scholarly journals Characteristics of the Population of the Rat Thymus CD68+ Cells in Response to Tautomeric Forms of Magnesium Orotate Introduction under Simulated Magnesium Deficiency

2019 ◽  
Vol 8 (1) ◽  
pp. 82-88
Author(s):  
N. N. Chuchkova ◽  
K. A. Tukmachova ◽  
M. V. Smetanina ◽  
O. M. Kanunnikova ◽  
V. G. Sergeev ◽  
...  
Keyword(s):  
Mechanical Activation ◽  
Cortical Area ◽  
Magnesium Deficiency ◽  
Intracellular Localization ◽  
Fluorescent Staining ◽  
Macrophage Population ◽  
Tautomeric Forms ◽  
Hematoxylin And Eosin ◽  
Magnesium Orotate

he aim of study was to identify a response of the thymus CD68+ cells population to the introduction of tautomeric forms of magnesium orotate under magnesium deficiency.Material and methods. Magnesium-deficient status in rats was simulated by furosemide loading (30 mg/kg 14 days); then the animals were injected magnesium orotate in the initial oxo-form (Magnerot©, Vervag, Germany) and hydroxy-form obtained by mechanical activation, dosage 50 mg/kg, for 14 days., The density of thymocytes and macrophages was calculated in the subcapsular zone, cortex and brain matter (stained with hematoxylin and eosin) in the thymus, macrophages were studied immunohistochemically using monoclonal CD68 antibodies (Clone PG-M1).Results. The oxo-form of the drug was shown to lead to an increase in the number of CD68+ cells in the cortex of thymus (1.7 compared with the control) and medulla of thymus (2.3 times), an increase in the intensity of immunofluorescence (22.5%), the appearance of the thymus lobule of large CD68+cells in the cortex and the medulla part with a clearly traced intracellular localization of granules. The introduction of mechanically activated hydroxyl-form was accompanied by an increase in the number of CD68+ macrophages in the cortical area (1.5 times), and cortico-medullary partition (in 1.4 times), but did not affect the cerebral area of the thymus. The overall intensity of fluorescent staining reduced to 27.87±2.2 relative units (RU).Conclusion. The tautomeric forms of magnesium orotate have a diverse effect on the macrophage population of the thymus: the introduction of the oxo-form is accompanied by a significant activating effect on the CD68+macrophages in contrast to the hydroxyl form having a moderate modulating effect.

scholarly journals Morphofunctional characterization of rat thymus mast cells after administration of magnesium orotate mechanically activated forms

2021 ◽  
Vol 25 (3) ◽  
pp. 248-255
Author(s):  
Natalya N. Chuchkova ◽  
Marina V. Smetanina ◽  
Alexei E. Shklyaev ◽  
Ksenia A. Pazinenko ◽  
Natalya V. Kormilina ◽  
...  
Keyword(s):  
Magnesium Deficiency ◽  
Argon Plasma ◽  
Atomic Emission ◽  
Immunomodulatory Activity ◽  
Drug Induced ◽  
Inductively Coupled ◽  
The People ◽  
Thymus Tissue ◽  
Magnesium Orotate

Relevance. The topicality of the work is determined by the wide spread of hypomagnesemia among the people, which makes it necessary to correct it. The aim of the work is to elucidate the cell-mediated response of the thymus mastocytic link to magnesium deficiency and its correction by the mechanoactivated form of magnesium orotate. Materials and Methods . Animals with drug-induced magnesium deficiency (administration of furosemide 30 mg/kg for 14 days) were administered either the initial preparation Magnerot (Magnerot, Vervag Pharma, Germany), or its mechanoactivated form. The level of magnesium in the blood was determined by test systems ARKREY (Japan). The concentration of magnesium in the thymus tissue was determined by the method of emission spectroscopy with inductively coupled (argon) plasma on an atomic emission spectrometer. Density of mastocytes and the indices of degranulation and granulolosis were calculated on paraffin sections of the thymus after coloration with toluidine blue. Results and Discussion . It was shown that furosemide administration the amount of magnesium decreased in the blood (from 1,750,08 to 0,9020,18 mmol/l, p0,05), but increased in the thymus (from 1,60,6 in the control to 3,71,2 mg/l); in the gland tissue, the number of mastocytes of morphotype A decreased and the number of mastocytes of morphotype D, after active degranulation, increased (by 7,1 times, p0,05). The type of mastocyte secretion in hypomagnesemia is represented by the merocrine variant. The administration of the initial magnesium orotate led to an increase in the concentration of magnesium in the blood to 1,150,25 mmol/l, which is 65,7% of the initial level, the amount of magnesium in the thymus remained elevated (3,41,1 mg/l), the number of actively degranulating cells (morphotype D) was increased. Mechanoactivated magnesium orotate restored the concentration of Mg2+ in the blood to 89,1% (1,560,18 mmol/l, p0,05) and decreased in the thymus (to 2,30,7 mg/l), restored the subpopulation of mastocytes saturated with heparin (type A), reduced the number of mastocytes of morphotype D. Conclusion . The mechanoactivated form of magnesium orotate has a normalizing effect on the population of thymic mastocytes, shows pronounced immunomodulatory activity, which allows us to consider it as a potential therapeutic agent for clinical testing in the complex therapy of hypomagnesemia and associated immunodeficiency.

