Increase In Proliferative Activity Research Articles

Abstract LB318: Exploring early precursor lesions of ovarian cancer: Insights from an in vitro model

Abstract Ovarian cancer ranks as the fifth leading cause of cancer-related deaths in women, surpassing other cancers of the female reproductive system. Recent epidemiological and molecular studies highlight the significance of high-grade serous carcinoma (HGSC), the most aggressive subtype of ovarian cancer, constituting nearly 96% of cases with mutations in p53. HGSC originates from fallopian tube secretory epithelial (FTSEC) cells through a progression of precursor lesions, ranging from p53 signatures to serous tubal intraepithelial carcinoma (STIC). The p53 signature involves a linear expansion of over 12 TP53 mutant FTSECs, morphologically normal and displaying no increase in proliferative activity. In this study, we established a model system for P53 signature by either overexpressing p53 mutations (R175H and R273H) or knocking down P53 expression in normal FTSECs maintained as 2D, 3D, or organoid cultures. We have characterized the cells using RNA seq, mass spectroscopy and functional assays. Functional assays revealed increased migration and proliferation, along with decreased apoptosis in response to the cytotoxic stimulus (cisplatin). Despite the inability of p53 signature cells to proliferate or migrate in vivo, our hypothesis posited that clonal expansion of progenitor cell with a p53 mutation maintains plasticity by preserving the stemness of one or more cells capable of responding to cues from the microenvironment. Our experiments revealed heterogeneity in the model system, reflecting the composition of p53 signature in vivo. We also observed that stem-like cells tend to organize into clusters. Notably, stem-like cluster cells bearing a p53 signature exhibit migratory behavior, whereas clusters lacking p53 do not demonstrate such mobility. Given that most high-grade serous carcinoma (HGSC) cases occur in postmenopausal women, we demonstrated induction of proliferation and migration of p53 mutant stem cells in response to stimuli from aging stromal cells (fibroblasts and autologous immune cells) and to changes in hormonal background in our model. Conclusion: This study illuminates a novel aspect of ovarian cancer development, underscoring the role of a distinct population of fallopian tube epithelial cells with stem-like properties within p53 signatures. Further investigations into the tumorigenicity of these cells hold promise for providing valuable insights into future therapeutic strategies. Citation Format: Yan E. Wang, Vera Levina, Deborah L. Galson, Anna E. Lokshin. Exploring early precursor lesions of ovarian cancer: Insights from an in vitro model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB318.

A non-immunotoxic exopolysaccharide with anticancer effect in colorectal cancer isolated from Lacticaseibacillus rhamnosus ACS5 strain

Recent studies demonstrated the potential of exopolysaccharides (EPS) produced by LAB (lactic acid bacteria), to be used in different fields. In our study, L-EPS-ACS5 from Lacticaseibacillus rhamnosus ACS5 strain isolated from Ayvalık Cunda Sepet Cheese, one of the traditional cheeses of Turkey, showed antiproliferative effect in a time-dependent manner in human colon cancer (HT-29) cell line compared to healthy control line (L929). This effect was also confirmed by colony formation analysis. Apoptotic changes were investigated at molecular level using qPCR, Western blot and it was determined that the relevant L-EPS induced apoptosis by increasing Bax, Caspase 3, and 9 expression while reducing the Bcl-2 expression. These changes were confirmed by observations done at nuclear and cellular morphology level using a fluorescence microscope (Hoescht 33258) and TEM. In addition, the immunostimulatory activity of the relevant L-EPS was evaluated in terms of proliferation, phagocytosis ability and nitric oxide (NO) production parameters in the RAW 264.7 macrophage cell line. The increase in proliferative activity was not significant, and in parallel with these results, the differences in the control and application groups as a result of 'phagocytic activity and NO production analyzes' were also not significant. However, L-EPS-ACS5 did not show immunotoxic/cytotoxic activity in macrophage cells (% viability >70). All these cellular and molecular evidences indicate that L-EPS-ACS5 may be a good alternative to synthetic anti-cancer agents with its anticancer effect and non-immunotoxic properties in colorectal cancer.

