RESUMO
The qualitative composition and zeta potential of magnetite nanoparticles (size 4.2±1.2 nm) obtained by co-precipitation method were determined by X-ray and diffraction dynamic light scattering. The zeta potential of Fe3O4 particles was -15.1±4.5 mV. The possibility of interaction of magnetite nanoparticles with human blood plasma proteins and hemoglobin as well as with erythrocyte membranes was demonstrated by spectrophotometry, electrophoresis, and fluorescence methods. No changes in the sizes of hemoglobin molecules and plasma proteins after their modification by Fe3O4 particles were detected. The possibility of modifying the structural state of erythrocyte membranes in the presence of magnetite nanoparticles was demonstrated by means of fluorescent probe 1-anilinonaphthalene-8-sulfonate.
Assuntos
Hemoglobinas , Nanopartículas de Magnetita , Humanos , Nanopartículas de Magnetita/química , Hemoglobinas/química , Membrana Eritrocítica/efeitos dos fármacos , Membrana Eritrocítica/química , Membrana Eritrocítica/metabolismo , Tamanho da Partícula , Proteínas Sanguíneas/química , Naftalenossulfonato de Anilina/química , Difração de Raios X , Óxido Ferroso-Férrico/química , Corantes Fluorescentes/químicaRESUMO
We studied the effect of amphotericin B (2.5×10-5 and 5.4×10-5 M) on osmotic resistance and surface cytoarchitectonics of donor blood erythrocytes. Antibiotic at a concentration of 2.5×10-5 M induced most pronounced changes in the studied parameters, which can be related to the specifics of the spatial organization of the cholesterol-amphotericin B complexes at different stoichiometric ratios of the components and their ability to pore formation in the membranes. Cholesterol binding to the polyene antibiotic and the appearance of perforations in the plasma membrane lead to accumulation of reversibly and irreversibly deformed cells and their hemolysis. The appearance of a large number of irreversibly deformed erythrocytes indicates an impaired ability to elastic deformation in the microcirculatory stream, which can lead to disruption of their functions in vivo and intravascular hemolysis.
Assuntos
Anfotericina B , Antibacterianos , Humanos , Anfotericina B/farmacologia , Anfotericina B/química , Antibacterianos/farmacologia , Microcirculação , Polienos/análise , Polienos/farmacologia , Eritrócitos , Colesterol , HemóliseRESUMO
Exposure of human lymphocytes to silver nanoparticles (2-12 nm) reduced viability of cells and RNA, reduced activities of lactate dehydrogenase, glutathione reductase, and cytosolic calcium, increased ROS content in cells, affected surface architectonics of cells and changed hydrophobicity and charge of their plasma membranes. Silver nanoparticles triggered the process of cellular death by the mechanism of ETosis that is accompanied by chromatin release into the extracellular environment and formation of extracellular traps.
Assuntos
Linfócitos/metabolismo , Nanopartículas Metálicas/química , Prata/química , Adolescente , Adulto , Cálcio/metabolismo , Armadilhas Extracelulares/metabolismo , Glutationa Redutase/metabolismo , Humanos , L-Lactato Desidrogenase/metabolismo , Masculino , Microscopia de Fluorescência , Espécies Reativas de Oxigênio/metabolismo , Adulto JovemRESUMO
The effect of UV-light (240-390 nm) in doses of 151 and 755 J/m2 on the expression of membrane markers CD5, CD19, CD20 in human peripheral blood B cells was studied by flow cytometry. In 24 h after exposure to UV light, we observed activation of processes accompanied by structural rearrangements of B-cell membranes leading to changes in the expression of receptor molecules: the content of of CD19 and CD20 increased due to activation of the synthesis of these proteins, while the content of CD5 decreased. The percentage of CD5+ cells decreased over 24 h after UV-irradiation of lymphocytes, while addition of autologous plasma to the incubation medium produced a photoprotective effect on CD5+ cells.
