The Expression of Markers of Acute Kidney Injury Kim1 and NGAL after Administration of High Doses of Lithium Carbonate in Mice with Engrafted Skin Melanoma B16.

The expression of marker proteins of acute kidney injury after administration of high doses of lithium carbonate was assessed to evaluate the possibility of lithium use in neutron capture therapy. In mice with implanted skin melanoma B16, the expression of Kim1 (kidney injury molecule 1) and NGAL (neutrophil gelatinase-associated lipocalin) proteins in the kidneys was evaluated immunohistochemically 15, 30, 90, 180 min, and 7 days after peroral administration of lithium carbonate at single doses of 300 and 400 mg/kg. An increase in the expression of the studied proteins was found in 30 and 90 min after administration of 400 mg/kg lithium carbonate, however, 7 days after the drug administration, the expression returned to the level observed in the control group. It can be suggested that single administration of lithium carbonate in the studied doses effective for lithium neutron capture therapy will not significantly affect the renal function.

Assessment of Autophagy in Tumor Cells of Human Skin Melanoma of Different Stages.

High levels of autophagy can increase the viability of tumor cells as well as their resistance to chemotherapy. Evaluation of the dynamics of autophagy processes at different stages of carcinogenesis can extend our understanding of melanoma pathogenesis to develop new therapeutic approaches. We performed a comparative study of tumor cell autophagy in stages II and III human skin melanoma. Tumor cells were characterized by high content of structures associated with autophagy (autophagosomes and autolysosomes). In stage III melanoma characterized by the presence of regional metastases in the lymph nodes, tumor cells showed higher expression of the autophagy marker protein LC3beta in comparison with stage II melanoma cells, which can indicate the involvement of autophagy processes in tumor progression and the formation of metastases in the lymph nodes.

Ultrastructural Reorganization of Endotheliocytes of Pulmonary Blood Capillaries in COVID-19.

The ultrastructural organization endotheliocytes of pulmonary blood capillaries in COVID-19 was studied on autopsy material using electron microscopy. Swelling of the cytoplasm and mitochondria with destruction of the cristae, dilation of the Golgi complex cisternae, a decrease in the volume density of the luminal and basal caveolae and free transport vesicles, an increase of the rough endoplasmic reticulum, as well as the presence of elements of coronavirus replication (reticulovesicular structures, zippered endoplasmic reticulum, electron-dense particles in the Golgi cisternae, and vacuoles with viral particles) were revealed. Further studies of the intracellular mechanisms used by the virus to replicate could help to develop antiviral drugs for the treatment of the new coronavirus infection.

The immunomodulatory activity of the betulonic acid based compound.

The immunomodulatory activity of a betulonic acid-based compound with furocoumarin (BABCF; 2-azido, 9-N-methylpiperazinomethyl oreozelone) has been investigated. Male C57BL/6 mice (aged 3 months) treated with the cytostatic agent cyclophosphamide (CP) and intact individuals served as experimental models. The expression of genes was studied in bone marrow (IL-12, IL-10, IL-1ß, TNF-α, TGF-ß, M-CSF, GM-CSF) or in the suspension of peritoneal cells (IL-12, IL-10; as the injection site). The surface markers of T-lymphocytes (CD3, CD4, and CD8) in fractions of venous blood mononuclear cells (MNCs) were determined by means of flow cytometry using antibodies. Histological and morphometric studies were performed to assess the impact of CP and BABCF on the thymus. BABCF caused a pronounced (about 3-fold) increase in relative expression of the GM-KSF gene. BABCF caused a local increase in the expression of IL-12 in the peritoneal cavity cells and restored the relative content of T-lymphocytes in the blood of CP-treated mice treated affecting mainly CD3⁺CD4⁺ lymphocytes. This substance reduced the tissue density of the thymic cortex and thymic medulla in CP-treated mice. Thus, results of this study suggest that BABCF exhibits a stimulating effect on the cellular link of immunity and promotes maintenance of the number of T-lymphocytes in the blood due to their migration from the central organs of the immune system.

Autophagy in Human Retinal Neurons in Glaucoma.