Postnatal Exposure to the Endocrine Disruptor Dichlorodiphenyltrichloroethane Affects Adrenomedullary Chromaffin Cell Physiology and Alters the Balance of Mechanisms Underlying Cell Renewal.

Dichlorodiphenyltrichloroethane (DDT) is a wide-spread systemic pollutant with endocrine disrupting properties. Prenatal exposure to low doses of DDT has been shown to affect adrenal medulla growth and function. The role of postnatal exposure to DDT in developmental disorders remains unclear. The aim of the present investigation is to assess growth parameters and the expression of factors mediating the function and renewal of chromaffin cells in the adult adrenal medulla of male Wistar rats exposed to the endocrine disruptor o,p'-DDT since birth until sexual maturation. The DDT-exposed rats exhibited normal growth of the adrenal medulla but significantly decreased tyrosine hydroxylase production by chromaffin cells during postnatal period. Unlike the control, the exposed rats showed enhanced proliferation and reduced expression of nuclear β-catenin, transcription factor Oct4, and ligand of Sonic hedgehog after termination of the adrenal growth period. No expression of pluripotency marker Sox2 and absence of Ascl 1-positive progenitors were found in the adrenal medulla during postnatal ontogeny of the exposed and the control rats. The present findings indicate that an increase in proliferative activity and inhibition of the formation of reserve for chromaffin cell renewal, two main mechanisms for cell maintenance in adrenal medulla, in the adult DDT-exposed rats may reflect a compensatory reaction aimed at the restoration of catecholamine production levels. The increased proliferation of chromaffin cells in adults suggests excessive growth of the adrenal medulla. Thus, postnatal exposure to DDT alters cell physiology and increases the risk of functional insufficiency and hyperplasia of the adrenal medulla.

ULTRASTRUCTURAL CHANGES IN RATS SKIN SEBOCYTES DURING EXPERIMENTAL CHRONODESTRUCTION

Purpose of the study ultrastructural changes in rat skin sebocytes during experimental chronodestruction. In the experiments, 22 white outbred male rats with a body weight of 170-220 g were used. The experimental animals, in accordance with the experimental design, were randomly divided into 3 groups: group 1 - intact (n = 8) - animals under standard fixed conditions lighting (12 hours light/12 hours dark); group 2 (n=8) – animals with simulated light deprivation in conditions of round-the-clock darkness (24-hour darkness) for 21 days; group 3 (n=6) – animals with simulated dark deprivation under conditions of round-the-clock lighting (24 hours light) for 21 days. For morphometric assessment, 30 non-overlapping fields of view were analyzed in each preparation at a magnification of 100,000. The number of mitochondria and lysosomes was calculated per 100 μm2 area of the cytoplasm of cells of a certain morphological type. Using the ImageScopeM application program, the average cross-sectional area of mitochondria (μm2) and the average relative electron density of their matrix were determined. During chronodestruction, significant ultrastructural changes are observed in sebocytes, which indicate a change in the functional activity of the cells. During dark deprivation, the functional activity of sebocytes increases, and this, in turn, leads to a significant change in their secretory activity. At the same time, light deprivation leads to an increase in proliferative activity in the sebaceous gland with a simultaneous change in the chemical composition of sebum (polymorphism of granules).

Does insulin make breast cancer cells resistant to doxorubicin toxicity?