Assuntos
Antígenos CD , Linfócitos B , Transfusão de Sangue Autóloga , Antígenos CD/metabolismo , Antígenos CD/efeitos da radiação , Antígenos CD19/metabolismo , Antígenos CD19/efeitos da radiação , Antígenos CD20/metabolismo , Antígenos CD20/efeitos da radiação , Linfócitos B/metabolismo , Linfócitos B/efeitos da radiação , Biomarcadores/metabolismo , Antígenos CD5/metabolismo , Antígenos CD5/efeitos da radiação , Membrana Celular/metabolismo , Membrana Celular/efeitos da radiação , Humanos , Imunidade Celular/efeitos da radiação , Imunoterapia/métodos , Raios Ultravioleta/efeitos adversosRESUMO
The effects of ROS on functional properties (cytotoxic activity, antibody-producing activity, TNFα synthesis, and free cytosol calcium level), membrane structure (by expression of some surface markers), and apoptosis of lymphocytic cells were estimated in the peripheral blood of healthy volunteers. 1O2, [Formula: see text], OHâ¢, and H2O2 mostly suppressed cytotoxic activity of lymphocytes against Ehrlich ascites carcinoma cells and inhibited IgG synthesis and expression of receptors and surface markers (Fc receptors, CD3, CD19, and CD56). The exposure of lymphocytes to H2O2 (10-6 M), 1O2, and OH⢠was followed by an increase in the level of a secondary messenger, intracellular calcium, in comparison with non-exposed cells. The presence of exogenous calcium in the medium for lymphocyte suspending induced an increase in the number of cells at early and late stages of apoptosis 6 h after exposure to H2O2 and 1O2 in comparison with lymphocytes incubated in Ca2+-free medium.
Assuntos
Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Adolescente , Adulto , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Células Cultivadas , Voluntários Saudáveis , Humanos , Peróxido de Hidrogênio/farmacologia , Radical Hidroxila/farmacologia , Masculino , Pessoa de Meia-Idade , Oxigênio Singlete/farmacologia , Adulto JovemRESUMO
Scanning electron microscopy study showed that exposure to CO for 60, 75, and 90 min induced heterogeneous changes in erythrocyte population. Increasing the duration of exposure of blood erythrocytes to CO was followed by the appearance of cells with morphological changes. The formation of discocytes with processes (≥1) was followed by the appearance of "deflated ball"-shaped erythrocytes. Moreover, CO modulated activity of glucose-6-phosphate dehydrogenase in human erythrocytes and disturbed their energy metabolism (suppressed lactate dehydrogenase activity in forward reaction and increased it in reverse reaction). A significant decrease in the coefficient of energy metabolism of erythrocytes (from 36±14 to 5.0±2.5 arb. units) reflected metabolic maladaptation induced by the exposure to CO.
Assuntos
Monóxido de Carbono/farmacologia , Metabolismo Energético , Eritrócitos/metabolismo , Adolescente , Adulto , Catálise , Eritrócitos/citologia , Feminino , Glucosefosfato Desidrogenase/metabolismo , Voluntários Saudáveis , Humanos , L-Lactato Desidrogenase/metabolismo , Masculino , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Propriedades de Superfície , Fatores de Tempo , Adulto JovemRESUMO
We investigated the mechanism of UV-radiation influence on trypsin in free and immobilized (on chitosan) states. The catalytic activity of free enzyme under the action of UV-light is subjected to changes to a greater extent than that of the immobilized one. We assume that the photoprotection effect of chitosan is caused for the following reasons: firstly, through interactaction with trypsin molecules chitosan forms a more photoresistant complex as compared to the native protein; secondly, chitosan probably binds the active photopro- ducts of a free radical nature, thus preventing oxidation (destruction) of several amino acids of the enzyme under its UV-radiation.
Assuntos
Quitosana/química , Enzimas Imobilizadas/química , Tripsina/química , Raios Ultravioleta/efeitos adversos , Aminoácidos/química , Aminoácidos/efeitos da radiação , Enzimas Imobilizadas/efeitos da radiação , Radicais Livres/química , Concentração de Íons de Hidrogênio , Oxirredução/efeitos da radiação , Proteção Radiológica , Tripsina/efeitos da radiaçãoRESUMO
We studied the effect of carbon monoxide (60-, 75-, and 90-min exposure) on the expression of antiapoptotic proteins (survivin and Bcl-2) in human blood lymphocytes in the presence of recombinant IL-2 in an apoptosis-inducing dose (0.1 ng/ml). Incubation of cells in atmosphere with carbon monoxide in the presence of recombinant IL-2 was accompanied by accumulation of Bcl-2 protein with simultaneous decrease of survivin content. It was concluded that carbon monoxide plays a role in the dysregulation of apoptosis of human blood lymphocytes Bcl-2 (i.e. CO inhibits the proapoptotic effect of recombinant IL-2).
