeĭstvii faktora differentsirovki HLDF ustanovleno, chto pokazateli patsientov s IKNT byli statisticheski znachimo bolee vysokimi po produktsii IL-1Ra, IL-17, IL-18 i VEGF i statisticheski znachimo bolee nizkimi po produktsii IL-6 po sravneniiu s NZ molochnoĭ zhelezy. Pri vozdeĭstvii HLDF na produktsiiu IL-18 ego soderzhanie okazalos' vyshe u patsientov vsekh kliniko-patologicheskikh grupp po sravneniiu s patsientami s NZ. Po sravneniiu s NZ tol'ko u patsientov grupp IA i IB pokazateli IL-6 pri vozdeĭstvii HLDF byli nizhe, a indutsirovannaia HLDF produktsiia IL-17 byla vyshe tol'ko u patsientov gruppy IA. Statisticheski znachimye razlichiia po indeksu vliianiia HLDF (IVHLDF), predstavliaiushchim soboĭ otnoshenie kontsentratsii tsitokinov v supernatante bioptata, pri vozdeĭstvii HLDF k spontannoĭ produktsii tsitokinov byli ustanovleny mezhdu NZ i IKNT po IVHLDF na produktsiiu IFN-γ, a takzhe po IVHLDF na produktsiiu IL-4 mezhdu patsientami pri otsutstvii i nalichii limfogennogo metastazirovaniia. IVHLDF na produktsiiu IL-6, IL-8 i TNF-α byl nizhe u patsientov gruppy IIA po sravneniiu s gruppoĭ IA, a IVHLDF na produktsiiu GM-CSF i MCP-1 byl nizhe u gruppy IIA po sravneniiu s pokazateliami gruppy III, krome togo, IVHLDF na produktsiiu MCP-1 byl nizhe u gruppy IIA po sravneniiu s NZ. Vliianie faktora differentsirovki HLDF na produktsiiu tsitokinov opukhol'iu i ee mikrookruzheniem razlichno dlia patsientov s NZ i IKNT. Faktor differentsirovki okazyval supressiruiushchee vliianie na produktsiiu IFN-γ u patsientov s IKNT i na produktsiiu IL-4 tol'ko u patsientov s limfogennym metastazirovaniem i na produktsiiu IL-6, IL-8, TNF-α, GM-CSF i MCP-1 u patsientov s IKNT gruppy IIA.

Breast cancer, in most cases, is a malignant neoplasm associated with infiltration of a tumor with the cells that form its microenvironment and produce various cytokines. The aim of the study was to evaluate the cytokine-producing function of tumor cells and their microenvironment in biopsy specimen of patients with invasive carcinoma of no special type and in patients with benign breast diseases. To assess the cytokine-producing activity of the tumor and its microenvironment, the index of polyclonal activators influence on cytokine production by biopsy specimens of patients with invasive carcinoma of no special type (group I) and in patients with benign breast tumors (group II) was calculated. Group II was further subdivided into group IIa, which included only patients with fibroadenoma, and group IIb, which included the patients with leaf-shaped fibroadenoma, fibroadenomatosis, fibrocystic mastopathy, intraductal papillomatosis, sclerosing adenosis and fibrocystic mastopathy with microcalcifications. The concentrations of IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1Ra, TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF, and MCP-1 were measured in tumor biopsy supernatants. When comparing groups I and II, higher indices of the polyclonal activators influence on the production of IL-17, IL-18 and TNF-α were observed in patients with benign diseases. Higher indices of the polyclonal activators influence on the production of IL-18, TNF-α, and IL-1β and the ratio of IL1β/IL1Ra were observed in patients with fibroadenoma as compared to those with invasive carcinoma of no special type. There were no significant differences in the indices of the polyclonal activators influence between groups I and IIb. This suggests the existence of changes in the mammary gland in patients of group IIb similar to those present in patients with invasive carcinoma of no special type. Higher indices of polyclonal activators influence on the production of IL-1β, as well as the ratio of IL1β/IL1Ra were observed in the patients of group IIa compared to the patients of group IIb. The results of the study identify the features of the cytokine-producing resource of tumor biopsy specimens in patients with invasive carcinoma of no special type and with benign breast tumors.Rak molochnoĭ zhelezy v bol'shinstve sluchaev iavliaetsia primerom zlokachestvennogo novoobrazovaniia, sviazannogo s infil'tratsieĭ opukholi kletkami, obrazuiushchimi mikrookruzhenie opukholi i produtsiruiushchimi razlichnye tsitokiny. Tsel'iu issledovaniia byla otsenka tsitokinprodutsiruiushcheĭ funktsii opukholevykh kletok i ikh mikrookruzheniia v bioptatakh opukholi patsientov s invazivnoĭ kartsinomoĭ nespetsificheskogo tipa (gruppa I) i u patsientov s nezlokachestvennymi zabolevaniiami molochnoĭ zhelezy (gruppa II). Bol'nye II gruppy byli razdeleny na gruppu IIa, v kotoruiu voshli patsienty tol'ko s fibroadenomoĭ, i gruppu IIb, v kotoruiu voshli ostal'nye patsienty s listovidnoĭ fibroadenomoĭ, fibroadenomatozom, fibrozno-kistoznoĭ mastopatieĭ, vnutriprotokovym papillomatozom, skleroziruiushchim adenozom i fibrozno-kistoznoĭ mastopatieĭ s mikrokal'tsinatami. Kontsentratsiiu IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1Ra, TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF i MCP-1 opredeliali v supernatante bioptatov opukholeĭ. Dlia otsenki tsitokinprodutsiruiushcheĭ aktivnosti bioptatov opukholi i ee mikrookruzheniia opredeliali indeks vliianiia poliklonal'nykh aktivatorov na produktsiiu tsitokinov. Pri sravnenii I i II grupp byli polucheny vysokie indeksy vliianiia poliklonal'nykh aktivatorov na produktsiiu IL-17, IL-18 i TNF-α u patsientov s nezlokachestvennymi zabolevaniiami. Bolee vysokie znacheniia indeksov vliianiia poliklonal'nykh aktivatorov na produktsiiu IL-18, TNF-α, i IL-1β i sootnosheniia IL1β/IL1Ra byli u patsientov s fibroadenomoĭ po sravneniiu s invazivnoĭ kartsinomoĭ nespetsificheskogo tipa. Statisticheski znachimykh razlichiĭ v gruppakh I i IIb po indeksam vliianiia poliklonal'nykh aktivatorov obnaruzheno ne bylo. Éto pozvoliaet predpolozhit' sushchestvovanie analogichnykh invazivnoĭ kartsinome nespetsificheskogo tipa izmeneniĭ v molochnoĭ zheleze u patsientov IIb gruppy. Pri sravnenii grupp IIa i IIb byli polucheny bolee vysokie znacheniia indeksa vliianiia poliklonal'nykh aktivatorov na produktsiiu IL-1β, a takzhe sootnosheniia IL1β/IL1Ra u patsientov s fibroadenomoĭ molochnoĭ zhelezy. Provedennoe issledovanie pozvolilo vyiavit' osobennosti tsitokinprodutsiruiushchego resursa bioptatov opukholeĭ u patsientov s invazivnoĭ kartsinomoĭ nespetsificheskogo tipa i s dobrokachestvennymi opukholiami molochnoĭ zhelezy.