The effects of insulin on the doxorubicin (Dox) sensitivity of breast cancer cell line MCF-7 and its Dox-resistant counterpart MCF-7/Dox were studied and glucose metabolism, content of essential minerals, and the expression of several microRNAs in these cells upon exposure to insulin and Dox were compared. Cell viability colorimetric assay, colorimetric enzymatic technique, flow cytometry, immunocytochemical techniques, inductively-coupled plasma atomic emission spectroscopy, and quantitative polymerase chain reaction were used in the study. We found that insulin in high concentration significantly suppressed Dox toxicity, especially in parental MCF-7 cell line. The increase in proliferative activity triggered by insulin in MCF-7 but not MCF-7/Dox cells occurred in the setting of the increased level of specific binding sites for insulin and increased glucose uptake. Insulin treatment of MCF-7 cells in low and high concentrations resulted in the increase of Mg, Ca, and Zn content while in DOX-resistant cells, only Mg content increased upon exposure to insulin. High concentration of insulin increased the expression of kinase Akt1, P-glycoprotein 1 (P-gp1) and DNA excision repair protein ERCC-1 in MCF-7 cells, while in MCF-7/Dox cells, Akt1 expression decreased, and cytoplasmic expression of P-gp1 increased. In addition, insulin treatment affected expression of miR-122-5p, miR-133a-3p, miR-200b-3p, and miR-320a-3p. The decreased manifestation of biological effects of insulin in Dox-resistant cells could be partly explained by the different patterns of energy metabolism in MCF-7 cells and their Dox-resistant counterpart.

Relative survival and cell adhesion of multipotent mesenchymal cells from primary teeth pulp on the surface membranes and sponges produced from collagen

To study the behavior of SHED cell culture on different types of materials for the regeneration of periodontal tissues with different porosity. Porous collagen material Fibro-Gide (Geitstlich Pharma AG, Switzerland), designed to increase the volume of the gum and the barrier collagen membrane Bio-Gide (Geitstlich Pharma AG, Switzerland) were studied in vitro on SHED cultures. As a control sample, a Spongostan sponge made of gelatin (Johnson & Johnson Medical, UK) with the most pronounced porosity and wettability was used. Acute cytotoxicity was determined using a screening method for assessing the number of living cells in a sample (MTT test). SHED cells were sown on the materials to study the attachment of cells to materials and their migration inside the samples. Before seeding, the cells were stained with vital fluorescent dye PKH26 (red fluorescent cell linker kit, Sigma, Germany) for further visualization. Using the MTT test it was shown that they do not have cytotoxic effects. At the same time by the 8th day of the experiment in the presence of Fibro-Gide and Bio-Gide the cells showed an increase in proliferative activity by 19% and 12%, respectively compared with the control group. The cells attached and spread out on the surface of the materials and migrated into the thickness of porous Fibro-Gide and Spongostan. The in vitro study showed that the most favorable material for SHED cell culture is the collagen material Fibro-Gide with sufficient porosity, elasticity and hydrophilicity. SHED cells attach to the collagen matrix and easily penetrate into the sample, filling the entire internal space, while the proliferative capacity of the cell culture increases.

Enhanced proliferation of oligodendrocyte progenitor cells following retrovirus mediated Achaete-scute complex-like 1 overexpression in the postnatal cerebral cortex in vivo.

The proneural transcription factor Achaete-scute complex-like 1 (Ascl1) is a major regulator of neural fate decisions, implicated both in neurogenesis and oligodendrogliogenesis. Focusing on its neurogenic activity, Ascl1 has been widely used to reprogram non-neuronal cells into induced neurons. In vitro, Ascl1 induces efficient reprogramming of proliferative astroglia from the early postnatal cerebral cortex into interneuron-like cells. Here, we examined whether Ascl1 can similarly induce neuronal reprogramming of glia undergoing proliferation in the postnatal mouse cerebral cortex in vivo. Toward this goal, we targeted cortical glia during the peak of proliferative expansion (i.e., postnatal day 5) by injecting a retrovirus encoding for Ascl1 into the mouse cerebral cortex. In contrast to the efficient reprogramming observed in vitro, in vivo Ascl1-transduced glial cells were converted into doublecortin-immunoreactive neurons only with very low efficiency. However, we noted a drastic shift in the relative number of retrovirus-transduced Sox10-positive oligodendrocyte progenitor cells (OPCs) as compared to glial fibrillary acidic protein (GFAP)-positive astrocytes. Genetic fate mapping demonstrated that this increase in OPCs was not due to Ascl1-mediated astrocyte-to-OPC fate conversion. Rather, EdU incorporation experiments revealed that Ascl1 caused a selective increase in proliferative activity of OPCs, but not astrocytes. Our data indicate that rather than inducing neuronal reprogramming of glia in the early postnatal cortex, Ascl1 is a selective enhancer of OPC proliferation.

