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1.
Cryobiology ; 113: 104789, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37924929

ABSTRACT

Values of cell membranes permeability coefficients for water and molecules of cryoprotective agents (CPAs) are the necessary characteristics for developing physical-mathematical models describing mass transfer processes through cell membranes in order to predict optimal cell cooling rates. We carried out a comparative analysis of the permeability coefficients of mouse oocyte membranes for molecules of water, ethylene glycol (EG), propane-1,2-diol (1,2-PD) and dimethyl sulfoxide (Me2SO), determined by applying the classical Kedem-Katchalsky model, which considers only the penetration of non-electrolyte molecules (water and CPA) through the membrane, and the model developed by us, which takes into account the transmembrane transfer of ions and the associated changes in the transmembrane electric potential. We shown that calculations based on the developed modified model provide lower values of the permeability coefficients of the oocyte membrane for water and CPA molecules. What is important that the obtained by our modified model permeability coefficients for water molecules do not depend on the type of cryoprotectant, while the application of the classical model both in our studies and works of other authors always gave different values of these coefficients in solutions with different cryoprotectants. Our modified model also makes it possible to determine the dynamics of the transmembrane electric potential of the cell under the conditions of transmembrane mass transfer and the duration of the membrane being influenced by the changes in electric potential, that is a parameter that can directly affect the viability of cells.


Subject(s)
Cryopreservation , Oocytes , Animals , Mice , Cell Membrane Permeability , Cryopreservation/methods , Cryoprotective Agents/pharmacology , Cryoprotective Agents/metabolism , Dimethyl Sulfoxide/pharmacology , Dimethyl Sulfoxide/metabolism , Ethylene Glycol/pharmacology , Ethylene Glycol/metabolism , Oocytes/metabolism , Permeability , Water/metabolism , Female
2.
Dokl Biol Sci ; 512(1): 354-359, 2023 Oct.
Article in English | MEDLINE | ID: mdl-38087026

ABSTRACT

The common gromwell Lithospermum officinale L. is a valuable medicinal plant that has been used in traditional medicine since ancient times. A method to quantify flavonoids in L. officinale leaves by differential spectrophotometry was developed taking advantage of the flavonoid reaction with aluminum chloride. The optimum duration of the reaction was determined, as well as the optimum volume-to-volume ratio between an aqueous ethanolic extract of L. officinale leaves and 2% aluminum chloride (aqueous ethanolic solution). Rutin was used as a standard. The method was validated in terms of specificity, linearity, precision, and accuracy and proved suitable for analytical purposes. The flavonoid content expressed in terms of rutin was found to exceed 2% of the absolutely dry weight in L. officinale leaves over different years of cultivation.


Subject(s)
Boraginaceae , Lithospermum , Flavonoids , Aluminum Chloride , Rutin , Plant Leaves , Plant Extracts
3.
Morfologiia ; 149(2): 11-5, 2016.
Article in Russian | MEDLINE | ID: mdl-30136788

ABSTRACT

The purpose of the present investigation was a comparative study of the effect of prenatal exposure to alcohol on the histological characteristics of neurons in the frontal cortex of the rats of different ages. The study was conducted on 175 outbred albino rats ­ the offspring of 25 females given a 15% solution of ethanol as a source of drinking throughout pregnancy. The cortex was examined at Days 2­90 after birth using histological, histochemical and morphometric methods. An increase (Days 2, 5), followed by the reduction (Days 10 and 90) of the thickness of the cortex and the size of neurons (Days 20­90) were detected, together with the decrease in the number of neurons in layer V of the cortex, reduction of the number of normochromic and an increase of the number of shrunken hyperchromic neurons and ghost cells in all study periods. Antenatal alcoholization was found to cause a variety of histological changes in the frontal cortex of rat brain in postnatal ontogenesis that had a long-term and progressive nature.