The relationship between the content of supernatant cytokines and the expression of non-specific type of markers of epithelial-mesenchymal transition markers in the presence (group II) and the absence of lymphogenous metastasis (group I) were studied in biopsy specimens of mammary invasive breast carcinoma. The concentrations of TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF, MCP-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β and IL-1Ra, as well as the expression of immunohistochemical (IHC) markers of the epithelial-mesenchymal transition - cadherin-E (CDH1), β-1 integrin (CD29) and type II collagen (CII) were assayed. Results have shown that patients of these groups statistically significantly differed in spontaneous production of IL-18 and G-CSF, in terms of the index of the effect of the polyclonal activator on G-CSF production. There was a correlation between the parameter of CII expression in tumor tissue and the production of cytokines by tumor biopsy specimens; it was characteristic of all patients with invasive carcinoma of a non-specific type, and correlations, both direct and reverse between the expression indices of CDH1, CD29 and cytokine production varied depending on the presence or the absence of lymphogenous metastasis. The study revealed the features of the correlation between the production of cytokines by the tumor, its microenvironment and the expression of IHC markers of the epithelial-mesenchymal transition in patients with invasive non-specific breast carcinoma in the presence and absence of lymphogenous metastasis.Izuchena vzaimosviaz' mezhdu soderzhaniem tsitokinov supernatantov i ékspressieĭ v bioptatakh invazivnoĭ kartsinomy molochnoĭ zhelezy nespetsificheskogo tipa markerov épitelial'no-mezenkhimal'nogo perekhoda pri nalichii (II gruppa) i otsutstvii (I gruppa) limfogennogo metastazirovaniia. Opredeliali kontsentratsiiu TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF, MCP-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β i IL-1Ra, a takzhe ékspressiiu immuno-gistokhimicheskikh (IGKh) markerov épitelial'no-mezenkhimal'nogo perekhoda – kadgerina-E (CDH1), β-1-integrina (CD29) i kollagena II tipa (CII). Po rezul'tatam issledovaniia patsienty étikh grupp statisticheski znachimo otlichalis' po spontannoĭ produktsii IL-18 i G-CSF, po indeksu vliianiia poliklonal'nogo aktivatora na produktsiiu G-CSF. Pri izuchenii vzaimosviazi mezhdu produktsieĭ tsitokinov supernatantami i ékspressieĭ v nikh IGKh markerov ustanovleno, chto korreliatsionnye sviazi mezhdu pokazatelem ékspressii CII v tkani opukholi i produktsieĭ tsitokinov bioptatami opukholi kharakterny dlia vsekh patsientov s invazivnoĭ kartsinomoĭ nespetsificheskogo tipa, a korreliatsionnye sviazi, kak priamye, tak i obratnye mezhdu pokazateliami ékspressii CDH1, CD29 i produktsieĭ tsitokinov, razlichalis' v zavisimosti ot nalichiia ili otsutstviia limfogennogo metastazirovaniia. Provedennoe issledovanie pozvolilo vyiavit' osobennosti korreliatsionnykh sviazeĭ mezhdu produktsieĭ tsitokinov opukhol'iu, ee mikrookruzheniem i ékspressieĭ IGKh – markerov épitelial'no-mezenkhimal'nogo perekhoda u patsientov s invazivnoĭ kartsinomoĭ molochnoĭ zhelezy nespetsificheskogo tipa pri nalichii i otsutstvii limfogennogo metastazirovaniia.

Investigation of the cytokine expression dynamics as well as the cytokine-producing potential of immune-competent cells allows extensive studies of their functional characteristics. mRNAs encoding a number of cytokine genes are relatively stable, thus their level may be used as a marker for assessing the levels of activation and proliferation of immunocompetent cells as well as for evaluating the cytokine-producing potential of immunocompetent cells. In our work, we assessed correlations between the levels of mRNA expression specific for IL-10, TNF- α , GM-CSF cytokines determined in a culture of differentiated macrophage U937 cells, and protein concentrations of the same cytokines as measured in supernates of U937 cell cultures, without and after exposure to polyclonal activators. The IL-10, TNF-α, GM-CSF mRNA expression was determined by real-time quantitative polymerase chain reaction. Protein concentrations of IL-10, TNF-α, GM-CSF cytokines in the culture supernatant of U937 cells were measured by an enzyme immunoassay. The use of an initially homogeneous cell culture in the study is convenient due to the identical conditions in all experimental variants. The most pronounced effect of polyclonal activators is exerted upon production of GM-CSF mRNA, as well as protein concentration of this cytokine in the cell culture supernatants, thus actually coinciding with RT-qPCR results. The TNF-α mRNA level decreased under the influence of polyclonal activators, whereas concentration of this cytokine was decreased in the cell supernate. The TNF-α protein in a culture medium did not reflect temporal changes in the cellular TNF-α mRNA expression, probably, due to potential decrease of cellular mRNA occurring by the feedback inhibitory mechanism. While the cytokines can accumulate and remain in the supernatant, the mRNA-related events leading to cytokine formation may be completed earlier. Therefore, the signalling pathways and cytokine release kinetics should be studied after establishing the time dependence at short time intervals, which may be individual for each cytokine. Thus, the results of a study using polyclonal activators suggest that polyclonal activators applied as mitogens, have a significant effect upon the concentration of secreted IL-10, TNFα and GM-CSF. In this case, polyclonal activators affect the levels of mRNA encoded by cytokine genes, thus indicating transcriptional mechanisms of its action. But, in view of the fact that the data are ambiguous, in order to achieve greater correspondence between the changes in the studied proteins and specific mRNAs, a detailed description of the time dependence is required for the changes in mRNA contents.

We performed clinical observations and laboratory examination of 22 patients with chronic pyelonephritis (chronic renal failure, CRF) and 30 healthy individuals. The patients with CRF were examined twice. The first group (Group I) included patients with exacerbation of the disease. The comparison series (Group II) was represented by the same patients who were examined 1.5-3 months after completion of treatment, without clinical exacerbation of chronic pyelonephritis (CPN). Laboratory signs of acute renal damage were not detectable in all the patients examined. Concentrations of VEGF, MCP-1, IL-8 and IL-18 were determined in urine samples of all examined persons by ELISA technique. Protein spectrum of urine was assessed in six patients from Group I, and in six cases of Group II by means of mass spectrometry, using Agilent 1100 chromatographic device, and LTQ-FT Ultra hybrid mass spectrometer. The results of parallel determination of urine proteins by the two methods have shown that the evolving CPN exacerbation is associated with local secondary immune deficiency at the level of renal tubular urothelium. Determination of urine proteome by means of mass spectrometry in exacerbating disease allows identify the proteins associated with damage to epithelial lining of renal tubules and development of local immune response.

The synthesis of inhibitors for oxidative stress-associated destructive processes based on 2H-imidazole-derived phenolic compounds affording the bifunctional 2H-imidazole-derived phenolic compounds in good-to-excellent yields was reported. In particular, a series of bifunctional organic molecules of the 5-aryl-2H-imidazole family of various architectures bearing both electron-donating and electron-withdrawing substituents in the aryl fragment along with the different arrangements of the hydroxy groups in the polyphenol moiety, namely derivatives of phloroglucinol, pyrogallol, hydroxyquinol, including previously unknown water-soluble molecules, were studied. The structural and antioxidant properties of these bifunctional 5-aryl-2H-imidazoles were comprehensively studied. The redox transformations of the synthesized compounds were carried out. The integrated approach based on single and mixed mechanisms of antioxidant action, namely the AOC, ARC, Folin, and DPPH assays, were applied to estimate antioxidant activities. The relationship “structure-antioxidant properties” was established for each of the antioxidant action mechanisms. The conjugation effect was shown to result in a decrease in the mobility of the hydrogen atom, thus complicating the process of electron transfer in nearly all cases. On the contrary, the conjugation in imidazolyl substituted phloroglucinols was found to enhance their activity through the hydrogen transfer mechanism. Imidazole-derived polyphenolic compounds bearing the most electron-withdrawing functionality, namely the nitro group, were established to possess the higher values for both antioxidant and antiradical capacities. It was demonstrated that in the case of phloroglucinol derivatives, the conjugation effect resulted in a significant increase in the antiradical capacity (ARC) for a whole family of the considered 2H-imidazole-derived phenolic compounds in comparison with the corresponding unsubstituted phenols. Particularly, conjugation of the polyphenolic subunit with 2,2-dimethyl-5-(4-nitrophenyl)-2H-imidazol-4-yl fragment was shown to increase ARC from 2.26 to 5.16 (104 mol-eq/L). This means that the considered family of compounds is capable of exhibiting an antioxidant activity via transferring a hydrogen atom, exceeding the activity of known natural polyphenolic compounds.

Introduction Breast cancer is a heterogeneous disease that has multiple molecular and morphological subtypes. Nonetheless, the relation between various molecular subtypes and functional characteristics of a tumor in terms of cytokine secretion remains unknown. Methods We studied spontaneous and mitogen-induced cytokine secretion by invasive breast carcinoma of no special type (IBC NST; cultured tumors and cultured peripheral blood cells), depending on a molecular tumor subtype (where “mitogens” means “polyclonal activators” (PA): phytohemagglutinin p, phytohemagglutinin M, concanavalin A, and Escherichia coli lipopolysaccharide). Enzyme-linked immunosorbent assays were used to determine concentrations of IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1Ra, TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF, and MCP-1 in culture supernatants of the tumors and peripheral blood cells. Results The luminal B HER2-positive molecular subtype of IBC NST was found to feature the highest spontaneous secretion of IL-6 and IL-8 and the highest mitogen-induced secretion of IL-6, IL-8, IL-1Ra, and TNF-α by tumors; the highest mitogen-induced secretion of IL-2, IL-6, IL-8, IL-1β, TNF-α, IFN-γ, and G-CSF by peripheral blood cells; and the highest cytokine-producing potential (the ratio of mitogen-induced to spontaneous secretion) of peripheral blood cells for the secretion of IL-6, IL-8, and IL-1Ra as compared to other molecular subtypes. The triple-negative subtype of IBC NST was characterized by the lowest cytokine-producing potential of tumors for the secretion of IL-6 and IL-8 as compared to other molecular subtypes as well as a lower “stimulation index of polyclonal activators” (calculated as (cytokine secretion after incubation with PA)/(spontaneous cytokine secretion)) for IL-18 secretion as compared to luminal subtypes. The XYZ correlated with a suppressive effect of PA on cytokine secretion by tumors of the triple-negative molecular subtype. Conclusion Therefore, our findings indicate that in IBC NST of luminal B HER2-positive and triple-negative molecular subtypes, the cytokine network has distinctive functional features.