Atypical thymomas with squamoid and spindle cell features: clinicopathologic, immunohistochemical and molecular genetic study of 120 cases with long-term follow-up

AbstractThymomas are rare tumors characterized by a broad range of morphologic appearances that can sometimes give rise to difficulties for classification. We have studied a series of 120 thymoma patients in whom the tumors were characterized by sheets of atypical epithelial cells with squamoid and/or spindle cell features. They occurred in 63 men and 57 women and presented as a discrete mass in the anterior mediastinum measuring 2–23 cm (mean: 8.2 cm). Patients' ages ranged from 14 to 86 years (mean: 57.8) and most had symptoms referable to a mass lesion. 20 patients had myasthenia gravis or other autoimmune disorder. 76 cases were characterized by a predominant population of round to polygonal tumor cells while 32 cases were characterized by atypical oval or spindle cells. 12 cases showed mixed features and 16 cases showed the development of thymic carcinoma arising from thymoma. All cases were positive for p40/p63 and cytokeratin AE1/AE3. 23 cases were positive for CD5 (25%), and 13 for CD117 (14%). MIB1 showed a significant increase in proliferative activity (mean = 11.6%). Next generation sequencing in 47 cases did not disclose any variants amenable to current targeted therapies. Clinical follow up ranging from 2 to 29 years showed a progressive increase in aggressive behavior and fatality rate with advancing stage. Overall survival was 87% at 5 years, 67% at 10 years, and 23% at 20 years. Completeness of resection and staging were the most significant parameters for survival. The more aggressive tumors followed a protracted clinical course with multiple recurrences and metastases over a long period of time (mean = 19.8 years from time of initial relapse to death). Atypical thymomas are a distinct category of thymic epithelial neoplasm characterized by a slowly progressive clinical course with increased potential for metastases, transformation to a higher-grade malignancy, and fatal outcome in some cases.

Rat liver ECM incorporated into electrospun polycaprolactone scaffolds as a platform for hepatocyte culture.

Liver disease is expanding across the globe; however, health-care systems still lack approved pharmaceutical treatment strategies to mitigate potential liver failures. Organ transplantation is the only treatment for liver failure and with increasing cases of liver disease, transplant programs increasingly cannot provide timely transplant availability for all patients. The development of pharmaceutical mitigation strategies is clearly necessary and methods to improve drug development processes are considered vital for this purpose. Herein, we present a methodology for incorporating whole organ decellularised rat liver ECM (rLECM) into polycaprolactone (PCL) electrospun scaffolds with the aim of producing biologically relevant liver tissue models. rLECM PCL scaffolds have been produced with 5w/w% and 10w/w% rLECM:PCL and were analyzed by SEM imaging, tensile mechanical analyses and FTIR spectroscopy. The hepatocellular carcinoma cell line, HepG2, was cultured upon the scaffolds for 14 days and were analyzed through cell viability assay, DNA quantification, albumin quantification, immunohistochemistry, and RT-qPCR gene expression analysis. Results showed significant increases in proliferative activity of HepG2 on rLECM containing scaffolds alongside maintained key gene expression. This study confirms that rLECM can be utilized to modulate the bioactivity of electrospun PCL scaffolds and has the potential to produce electrospun scaffolds suitable for enhanced hepatocyte cultures and in-vitro liver tissue models.