Subject(s)
Alcohol-Induced Disorders, Nervous System/pathology , Fetal Alcohol Spectrum Disorders/pathology , Frontal Lobe/pathology , Prenatal Exposure Delayed Effects/pathology , Alcohol-Induced Disorders, Nervous System/metabolism , Animals , Female , Fetal Alcohol Spectrum Disorders/metabolism , Frontal Lobe/metabolism , Pregnancy , Prenatal Exposure Delayed Effects/metabolism , Rats
4.
Morfologiia ; 148(6): 64-9, 2015.
Article in Russian | MEDLINE | ID: mdl-27141588

ABSTRACT

The neural apparatus and the endocrine part of the pancreas was studied in Wistar rats aged 3-4 and 19 months (n = 24) using the immunohistochemical reactions for synaptophysin (Syn), tyrosine hydroxylase (TH) and protein gene product 9.5 (PGP 9.5). Since Syn and PGP 9.5 are highly selective in detection of pancreatic islet (PI) endocrinocytes, it was possible to examine their topography and density in all parts of the pancreas. It was found that in rats aged 19 months, the total number of PI was decreased as compared to that in young animals. The study of PI size distribution has shown that the number of large islets decreased with age. Young animals showed rich innervation of the pancreas which was represented by three nerve plexuses: the first was a broadly-looped one, formed by small nerve trunks and bundles of unmyelinated and myelinated nerve fibers, the second consisted of thin bundles of postganglionic axons and microganglia, and the third (main terminal plexus) was formed by axons with varicosities and synapses of "en passant" type. In aged rats, marked degenerative changes in the neurons of intramural ganglia, nerve trunks and bundles were noted together with the reduction or complete absence of Syn- and TH-positive efferent parasympathetic and sympathetic terminals around blood vessels, excretory ducts, denervation of the exocrine and endocrine parts of the pancreas. Innervation disturbances in some lobules were accompanied by small inflammatory perivascular infiltrates.


Subject(s)
Aging/pathology , Islets of Langerhans/innervation , Islets of Langerhans/pathology , Aging/metabolism , Animals , Ganglia, Autonomic/metabolism , Ganglia, Autonomic/ultrastructure , Immunohistochemistry , Islets of Langerhans/metabolism , Nerve Fibers/metabolism , Nerve Fibers/ultrastructure , Presynaptic Terminals/metabolism , Presynaptic Terminals/ultrastructure , Rats, Wistar , Synaptophysin/metabolism , Tyrosine 3-Monooxygenase/metabolism
5.
Morfologiia ; 147(2): 49-53, 2015.
Article in Russian | MEDLINE | ID: mdl-26234040

ABSTRACT

The uterine tissues of female rats (n=30) with a scarred myometrium were examined by methods of light microscopy after the delivery. 1.5-2 months after the delivery no significant differences in the parameters of blood and lymph flow in the deep layers of the endometrium, myometrium and the myometrial scar tissue were found between the intact rats, nulliparous rats with a scarred uterus, rats that gave birth after laparotomy only and those that gave birth under the conditions of myometrial scar. In the course of pregnancy and labor activity, the damage of the tissues was observed not in the uterine scar proper, but at its borders with the myometrium. This is supported by the old hemorrhages and lymphostasis phenomena, greater number of lymphocytes, neutrophils, monocytes, macrophages and erythrocytes. In determining the indications and contraindications to vaginal childbirth in women with scarred uterus it is necessary to examine not only the scar proper, but also its border with the myometrium. The myometrial scar by itself, is not an absolute contraindication to vaginal delivery, the natural delivery is feasible in the absence of cavities with liquid and hemorrhages in the tissues of the uterine scar and at its border with myometrium.


Subject(s)
Cicatrix , Myometrium , Parturition/physiology , Postpartum Period/physiology , Pregnancy/physiology , Animals , Erythrocytes/cytology , Erythrocytes/metabolism , Female , Leukocytes/cytology , Leukocytes/metabolism , Myometrium/cytology , Myometrium/physiology , Rats , Rats, Wistar
6.
Article in Russian | MEDLINE | ID: mdl-24960996

ABSTRACT

The article emphasizes that, the protection of health of mother and child is actual especially in the present conditions considering demographic characteristics of particular territory. Hence, the development of optimal strategy in system of rendering of obstetrics and perinatal care and organization of operation of obstetrics institutions are the most important issues of modern obstetrics. The analysis is presented concerning conditions and main directions of optimization of organizational technologies in the system of obstetrics of the Ryazan oblast. The purpose and tasks of mechanism of optimization of rendering obstetrics and perinatal care are determined.