To date, data on the presence of adenoviral receptors in fish are very limited. In the present work, we used mouse recombinant adeno-associated viral vectors (rAAV) with a calcium indicator of the latest generation GCaMP6m that are usually applied for the dorsal hippocampus of mice but were not previously used for gene delivery into fish brain. The aim of our work was to study the feasibility of transduction of rAAV in the mouse hippocampus into brain cells of juvenile chum salmon and subsequent determination of the phenotype of rAAV-labeled cells by confocal laser scanning microscopy (CLSM). Delivery of the gene in vivo was carried out by intracranial injection of a GCaMP6m-GFP-containing vector directly into the mesencephalic tegmentum region of juvenile (one-year-old) chum salmon, Oncorhynchus keta. AAV incorporation into brain cells of the juvenile chum salmon was assessed at 1 week after a single injection of the vector. AAV expression in various areas of the thalamus, pretectum, posterior-tuberal region, postcommissural region, medial and lateral regions of the tegmentum, and mesencephalic reticular formation of juvenile O. keta was evaluated using CLSM followed by immunohistochemical analysis of the localization of the neuron-specific calcium binding protein HuCD in combination with nuclear staining with DAPI. The results of the analysis showed partial colocalization of cells expressing GCaMP6m-GFP with red fluorescent HuCD protein. Thus, cells of the thalamus, posterior tuberal region, mesencephalic tegmentum, cells of the accessory visual system, mesencephalic reticular formation, hypothalamus, and postcommissural region of the mesencephalon of juvenile chum salmon expressing GCaMP6m-GFP were attributed to the neuron-specific line of chum salmon brain cells, which indicates the ability of hippocampal mammal rAAV to integrate into neurons of the central nervous system of fish with subsequent expression of viral proteins, which obviously indicates the neuronal expression of a mammalian adenoviral receptor homolog by juvenile chum salmon neurons.

The considerable post-traumatic brain recovery in fishes makes them a useful model for studying the mechanisms that provide reparative neurogenesis, which is poorly represented in mammals. After a mechanical injury to the telencephalon in adult fish, lost neurons are actively replaced due to the proliferative activity of neuroepithelial cells and radial glia in the neurogenic periventricular zone. However, it is not enough clear which signaling mechanisms are involved in the activation of adult neural stem cells (aNSC) after the injury (reactive proliferation) and in the production of new neurons (regenerative neurogenesis) from progenitor cells (NPC). In juvenile Pacific salmon, the predominant type of NSCs in the telencephalon are neuroepithelial cells corresponding to embryonic NSCs. Expression of glutamine synthetase (GS), a NSC molecular marker, was detected in the neuroepithelial cells of the pallium and subpallium of juvenile chum salmon, Oncorhynchus keta. At 3 days after a traumatic brain injury (TBI) in juvenile chum salmon, the GS expression was detected in the radial glia corresponding to aNSC in the pallium and subpallium. The maximum density of distribution of GS+ radial glia was found in the dorsal pallial region. Hydrogen sulfide (H2S) is a proneurogenic factor that reduces oxidative stress and excitotoxicity effects, along with the increased GS production in the brain cells of juvenile chum salmon. In the fish brain, H2S producing by cystathionine β-synthase in neurogenic zones may be involved in maintaining the microenvironment that provides optimal conditions for the functioning of neurogenic niches during constitutive neurogenesis. After injury, H2S can determine cell survivability, providing a neuroprotective effect in the area of injury and reducing the process of glutamate excitotoxicity, acting as a signaling molecule involved in changing the neurogenic environment, which leads to the reactivation of neurogenic niches and cell regeneration programs. The results of studies on the control of the expression of regulatory Sonic Hedgehog genes (Shh) and the transcription factors Paired Box2 (Pax2) regulated by them are still insufficient. A comparative analysis of Pax2 expression in the telencephalon of intact chum salmon showed the presence of constitutive patterns of Pax2 expression in neurogenic areas and non-neurogenic parenchymal zones of the pallium and subpallium. After mechanical injury, the patterns of Pax2 expression changed, and the amount of Pax2+ decreased (p &lt, 0.05) in lateral (Dl), medial (Dm) zones of the pallium, and the lateral zone (Vl) of the subpallium compared to the control. We believe that the decrease in the expression of Pax2 may be caused by the inhibitory effect of the Pax6 transcription factor, whose expression in the juvenile salmon brain increases upon injury.

Fish are a convenient model for the study of reparative and post-traumatic processes of central nervous system (CNS) recovery, because the formation of new cells in their CNS continues throughout life. After a traumatic injury to the cerebellum of juvenile masu salmon, Oncorhynchus masou, the cell composition of the neurogenic zones containing neural stem cells (NSCs)/neural progenitor cells (NPCs) in the acute period (two days post-injury) changes. The presence of neuroepithelial (NE) and radial glial (RG) neuronal precursors located in the dorsal, lateral, and basal zones of the cerebellar body was shown by the immunohistochemical (IHC) labeling of glutamine synthetase (GS). Progenitors of both types are sources of neurons in the cerebellum of juvenile O. masou during constitutive growth, thus, playing an important role in CNS homeostasis and neuronal plasticity during ontogenesis. Precursors with the RG phenotype were found in the same regions of the molecular layer as part of heterogeneous constitutive neurogenic niches. The presence of neuroepithelial and radial glia GS+ cells indicates a certain proportion of embryonic and adult progenitors and, obviously, different contributions of these cells to constitutive and reparative neurogenesis in the acute post-traumatic period. Expression of nestin and vimentin was revealed in neuroepithelial cerebellar progenitors of juvenile O. masou. Patterns of granular expression of these markers were found in neurogenic niches and adjacent areas, which probably indicates the neurotrophic and proneurogenic effects of vimentin and nestin in constitutive and post-traumatic neurogenesis and a high level of constructive metabolism. No expression of vimentin and nestin was detected in the cerebellar RG of juvenile O. masou. Thus, the molecular markers of NSCs/NPCs in the cerebellum of juvenile O. masou are as follows: vimentin, nestin, and glutamine synthetase label NE cells in intact animals and in the post-traumatic period, while GS expression is present in the RG of intact animals and decreases in the acute post-traumatic period. A study of distribution of cystathionine &beta, synthase (CBS) in the cerebellum of intact young O. masou showed the expression of the marker mainly in type 1 cells, corresponding to NSCs/NCPs for other molecular markers. In the post-traumatic period, the number of CBS+ cells sharply increased, which indicates the involvement of H2S in the post-traumatic response. Induction of CBS in type 3 cells indicates the involvement of H2S in the metabolism of extracellular glutamate in the cerebellum, a decrease in the production of reactive oxygen species, and also arrest of the oxidative stress development, a weakening of the toxic effects of glutamate, and a reduction in excitotoxicity. The obtained results allow us to consider H2S as a biologically active substance, the numerous known effects of which can be supplemented by participation in the processes of constitutive neurogenesis and neuronal regeneration.