Role of mTORC1 activity during early retinal development and lamination in human-induced pluripotent stem cell\u2010derived retinal organoids

Retinal organoids derived from human-induced pluripotent stem cells (hiPSC) are powerful tools for studying retinal development as they model spatial and temporal differentiation of retinal cell types. Vertebrate retinal development involves a delicate and coordinated process of retinal progenitor cell (RPC) differentiation, and the mammalian target of rapamycin complex 1 (mTORC1) has been reported to play a significant role in this complex process. Herein, using hiPSC-derived retinal organoids, we identify the time-dependent role of mTORC1 in retinal development, specifically in retinal ganglion cell (RGC) differentiation and the retinal lamination process, during the early stages of retinal organoid (RO) development. mTORC1 activity in ROs was the highest at 40 days of differentiation. MHY1485-induced hyperactivation of mTORC1 during this period resulted in a significant increase in the overall size of ROs compared to the untreated controls and rapamycin-treated Ros; there was also a marked increase in proliferative activity within the inner and outer layers of ROs. Moreover, the MHY1485-treated ROs showed a significant increase in the number of ectopic RGCs in the outer layers (indicating disruption of retinal laminar structure), with robust expression of HuC/D-binding proteins in the inner layers. These results demonstrate that mTORC1 plays a critical role in the development of hiPSC-derived ROs, especially during the early stages of differentiation.

BUCCAL CELL MICRONUCLEI AMONG PATIENTS WITH ORAL LEUKOPLAKIA.

The aim: The purpose of research was to evaluate the efficiency of micronucleus test in buccal cells for the diagnosis of oral leukoplakia. Materials and methods: We have conducted a comprehensive clinical and laboratory examination of 155 patients with oral leukoplakia. It was conducted histological examination leukoplakia mucosal sites, to assess the buccal epithelium cell micronucleus test was carried out. Results: Histological evaluation of the material was made according to the classification of leukoplakia WHO (2005). They are established 10 (14%) sites unmodified mucosa, 10 (14%) of the samples hyperkeratosis without atypia, 14 (19%) biopsies hyperkeratosis SIN1, 15 (21%) of hyperkeratosis SIN2, 10 (14%) -- SIN3 and 13 (18 %) of the cases of squamous cell carcinoma. Micronuclei, whose appearance is caused by violation of differentiation of epithelial cells, were found in patients with leukoplakia, the detection of micronuclei almost equally high as in patients with leukoplakia SIN2, and with SIN3 (a difference of 1.3 times (p <0,05, rxy = + 0.271)), and consequently the probability of occurrence of tumoral diseases of the oral mucosa or malignancy existing large. Conclusions: Thus, on the background of the general increase in proliferative activity of epithelial cells with increasing SIN, for each treatment group revealed the appearance of micronuclei in buccal cells. And the frequency of micronuclei and the fourth type of increases with hyperplasia, indicating an increase in the likelihood of malignancy and cancer of the oral mucosa in patients with leukoplakia SIN3.

Proliferative activity of the epithelium in squamous epithelial lesions of the cervix

As a criterion for precancerous changes in the stratified squamous epithelium of the cervix, its proliferative activity, studied using monoclonal antibodies PC-10 to the antigen of proliferating cell nuclei (PCNA), is considered. The results of the studies showed that patients with a low degree of squamous intraepithelial lesion are characterized by weak proliferative activity, and for patients with a high degree of lesion it is moderate and pronounced. An increase in proliferative activity is a prognostic factor that determines long-term persistence and the likely progression of the lesion.

IN VITRO STUDY ACTIVITY PROLIFERATIVE FIBROBLAST HUMAN SUBMITTED TO TREATMENT ZINCUM METALLICUM AND CALENDULA OFFICINALIS IN ULTRA DILUTIONS HOMEOPATHIC

Cicatrization can be divided into three phases: inflammation, fibroblastic, maturation and remodeling [1]. The extracellular matrix may be replaced by a stronger and more elastic connective tissue. In a scar collagen is the major component of the mature connective tissue [2]. In homeopathic area, the greater is the investigated segment ultra dilutions [6]. However, little research has been done to explore the effect of dynamized drugs in in vitro cell culture [4]. Using the Zincum metallicum 6CH and Calendula officinalis 6CH applied independently in different concentrations in fibroblast cultures sought to determine the increase in proliferative activity using techniques such as IC50, MTT, flow cytometry and quantification of collagen. As expected from the literature, ie both homeopathic according to the literature are used for treatments that Require skin healing, both showed increased proliferative activity, having Calendula most cellular response, presenting as cell cycle stimulating checked via flow cytometry.