Assuntos
Apoptose/efeitos dos fármacos , Monóxido de Carbono/farmacologia , Proteínas Inibidoras de Apoptose/genética , Interleucina-2/farmacologia , Linfócitos/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/genética , Apoptose/genética , Regulação da Expressão Gênica , Humanos , Proteínas Inibidoras de Apoptose/antagonistas & inibidores , Proteínas Inibidoras de Apoptose/metabolismo , Interleucina-2/antagonistas & inibidores , Linfócitos/citologia , Linfócitos/metabolismo , Cultura Primária de Células , Proteínas Proto-Oncogênicas c-bcl-2/agonistas , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas Recombinantes/farmacologia , Transdução de Sinais , SurvivinaRESUMO
The influence of UV-light (240-390 nm) at dozes of 151 and 755 J/m2 on the content of membrane markers of lymphocytes using the method of flow cytometry was investigated. It was demonstrated that during incubation of UV-irradiated lymphocytes the change of their populational and sub-populational composition occurs. Expression of complexes of CD3, CD 19,.CD8, CD 16, CD25 and CD95 increased. This increase was caused mainly by de novo synthesis. UV-light had immunostimulating effect on CD8+ T-lymphocyte population. Together with the increase of cytotoxic cells and NK-cells, activation of lymphocytes (increased amount of CD25+ and CD95+ cells) took place. Amount of cells undergone apoptosis or necrosis increased proportionally to the dosage. These changes were more expressed during incubation of lymphocytes in nutrition medium without autological blood serum, e.g. under deficiency of growth factors and antioxidants.
Assuntos
Antígenos CD/efeitos da radiação , Apoptose/efeitos da radiação , Biomarcadores/sangue , Linfócitos/efeitos da radiação , Antígenos CD/sangue , Doadores de Sangue , Citometria de Fluxo , Humanos , Células Matadoras Naturais/metabolismo , Células Matadoras Naturais/efeitos da radiação , Linfócitos/metabolismo , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/efeitos da radiação , Raios UltravioletaRESUMO
The spectral and oxygen-binding characteristics of human intracellular hemoglobin in the presence of nitroglycerin at concentrations of 5 ng/mL and 5 µg/mL have been studied. Short incubation (20 min) of erythrocytes with the drug led increasing hemoglobin affinity to oxygen and weakening of cooperative interactions in hemoprotein molecules. As a result, the amount of O(2) supplied to tissues in the process of gas exchange decreased by 23.96% (5 ng/mL) and 26.68% (5 µg/ml), p < 0.05. Incubation of cells for 24 h resulted in oxidation of the heme iron atom, accumulation of methemoglobin, and partial hemolysis. Nitroglycerin reduces the intensity of oxidative processes. However, no dependence of the degree of changes in the physical and chemical properties of hemoglobin on the concentration of nitroglycerin was found.
Assuntos
Eritrócitos/metabolismo , Nitroglicerina/farmacologia , Oxigênio/metabolismo , Oxiemoglobinas/metabolismo , Relação Dose-Resposta a Droga , Eritrócitos/química , Humanos , Oxigênio/química , Oxiemoglobinas/química , Análise EspectralRESUMO
UV-induced changes in the catalytic activity and radiuses of inulinases molecules from various producers (plants, fungy, yeast) are studied. It is established that specific enzymes activity and the sizes of inulinases molecules from Helianthus tuberosus and Kluyveromyces marxianus under the influence of UV-light in the ranges of doses 4530-6040 and 755-6040 J/m2, respectively, are subjected to changes more than structural and functional characteristics of inulinase fromAspergillus niger. It is probably connected with lower contents in it of aromatic amino acids such as tyrosine and phenylalanine. The most expressed loss of functional properties of inulinase from Helianthus tuberosus can be caused by the'existence of significantly more numbers of cysteine in plant fructan-exohydrolases in relation to microbic enzymes. A scheme for the stages of response of inulinases of various origins on the influence of UV-light in a certain range of radiation doses is offered.