Natalia V Yurina,1 Tatiana A Ageeva,2 Aleksandr M Goryachkin,2 Nikolay A Varaksin,3 Tatiana G Ryabicheva,3 Alexandr A Ostanin,4 Elena R Chernykh,4 Alexander V Romashchenko,5 Anastasia S Proskurina,5 Sergey Bogachev,5 Alexey V Purtov6 1Autonomous Non-Profit Organization “Regional Center for High Medical Technologies”, Novosibirsk, Russia; 2Novosibirsk State Medical University, Novosibirsk, Russia; 3JSC “Vector-Best”, Novosibirsk, Russia; 4Research Institute of Fundamental and Clinical Immunology, Novosibirsk, Russia; 5Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia; 6LLC “Angiopharm Laboratory”, Novosibirsk, RussiaCorrespondence: Sergey Bogachev Email labmolbiol@mail.ruPurpose: The purpose of this study was to assess the capability of recombinant angiogenin isolated from Pichia pastoris yeasts to stimulate regenerative processes in the dermis of experimental animals.Patients and Methods: Wistar rats were administered with recombinant angiogenin intracutaneously. Morphological examination of the skin and the assessment of the proliferative activity of the epidermal cells were carried out. Additionally, cytokine production by human whole blood cells exposed to angiogenin was analyzed ex vivo.Results: Administration of angiogenin stimulates collagen fiber formation and angiogenesis. This stimulation is tightly associated with an increase in the number of fibroblasts, an increased numerical density of dermal blood vessels and an increased density of collagen fibers; also, it activates the proliferation of basal cells. Angiogenin induces the production of MCP, IL-8, IL-6, IL-1β, TNF-α, IL-10, TGF-β, and VEGF by blood cells.Conclusion: The results obtained indicate a broad spectrum of actions of recombinant angiogenin during regenerative processes in the basal layer of the dermis.Keywords: intracutaneous injections, pathomorphological analysis, cytokines, regeneration

The research looks at concentrations of proteins and cytokines in blood cell supernatants as well as expression of histological markers of atypical cells in the breast diseases. Also, it examines the correlation between concentrations of proteins and cytokines and histological markers of atypical cells. It is found that concentration of histidine rich glycoprotein in blood cell supernatants in the invasive carcinoma of no special type is higher compared to benign breast diseases. Furthermore, the research observes the following statistical relationships: negative correlation between concentrations of estrogen receptor-α and expression of integrin β-1; positive correlation between concentrations of histidine rich glycoprotein and expression of type II collagen; positive correlation between concentrations of e-cadherin and expression of e-cadherin; and positive correlation between concentrations of plasminogen activator inhibitor-1 and expression of type II collagen. Cytokines production by blood cells is identified as Index of the influence of polyclonal activators (combination of mitogens). The research shows the difference in the Index of influence on IL-1RA and TNF-α production between benign breast diseases with high-risk and low-risk of malignant transformation, and between benign breast diseases with high-risk of malignant transformation and breast cancer with lymph node metastasis. The Index of the influence of polyclonal activators on IL-1RA production gets higher as disease prognosis worsens. As blood and tumor samples show, patients with benign breast diseases have negative correlation between index of the influence of polyclonal activators on IL-1RA production and expression of type II collagen. Additionally, patients with breast cancer, as well as breast cancer with lymph node metastasis have positive correlation between expression of e-cadherin and spontaneous and polyclonal activator-stimulated IL-1RA production. The results of the research showed that concentrations of histidine rich glycoprotein and IL-1RA in blood cells supernatant and expression of e-cadherin can be used as markers for early diagnosis of benign breast diseases with risk of malignant transformation.

Introduction.Occupational exposure to dust in concentrations sometimes exceeding allowable norms, infl uence of associated hazards (irritating gases, toxic chemicals, unfavorable microclimate at workplace, heavy physical work) cause occupational and occupationally conditioned bronchopulmonary diseases and lower work capacity in workers with main occupations of refractory materials production.Objective.To study functional state of respiratory system for diagnosis of early disorders of pulmonary ventilation and gases exchange in workers of moulded refractory materials production.Materials and methods.Prospective randomized study included apparently healthy male workers (n = 61) of refractory materials plant producing chamott e-silica and spinel-periclase-carbon refractories. Clinic of Ekaterinburg medical research center in 2017–2018 provided examination of carriers (n=21) in spinel-periclase-carbon refractories production shop and pressmen (n=40) of moulding area in chamott e-silica refractories production, aged 27 to 60 years, with length of service in hazardous conditions from 4 to 37 years. Bodyplethysmography helped to assess general lung capacity (GLC), residual lung volume, ratio of residual lung volume to general lung capacity, functional residual lung capacity, bronchial resistance and diff usion lung ability by carbon oxide via single inspiration method. For nonventilated lung volume, the authors used ∆ GLC value that is a diff erence between GLC values measured via bodyplethysmography and via helium dilution in single inspiration maneuver.Results.Obstructive syndrome (6.5%) was a main type of ventilation disorders among the examinees, and equally frequent among the workers engaged into spinel-periclase-carbon refractories production (9.5%) and in those engaged into chamott esilica refractories production (5%). Th e workers engaged into spinel-periclase-carbon refractories production had obstructive syndrome associated with lung hyperinfl ation, and those engaged into chamott e-silica refractories production had also a tendency to restrictive disorders. Lung gases exchange disorders were seen in one third of the examinees, equally frequent in both workers engaged into spinel-periclase-carbon refractories production and those engaged into chamott e-silica refractories production, manifested in 2 variants: lower diff usion lung capacity (fi rst variant) and lower diff usion lung capacity with increased ∆ GLC (second variant).Conclusion.Th e study results prove necessity of bodyplethysmography and diff usion lung capacity diagnosis to reveal perfusion and ventilation disorders at early stages in workers engaged into spinel-periclase-carbon refr actories production over 8 years and in those engaged into chamott e-silica refr actories production over 12 years.

Expression of cystathionine β-synthase (CBS) in the brain of adult trout under normal conditions and 1 week after an eye injury was assessed using Western blot analysis. The study of CBS distribution in the brain of intact trout and after a mechanical eye injury in the telencephalon, tectum, cerebellum, and brainstem was carried out by the method of immunoperoxidase labeling on free-floating sections. The results of the study showed an increase in CBS expression in different brain divisions after an eye injury. In the visual projection center of the brain (tectum), radial glia cells expressing CBS were revealed after the injury. The emergence of CBS+ heterogeneous radial glia in the trout tectum after eye injury indicates the ability of the reactive neural stem cells (NSC) to synthesize hydrogen sulfide. CBS+ fibers and cells were found in the proliferative zones of cerebellum (valvula) and telencephalon (ventral region). In the intact trout tegmentum, CBS expression was observed in large neurons of the dorsal region and in the proliferative zones. After the eye injury, reactive neurogenic niches appeared in the brainstem, and no CBS-immunopositivity was detected in the periventricular zone. The increased expression of H2S-producing CBS enzyme after a traumatic impact can probably be explained by the neuroprotective functions of hydrogen sulfide, which are implemented in the matrix zones of the brain associated with reparative neurogenesis. A cultivation of trout brain cells showed the formation of neurosphere-like complexes, a part of which exhibited CBS-immunopositivity in the monolayer.

Processes of proliferation and constitutive neuro- and gliogenesis in the mesencephalic tegmentum of intact juvenile chum salmon, Oncorhynchus keta, and at 3 days after a traumatic injury were studied by immunohistochemistry (IHC) labeling of PCNA, HuCD, and GFAP. In the chum tegmentum, the proliferative activity was revealed both in separate cells and in small cell clusters of the periventricular zone (PVZ). The presence of constitutive neurogenic zones provides the processes of persistent brain growth. After damage to the tegmentum, proliferation in PVZ is activated, the constitutive neurogenic zones reactivate, reactive neurogenic niches form in the parenchyma, and the proliferative activity is also initiated in the centers of secondary proliferation (basal tegmentum). It was first found that traumatic damage to the tegmentum leads to accelerated differentiation of neurons in the subventricular zone (SVZ) and dorsomedial tegmentum as well as to the appearance of HuCD+ cells with the ependymo- and radioglial phenotype in SVZ, which are absent in intact animals. It was first shown that the local foci of posttraumatic neurogenesis, located in the reticular formation parenchyma, and the zones of posttraumatic gliosis, which contribute to a more efficient process of cell migration to the injury area, are formed as a result of tegmentum damage. The data obtained provide new information on the constitutive biology of neural stem cells and their involvement in brain regeneration.