Subject(s)
Delivery of Health Care/organization & administration , Infant Welfare , Maternal Welfare , Perinatal Care/organization & administration , Female , Humans , Infant, Newborn , Obstetrics/organization & administration , Pregnancy , Russia
7.
PLoS One ; 19(5): e0301000, 2024.
Article in English | MEDLINE | ID: mdl-38805476

ABSTRACT

As imaging techniques rapidly evolve to probe nanoscale genome organization at higher resolution, it is critical to consider how the reagents and procedures involved in sample preparation affect chromatin at the relevant length scales. Here, we investigate the effects of fluorescent labeling of DNA sequences within chromatin using the gold standard technique of three-dimensional fluorescence in situ hybridization (3D FISH). The chemical reagents involved in the 3D FISH protocol, specifically formamide, cause significant alterations to the sub-200 nm (sub-Mbp) chromatin structure. Alternatively, two labeling methods that do not rely on formamide denaturation, resolution after single-strand exonuclease resection (RASER)-FISH and clustered regularly interspaced short palindromic repeats (CRISPR)-Sirius, had minimal impact on the three-dimensional organization of chromatin. We present a polymer physics-based analysis of these protocols with guidelines for their interpretation when assessing chromatin structure using currently available techniques.


Subject(s)
Chromatin , DNA , Formamides , In Situ Hybridization, Fluorescence , Formamides/chemistry , In Situ Hybridization, Fluorescence/methods , DNA/chemistry , Chromatin/chemistry , Chromatin/genetics , Nucleic Acid Denaturation , Animals
8.
Morfologiia ; 143(3): 77-80, 2013.
Article in Russian | MEDLINE | ID: mdl-24020190

ABSTRACT

In this paper we describe the modern immunohistochemical method detecting the neuronal marker PGP 9.5. The application of this method for the study of the innervation of rat and human heart, the detailed examination of the topography of the cardiac nervous apparatus, as well as their changes in pathological states, is demonstrated. Structural organization and the nature of the terminal branches of nerve apparatus suggest that they have afferent function. Protocol of the reaction demonstrating PGP 9.5 on paraffin sections is presented.


Subject(s)
Heart/innervation , Immunohistochemistry/methods , Myocardium/chemistry , Ubiquitin Thiolesterase/analysis , Animals , Humans , Nerve Fibers/chemistry , Rats
9.
Article in Russian | MEDLINE | ID: mdl-24027840

ABSTRACT

The article presents the results of expertise of 506 appointment cards of chronic patients consultation to physicians-specialists in oblast polyclinic. The analysis established the social demographic characteristics of patients, advanced cases, percentage of ungrounded and untimely appointments, scope of care provided in the oblast curative preventive establishment and need of patients in continuation of treatment at place of residence. The conclusions and recommendations make it possible to optimize the selection of patients referred to oblast polyclinic and to enhance the quality of medical care of rural population.


Subject(s)
Delivery of Health Care/statistics & numerical data , Health Services Needs and Demand/statistics & numerical data , Referral and Consultation/statistics & numerical data , Adolescent , Adult , Female , Humans , Male , Middle Aged , Patient Selection , Quality of Health Care , Rural Health Services/organization & administration , Rural Health Services/standards , Russia , Young Adult
10.
J Biol Chem ; 285(14): 10806-21, 2010 Apr 02.
Article in English | MEDLINE | ID: mdl-20106984

ABSTRACT

Insulin fibrillation provides a model for a broad class of amyloidogenic diseases. Conformational distortion of the native monomer leads to aggregation-coupled misfolding. Whereas beta-cells are protected from proteotoxicity by hexamer assembly, fibrillation limits the storage and use of insulin at elevated temperatures. Here, we have investigated conformational distortions of an engineered insulin monomer in relation to the structure of an insulin fibril. Anomalous (13)C NMR chemical shifts and rapid (15)N-detected (1)H-(2)H amide-proton exchange were observed in one of the three classical alpha-helices (residues A1-A8) of the hormone, suggesting a conformational equilibrium between locally folded and unfolded A-chain segments. Whereas hexamer assembly resolves these anomalies in accordance with its protective role, solid-state (13)C NMR studies suggest that the A-chain segment participates in a fibril-specific beta-sheet. Accordingly, we investigated whether helicogenic substitutions in the A1-A8 segment might delay fibrillation. Simultaneous substitution of three beta-branched residues (Ile(A2) --> Leu, Val(A3) --> Leu, and Thr(A8) --> His) yielded an analog with reduced thermodynamic stability but marked resistance to fibrillation. Whereas amide-proton exchange in the A1-A8 segment remained rapid, (13)Calpha chemical shifts exhibited a more helical pattern. This analog is essentially without activity, however, as Ile(A2) and Val(A3) define conserved receptor contacts. To obtain active analogs, substitutions were restricted to A8. These analogs exhibit high receptor-binding affinity; representative potency in a rodent model of diabetes mellitus was similar to wild-type insulin. Although (13)Calpha chemical shifts remain anomalous, significant protection from fibrillation is retained. Together, our studies define an "Achilles' heel" in a globular protein whose repair may enhance the stability of pharmaceutical formulations and broaden their therapeutic deployment in the developing world.


