Polymorphism of Transforming Growth Factor TGFB and Its Receptors TGFBRI, TGFBRII, TGFBRIIII Genes in Western Siberia Patients with Open-Angle Glaucoma.

Polymorphism of genes of transforming growth factor TGFB and its receptors (TGFBRI, TGFBRII, and TGFBRIIII) in patients with primary open-angle glaucoma was analyzed. The frequency of the TGFBRII CC genotype in patients is increased relative to the control group (OR=6.10, p=0.0028). Heterozygosity in this polymorphic position is reduced (OR=0.18, p=0.0052). As the effects of TGF-ß is mediated through its receptors, we analyzed complex of polymorphic variants of the studied loci in the genome of patients. Two protective complexes consisting only of receptor genes were identified: TGFBRI TT:TGFBRII CG (OR=0.10, p=0.02) and TGFBRII CG:TGFBRIII CG (OR=0.09, p=0.01). The study showed an association of TGFBRII polymorphism with primary open-angle glaucoma and the need to study functionally related genes in the development of the disease, which should contribute to its early diagnosis and prevention.

Studying the Proliferative Activity of Human Corneal Stromal Cell Populations.

The proliferative activity of populations of stromal cells (fibroblasts) obtained from human corneal lenticles under conditions of their differentiation into keratocytes was studied. It was shown that during differentiation, the number of dividing fibroblasts and the frequency of divisions, and motor activity of these cells (speed of movement along the cell trajectory and the length of the trajectory) sharply decreased. These findings indicate a decrease in the proliferative activity of fibroblasts under conditions of their differentiation and transformation into keratocytes. A period of 17 days is sufficient for differentiation of corneal fibroblasts into keratocytes.

Effect of Conditioned Media of Limbal Epithelial Cells and Corneal Stromal Cells on Functional Activity of Limb Mesenchymal Stem Cells.

We studied the effect of conditioned media from limbal epithelial stem cells, fibroblasts, and corneal keratocytes on the functional activity of human limbal mesenchymal stem cells. It was shown that the conditioned media from limbal epithelial stem cells reduced proliferative activity and inhibited migration of limbal mesenchymal stem cells. In the conditioned media of limbal epithelial stem cells, increased concentrations of VEGF and TNFα and reduced concentration of BDNF, vimentin, and fibronectin were found. The conditioned medium from corneal stromal cells did not affect functional activity of mesenchymal stem cells in the limbus. These data contribute to the understanding of the interaction of cells in the limbal niche and with corneal cells essential for the maintenance of the cellular homeostasis in the cornea.

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.

Morphofunctional Properties of Corneal Stromal Cells.

Human corneal stromal cells were isolated by enzymatic digestion from a new source, lenticules obtained during laser vision correction by the ReLEx SMILe method. The resulting culture was mainly presented by fibroblast-like cells with a phenotype CD90-/CD73+/CD105+/keratocan-/lumican-/ALDH1A1+ that differentiate into keratocytes in a specialized medium. The concentration of fetal calf serum-derived growth factors affects the rate of proliferation, production of erythropoietin and brain neurotrophic factor by corneal fibroblasts, and to a lesser extent, their migration activity and production of extracellular matrix components. Thus, the high functional potential of fibroblast-like cells isolated from stromal lenticles can be used to develop cell technologies in ophthalmology.

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.

[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.

[Differential diagnosis and treatment of Chandler iridocorneal endothelial syndrome]. / Differentsial'naia diagnostika i lechenie iridokorneal'nogo éndotelial'nogo sindroma Chandlera.

The article presents a profound clinical, instrumental and morphological analysis of a family case of iridocorneal endothelial syndrome (Chandler syndrome) notable for atypical aggressive bilateral course and unusual hereditary manifestation. Five stages of the pathological process have been suggested.

[Multi-level analysis of the prevalence of pseudoexfoliative syndrome and pseudoexfoliative glaucoma]. / Vozmozhnosti i otsenka éffektivnosti gipotenzivnoi terapii pri psevdoéksfoliativnom sindrome.

Epidemiological investigation of pseudoexfoliative syndrome (PEX) as a glaucoma predictor is necessary for antiglaucoma treatment planning. AIM: To study the prevalence of PEX and pseudoexfoliative glaucoma (PXFG) among healthy people, naïve patients, and patients under observation as well as to evaluate the hypotensive efficacy of bimatoprost/timolol fixed combination (FCBT) over 4 months in PXFG. MATERIAL AND METHODS: Epidemiological analysis, ophthalmological examination, statistical analysis. RESULTS: The prevalence of PEX among healthy Novosibirsk and Novosibirsk region citizens is 19.9%. In ophthalmologically compromised population and first-time patients, the prevalence of PEX is 24%, of which PXFG constitutes 57.2%. Retrospectively, the prevalence of PXFG in PEX patients who were treated and followed up appeared to be as high as 60.8%. PXFG also accounts for 70% of all open-angle glaucoma cases in the Novosibirsk region, which is much higher than the corresponding rates in the European part of Russia. A 4-month FCBT therapy in 5 groups of PXFG patients (no treatment, monotherapy, another fixed combination, two drugs, three drugs) has yielded an evident hypotensive effect in most patients. The decrease in intraocular pressure was clinically significant in all groups. CONCLUSION: There has been found a high prevalence of PEX and PXFG in the Novosibirsk region. FCBT has demonstrated high hypotensive efficacy at all stages of PXFG and was beneficial even in patients treated with other medications or fixed combinations and in refractory cases.

[Cytokine gene polymorphisms in patients with age-related macular degeneration].