In contrast to astrocytes in mammals, fish astrocytes promote axon regeneration after brain injury and actively participate in the regeneration process. Neurolin, a regeneration-associated, Zn8-labeled protein, is involved in the repair of damaged optic nerve in goldfish. At 1 week after unilateral eye injury, the expression of neurolin in the optic nerve and chiasm, and the expression of Pax6 that influences nervous system development in various brain regions in the rainbow trout (Oncorhynchus mykiss) were detected. Immunohistochemical staining revealed that the number of Zn8+ cells in the optic nerve head and intraorbital segment was obviously increased, and the increase in Zn8+ cells was also observed in the proximal and distal parts of injured optic nerve. This suggests that Zn8+ astrocytes participate in optic nerve regeneration. ELISA results revealed that Pax6 protein increased obviously at 1 week post-injury. Immunohistochemical staining revealed the appearance of Pax6+ neurogenic niches and a larger number of neural precursor cells, which are mainly from Pax6+ radial glia cells, in the nuclei of the diencephalon and optic tectum of rainbow trout (Oncorhynchus mykiss). Taken together, unilateral eye injury can cause optic nerve reaction, and the formation of neurogenic niches is likely a compensation phenomenon during the repair process of optic nerve injury in rainbow trout (Oncorhynchus mykiss).

At present, only ductal carcinoma in situ is included into the group of precancerous lesions of mammary ducts, according to International Agency for the Study of Cancer. However, based on recent publications, in addition to ductal carcinoma in situ, sclerosing adenosis, intraductal proliferative lesions and radial scar may be also attributed to precancerous changes. A variety of both benign and malignant events in mammary gland, the features of neoplastic growth and age of the patients require new approaches to study of carcinogenic events in mammary gland. As based on the known role of cytokines in genesis of malignancies, the aim of the study was to evaluate the cytokine-producing resource of immunocompetent blood cells in malignant, benign and precancerous mammary disorders. To assess the cytokine-producing resource of immunocompetent blood cells in the patients, we studied quantitative effects of polyclonal activators upon production of cytokines by immunocompetent blood cells of patients with invasive ductal cancer representing a histological type of adenocarcinoma (group I), and patients with non-malignant breast neoplasias (group II). At subsequent step, the patients with non-malignant neoplasms of the breast were divided into a subgroup of patients with only fibroadenoma and mastopathy (group III), and a group which included patients with precancerous diseases, i.e., sclerosing adenosis and interductal proliferates (group IV). Concentrations of IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1ra, TNFα, IFNγ, G-CSF, GM-CSF, VEGF, and MCP-1 were determined by solid-phase enzyme immunoassay. When comparing groups I and II, we revealed higher influence of polyclonal activators upon production of G-CSF and GM-CSF in patients with invasive ductal cancer. When comparing the influence of polyclonal activation for cytokine production in patients of I and III groups, higher values were registered in patients with invasive ductal cancer (production of IL-2, G-CSF, and GM-CSF), and in patients with fibroadenoma and mastopathy (IL-18, and TNFαproduction). When comparing patients of groups I and IV, higher indexes of the polyclonal activator effects were found only for IL-1ra, G-CSF, and VEGF production in invasive ductal cancer. When comparing the indexes of polyclonal activator influence upon cytokine production of groups III and IV, higher values were obtained in patients with benign changes for the following cytokines: IL-8, IL-18, IL-1β, IL-1ra and TNFα, in contrast to patients with sclerosing adenosis and proliferates. The lower indexes of polyclonal activating effects upon the production of a number of cytokines in patients with precancerous changes, as compared to patients with malignant and benign breast tumors, do not indicate a decreased functional activity of immunocompetent blood cells. However, those may be due to high level of spontaneous cytokine production in sclerosing adenosis and interductal proliferates.

The distribution of the Pax2+ transcription factor in the optic nerve after a mechanical eye injury on the side of damage and in the contralateral nerve has been studied in the trout Oncorhynchus mykiss. It has been found that injury of the optic nerve in this fish species causes Pax2+ reactive astrocytes involved in the initial stages of optic nerve axon regeneration to increase in number, especially in the area of the head and the proximal part of the optic nerve. As the optic nerve in trout is damaged, a significant growth of the heterogeneous population of Pax6+ cells occurs in the brain divisions that have direct retinal inputs, diencephalon, and optic tectum. A part of the Pax6+ cells have an undifferentiated phenotype and are a component of reactive neurogenic niches located in the periventricular zone and parenchymal regions of the brain. Another population of Pax6+ cells has the radial glial phenotype and appears as a result of activation of the constitutive neurogenic domains also within the newly formed reactive neurogenic niches. Thus, due to the optic nerve injury, a pronounced neurogenic response associated with the appearance of reactive neurogenic niches and radial glia arises both in the brain divisions with direct retinal projections and in those lacking the retinal projections as well as in remote regions. The results obtained indicate that the damage to the optic nerve causes an increased reactive neurogenesis in the brain of adult trout.

A series of novel water-soluble azaheterocyclic derivatives of nido-carborane bearing quinoxaline, 2H-imidazole or 1,2,4-triazine moieties were first synthesized in 82–91% yields. The structures of these boron-enriched compounds were confirmed by the data of NMR, IR spectroscopy, and mass spectrometry. To access the toxicity level for these organoboron compounds, the cytotoxicity indexes (IC50) were determined using by the MTT test on both human glioblastoma cell A-172 (IC50 = 150–243 μM) and human embryonic lung cells (IC50 = 424–944 μM) lines. The obtained preliminary results from in vitro analysis enable the synthesized water-soluble azaheterocyclic carboranes to be considered as challenging candidates in the design of agents for boron-neutron capture therapy (BNCT) of malignant tumors.

Comparative and combined damaging effects of NiO and Mn3O4 nanoparticles were estimated on cultures of several established human cell lines. The cytotoxicity indices used were: (a) reduction in cellular dehydrogenase activity, (b) decrease in the ATP-content, (c) for SH-SY5Y cells also decrease in the tyrosine hydroxylase content. The combined cytotoxicity was modeled using the Response Surface Methodology. When assessing the stability of metal oxide nanoparticles (MeO-NPs) in cultural media used by us, we found that the addition of the fetal bovine serum (FBS) to them renders NiO-NPs and, to even greater extent, Mn3O4-NPs exponentially slow soluble while without FBS their dissolution was virtually undetectable. At the same time, sedimentation of these MeO-NPs noticeably slowed down in the presence of the same FBS. We have found dependence of cell damage on concentrations of MeO-NPs and higher cytotoxicity of Mn3O4-NP compared with NiO-NP. Thus, comparative assessment of the NPs unspecific toxicity obtained in our animal experiments was reproduced by the "in vitro" tests. However, with respect to manganese-specific brain damage "in vivo" discovered previously, present experiments on neurons "in vitro" showed only a certain enhancing effect of Mn3O4-NP on the action of NiO-NP, but the role of NiO-NP in the combination prevailed.

It is known that a relationship exists between patient’s age and pathogenesis of malignant tumors associated with ability of tumor and its microenvironment to secrete cytokines that may play an important role in carcinogenesis. The aim of present study was a comparative analysis of cytokine production by peripheral blood cells and tumor samples when exposed to polyclonal activators (PA) in patients of different age groups with invasive ductal carcinoma. Peripheral blood samples of 54 women with invasive ductal carcinoma were under study. The patients were divided into the following groups: I, 40-60 years old; II, 61 year and older. Metastases into lymph nodes were revealed at the moment of study in 13 patients (42%) from group I. By contrast, only 3 of 23 (10%) patients with metastases were revealed in group II. Fisher’s ratio test showed significant differences between occurrence of metastases in lymph nodes, and patient age (p = 0.034). In both groups of patients, there was an increase of IL-6, IL-8, IL-10, IL-17, IL-18, IL-1ra, G-CSF, GM-CSF and VEGF concentrations in tumor supernates and its microenvironment, when compared with the production of cytokines by blood cells, thus indicating to high functional activity of tumor together with its microenvironment. Effect of PA led to increase of IL-2, IL-10, IL-17, IL-1β, IL-1ra, TNFα and IFNγ concentrations in blood cells supernatant which suggested their high ability to produce cytokines. In the group of patients who were 40-60 years old, the stimulatory index of PA (SIPA) for IFNγ production was several times higher than similar index in patients aged 61 years or more. Increase of SIPA for all cytokines in blood cells supernates, in comparison to SIPA of tumor supernates, could be explained by initially high levels of spontaneous cytokine production by the tumor and its microenvironment that are able to promote tumor growth. Therefore, effect of PA upon tumor tissue was not so expressed as its effect upon immunocompetent blood cells. Both tumor and microenvironment maintained high ability of IL-18 production, thus confirming its important role in carcinogenesis. In summary, these findings suggest that circulating immunocompetent cells may serve as an additional factor providing tumor progression, due to their high ability to secrete cytokines.