Subject(s)
Amyloid/chemistry , Diabetes Mellitus, Experimental/metabolism , Drug Design , Insulin/chemistry , Insulin/pharmacology , Amyloid/metabolism , Animals , Antibiotics, Antineoplastic/toxicity , Crystallography, X-Ray , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/drug therapy , Humans , Magnetic Resonance Spectroscopy , Male , Protein Structure, Secondary , Rats , Rats, Inbred Lew , Receptor, IGF Type 1/metabolism , Streptozocin/toxicity
11.
Lik Sprava ; (1-2): 21-9, 2011.
Article in English | MEDLINE | ID: mdl-21954632

ABSTRACT

More then 25 years after the Chernobyl accident, a higher prevalence of bronchial hyperreactivity, reduced lung function, and increased levels of free radicals in exhaled breath condensates (EBC) were observed in children residing in radioactive contaminated territories. Comparing children with different residential radiation background, this study investigated fatty acids of EBC using gas liquid chromatography, counts of B-lymphocyte antigen CD19 in T-cell (CD3) and phagocytotic activity of neutrophils in blood samples. Regarding EBC, we demonstrate that lipid peroxidation was activated, antioxidant properties of pulmonary surfactant were decreased, were detected metabolic disorders of essential fatty acids at the stage of bioregulators-eicosanoids formation. Regarding the immune function of blood cells, we found a decrease of the proportions of CD3+ 19- and CD3- 19+ lymphocyte subpopulations and an unbalance of their numbers. Also the phagocytotic activity of neutrophils was reduced in higher exposed children. Children living in the radioactive contaminated territories have more alterations of surfactant properties and immune activities, which may contribute to an increased risk of respiratory problems. This research was supported by grants from the U.S. Civilian Research and Development Foundation (UKB1-2929-KV-08).


Subject(s)
B-Lymphocytes/immunology , Body Fluids , Bronchial Hyperreactivity/immunology , Chernobyl Nuclear Accident , Neutrophils/immunology , Respiratory System , T-Lymphocytes/immunology , Adolescent , Antigens, CD19/analysis , Antigens, CD19/immunology , B-Lymphocytes/cytology , Body Fluids/chemistry , Body Fluids/cytology , Bronchial Hyperreactivity/blood , Bronchial Hyperreactivity/epidemiology , Bronchial Hyperreactivity/physiopathology , CD3 Complex/analysis , CD3 Complex/immunology , Case-Control Studies , Child , Exhalation , Fatty Acids/analysis , Female , Free Radicals/analysis , Free Radicals/metabolism , Gas Chromatography-Mass Spectrometry , Humans , Lipid Peroxidation , Male , Neutrophils/cytology , Nuclear Reactors , Phagocytosis/immunology , Prevalence , Respiratory Function Tests , Respiratory System/immunology , Respiratory System/physiopathology , T-Lymphocytes/cytology , Ukraine
12.
J Gastrointest Surg ; 25(2): 428-435, 2021 02.
Article in English | MEDLINE | ID: mdl-32026333

ABSTRACT

BACKGROUND: Nodal involvement has been identified as one of the strongest prognostic factors in patients with nonfunctional pancreatic neuroendocrine tumors (NF-PanNETs). Sufficient lymphadenectomy and evaluation is vital for accurate staging. The purpose of this study was to identify the optimal number of examined lymph nodes (ELN) required for accurate staging. METHODS: The SEER database was used to identify patients with resected NF-PanNETs between 2004 and 2014. The distributions of positive lymph nodes (PLN) ratio and total lymph nodes were used to develop a mathematical model. The sensitivity of detecting nodal disease at each cutoff of ELN was estimated and used to identify the optimal cutoff for ELN. RESULTS: A total of 1098 patients were included in the study of which 391 patients (35.6%) had nodal disease. The median ELN was 12 (interquartile range [IQR]: 7-19.5), and the median PLN was 2 (IQR: 1-4) for patients with nodal disease. With an increase in ELN, the sensitivity of detecting nodal disease increased from 12.0% (ELN: 1) to 92.2% (ELN: 20), plateauing at 20 ELN (< 1% increase in sensitivity with an additional ELN). This sensitivity increase pattern was similar in subgroup analyses with different T stages. CONCLUSIONS: The sensitivity of detecting nodal disease in patients with NF-PanNETs increases with an increase in the number of ELN. Cutoffs for adequate nodal assessment were defined for all T stages. Utilization of these cutoffs in clinical settings will help with patient prognostication and management.