scholarly journals Tautomers of the Orotate Anion Have Anti-Inflammatory Effects in the Correction of Drug-Induced Hepatitis in Rats

2021 ◽  
pp. 366-373
Author(s):  
Kseniya A. Pazinenko ◽  
◽  
Natalʼya N. Chuchkova ◽  
Marina V. Smetanina
Keyword(s):  
Orotic Acid ◽  
Intervention Group ◽  
Biological Research ◽  
Control Group ◽  
Drug Induced ◽  
Tautomeric Forms ◽  
Inflammatory Phenotype ◽  
Hematoxylin And Eosin ◽  
Anti Inflammatory ◽  
Initial Drug

The purpose of this paper was to conduct a comparative experimental study of the anti-inflammatory effects of tautomeric forms of orotic acid in the correction of drug-induced hepatitis in rats. Materials and Methods. A total of 40 rats (Rattus norvegicus Berk.) were randomly divided into 5 groups: control group (n = 10); intervention group with drug-induced hepatitis (n = 15); animals with drug-induced hepatitis who were injected with orotic acid (ОА) tautomers: initial oxo-form (n = 5), hydroxy-form (n = 5) and dihydroxyform (n = 5) at a dose of 0.5 g/kg body weight a day in the course of 14 days. The tautomers were obtained by mechanical activation in a planetary ball mill AGO-2C for 1 (hydroxy-form) and 6 (dihydroxyform) hours. In the blood of animals of all experimental groups, the content of leukocytes, granulocytes, lymphocytes, and monocytes was determined. Liver sections were stained with hematoxylin and eosin to assess the histo- and cytostructure of the tissues, and immunohistochemically using a set of monoclonal antibodies to detect the expression of the CD68+ macrophage marker. Results. It was found that when correcting drug-induced hepatitis with the hydroxy-form of OA, the severity of leuko- and monocytosis decreased and the number of lymphocytes was restored. The number of CD68+ macrophages with a pro-inflammatory phenotype decreased in the group receiving the hydroxy-form of OA (by a factor of 1.32; p = 0.019), but remained unchanged at the administration of oxo- and dihydroxy-forms of OA. The intensity of reaction product expression decreased by a factor of 1.5 in the group administered with the initial drug and by a factor of 1.9 in animals administered with mechanically activated drugs (p = 0.0001). Thus, the obtained data indicate a pronounced anti-inflammatory activity of the hydroxyform of OA, which can substantiate its use as a hepatoprotective agent. For citation: Pazinenko K.A., Chuchkova N.N., Smetanina M.V. Tautomers of the Orotate Anion Have Anti-Inflammatory Effects in the Correction of Drug-Induced Hepatitis in Rats. Journal of Medical and Biological Research, 2021, vol. 9, no. 4, pp. 366–373. DOI: 10.37482/2687-1491-Z074

scholarly journals Voltage-dependent anion channels 1 and 2 are expressed in porcine oocytes

2009 ◽  
Vol 30 (3) ◽  
pp. 193-200 ◽  
Author(s):  
María Carolina Cassará ◽  
Viviana Andrea Menzel ◽  
Klaus-Dieter Hinsch ◽  
Christine Wrenzycki ◽  
Elvira Hinsch
Keyword(s):  
Protein Interactions ◽  
Cortical Area ◽  
Intracellular Localization ◽  
Germinal Vesicle ◽  
Anion Channel ◽  
Confocal Laser ◽  
Protein Protein Interactions ◽  
Voltage Dependent Anion Channel ◽  
Voltage Dependent ◽  
Porcine Oocyte

The eukaryotic VDAC (voltage-dependent anion channel) is a pore-forming protein originally discovered in the outer membrane of mitochondria. It has been established as a key player in mitochondrial metabolism and ion signalling. In addition, in recent years, it has also been proposed that VDAC is present in extra-mitochondrial membranes, and it has been related to cytoskeletal structures. However, little is known about the presence and intracellular localization of VDAC subtypes in mammalian gametes. In the present study, we confirm the synthesis of VDAC1 and 2 subtypes in GV (germinal vesicle) and MII (meiosis II) stage porcine oocytes as well as their protein expression. A shift in the abundance of immunoreactive 32 kDa VDAC protein between GV and MII stage oocytes was observed with anti-VDAC2 antibody. Furthermore, subcellular localization by confocal laser microscopy demonstrated fluorescent labelling of VDAC1 over the entire oocyte surface, suggesting the presence of VDAC1 in the porcine oocyte plasma membrane and around the cortical area. Anti-VDAC2 immunostaining yielded ring-like clusters of structures distributed on the cortical area in some GV, but not in MII, stage oocytes. These results are the first data obtained for VDAC in mammalian female gametes and provide the basis for studying protein–protein interactions, distribution and possible functions of VDAC subtypes during maturation and fertilization of mammalian oocytes.