Objectives: Studying the influence of the method of selecting a control group in the radon case-control study on assessment of dose-effect dependence. Study the effect of additional additive or multiplicative risk factors on the results of the study. Materials and Methods: Simulation of a large-scale radon epidemiological case-control study. Analyzing of the influence of the representative value of the comparison subgroup on the results of calculating the odds ratio of the incidence of radiation-induced lung cancer. Analysis of different variants of standardization of the control group on the correctness of the dose-effect dependence. Development of methods for identifying risk factors that affect the incidence of lung cancer and determining the mechanism of their influence (additive or multiplicative). Results: It is shown that the insufficient representativeness of the comparison subgroup used in calculating the odds ratio can significantly distort the estimates of dose-effect dependence. In the presence of factors (sex, smoking, etc.), correlating both with the incidence of lung cancer and of radon concentration, complete standardization of such factors is necessary. Multiplicative risk factors without such correlation do not require additional standardization in the selection of a control group. Calculation of the odds ratio for various risk factors in subgroups differing in the levels of spontaneous cancer incidence allows to determine the significance of this factor for the incidence of lung cancer, and to establish the preferential mechanism of the influence of this factor (multiplicative or additive). Conclusions. When carrying out the radon case-control study, it is necessary to identify risk factors that correlate both with the incidence of lung cancer, and with the radon concentration, to assess the mechanism of action of these factors and to made the complete standardization of the control group for them.

A superficially located periventricular proliferative area with PCNA-immunopositive (PCNA+) cells, which corresponds to the pallial periventricular zone (PVZ) of other fish species, including its dorsal, lateral, and medial compartments, is discovered in the telencephalon of the juvenile masu salmon Oncorhynchus masou. The PCNA+ cells are also identified in the parenchyma of the masu salmon intact brain, and their maximum concentration is observed in the medial zone. After a mechanical injury, the zones of induced neurogenesis—neurogenic niches and sites of secondary neurogenesis surrounded by radial glial fibers—appear in the masu salmon telencephalon. The PVZ of the juvenile masu salmon pallium contains clusters of undifferentiated HuCD-immunopositive (HuCD+) neurons. A change in the HuCD+ cell topography is observed in the mechanically injured masu salmon telencephalon, namely, neurogenic niches in the lateral zone and an increase in the cell distribution density and cell migration patterns in the medial zone. A high level of persistent neurogenesis is characteristic of the juvenile masu salmon brain.

The objective of this work was to study proliferation processes and the role of glia and neural stem cells in the event of injurious action on cerebellum of masu salmon’s (Oncorhynchus masou) juvenile. Using the immunoperoxidase staining of the glial fibrillary acidic protein (GFAP) and proliferating cells nuclear antigen (PCNA), processes of proliferation and gliogenesis after mechanical trauma of cerebellum of cherry salmon’s (Oncorhynchus masou) juvenile were studied. After the trauma, the intensity of proliferation and migration processes varies in different zones. Proliferation processes decrease after the trauma in lateral and basal zones, and migration increases. In the dorsal zone, on the contrary, migration processes significantly decrease and proliferation increases. In the dorsal matrix zone of a cerebellum, intense cell proliferation was detected. In the dorsal, lateral, and basal zone of the molecular layer of cerebellum after traumatic damage, neurogenic niches containing PCNA and cells, as well as a heterogeneous population of PCNA-cells, were identified. At the location of neurogenic niches, fibers of radial glia and small single intensely or moderately labeled GFAP cells were discovered. As a result of damaging action, GFAP+ fibers of radial glia, which form differently directed radially oriented bundles, appeared in the dorsal matrix zone. Such structural formations have not been discovered in intact animals. We suppose that, after the trauma, structural reconstruction connected with partial spatial reorientation of the radial glia fibers and formation of specific directions for cells formed in this zone occurs in the dorsal matrix zone. As a result of the trauma, in masu salmon’s cerebellum, elements of the radial glia, including both cells possessing typical morphology and cell fragments presented as long radially oriented processes or cell body containing initial fragments of radial fibers, appeared.

In recent years, the concept of formation of a sufficiently autonomous cytokine network in a malignant tumour has emerged. In this regard, the data on the role of this network and its signalling pathways in the process of metastasis are an interesting topic. The aim of this study was to evaluate the in vitro cytokine-producing potential of mammary adenocarcinoma (MAC; and cells of its microenvironment) from patients with or without metastases in regional lymph nodes (LNs). By enzyme-linked immunosorbent assays of culture supernatants, we studied the cytokine production by biopsy samples of MAC: spontaneous and stimulated by polyclonal activators (PAs: phytohaemagglutinin, concanavalin A and lipopolysaccharide). The levels of spontaneous production of interleukin (IL)-10 and granulocyte colony-stimulating factor (G-CSF) and the amounts of IL-2, IL-10, G-CSF and monocyte chemoattractant protein-1 (MCP-1) produced during stimulation by PAs, as well as the index of stimulation by polyclonal activators (ISPA) for IL-2 production, were lower for MAC with LN metastasis than for MAC without LN metastasis. The levels of spontaneous production of IL-2 and interferon (IFN)-γ and the ISPA for granulocyte–macrophage colony-stimulating factor (GM-CSF) production were higher for MAC with LN metastasis. There were only three pairwise correlations between the produced cytokines that were specific to MAC with LN metastasis: IL-2 and IFN-γ, IL-6 and GM-CSF, and IL-8 and GM-CSF. There were 10 pairwise correlations between the produced cytokines that were specific to nonmetastasising MAC: IL-6 and IL-10, IL-6 and MCP-1, IL-8 and IL-10, IL-8 and MCP-1, IL-10 and G-CSF, IL-10 and MCP-1, IFN-γ and MCP-1, MCP-1 and G-CSF, G-CSF and IL-1Ra, and GM-CSF and tumour necrosis factor (TNF)-α. Our data indicate that metastatic tumours show desynchronisation of many pathways of induction and synthesis of cytokines that are characteristic of nonmetastatic tumours.

The aim of this research was to study cytokine production by blood immune cells, tumor, and its microenvironment, and characterize extracellular matrix of patients with invasive ductal carcinoma of no special type and lymphatic metastases. Spontaneous and polyclonal activators stimulated production of cytokines by blood immune cells, tumor and its microenvironment were studied in 95 patients with invasive ductal carcinoma of no special type. The concentration of IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1Ra, TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF and MCP-1 was determined by the solid-phase enzyme-linked immunosorbent assay. The condition of fibrous component and presence of neutral glycoproteins and sulfated glycosaminoglycans were evaluated during the research of extracellular matrix. Regional lymphatic metastases were detected in 35 of 95 patients. It was shown that in the presence or absence of lymphatic metastases index of polyclonal activators influence on the production of cytokines by blood immune cells was different for IL-6, IL-8, and IL-1β; while in the case of cytokine production by tumor and its microenvironment the index of influence was different for IL-2 and IL-17. The presence of lymphatic metastases corresponded with the rise of cytokines spontaneous production, while the absence of lymphatic metastases corresponded with the rise of cytokines production stimulated by polyclonal activators. The value of indices of polyclonal activators influence on the production of cytokines by blood immune cells pointed to the highly stimulating effect of polyclonal activators while the value of indices of polyclonal activators influence on cytokines production by tumor and its microenvironments pointed to the low and sometimes even absent effect of polyclonal activators. Basing on these data we propose a ratio of indices of polyclonal activators influence for the better evaluation of the probability of lymphatic metastases during preoperative period. After characterizing extracellular matrix we found out a point threshold, which, in 100% of cases, predicted the presence of lymphatic metastases basing on the condition of extracellular matrix. Using the data acquired, we are proposing a risk group for metastasis among women with no lymphatic metastases in the moment of check-up.Issledovali spontannuiu i stimulirovannuiu poliklonal'nymi aktivatorami produktsiiu tsitokinov immunokompetentnymi kletkami krovi, opukhol'iu i ee mikrookruzheniem, a takzhe sostoianie vnekletochnogo matriksa u 95 patsientok s invazivnoĭ kartsinomoĭ molochnoĭ zhelezy nespetsificheskogo tipa pri limfogennom metastazirovanii. S pomoshch'iu immunofermentnogo analiza opredeliali kontsentratsiiu IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1Ra, TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF i MCP-1. Pri izuchenii vnekletochnogo matriksa otsenivali sostoianie voloknistogo komponenta, soderzhanie neĭtral'nykh glikoproteinov i sul'fatirovannykh glikozaminoglikanov. Metastazy v regionarnye limfaticheskie uzly prisutstvovali u 35 iz 95 patsientok. Issledovanie pokazalo, chto pri nalichii ili otsutstvii limfogennogo metastazirovaniia indeks vliianiia poliklonal'nykh aktivatorov na produktsiiu tsitokinov immunokompetentnymi kletkami krovi razlichalsia po trem pokazateliam (IL-6, IL-8 i IL-1β), a indeks vliianiia poliklonal'nykh aktivatorov na produktsiiu tsitokinov kletkami opukholi i ee mikrookruzheniia po dvum (IL-2 i IL-17). Nalichiiu limfogennykh metastazov sootvetstvovalo povyshenie spontannoĭ produktsii tsitokinov, a ikh otsutstviiu – povyshenie produktsii, stimulirovannoĭ poliklonal'nymi aktivatorami. Znacheniia indeksov vliianiia poliklonal'nykh aktivatorov na produktsiiu tsitokinov immunokompetentnymi kletkami krovi svidetel'stvovali o vysokom stimuliruiushchem éffekte poliklonal'nykh aktivatorov, a znacheniia indeksov vliianiia poliklonal'nykh aktivatorov na produktsiiu tsitokinov kletkami opukholi i ee mikrookruzheniia – o nizkom i dazhe otsutstvuiushchem éffekte poliklonal'nykh aktivatorov. Na osnovanii poluchennykh dannykh bylo sostavleno sootnoshenie indeksov vliianiia poliklonal'nykh aktivatorov dlia otsenki veroiatnosti limfogennogo metastazirovaniia v dooperatsionnom periode. Pri otsenke sostoianiia vnekletochnogo matriksa byl vyiavlen porog, kotoryĭ v 100% sluchaev pokazal sootvetstvie sostoianiia vnekletochnogo matriksa nalichiiu metastazov v regionarnykh limfaticheskikh uzlakh. Na osnovanii poluchennykh rezul'tatov byla predlozhena gruppa riska po metastazirovaniiu sredi zhenshchin, ne imeiushchikh na moment issledovaniia metastazov v limfaticheskikh uzlakh.