Subject(s)
Neuroendocrine Tumors , Pancreatic Neoplasms , Humans , Lymph Node Excision , Lymph Nodes/pathology , Lymph Nodes/surgery , Lymphatic Metastasis , Neoplasm Staging , Neuroendocrine Tumors/surgery , Pancreatic Neoplasms/pathology , Pancreatic Neoplasms/surgery , Prognosis
13.
Probl Radiac Med Radiobiol ; 26: 449-463, 2021 Dec.
Article in English, Ukrainian | MEDLINE | ID: mdl-34965566

ABSTRACT

OBJECTIVE: to determine the influence of hereditary predisposition, polymorphism of GSTT1, GSTM1, GSTP1 genes andenvironmental factors on the development of bronchial asthma in children - residents of radioactively contaminat-ed areas. MATERIALS AND METHODS: School-age children-residents of radioactively contaminated areas with bronchial asthma,and those without clinical signs of respiratory pathology were examined. Genetic, medical, biological and social riskfactors were determined based on the study of anamnestic data and medical records. Ventilation lung capacity wasassessed by the method of computer spirometry. Molecular genetic studies were carried out using polymerase chainreaction (PCR) and restriction fragment length polymorphism (RFLP) for further analysis. RESULTS: Molecular genetic studies of the distribution of genotypes and frequencies of polymorphic variants of thegenes GSTT1, GSTM1, GSTP1 were performed in children living under long-term intake of 137Cs by food chains. It wasfound that in children with BA the tendency to frequency of the deletion variant of the GSTT1 and GSTM1 genes incomparison with children without bronchial and pulmonary pathology was increased. The study of distributing theGSTP1 A313G gene polymorphic variants revealed in children with BA a significant increase in the frequency of AG-genotype, compared with the data of reference group. Adverse factors that increase the risk of developing bron-choobstructive disorders and the probability of their implementation in the form of bronchial asthma in children -residents of RCA have been identified. It is established that among them the leading role is played by hereditarypredisposition to this disease. On the part of the child, such negative factors were unfavorable conditions of fetaldevelopment, the presence of signs of exudative-catarrhal diathesis, manifestations of allergies and frequent respi-ratory diseases from the first months of life. It was found that the risk of developing BA was significantly increasedin children with the GSTT1 and GSTM1 gene deletion genotypes; an increased risk of developing BA in children witha combination of the GSTP1 A313G gene polymorphism with deletion polymorphism of the GSTT1 or GSTM1 gene wasdetermined. Сonclusion. Оne of the leading mechanisms, due to which there is a realization of hereditary predisposition tobronchial asthma in children living under constant intake of radionuclides with a long half-life, is the polymorphismof certain glutathione-S-transferase genes, namely, GSTT1, GSTM1 and A313G gene deletion polymorphism and GSTP1gene polymorphism.


Subject(s)
Asthma/genetics , Asthma/physiopathology , Cesium Radioisotopes/adverse effects , Chernobyl Nuclear Accident , Genetic Predisposition to Disease , Glutathione Transferase/genetics , Polymorphism, Genetic/radiation effects , Radiation Exposure/adverse effects , Adolescent , Asthma/epidemiology , Case-Control Studies , Child , Female , Genotype , Glutathione S-Transferase pi/genetics , Glutathione S-Transferase pi/metabolism , Humans , Male , Risk Factors , Ukraine/epidemiology
15.
Cancer Res ; 80(22): 4998-5010, 2020 11 15.
Article in English | MEDLINE | ID: mdl-33023947