scholarly journals Cellular and molecular events of inflammation induced transdifferentiation (EMT) and regeneration (MET) in mesenteric mesothelial cells

2020 ◽  
Vol 69 (12) ◽  
pp. 1173-1179
Author(s):  
Viktória Zsiros ◽  
Anna L. Kiss
Keyword(s):  
Intracellular Localization ◽  
Mesothelial Cells ◽  
Lysosomal Degradation ◽  
Environmental Stimulus ◽  
Macrophage Population ◽  
Molecular Events ◽  
Late Endosomes ◽  
Early Endosomes ◽  
Signaling Organelles ◽  
Intracellular Compartmentalization

Abstract In this review we summarize the cellular and molecular events of inflammation induced epithelial-to-mesenchymal (EMT) and mesothelial-to-macrophage transition (MET) during regeneration. Since the receptor transmits the environmental stimulus, downregulating or upregulating the process on an epigenetic level, the intracellular localization of receptors (signaling organelles: early endosomes or lysosomal degradation: late endosomes) plays a crucial role in the signaling events regulating inflammation and regeneration. Therefore, we focused on the internalization of the receptors as well as the intracellular compartmentalization of signaling molecules during EMT and MET. The review draws the reader’s attention to the plasticity of mesothelial cells and supports the idea that during inflammation an ambient macrophage population might derive from mesothelial cells.

The Morphology of the Rat Kidney Following Folic Acid Administration

1970 ◽  
Vol 28 ◽  
pp. 200-201
Author(s):  
Aline Byrnes ◽  
Elsa E. Ramos ◽  
Minoru Suzuki ◽  
E.D. Mayfield
Keyword(s):  
Folic Acid ◽  
De Novo ◽  
Specific Activity ◽  
Rat Kidney ◽  
Present Report ◽  
Intracellular Localization ◽  
Male Rats ◽  
Renal Hypertrophy ◽  
Electron Microscopic ◽  
Biochemical Alterations

Renal hypertrophy was induced in 100 g male rats by the injection of 250 mg folic acid (FA) dissolved in 0.3 M NaHCO3/kg body weight (i.v.). Preliminary studies of the biochemical alterations in ribonucleic acid (RNA) metabolism of the renal tissue have been reported recently (1). They are: RNA content and concentration, orotic acid-c14 incorporation into RNA and acid soluble nucleotide pool, intracellular localization of the newly synthesized RNA, and the specific activity of enzymes of the de novo pyrimidine biosynthesis pathway. The present report describes the light and electron microscopic observations in these animals. For light microscopy, kidney slices were fixed in formalin, embedded, sectioned, and stained with H & E and PAS.

The Immuno-Electron Microscopic Localization of the Mo-Fe Protein Component of Nitrogenase in Cells of Azotobacter Vinelandii

1973 ◽  
Vol 31 ◽  
pp. 574-575
Author(s):  
J. T. Stasny ◽  
R. C. Burns ◽  
R. W. F. Hardy
Keyword(s):  
Cellular Localization ◽  
Direct Evidence ◽  
Azotobacter Vinelandii ◽  
Intracellular Localization ◽  
Protein Component ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Fe Protein ◽  
Intracytoplasmic Membranes

Structure-functlon studies of biological N2-fixation have correlated the presence of the enzyme nitrogenase with increased numbers of intracytoplasmic membranes in Azotobacter. However no direct evidence has been provided for the internal cellular localization of any nitrogenase. Recent advances concerned with the crystallizatiorTand the electron microscopic characterization of the Mo-Fe protein component of Azotobacter nitrogenase, prompted the use of this purified protein to obtain antibodies (Ab) to be conjugated to electron dense markers for the intracellular localization of the protein by electron microscopy. The present study describes the use of ferritin conjugated to goat antitMo-Fe protein immunoglobulin (IgG) and the observations following its topical application to thin sections of N2-grown Azotobacter.