Immunohistochemical and ultrastructural analysis revealed signs of structural alterations in neurons and autophagy in all layers of the human retina at the end-stage glaucoma. The most pronounced destructive changes associated with swelling and destruction of mitochondria, endoplasmic reticulum, and Golgi apparatus, as well as structural signs of impaired synaptic activity and apoptosis were noted in ganglion, bipolar, and amacrine neurons. In the structure of photoreceptor cells, alone with destructive processes associated with structural alterations of rods and cones in the outer membrane discs, as well as swelling of organelles, we observed processes aimed at the maintenance of cell homeostasis. Structural signs of autophagy (mainly mitophagy) and changes of the ultrastructural organization in rod neurons were more pronounced than in cones.

Ultrastructure of Endothelial Cells of Myocardial Capillaries in Burn Septicotoxemia.

We studied ultrastructure and vesicular structures in endothelial cells of myocardial micro-vessels in burn patients. Electron microscopy revealed a significant decrease in volume density of vesicular structures in the endotheliocytes of myocardial capillaries in patients with burn septicotoxemia. The observed structural signs of endothelial dysfunction revealed in this category of patients can be a promising area for further research and for the development of methods of pathogenetic correction of myocardial disorders in the case of burn injury.

In Vitro Effects of Lithium Carbonate on Cell Cycle, Apoptosis, and Autophagy in Hepatocellular Carcinoma-29 Cells.

We studied the effects of lithium carbonate on the cell cycle, apoptosis, and autophagy in hepatocellular carcinoma-29 cells (HCC-29) in vitro. Flow cytofluorometry analysis revealed accumulation of G2/M-phase HCC-29 cells and increase in the number of apoptotic cells in 48 h after administration of 5 mM lithium carbonate. Induction of autophagy in HCC-29 cells was detected by transmission electron microscopy and immunofluorescence staining. Thus, lithium carbonate produces an antitumor effect by arresting cell cycle in the G2/M-phase and induction of apoptosis and autophagy in HCC-29 cells, which confirms the lithium potential as a promising drug for the treatment of hepatocellular carcinoma.

[The morphology of the connective tissue matrix and lymphatic bed of the external genitalia in primary massive localized lymphedema]. / Morfologiya soedinitel'notkannogo matriksa i limfaticheskogo rusla naruzhnykh polovykh organov pri pervichnoi massivnoi lokalizovannoi limfedeme.

OBJECTIVE: To identify the structural organization of connective tissue and vascular bed of the external genitalia in primary massive localized lymphedema. To analyze a clinical case of the development of massive localized lymphedema of the external genitalia concurrent with primary lymphedema of the lower limb in a patient with a normal body mass index. MATERIAL AND METHODS: The samples obtained by resection during volume reduction surgery were used to study the morphological features of the lymphatic bed and extracellular matrix of the scrotal skin and testicular dartos versus the samples without pathological changes. Biological samples were processed using standard techniques for histological, immunohistochemical and ultrastructural analyses. Lymphatic vessels were differentiated using the molecular lymphatic endothelial marker Podoplanin. RESULTS: In lymphedema, there was an increase in the thickness of all scrotal skin layers, a decrease in the volumetric lymphatic vessel density, an expansion of the interstitial spaces, and a change in the structure of collagen fibers that were homogenized, loosened, and swollen and did not form a three-dimensional network. The testicular dartos exhibited intermuscular fibrosis, expansion of the interstitial spaces, and perivascular leukocytic infiltration. CONCLUSION: Histological and immunohistochemical analyses revealed changes in the structural organization of the connective tissue matrix and lymphatic bed of the scrotal skin and testicular dartos in long-standing massive localized lymphedema. The feature of the described clinical case was the absence of signs of chronic inflammation and severe diffuse fibrosis in primary scrotal massive localized lymphedema in a patient with a normal body mass index.

Morphological Analysis of Blood Capillaries and Transport Function of Endothelial Cells in Hepatocellular Carcinoma-29.