ABSTRACT

Intratumoral hypoxia occurs in 90% of solid tumors and is associated with a poor prognosis for patients. Cancer cells respond to hypoxic microenvironments by activating the transcription factors, hypoxia-inducible factor 1 (HIF1) and HIF2. Here, we studied the unique gene expression patterns of 31 different breast cancer cell lines exposed to hypoxic conditions. The EGFR, a member of the ErbB (avian erythroblastosis oncogene B) family of receptors that play a role in cell proliferation, invasion, metastasis, and apoptosis, was induced in seven of the 31 breast cancer cell lines by hypoxia. A functional hypoxia response element (HRE) was identified, which is activated upon HIF1 binding to intron 18 of the EGFR gene in cell lines in which EGFR was induced by hypoxia. CpG methylation of the EGFR HRE prevented induction under hypoxic conditions. The HRE of EGFR was methylated in normal breast tissue and some breast cancer cell lines, and could be reversed by treatment with DNA methyltransferase inhibitors. Induction of EGFR under hypoxia led to an increase in AKT, ERK, and Rb phosphorylation as well as increased levels of cyclin D1, A, B1, and E2F, and repression of p21 in an HIF1α-dependent manner, leading to cell proliferation and migration. Also, increased EGFR expression sensitized cells to EGFR inhibitors. Collectively, our data suggest that patients with hypoxic breast tumors and hypomethylated EGFR status may benefit from EGFR inhibitors currently used in the clinic. SIGNIFICANCE: Hypoxia sensitizes breast cancer cells to EGFR inhibitors in an HIF1α- and a methylation-specific manner, suggesting patients with hypoxic tumors may benefit from EGFR inhibitors already available in the clinic. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/80/22/4998/F1.large.jpg.


Subject(s)
Antineoplastic Agents/pharmacology , Breast Neoplasms/metabolism , DNA Methylation , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Tumor Hypoxia/physiology , Agammaglobulinaemia Tyrosine Kinase/metabolism , Animals , Basic Helix-Loop-Helix Transcription Factors/metabolism , Breast Neoplasms/drug therapy , Breast Neoplasms/genetics , Cell Line, Tumor , Cell Movement , Cell Proliferation , CpG Islands , Cyclin D1/metabolism , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Cytosine/metabolism , ErbB Receptors/antagonists & inhibitors , ErbB Receptors/genetics , Female , Genes, erbB-1 , HSP70 Heat-Shock Proteins/metabolism , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , MAP Kinase Signaling System , Methyltransferases/antagonists & inhibitors , Mice , Phosphorylation , Retinoblastoma Protein/metabolism , Signal Transduction/physiology
16.
Probl Radiac Med Radiobiol ; 25: 531-542, 2020 Dec.
Article in English, Ukrainian | MEDLINE | ID: mdl-33361859

ABSTRACT

OBJECTIVE: to determine the relationship between polymorphisms of glutathione S-transferase gene family andbronchial hyperreactivity in children living in radioactively contaminated areas. MATERIALS AND METHODS: School age children-residents of radioactively contaminated areas (RCA), without clinicalsigns of respiratory pathology were examined. Molecular genetic studies were carried out by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) for further analysis. The GSTT1, GSTM1 gene deletion polymorphism was investigated using multiplex PCR. PCR and PCR-RFLP analyses were performed in the studyof the GSTP1 gene A313G polymorphism. The ventilation lung capacity was examined by the pneumotachographicmethod according to the analysis of «the flow-volume¼ loop. The pharmacologic inhalation test with bronchodilator drug, affecting the ß2-adrenergic lung receptors was used to detect the early changes in the ventilation lungcapacity - the bronchial hyperreactivity (latent and nonlatent bronchospasm). RESULTS: Molecular genetic studies showed that the GSTM1 gene deletion genotype and the GSTP1 gene A313G polymorphism were found significantly more often in the subgroup of children with bronchial hyperreactivity living inRCA than in children without bronchial hyperreactivity and children of the control group. The frequency of GSTT1deletion polymorphism did not have a statistically significant difference in all subgroups. CONCLUSIONS: The GSTM1 gene deletion polymorphism and the GSTP1 gene A313G genotype may be a risk factor fordeveloping bronchial hyperreactivity in children living under adverse environmental conditions, including radioactively contaminated areas.