Objective: The aim of this study was to evaluate the relationship between cytokine production, GM-CSF receptor (CSF2RA), and IL-1 receptor (IL1R2) expression in mammary adenocarcinoma and their association with it histopathological parameters and lymph node metastasis. Methods: We analyzed tumor biopsy samples (cultured in vitro) from 50 women (aged 43-75) with invasive ductal mammary adenocarcinomas. Enzyme-linked immunosorbent assay method the concentrations of interleukin 2, interleukin 6, interleukin 8, interleukin 10, interleukin 17, interleukin 18, interleukin 1β, interleukin 1Ra, tumor necrosis factor α, interferon γ, granulocyte colony-stimulating factor, granulocyte macrophage colony-stimulating factor, and vascular endothelial growth factor A were determined in culture supernatants. The expression of CSF2RA and IL1R2 in tumor biopsy was evaluated by immunohistochemical method. Results: We showed that the “cytokine profile” of a tumor (the ability of tumor cells and its microenvironment to produce different cytokines) is very individual. It has been shown that the features of the cytokine profile of the mammary adenocarcinoma are important for the formation and realization of the metastatic potential of the mammary adenocarcinoma. We found correlations between some histopathological parameters of mammary adenocarcinoma and coefficients KGM-CSF/CSF2RA and KIL-1β/IL1R2, which are the ratios of concentrations of granulocyte macrophage colony-stimulating factor and interleukin -1β to expression of CSF2RA and IL1R2, respectively. KGM-CSF/CSF2RA positively correlated with highly differentiated cells, and KIL-1β/IL1R2 positively correlated with the number of mitoses, poorly differentiated cells, and a number of lymph nodes with metastases. KGM-CSF/CSF2RA positively correlated with the concentrations of interleukin 6, interleukin 8, interleukin 1Ra, and granulocyte colony-stimulating factor. KIL-1β/IL1R2 positively correlated with concentrations of interleukin 1β and interferon γ and negative correlated with the concentrations of vascular endothelial growth factor A and tumor necrosis factor α. It is shown that KIL-1β/IL1R2 can be considered as a prognostic indicator predicting the probability of mammary adenocarcinoma metastasis to regional lymph nodes. Conclusions: The ratios of granulocyte macrophage colony-stimulating factor and interleukin 1β cytokines, produced in tumor, to the expression of CSF2RA and IL1R2 depend on levels of interleukin 6, interleukin 8, tumor necrosis factor α, interferon γ, granulocyte colony-stimulating factor, and vascular endothelial growth factor A and are important factors affecting the progression and metastasis of the breast cancer.

In the cerebellum of juvenileOncorhynchus masou, proliferating BrdU+ and HuCD+ cells and constitutive neurogenic niches were detected in different zones; the largest number of labeled cells were found in the dorsal part of the molecular layer and the dorsal matrix zone (DMZ). Cells labeled with glutamine synthetase (GS) and radial glia were also present in the intactO. masoucerebellum. The most intensive proliferation was detected in the rostral part of cerebellum. This part is assumed to contain active zones of constitutive neurogenesis. After an injury inflicted to the cerebellum, the number of BrdU+ and HuCD+ cells increased significantly. The number of BrdU+ cells after this type of injury was much greater than after a telencephalon trauma. A quantitative analysis revealed that after the cerebellum injury the proliferative activity in the caudal part of CCb is increased compared to that in the control. A reactivation of neurogenic and neuroepithelial niches and their transformation into reactive neurogenic domains, with an increased distribution density of intensely labeled HuCD+ cells of different types, were observed. The increase in the number of HuCD+ differentiated cells in the basal area suggests that the processes of neuronal differentiation are intensified in the cerebellum of juvenileO. masouafter injury. The number of GS positive cells (GS+) and fibers increased in all the zones of cerebellum. The most intensive astrocytic response was noted in the dorsal part of cerebellum. The data of the enzyme immunoassay confirm the multiple variations in the level of GS after a traumatic injury to cerebellum inO. masou.

Interrelations between cytokines, produced by invasive ductal carcinoma (IDC) and fibroadenoma (FA) of the breast, and angiogenic growth factor VEGF-A, expressed in IDC and FA, were investigated. The analysis of the cytokine profiles of IDC and FA was performed by cultivation of tumor biopsy specimens in vitro. Testing of the cytokine-producing reserve of the tumors for production of VEGF-A was conducted by culturing samples of IDC and FA in a medium containing polyclonal activator (a complex of phytohemagglutinin, concanavalin A, and lipopolysaccharide). Levels of cytokines and growth factors (IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1Ra, TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF-A) and MCP-1 (monocyte chemoattractant protein-1) in tumor supernatants were determined by an ELISA. Expression of VEGF-A was analyzed in tumor biopsy specimens by immunohistochemical analysis. In the IDC supernatants, the concentrations of IL-17, IL-18, and IFN-γ were higher and the concentrations of IL-10 and MCP-1 were lower in comparison with the FA supernatants. We observed negative correlations between the macrophage infiltration and VEGF-A concentration in the IDC supernatants (r = −0.508; P = 0.011) and between VEGF-A expression and the IDC vascularization degree (r = −0.423, P = 0.039). Spontaneous expression of VEGF-A in samples of IDC significantly exceeded the VEGF-A expression in FA. There was no difference between IDC and FA in VEGF-A expression after treatment with the polyclonal activators. Our results indicate that greater malignancy may have a paradoxical effect that is controlled by cytokines and characterized by weakening of tumor angiogenesis during overproduction of VEGF-A. These findings point to complex mechanisms of positive and negative regulation of tumor angiogenesis by cytokines that are produced by the tumor and by cells in its microenvironment, whose cytokine profiles may change at different stages of tumor progression.

Background: A high level of soluble thrombomodulin (sTM) is associated with a lower risk of thrombosis but can cause severe bleeding after operations. Deep vein thrombosis (DVT) and blood loss are serious threats after orthopedic surgery. The aim of our pilot study was to evaluate the effect of the preoperative level of sTM on coagulation and inflammation as well as the blood loss and the development of symptomatic DVT after total large joint replacement. Methods and Results: In all patients (n=50) who underwent total hip or knee replacement, sTM, PrC, D-dimer, vWF, CRP, and platelets were determined before and after the operation. According to the preoperative sTM level, patients were divided into 2 groups: the thrombomodulin low (TML) group (n=25) and thrombomodulin high (TMH) group (n=25). The concentration of sTM was 4.4 [3.4, 4.7] ng/ml in the TML-group and 8.7[7.3, 10.6] ng/ml in the TMH-group. After surgery, D-dimer, vWF, platelet count and CRP were higher and total blood loss was lower in the TML group. In the TML-group, a symptomatic DVT was detected in 3(12%) patients; in the TMH-group, a symptomatic DVT was identified only in 1(4%) case. Conclusion: These findings support the important role of sTM in coagulation, inflammation, bleeding, and presumably in venous thrombosis after major orthopedic surgery.