Comparative Evaluation of Osteogenic Potential of Conventional Glass-ionomer Cement with Chitosan-modified Glass-ionomer and Bioactive Glass-modified Glass-ionomer Cement An In vitro Study.

Aim:The aim of this study was to compare the osteogenic potential of conventional glass-ionomer cement (GIC) with chitosan-modified GIC (CH-GIC) and bioactive glass-modified GIC (BAG-GIC) as a function of time in varying proportions.Materials and Methods:CH-GIC was prepared by adding 10 v/v% (Group II) and 50 v/v% (Group III) CH to the commercial liquid of GIC. BAG-GIC was prepared by the addition of 10 wt% (Group IV) and 30 wt% (Group V) of BAG to the GIC powder. Conventional GIC was kept as Group I. Nine round-shaped samples measuring 2 mm thick and 5 mm in diameter were prepared for every experimental material. Human osteosarcoma cells were cultured and cell proliferation was assessed at 24, 48, and 72 h using 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay, and cell differentiation was assessed at 7,14, and 21 days using alkaline phosphatase (ALP) assay. All experiments were done in triplicate. The data obtained were analyzed using one-way analysis of variance and Tukey honestly significant difference post hoc multiple comparisons at 0.05 level significance.Results:Cell culture studies showed a significant increase in proliferative activity and ALP activity in Group II, III, IV, and V than Group I at all-time intervals (P < 0.05). There was no statistically significant difference in osteogenic potential between CH-GIC and BAG-GIC groups.Conclusion:The osteogenic potential was significantly higher in CH-GIC and BAG-GIC compared to conventional GIC.

Gpr37l1/prosaposin receptor regulates Ptch1 trafficking, Shh production, and cell proliferation in cerebellar primary astrocytes.

Mammalian cerebellar astrocytes critically regulate the differentiation and maturation of neuronal Purkinje cells and granule precursors. The G protein-coupled receptor 37-like 1 (Gpr37l1) is expressed by Bergmann astrocytes and interacts with patched 1 (Ptch1) at peri-ciliary membranes. Cerebellar primary astrocyte cultures from wild-type and Gpr37l1 null mutant mouse pups were established and studied. Primary cilia were produced by cultures of both genotypes, as well as Ptch1 and smoothened (Smo) components of the sonic hedgehog (Shh) mitogenic pathway. Compared to wild-type cells, Gpr37l1-/- astrocytes displayed striking increases in proliferative activity, Ptch1 protein expression and internalization, intracellular cholesterol content, ciliary localization of Smo, as well as a marked production of active Shh. Similar effects were reproduced by treating wild-type astrocytes with a putative prosaptide ligand of Gpr37l1. These findings indicate that Gpr37l1-Ptch1 interactions specifically regulate Ptch1 internalization and trafficking, with consequent stimulation of Shh production and activation of proliferative signaling.