An immunocytochemical study of maternal immunoglobulin localization in the suckling pig intestine

1990 ◽  
Vol 48 (3) ◽  
pp. 922-923
Author(s):  
László G. Kömüves ◽  
Donna S. Turner ◽  
Kathy S. McKee ◽  
Buford L. Nichols ◽  
Julian P. Heath
Keyword(s):  
Sodium Ethoxide ◽  
Intracellular Localization ◽  
Protein A ◽  
Tween 20 ◽  
Intestinal Epithelial ◽  
Immunocytochemical Study ◽  
Fish Skin ◽  
Newborn Piglets ◽  
Rabbit Igg ◽  
Fish Skin Gelatin

In this study we used colloidal gold probes to detect the intracellular localization of colostral immunoglobulins in intestinal epithelial cells of newborn piglets.Tissues were obtained from non-suckled newborn and suckled piglets aged between 1 hour to 1 month. Samples were fixed in 2.5 % glutaraldehyde, osmicated and embedded into Spurr’s resin. Thin (80 nm) sections were etched with 5% sodium ethoxide for 5 min, washed and treated with 4 % sodium-m-periodate in distilled water for 30 min. The sections were then first incubated with blocking buffer (2 % BSA, 0.25 % fish skin gelatin, 0.5 % Tween 20 in 10 mM Trizma buffer, pH=7.4 containing 500 mM NaCl) for 30 min followed by the immunoreagents diluted in the same buffer, 1 hr each. For the detection of pig immunoglobulins a rabbit anti-pig IgG antiserum was used followed by goat anti-rabbit IgG-Au10 or protein A-Au15 probes.

Intraneuronal accumulations of lysosomes in the cerebellum of rats and dogs after dosing with MDL-832

1990 ◽  
Vol 48 (3) ◽  
pp. 830-831
Author(s):  
S.D. Barnard ◽  
S.D. Warner
Keyword(s):  
Brown Pigment ◽  
Beagle Dogs ◽  
Sprague Dawley ◽  
Gelatin Capsules ◽  
Pigment Granules ◽  
Sprague Dawley Rats ◽  
Hematoxylin And Eosin ◽  
Dose Levels

1, 2, 9, 10-tetramethoxyaporphine phosphate (MDL-832) was once considered a potential human antitussive. MDL-832 was administered orally in the diets of Sprague-Dawley rats at dose levels of 0, 5, 10, 20, 40, 80 and 160 mg/kg/day for 3 and 6 months and in gelatin capsules to Beagle dogs at 0, 5, 10, 15, 30 and 60 mg/kg/day for 3, 6 and 12 months. Histopathologic examinations of hematoxylin and eosin-stained cerebellar sections revealed intracytoplasmic brown pigment accumulations in large fusiform neurons (presumably the motor type) of the pons. The pigment granules were found to be PAS-positive, non-acid fast, iron-free, Sudan B-positive and fuchsinophilic. Intraneuronal pigment accumulations were seen in rats after 3 months of treatment at 80 mg but not at 40 mg and after 6 months at 20 mg but not at 10 mg. For dogs the effect was observed after 3 months at 60 mg but not at 30 mg and after 12 months at 10 mg but not at 5 mg.

Immunoferritin Labelling of Murine Leukemia Virus Antigens in thin Sections

1980 ◽  
Vol 38 ◽  
pp. 508-509
Author(s):  
Ray A. Weigand ◽  
Gregory C. Varjabedian
Keyword(s):  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Intracellular Localization ◽  
Uranyl Acetate ◽  
Antibody Technique ◽  
Thin Sections ◽  
Tumor Virus ◽  
Labelling Procedure ◽  
Unlabelled Antibody ◽  
Murine Leukemia

We previously described the intracellular localization of murine mammary tumor virus (MuMTV) p28 protein in thin sections (1). In that study, MuMTV containing cells fixed in 3% paraformaldehyde plus 0.05% glutaraldehyde were labelled after thin sectioning using ferritin-antiferritin in an unlabelled antibody technique. We now describe the labelling of murine leukemia virus (MuLV) particles using the unlabelled antibody technique coupled to ferritin-Fab antiferritin. Cultures of R-MuLV in NIH/3T3 cells were grown to 90% confluence (2), fixed with 2% paraformaldehyde plus 0.5% glutaraldehyde in 0.1 M cacodylate at pH 7.2, postfixed with buffered 17 OsO4, dehydrated with a series of etha-nols, and embedded in Epon. Thin sections were collected on nickel grids, incubated in 107 H2O2, rinsed in HEPES buffered saline, and subjected to the immunoferritin labelling procedure. The procedure included preincubation in 27 egg albumin, a four hour incubation in goat antisera against purified gp69/71 of MuLV (3) (primary antibody), incubation in F(ab’)2 fragments of rabbit antisera to goat IgG (secondary antibody), incubation in apoferritin, incubation in ferritin-Fab ferritin, and a brief fixation with 2% glutaraldehyde. The sections were stained with uranyl acetate and examined in a Siemens IA electron microscope at an accelerating voltage of 60 KV.

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