Subject(s)
Bronchial Hyperreactivity/genetics , Chernobyl Nuclear Accident , Genetic Predisposition to Disease , Glutathione Transferase/genetics , Radiation Exposure/adverse effects , Adolescent , Alleles , Bronchial Hyperreactivity/etiology , Bronchial Hyperreactivity/physiopathology , Bronchial Provocation Tests , Case-Control Studies , Child , Female , Gene Expression , Gene Frequency , Humans , Male , Polymorphism, Restriction Fragment Length , Radiation, Ionizing , Respiratory Function Tests , Ukraine/epidemiology
17.
Nat Commun ; 10(1): 4862, 2019 10 24.
Article in English | MEDLINE | ID: mdl-31649238

ABSTRACT

Hypoxia is known to be detrimental in cancer and contributes to its development. In this work, we present an approach to fate-map hypoxic cells in vivo in order to determine their cellular response to physiological O2 gradients as well as to quantify their contribution to metastatic spread. We demonstrate the ability of the system to fate-map hypoxic cells in 2D, and in 3D spheroids and organoids. We identify distinct gene expression patterns in cells that experienced intratumoral hypoxia in vivo compared to cells exposed to hypoxia in vitro. The intratumoral hypoxia gene-signature is a better prognostic indicator for distant metastasis-free survival. Post-hypoxic tumor cells have an ROS-resistant phenotype that provides a survival advantage in the bloodstream and promotes their ability to establish overt metastasis. Post-hypoxic cells retain an increase in the expression of a subset of hypoxia-inducible genes at the metastatic site, suggesting the possibility of a 'hypoxic memory.'


Subject(s)
Reactive Oxygen Species/metabolism , Spheroids, Cellular/metabolism , Transcriptome , Tumor Hypoxia/genetics , Animals , Cell Line, Tumor , Female , Humans , MCF-7 Cells , Mice , Mice, Transgenic , Neoplasm Metastasis/genetics , Optical Imaging , Phenotype , Prognosis , Tumor Microenvironment
18.
Probl Radiac Med Radiobiol ; 24: 480-492, 2019 Dec.
Article in English, Ukrainian | MEDLINE | ID: mdl-31841489

ABSTRACT

OBJECTIVE: to evaluate the influence of processes of lipid peroxidation, and antioxidant protection on the function of external respiration in children-residents of radioactive contaminated territories. MATERIALS AND METHODS: There were examined children of school age, inhabitants of radioactive contaminated ter- ritories (RCT) without respiratory and pulmonary pathology, and patients with bronchial asthma (BA). Examination of the ventilation lung function was performed by the method of pneumotachography according to the analysis of the «flow-volume¼ loop. End products of lipid peroxidation (LPO), reacting with thiobarbituric acid (malondialde- hyde), enzymes-antioxidants - catalase, superoxide dismutase, glutathione peroxidase, glutathione transferase were studied to determine the signs of oxidative stress. RESULTS: Examinations of ventilation lung function in children-residents of RCT without respiratory and pulmonary pathology showed no significant deviations of its parameters beyond the limits of physiological fluctuations; a decrease in bronchial patency at different levels of the bronchial tree was established in children with BA. An increased incidence of bronchial hyperreactivity was noted in both the patients with BA and those who did not have bronchopulmonary pathology. The absence of an increase in the content of LPO products (malondialdehyde) in blood serum of children-residents of RCT with increased activity of catalase and group of glutathione enzymes indi- cates the sufficiency of compensatory possibilities of antioxidant protection. CONCLUSIONS: Studies of correlation between the function of external respiration and markers of oxidative stress determined that the indices of bronchial patency directly correlate with the activity of enzymes-antioxidants of the glutathione group and inversely - with the content of the LPO products in children-residents of RCT. The frequency of bronchospasm inversely correlated with the activity of glutathione group antioxidants. There are inverse correla- tion of 137Cs content in the body with the activity of glutathione transferase and glutathione peroxidase.


Subject(s)
Antioxidants/metabolism , Asthma/diagnosis , Bronchial Spasm/diagnosis , Chernobyl Nuclear Accident , Adolescent , Asthma/blood , Asthma/physiopathology , Biomarkers/blood , Bronchial Spasm/blood , Bronchial Spasm/physiopathology , Case-Control Studies , Catalase/blood , Cesium Radioisotopes/analysis , Child , Female , Glutathione Peroxidase/blood , Glutathione Transferase/blood , Humans , Lipid Peroxidation , Lung/metabolism , Lung/physiopathology , Male , Malondialdehyde/blood , Oxidative Stress , Radiation Exposure , Radioactive Fallout , Respiration/radiation effects , Respiratory Function Tests , Superoxide Dismutase/blood , Ukraine
19.
Mol Cancer Res ; 16(12): 1889-1901, 2018 12.
Article in English | MEDLINE | ID: mdl-30037853