Study on Hemostimulating Properties of Granulocyte-Macrophage Colony Stimulating Factor

The hemostimulating properties of granulocyte-macrophage colony-stimulating factor (GM-CSF) make possible its clinical use in alleviating side effects of anti-cancer radio- and chemotherapy, in bone marrow transplantation, and in the treatment of some primary immunodeficiency conditions associated with leukopenia. The State Research Center of Virology and Biotechnology “Vector” of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing has developed a high-performance technology for production of recombinant human GM-CSF (rhGM-CSF) based on a recombinant E. coli strain.The aim of the study was to assess hemostimulating activity of the rhGM-CSF preparation obtained using the new developed technology, as observed in cell culture and in the mice model of myelosuppression induced by cyclophosphamide administration.Materials and methods: in vitro evaluation of rhGM-CSF hemostimulating activity was performed by MTT assay in the commercial HL-60 promyelocytic leukemia cell culture with preliminary suppression of cell growth rate by adding a low concentration of dimethyl sulfoxide to the medium. In vivo studies were carried out in CBA/CaLac mice with cyclophosphamide-induced myelosuppression. The hemostimulating properties of the drug were evaluated after subcutaneous administration of 1–175 µg/kg doses for 4–5 days, following administration of a cytostatic agent. The total number of leukocytes and the content of their morphological forms were determined in blood samples taken at different time points after the drug administration. The statistical processing of the experimental data was based on analysis of variance using Statgraphics v. 5.0 software.Results: the proliferative activity of HL-60 cells incubated with the rhGM-CSF preparation for 72 hours was shown to be dose-dependent. The highest values of the increase in proliferative activity associated with an increase in the drug dose were observed in the concentration range from 0.04 to 0.64 ng/mL (proliferative activity increased by 11–18% when the dose was increased twofold). The experiments in mice demonstrated a two-phase pattern of the dose-dependent effect. The drug showed the highest hemostimulating effect at the dose of 90 µg/kg.Conclusions: the rhGM-CSF preparation obtained using the new developed technology has a pronounced hemostimulating activity confirmed by both in vitro and in vivo test systems.

Immunotropic Properties of an Experimental Synthetic Selenium-Organic Compound.

We studied immunotropic properties of synthetic selenium-organic preparation 2,6-dipyridinium-9-selenabicyclo[3.3.1]nonyl dibromide (974zh). The experimental preparation reduced the cAMP/cGMP ratio, which indicated an increase in proliferative activity of cells of immunocompetent organs (thymus and spleen) in experimental animals. It was shown that 974zh intensified the immune response to Yersinia pestis EV thereby increasing the resistance to the plague agent.

The expression of macrophage migration inhibitory factor in the non-small cell lung cancer.

ObjectiveThe objective was to investigate the expression of macrophage migration inhibitory factor (MIF) in non-small cell lung cancer (NSCLC), as well as the effects of macrophage MIF on tumor cells.MethodsThe human NSCLC cell strains H358 and H524 were selected as research objects. The Real-Time Polymerase Chain Reaction (RT-PCR) and Western Blot were utilized to detect the expression levels of MIF in human NSCLC cell strains. The lentiviral plasmid was utilized for MIF-mRNA interference. The expression levels of MIF before and after transfection were compared. The cell strains were cultured and proliferated for cell count and comparison.ResultsH358 showed MIF high expression while H524 showed MIF low expression. Once the H358 cells were constructed as silent MIF expression, compared with the original H358 cells, the difference was statistically significant. Once the H524 cells were constructed as high MIF expression, compared with original H524 cells, the difference was statistically significant. Being cultured for respective 3, 5, and 7 days, the transfected H358 cells showed a significant decrease in proliferative activity compared with original H358 cells, while the transfected H524 cells showed a significant increase in proliferative activity compared with original H524 cells.ConclusionMIF has high expression in H358 cells while low expression in H524 cells. The expression of MIF could enhance the proliferative activity of NSCLC tumor cells.