We investigated by the optical microscopy some cytological characteristics of the bronchoalveolar lavage fluid cell population 24 h after intratracheal instillation of microscale MnO2 and BaCrO4 particles (separately or together at two different doses) into the lungs of Wistar rats. Besides, the cytotoxicity of both particulates for rat peritoneal macrophages in vitro was assessed by the trypan blue exclusion test and proved significant. They were found to evoke a typical dose-dependent pulmonary phagocytosis response usually observed under inhalation or intratracheal impacts of low-soluble mineral and metal particles. A significant shift in the above mentioned cell population toward the prevalence of neutrophllic leukocytes (NL) over alveolar macrophages (AM) proved once more to be the most characteristic feature of this response. Although the particle load of a unit AM was always higher than that of a unit NL, the collective contribution of the recruited NLs to the total particles internalization by both AMs and NLs together was quite significant. This fact confirms that NL recruitment is an important auxiliary mechanism of the cytotoxic particle elimination from lungs compensating for the macrophage damage caused by them. Well adjusted functioning of this compensatory mechanism was additionally demonstrated by isobolographic analysis based on the Response Surface Methodology. On the other hand, this analysis confirmed that the type of combined toxicity depends on a particular effect this type is assessed for and on the effect’s dose-dependent level.

The aim of the study was to investigate a relationship between cytokine-producing reserve of invasive ductal cancer cells and its microenvironment, and cytokine-producing reserve of immunocompetent blood cells (IBC), as well as with histopathological and immunohistochemical characteristics of breast cancer. Using ELISA method we investigated the spontaneous and stimulated with polyclonal activators (PA) cytokine-producing reserve of IBC and biopsy specimens from invasive ductal cancer (adenocarcinoma) in 34 women. Appropriate values were expressed by the Influence Index of polyclonal activators (IIPA) upon cytokine production (IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1ra, TNFα, IFNγ, G-CSF, GM-CSF, VEGF-A, MCP-1). In tumor biopsies, we studied expression of VEGF-A, estrogen receptor, progesteron receptor and pro-proliferation marker Ki-67 by means of immunohistochemical method. Activation values of blood IBC in most cases, except of IL-18, IL-1β and MCP-1, were higher than appropriate effects upon cytokine production by tumor tisuues. Meanwhile, the IIPA activation index upon IL-18 (a proinflammatory and prooncogene cytokine) production by tumor cells and its microenvironment proved to be elevated, as compared to appropriate IIPA by the blood IBC. Statistical studies showed a direct correlation between IIPA and cytokine production in tumor supernates, IIPA of VEGF-A expression in tumor tissue, with pathohistological parameters and expression of estrogen and progesterone receptors and Ki-67 proliferation marker. A high positive correlation was obtained between IIPA TNFα production by the tumor tissue, and degree of tumor vascularization. We have revealed a negative correlation between IIPA for IL-6, MCP-1 and Ki-67 marker of cell proliferation. A direct correlation was found between IIPA values for IL-1ra/IL-1β production ratios in blood cells, and IIPA for VEGF-A expression in adenocarcinoma tissues, thus indicating to probable connections between IL-1ra production by IBC and VEGF-A expression. We have discerned several intersecting, diverging, and even circle-closed correlative chains of correlation, e.g.: VEGF-A (immunohistochemical marker of IIPA expression) – IL-10 and IL-8 (IIPA for supernate products); MCP-1, IL-6, and IL-10 (IIPA for supernate products)– VEGF-A (immunohistochemical indicator of expression for IIPA). The results of this study suggest a relationship between cytokine-producing reserves of the tumor identified by appropriate PA correlations with the processes occurring in malignant tumors, as assessed by histopathological and immunohistochemical parameters. The data are indicative for some complex mechanisms mediated by cytokines which provide invasive growth of malignant tumor.

Dependence of cytokine pattern in the tumor supernatant obtained after cultivation of biopsy samples-on the patients' age was evaluated among patients with invasive ductal carcinoma of the breast. An increase in VEGF and IL-6 production in a group of younger patients was observed. An increase only in interferon γ concentration was revealed in the supernatants of the tumor after addition of polyclonal activators to the culture medium. This result indicates likely secretion of interferon γ in younger patients. The relation among the production of angiogenic factors by tumor cells, age of the patients, and presence or absence of lymph node metastases shows that in such studies, patients have to be stratified by age.

A study of the lateral pallium in zebrafish and the visual tectum of the medaka revealed a population of adult neuroepithelial (NE) cells supported from the early stage of development to various postembryonic stages of ontogenesis. These data emphasize the importance of non-radial glial stem cells in the neurogenesis of adult animals, in particular fish. However, the distribution, cell cycle features, and molecular markers of NE cells and glial progenitors in fish are still poorly understood at the postembryonic stages of ontogenesis. Fetalization predominates in the ontogenetic development of salmon fish, which is associated with a delay in development and preservation of the features of the embryonic structure of the brain during the first year of life. In the present work, we studied the features of proliferation and the migration of neuronal precursors in the pallial proliferative zone of juvenile Oncorhynchus masou. The aim of the study is a comparative analysis of the distribution of glial-type aNSCs markers, such as vimentin and glial fibrillar acid protein GFAP, as well as the proliferation marker BrdU and migratory neuronal precursor doublecortin, in the pallial zone of the intact telencephalon in juvenile O. masou normal and after mechanical injury. The immunohistochemical IHC labeling with antibodies to vimentin, GFAP and doublecortin in the pallium of intact fish revealed single, small, round and oval immunopositive cells, that correspond to a persistent pool of neuronal and/or glial progenitors. After the injury, heterogeneous cell clusters, radial glia processes, single and small intensely labeled GFAP+ cells in the parenchyma of Dd and lateral part of pallium (Dl) appeared, corresponding to reactive neurogenic niches containing glial aNSCs. A multifold increase in the pool of Vim+ neuronal precursor cells (NPCs) resulting from the injury was observed. Vim+ cells of the neuroepithelial type in Dd and Dm and cells of the glial type were identified in Dl after the injury. Doublecortine (Dc) immunolabeling after the injury revealed the radial migration of neuroblasts into Dm from the neurogenic zone of the pallium. The appearance of intensely labeled Dc+ cells in the brain parenchyma might indicate the activation of resident aNSCs as a consequence of the traumatic process.

Currently, the role of cytokines in the tumor progression, including breast cancer, is universally recognized. At the same time, there are still many questions concerning the role of individual cytokines and receptors for cytokines in various morphogenetic processes underlying the tumor progression. The objective of this work was to study cytokine production and vascular endothelial growth factor (VEGF)-R2 and VEGF-R1 expression by mammary adenocarcinoma (MAC) and the correlations with histopathological parameters of malignant tumors. The object of the study was cultured tumor biopsy samples from 47 women aged 43–75 years with invasive ductal carcinoma, which was classified as grade II–III adenocarcinoma. It was shown that the cytokine profiles of the supernatants of MAC samples from patients differ greatly. A correlation between the levels of VEGF-R2 and tumor necrosis factor (TNF)-R1 expression was observed. Correlations were also revealed during analysis of the relations of histopathological MAC indicators with KVEGF-R2/VEGF-A and KTNF-R1/TNF-α coefficients, which are equal, respectively, to the ratio of expression values of receptors VEGF-R2 and TNF-R1 to the concentrations of the relevant cytokines (VEGF-A and TNF-α) in the culture supernatants of the same MAC samples. A direct correlation was identified between KVEGF-R/VEGF-A and some histopathological MAC characteristics: proportion of cells undergoing mitosis or pathological mitosis in MAC and poorly differentiated cells. KVEGF-R2/VEGF-A directly correlated with the concentration in supernatant interleukin (IL)-18 and interferon (IFN)-γ. KTNF-R1/TNF-α was inversely correlated with the concentration in supernatant of IL-1Ra, IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF). The data obtained show that the high-level production of IL-18 and IL-1β by MAC, overexpression of VEGF-R2 in tumor (at relatively low VEGF-A production), and the high level of IFN-γ production are attributed factors contributing to the formation of a population of low-grade cells in the tumor. The factors regulating the population of moderately differentiated cells in the tumor are referred to as IL-1Ra, IL-8, and GM-CSF.