Assuntos
Aspergillus niger/efeitos da radiação , Glicosídeo Hidrolases/metabolismo , Helianthus/efeitos da radiação , Kluyveromyces/efeitos da radiação , Monitoramento de Radiação/métodos , Raios Ultravioleta , Aspergillus niger/enzimologia , Relação Dose-Resposta à Radiação , Glicosídeo Hidrolases/química , Helianthus/enzimologia , Kluyveromyces/enzimologia , Tolerância a RadiaçãoRESUMO
The flow-cytometric analysis of condition of human peripheral blood lymphocytes in dynamics of development of the apoptosis induced by exposure to UV-light (240-390 nanometers) at a dose of 1510 J/m2 was carried out. Superficial architectonics and changes of the level of a membrane potential and functional activity of succinatedehydrogenase of mitochondrions are studied. The contribution of extracellular calcium to the processes of lymphocyte cellular death is revealed. The ideas about dynamics of the intracellular events leading to the death of photomodified lymphocytes 1-6 h after their UV-radiation exposure are specified and added on the basis of the analysis of our new and already published results.
Assuntos
Apoptose/efeitos da radiação , Fragmentação do DNA/efeitos da radiação , Linfócitos/efeitos da radiação , Raios Ultravioleta , Cálcio/metabolismo , Técnicas de Cultura de Células , Células Cultivadas , Meios de Cultura/química , Relação Dose-Resposta a Droga , Citometria de Fluxo , Humanos , Linfócitos/metabolismo , Linfócitos/patologia , Linfócitos/ultraestrutura , Mitocôndrias/enzimologia , Mitocôndrias/efeitos da radiação , Mitocôndrias/ultraestrutura , Succinato Desidrogenase/metabolismo , Fatores de TempoRESUMO
The computer model for a dimer of inulinase from Aspergillus ficuum is designed. The permolecular organization of inulinase from Aspergillus niger is experimentally investigated. The question about the role of various inulinase forms in manifestation of their functional activity is discussed. It is shown, that in the process of inulinase dimerization when contacts between monomeric forms of the enzyme are formed, a key role belongs to the nonpolar amino acid residues.
Assuntos
Aminoácidos/química , Aspergillus/química , Proteínas Fúngicas/química , Glicosídeo Hidrolases/química , Inulina/química , Aspergillus/enzimologia , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Multimerização Proteica , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Eletricidade Estática , TermodinâmicaRESUMO
The decomposition pathways of hemoglobin UV absorption spectrum into the absorption spectra of the protein and non-protein components are proposed and substantiated by means of an additive model. We have established that the heme component has an absorption band with a maximum at λ(max) = 269.2 nm (ε = 97163) and the apoprotein component has an absorption band with a maximum at λ(max) = 278.4 nm (ε = 48669) for the wavelength range from 240.0 to 320.0 nm. An integral relative proportion of absorption for the heme fraction (78.8%) and apoprotein (21.2%) in the investigating wavelength range is defined.
Assuntos
Aminoácidos/química , Apoproteínas/química , Hemoglobinas/química , Modelos Teóricos , Análise Espectral , Raios UltravioletaRESUMO
The effects of doxycycline on the morphology and function of donor erythrocytes were studied by scanning electron microscopy. The antibiotic in concentrations of 7.8 × 10(-5) and 7.8 × 10(-6) mol/liter induced heterogeneous changes in the surface architectonics in the erythrocyte population. These data indicated stimulation of erythrocyte "aging" under the effect of doxycycline.
Assuntos
Antibacterianos/farmacologia , Forma Celular/efeitos dos fármacos , Doxiciclina/farmacologia , Eritrócitos/ultraestrutura , Eritrócitos/efeitos dos fármacos , Humanos , Microscopia Eletrônica de VarreduraRESUMO
It is established that ion exchange resins AV-17-2P, KU-2, AV-16-GS, AM 21A, IMAC-HP, PUROLITE and fiber VION KN-1 can be applied as carriers for inulinase immobilization. The analysis of IR spectra for an enzyme, carriers and heterogeneous enzyme preparations showed that inulinase binding to matrices of various carriers occurs in general through electrostatic interactions. It is assumed that the mechanisms of interaction between inulinase from Kluyveromyces marxianus and the matrices of cation- and anion exchange polymers differ essentially from each other: different sites of protein molecule take part in adsorption that causes various conformational reorganizations in an enzyme molecule.