Hepatocellular carcinoma is characterized by the pronounced vascularization, high levels of vascular endothelial growth factors, and intensive angiogenesis. Caveola-mediated endocytosis and clathrin-dependent endocytosis contribute to internalization of various growth factors, thus affecting blood supply to the tumor and neoangiogenesis. We performed a quantitative analysis of blood vessels in the tumor and evaluated the basal levels of endocytosis-associated vesicular structures in the endothelial cells of the hepatocellular carcinoma-29. The numbers of blood vessels and clathrin-coated vesicles in endothelial cells in the tumor tissue were significantly higher than in normal tissue. The association between clathrin-dependent endocytosis in the endotheliocytes of the capillaries and tumor angiogenesis can provide the basis for the development of new molecular targets for antiangiogenic antitumor therapy.

Ultrastructural Organization of Uveal Melanoma Stromal Cells.

Typical blood capillaries and vessels in uveal melanoma were shown and different types of uveal melanoma stromal cells were determined by electron microscopy and immunohistochemical analysis. Macrophages, fibroblasts of varying degrees of differentiation and endothelial-like cells with numerous caveolae in the cytoplasm were found in the channels of the extracellular matrix surrounding accumulations of tumor cells. The presence local structures positively stained for markers of the blood and lymphatic vessels (CD31 and podoplanin) in channels of the extracellular matrix suggests that the described endothelial-like cells can be the structural basis for blood and lymphatic vessels of the tumor.

Cytological Characteristics of a Heterogeneous Population of Hepatocellular Carcinoma-29 Cells after Injection of Lithium Carbonate in the Experiment.

Five cytological types of hepatocellular carcinoma-29 (G-29) grown in the muscle tissue of the thigh of experimental animals were identified by transmission electron microscopy; 89% of these were poorly differentiated type I-III cells. Lithium in a concentration of 20 mM produced a damaging effect on poorly differentiated G-29 cells: the number of cells with zones of intracellular component destruction and volume density of these zones increased, while volume density of cisterns of endoplasmic reticulum decreased. These results suggest that lithium carbonate can cause destructive changes in the heterogeneous population of G-29 cells during in vivo tumor development.

Ultrastructural Changes in Hepatocellular Carcinoma-29 Cells after Treatment with Lithium Carbonate.

We studied the effect of lithium carbonate on hepatocellular carcinoma-29 cells in CBA male mice after injection in a dose of 20 mM along the tumor periphery. Transmission electron microscopy revealed a decrease in the volume density of the granular endoplasmic reticulum in the cell cytoplasm and an increase in the total numerical and volume density of autophagosomes and autolysosomes and zones of destruction of intracellular organelles. The ability of lithium carbonate to activate intracellular degradation processes in tumor cells and to stimulate cell death can be used to develop new combined strategies in the chemotherapy for hepatocellular carcinoma.

Therapeutic Potential of Biomedical Cell Product in DSS-Induced Inflammation in the Small Intestine of C57Bl/6J Mice.

We studied the effect of intravenous administration of biomedical cell product (bone marrow mesenchymal stem cells and media conditioned by these cells) on histological picture of the small intestine in mice with DSS-induced inflammation. It was shown that biomedical cell product promoted recovery of the intestinal mucosa in DSS-induced inflammation.

Influence of Apoptotic Bodies and Apoptotic Microvesicles on NO Production in Macrophages.

We studied the effect of extracellular vesicular particles generated during apoptosis by macrophages of M0, M1 and M2 phenotypes on spontaneous and LPS-stimulated production of NO. The fractions of apoptotic bodies and apoptotic microvesicles were obtained in the primary cultures of peritoneal macrophages undergoing apoptosis. The effect of these microparticles on LPS-induced proinflammatory response of recipient macrophages critically depends on the initial phenotype of "donor" macrophages. Microvesicles and especially apoptotic bodies from M1 macrophages stimulate basal NO production. LPS stimulation of these macrophages preincubated with apoptotic bodies was not followed by further growth of NO production; in macrophages preincubated with microvesicles, LPS even suppressed NO production. Apoptotic microparticles obtained from M2 macrophages produced little effect on the basal production of NO. LPS stimulation of macrophages-recipients preincubated with microparticles from M2 macrophages did not enhance NO production. Incubation of macrophages with apoptotic microparticles induces the formation of endotoxic tolerance.

Autophagy in Hepatocytes during Distant Tumor Growth.