ABSTRACT

Intratumoral hypoxia has been associated with invasion, metastasis, and treatment failure, prompting the need for a global characterization of the response to hypoxic conditions. The current study presents the results of a large-scale RNA sequencing (RNA-seq) effort, analyzing 31 breast cancer cell lines representative of breast cancer subtypes or normal mammary epithelial (NME) cells exposed to control tissue culture conditions (20% O2) or hypoxic conditions (1% O2). The results demonstrate that NME have a stronger response to hypoxia both in terms of number of genes induced by hypoxia as well as level of expression. A conserved 42-gene hypoxia signature shared across PAM50 subtypes and genes that are exclusively upregulated in Luminal A, Luminal B, and normal-like mammary epithelial cells is identified. The 42-gene expression signature is enriched in a subset of basal-like cell lines and tumors and differentiates survival among patients with basal-like tumors. Mechanistically, the hypoxia-inducible factors (HIF-1 and/or HIF-2) mediate the conserved hypoxic response. Also, four novel hypoxia-regulated and HIF-1-responsive genes were identified as part of the conserved signature. This dataset provides a novel resource to query transcriptional changes that occur in response to hypoxia and serves as a starting point for a clinical assay to aid in stratifying patients that would benefit from hypoxia-targeted therapies, some of which are currently in clinical trials. IMPLICATIONS: RNA-seq of 31 breast cancer cells exposed to control or hypoxic conditions reveals a conserved genomic signature that contains novel HIF-regulated genes and is prognostic for the survival of patients with triple-negative breast cancer.


Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Breast Neoplasms/genetics , Gene Expression Profiling/methods , Hypoxia-Inducible Factor 1/genetics , Sequence Analysis, RNA/methods , Breast Neoplasms/pathology , Cell Hypoxia , Cell Line, Tumor , Cell Survival , Female , Gene Expression Regulation, Neoplastic , Gene Regulatory Networks , Humans , Prognosis , Survival Analysis
20.
Probl Radiac Med Radiobiol ; 23: 359-372, 2018 Dec.
Article in English, Ukrainian | MEDLINE | ID: mdl-30582857

ABSTRACT

Objective was to determine the content of sodium (Na+), potassium (K+), calcium (Ca2+), and magnesium (Mg2+) elec-trolytes in non-stimulated mixed saliva of children, with disorders of autonomous nervous system (ANS), who wereborn and are permanently residing at radioactive contaminated territories after application of intermittent normo-baric hypoxia (INH) of sanogenic level.Patientes and methods. The children (41 boys and 62 girls) aged 6-17 years were examined. All they were dividedinto three groups: the control group consisted of 30 persons (group I); the comparison group - 30 person (groupII); the main group - 43 patients (group III). The collection of anamnesis, patient complaints, clinical and labora-tory examinations were included into the studied program. The content of electrolytes in oral fluids was determinedby the atomic absorption method. 10 seances of INH with a hypoxic component of 12 % oxygen in nitrogen wereused by us.Results. It was shown that the content of electrolytes in non-stimulated mixed saliva had a multidirectional signi-ficance in different age groups after INH: in examined children of primary school age (6-11 years), the Na+ concen-tration was significantly increased by 0.8 mmol/l, K+ concentration was decreased by 3 mmol/l, Ca2+ concentrationwas decreased by 1.07 mmol/l and in children of senior school age (12-17 years) - Na+, and Ca2+ concentrations weredecreased by 2 mmol/l and 0.17 mmol/l, respectively.Сonclusions. The obtained results allow to recommend the INH seances for addition to basic treatment of childrenwith disorders of autonomous nervous system, who were born and are permanently residing at radioactive contam-inated territories of Ukraine.


Subject(s)
Autonomic Nervous System Diseases/therapy , Chernobyl Nuclear Accident , Hypoxia/metabolism , Oxygen Inhalation Therapy/methods , Radiation Exposure/adverse effects , Radiation, Ionizing , Saliva/chemistry , Adolescent , Age Factors , Autonomic Nervous System Diseases/etiology , Autonomic Nervous System Diseases/metabolism , Autonomic Nervous System Diseases/physiopathology , Calcium/metabolism , Case-Control Studies , Cations, Divalent , Cations, Monovalent , Child , Electrolytes/metabolism , Female , Humans , Magnesium/metabolism , Male , Potassium/metabolism , Sodium/metabolism , Spectrophotometry, Atomic , Ukraine
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