Assuntos
Proteínas Fúngicas/química , Glicosídeo Hidrolases/química , Resinas de Troca Iônica/química , Kluyveromyces/enzimologia , Absorção Fisico-QuímicaRESUMO
The article reviews the results of the studies of marker parameters (indicators) of various pathways and mechanisms of apoptosis of lymphocytes in donor peripheral blood induced by UV light (240-390 nm) in doses of 151, 1510, and 3020 J/m2. The article analyses the processes of DNA fragmentation, distortion of the structural asymmetry of the cell membranes, changes in the degree of DNA damage (single-strand breaks), transcriptional factor Ñ53, cytochrome Ñ, Fas receptors (CD95), caspase-3, caspase-8, and caspase-9, reactive oxygen species, and calcium ions in UV modified cells. The study determined that programmed cell death of lymphocytes after UV irradiation with 1510 J/m2 involves the Ñ53-dependent pathway of the nuclear mechanism, as well as receptor-mediated caspase mechanism, mitochondrial mechanism, and the mechanism associated with the defects in calcium homeostasis. Cell death is mediated by reactive oxygen and calcium ions. The article suggests a scheme of possible intracellular events resulting in the apoptotic death of lymphocytes after UV irradiation.
RESUMO
In today's world, there is a wide array of materials engineered at the nano- and microscale, with numerous applications attributed to these innovations. This review aims to provide a concise overview of how nano- and micromaterials are utilized for enzyme immobilization. Enzymes act as eco-friendly biocatalysts extensively used in various industries and medicine. However, their widespread adoption faces challenges due to factors such as enzyme instability under different conditions, resulting in reduced effectiveness, high costs, and limited reusability. To address these issues, researchers have explored immobilization techniques using nano- and microscale materials as a potential solution. Such techniques offer the promise of enhancing enzyme stability against varying temperatures, solvents, pH levels, pollutants, and impurities. Consequently, enzyme immobilization remains a subject of great interest within both the scientific community and the industrial sector. As of now, the primary goal of enzyme immobilization is not solely limited to enabling reusability and stability. It has been demonstrated as a powerful tool to enhance various enzyme properties and improve biocatalyst performance and characteristics. The integration of nano- and microscale materials into biomedical devices is seamless, given the similarity in size to most biological systems. Common materials employed in developing these nanotechnology products include synthetic polymers, carbon-based nanomaterials, magnetic micro- and nanoparticles, metal and metal oxide nanoparticles, metal-organic frameworks, nano-sized mesoporous hydrogen-bonded organic frameworks, protein-based nano-delivery systems, lipid-based nano- and micromaterials, and polysaccharide-based nanoparticles.
RESUMO
Structural and functional state of human blood lymphocytes after exposure to UV light (240-390 nm) in doses of 151-1359 J/m(2) was studied by methods of laser flow cytofluorometry, indirect immunofluorescence, and fluorescent probes. Using a combination of these methods, we have showed that UV light in the specified doses induced changes in the surface phenotype of T cells: stimulation or suppression of the expression of antigen-recognizing receptor complex molecules (CD3, CD4, and CD8 markers) and their redistribution on the surface of immunocompetent cells (capping effect) with the formation of receptor clusters of various types.
Assuntos
Linfócitos B/efeitos da radiação , Complexo CD3/genética , Antígenos CD4/genética , Antígenos CD8/genética , Membrana Celular/efeitos da radiação , Linfócitos T/efeitos da radiação , Naftalenossulfonato de Anilina , Linfócitos B/imunologia , Linfócitos B/metabolismo , Complexo CD3/imunologia , Antígenos CD4/imunologia , Antígenos CD8/imunologia , Membrana Celular/metabolismo , Células Cultivadas , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Corantes Fluorescentes , Expressão Gênica/efeitos da radiação , Humanos , Espectrometria de Fluorescência , Linfócitos T/imunologia , Linfócitos T/metabolismo , Raios UltravioletaRESUMO
We studied the effect of UV light (240-390 nm) in doses of 75.5-4530 J/m(2) on functional properties and level of intracellular messengers (cAMP, Ca(2+), NO) in lymphocytic cells of the peripheral blood. Correcting effects of UV light in a dose of 151 J/m(2) on cytotoxic activity of lymphocytes against Ehrlich ascetic carcinoma and calcium concentration in lymphocyte cytosol were observed. We found that UV light reduces the cAMP content and increases NO concentration (151-4530 J/m(2)) in photomodified cells. The involvement of adenylate cyclase and phosphoinositide signaling pathway into the mechanisms of modulation of functional properties of lymphocytes after exposure to UV light was demonstrated.