Structural changes in the liver of CBA mice were studied during the development of experimental hepatocarcinoma-29 inoculated into the hip. A decrease in the volume density of hepatocyte cytoplasm, mitochondria, endoplasmic reticulum, and lipid inclusions and an increase in the volume density of lysosomal structures during tumor growth were observed. All stages of intracellular autophagy were recorded by the method of electron microscopy. These stages included the appearance of autophagosomes, autophagolysosomes, and secondary lysosomes in the hepatocyte cytoplasm. Fragments of cytoplasm, glycogen rosettes, mitochondria, and fragments of endoplasmic reticulum with ribosomes were found in autophagosomes. The obtained data indicate the development of non-selective autophagy in the liver during distant tumor growth in aimed at the maintenance of intracellular homeostasis in hepatocytes and energy and trophic homeostasis of organism.

[Lymphatic system of the eye]. / Limfaticheskaia sistema glaza.

Existence of lymphatic outflow of intraocular fluid is still an open question. Identification of the lymphatic capillaries and vessels in various human organs in normal and pathological conditions became possible with discovery of endotheliocyte markers for lymphatic vessels. However, the available information on the presence of lymphatic structures in the human eye is inconsistent and uncertain. The data on lymphatic drainage of the eye is of interest, particularly because it may help understand glaucoma pathogenesis, mechanisms of development of eye inflammatory diseases, and develop pathogenetic therapies. The article reviews literature and presents the authors' own views on lymphatic drainage of the human eye.

Hypolipidemic effect of mannans from C. albicans serotypes a and B in acute hyperlipidemia in mice.

Mannans, which are biological macromolecules of polysaccharide origin and function as immunomodulators, have been shown to stimulate macrophages in vivo by interaction with the mannose receptor. Thus, they can be used to stimulate macrophages in order to effectively remove circulating atherogenic lipoproteins. Our primary aim was to evaluate the hypolipidemic potential of mannans from C. albicans serotype A (mannan A) and serotype B (mannan B) in a murine model of hyperlipidemia. Mannan A and mannan B were shown to significantly (p<0.05) stimulate both the proliferation (p <0.05) and nitric oxide production of murine peritoneal macrophages in vitro. Pre-treatment of CBA/Lac mice with mannan A prior to induction of hyperlipidemia significantly (p<0.001) reduced serum atherogenic LDL-cholesterol, total cholesterol, and triglycerides. Mannan B exhibited a similar, but more potent, hypolipidemic effect. Electron microscopic analysis of liver revealed a significant (p<0.001) decrease in the volume of lipid droplets when hyperlipidemic mice were pretreated by both mannans. In conclusion, our findings would suggest that both polysaccharide-based biological macromolecules evaluated in the present study, specifically, the natural immunomodulators (mannans A and B), appeared to function as effective lipid-lowering macromolecules, which could potentially serve as adjunct therapy to more conventional hypolipidemic medications such as a statin drug.

Prospect of Using Cell Product for the Therapy of Skin Defects in Diabetes Mellitus.

The effectiveness of autologous cell product in the therapy of skin burn wounds was studied in C57B1/6 male mice against the background of streptozotocin-induced diabetes mellitus. In animals with and without modeled diabetes mellitus, significant decrease in skin defect area was observed after single administration of the cell product (bone marrow multipotent mesenchymal stromal cells, fibroblasts or media conditioned by these cells).

Toxic Effects of Nanostructured Silicon Dioxide on Multipotent Mesenchymal Stromal Cells.

We studied the effect of nanostructured silicon dioxide particles on functional activity and ultrastructure of rat multipotent mesenchymal stem cells. A dose-dependent decrease in cell adhesion to plastics and their proliferative potential, accumulation of apoptotic cells, a decrease in the electron density of cell cytoplasm due to swelling and degenerative changes of organelles, and intracellular accumulation of nanostructured of silicon dioxide particles were observed.

[Concurrence of acute graft rejection and polyomavirus nephropathy: A clinical case]. / Klinicheskoe nabliudenie sochetaniia ostrogo ottorzheniia transplantata s poliomavirusnoi nefropatiei.

The paper describes a case of diagnosing acute renal graft rejection concurrent with polyomavirus nephropathy. Histochemical and electron microscopic methods were used to examine biopsy specimens, which showed morphological changes occurring in the allograft, the ultrastructural characteristics of polyomavirus and the features of its spread in kidney tissue structures.