AIM: To establish possible association between age-related macular degeneration (AMD) and cytokine genotype polymorphisms; for that promoter regions of a number of cytokine genes were studied, namely, TNF-A863C, TNF-A308G, TNF-A238G, IL1ß-C-31T, IL4-C590T, IL6-C174G, and IL10A-1082G. MATERIAL AND METHODS: A total of 102 AMD and 100 non-AMD participants in the same age range were examined at the Novosibirsk branch of the Academician S.N. Fyodorov IRTC «Eye Microsurgery¼. In all cases restriction fragment length polymorphism analysis was performed. RESULTS: The frequency of TNF-308 GA and IL10-1082 GG genotypes was higher in AMD patients than in the controls, while the TNF-308 АA minor genotype has proved protective against the disease. Moreover, AMD has been found to be positively related to 48 genotypic combinations and negatively - to 32 (with specificity as high as 98-99%). The odds ratio was also higher in combination analysis as compared to monogenotypes. CONCLUSION: Complex genetic analysis of cytokines gives an idea of the balance of proinflammatory and anti-inflammatory cytokines in AMD and, therefore, plays an important role in studying the disease pathogenesis and exploring therapeutic options.

[Characteristics of exosomes andmicroparticles discovered in human tears].

Exosomes represent a sort of extracellular vesicles, which transfer molecular signals in organism and possess markers of producing cells. Our study was aimed at search of exosomes in the tears of healthy humans, confirmation of their nature and examination of exosome morphological and molecular-biological characteristics. The tears (110-340 ml) were collected from 24 healthy donors (aged 46-60 years); individual probes were centrifuged at 20000 g for 15 min to pellet cell debris. The supernatants were examined in electron microscope using negative staining; and they were also used for isolation and purification of the exosomes by filtration (100 nm pore-size) and double ultracentrifugation (90 min at 100000 g, 4°C). The "pellets" were subjected to electron microscopy, immunolabeling. The RNA and DNA were isolated from the samples, and their sizes were evaluated by capillary electrophoresis, the concentration and localization of nucleic acids were determined. Sequencing of DNA was performed using MiSeq ("Illumina", USA), data were analyzed using CLC GW 7.5 ("Qiagen", USA). Sequences were mapped on human genome (hg19). Electron microscopy revealed in supernatants of the tears cell debris, spherical microparticles (20-40 nm), membrane vesicles and macromolecular aggregates. The "pellets" obtained after ultracentrifugation, contained microparticles (17%), spherical and cup-shaped EVs (40-100 nm, 83%), which were positive for CD63, CD9 and CD24 receptors (specific markers of exosomes). Our study showed presence of high amount of exosomes in human tears, and relation of the exosomes with RNA (size less than 200 nt) and DNA (size was 3-9 kb). Sequencing of the DNA showed that about 92% of the reads mapped to human genome.

Proliferative and inflammatory factors in the vitreous of patients with proliferative diabetic retinopathy.

PURPOSE: The purpose was to measure the concentrations of various cytokines and growth factors (including vascular endothelial growth factor [VEGF] and pigment epithelium-derived factor [PEDF]) in the vitreous of patients with proliferative diabetic retinopathy (PDR) and to investigate interaction between inflammatory and proliferative factors in the genesis of PDR. Materials and Methods : Vitreous samples from 32 eyes with PDR and 25 eyes without diabetes mellitus and signs of DR (control) were collected. Vitreous concentrations of VEGF, PEDF, monocyte chemotactic protein-1 (MCP-1), interleukin-4 (IL-4), IL-6, IL-8, IL-10, IL-17A, and secretory immunoglobulin A (sIgA) were simultaneously measured using enzyme-linked immunoassay. Results : Vitreous levels of VEGF, PEDF, IL-17A, IL-6, IL-8, IL-4, and sIgA were significantly (Π < 0.05) higher in eyes with PDR compared to control. The concentration of VEGF was more than 17-times higher than in control, and the concentration of PEDF was not changed oppositely and was also higher (1.45-times) compared to control, that may indicate disturbances of compensatory mechanisms in angiogenesis regulation in PDR. Significant (Π < 0.05) positive correlations were observed between vitreous concentrations of VEGF and IL-17ΐ (r = 0.45), VEGF and IL-8 (r = 0.48), VEGF and IL-4 (r = 0.51), PEDF and IL-17ΐ (r = 0.48), PEDF and IL-8 (r = 0.59), MCP-1 and PEDF (r = 0.72), MCP-1 and IL-8 (r0 = 0.45), IL-4 and IL-17ΐ (r = 0.65), IL-4 and IL-8 (r = 0.71), IL-8 and IL-17ΐ (r = 0.59). CONCLUSIONS: Significantly raised levels of inflammatory and proliferative factors and numerous positive correlations between them may demonstrate a significant role of activation of vascular proliferation and local inflammation in the pathogenesis of PDR.

[THE STRUCTURE OF LYMPHATIC CAPILLARIES OF THE CILIARY BODY OF THE HUMAN EYE].

Using light microscopy, immunohistochemistry and electron microscopy, the structural organization of interstitial spaces and vessels of the ciliary body of the human eye (n = 5) were studied. The ciliary body was found to contain wide interstitial spaces--tissue clefts bound by collagen fibers and fibroblasts. Organ-specific lymphatic capillaries were also demonstrated in the ciliary body. According to the present findings and the lymphatic region concept, the first 2 elements of the lymphatic region of the eye were described: tissue clefts--prelymphatics and lymphatic capillaries of the ciliary body. The third element of the lymphatic region are the lymph nodes of the head and neck.