Analysis of expression of molecular marker KI-67 in patients with laryngeal cancer for regional metastases and recurrences

Background: Laryngeal cancer is a common localization of squamous cell carcinoma of the head and neck. The main factor influencing the survival of patients is the presence of regional metastases. The malignant degeneration of a cell is always accompanied by disruption of the cell cycle, in particular a sharp increase in proliferative activity. The molecular marker Ki-67 is used to investigate the proliferative activity of tumor cells. Aim of the study: To study the informative content of immunohistochemical investigation and to determine the expression of the molecular marker Ki-67 in patients with laryngeal cancer, prior to the prediction of regional metastasis and relapse. Materials and Methods: 70 patients with cancer, stage III-IV (T3-4N0-3M0) clinical group II were under observation. Of these, 39 patients from the main group were with regional laryngeal cancer metastases, and 31 patients in the comparison group – without revealed regional metastases. Patients' age ranged from 33 to 74 years. The average age was 61.9 years. Male patients made up the absolute majority of 99% No = 69. All patients were histologically diagnosed with squamous cell carcinoma. Monoclonal antibodies to Ki67 (TermoScientific, USA) were used as the primary molecular markers. The proliferation index (PI) was rated as low in the range of 0-30% and as high in more than 30% of positive cells with an intranuclear label. Results: The study of molecular markers Ki-67 showed its expression at 69 [60.2; 77.5]% (Me [25%; 75%]) for the squamous cell laryngeal cancer group, without metastases – 60 [29; 70.5]%, for the group with metastases – 72 [67; 80]% (P &lt;0.001). In statistical data processing, the AUC value (0.777) was set for Ki-67 for metastasis and showed an increased degree of expression for laryngeal cancer metastasis. Ki-67 has a good diagnostic value and can be used as an additional method for predicting regional laryngeal cancer metastases. However the practical use of this marker as a prognostic one for determining the volume of surgery on the lymphatic structures of the neck requires further investigation. It should be noted that this study did not reveal the link of Ki-67 AUC (0.588) with the degree of differentiation of laryngeal cancer and its recurrence, but more in-depth study of molecular markers in the future may reveal other patterns of tumor growth and be used in the prognosis and personalization of treatment. Conclusion: Expression of Ki-67 allowed to accurately predict tumor metastasis. A 1% increase in Ki-67 expression resulted in an 8.3% increase in the chance of tumor metastasis (95% CI - 1.043-1.142), and the age dependence on metastasis is not reliable. Determination of expression of Ki-67 does not accurately predict the possibility of recurrence of laryngeal cancer (p=0.27; AUC (95% CI) – 0.588 (0.414-0.763)) herewith age dependence of chance of a recurrence was not revealed.

Morphofunctional rearrangements of adrenal cortex of adult rats at the experimental microelementosis

Xenobiotics, including salts of heavy metals, determine the development of pathology of individual organs and systems of the body. Adrenal hormones occupy a key place in the regulation and maintenance of the organism basic functions. The study of morphological, biochemical and immunohistochemical changes in the adrenal cortex of pubescent male rats at exposure to the complex of heavy metal salts remains an actual aspect of modern morphology. The experiment involved 24 white adult male rats weighing 250–300 g, aged 7–8 months. The animals of the experimental group consumed ordinary drinking water saturated with a combination of heavy metal salts (Zn, Cu, Fe, Mg, Cr) for 60 days. Histological, biochemical (determination of the content of hormones COR – cortisol, DHS – dehydroepiandrosterone sulfate) and immunohistochemical (determination of the expression of proliferation marker Ki-67) methods were used. Long-term consumption of the combination of heavy metals salts by mature rats leads to thickening of the stromal component of the gland, the rupture of capillaries and the violation of rheological properties of blood. Chromatin condensation, its marginal location, and initial stages of necrobiotic changes have been found in the spongiocyte nuclear apparatus. For endocrinocytes of the zona glomerulosa and zona reticularis of the gland, a slight increase in proliferative activity has been revealed, while cells of the zona fasciculata were non-reactive. These results correlate with the results of biochemical research, according to which hormones of the zona reticularis – DHS were actively involved in the formation of adaptive reactions in the body of experimental animals at the 60-day period of administration of the complex of heavy metals salts. The results of complex studies indicate a weakening of the secretory activity of cells of the zona fasciculata, which adversely affects the development of compensatory-adaptive processes in the body and the course of the general adaptation syndrome in response to the action of the damaging agent.