The levels of erythropoietin (EPO) in biological fluids were studied on 43 patients with diabetes mellitus type I and II who suffered from a variety of clinical manifestations of heavy proliferative diabetic retinopathy (PDR) which incuded stable PDR, 12 patients with rhegmatogenous retinal detachment and 15 healthy individuals The correlation of EPO levels with the clinical features of heavy PDR was noted only at the local level (intravitreally). The highest intraocular levels of EPO were associated with severe features of the disease, abundant vascularizarion of fibroglial tissue, and intraoperative hemorrhagic complications, which corroborates the theory of active participation of locally secreted EPO in ocular neoangiogenesis, including the progression of PDR // Russian Ophthalmological Journal, 2016; 2: 55-60. doi: 10.21516/2072-0076-2016-9-2-55-60 .

Studying immunological aspects of pathogenesis of retinopathy of prematurity (ROP) helps develop new approaches to prevention, rational diagnosis and increase of the effectiveness of treatment of the disease. Purpose: to analyze the level of growth factors in blood serum of premature infants belonging to the group of ROP risk at the pre-clinical stage and find out how this level is related to further development and course of the disease. Material and methods. 85 prematurely born infants of the ROP risk group were examined. The average gestational age at birth was 27.7 ± 2.2 weeks, the average birth weight - 1086.1 ± 266.1 g. The examination included dynamic ophthalmoscopy and determination of the content of VEGF-A in blood serum (using a flow cytometer BD FACS Canto II) and TGF- β1, IGF-I, IGF-II (by enzyme-linked immunosorbent assay (ELISA) using a c-test system Bender MedSystems, Austria). Results. During the first ophthalmoscopic examination, i.e. before the clinical manifestation of the disease, blood serum of children who later developed ROP that subsequently required laser coagulation of avascular retinal areas, was found to have relatively higher concentrations of VEGF-A (above 1300 pg/ml) and IGF-II (above 140 pg/ml) and lower IGF-I concentrations (below 24 pg/ml) and TGF- β1 (below 8000 pg/ml) as compared with the groups of "safe" children. A dynamic study of VEGF-A content and, simultaneously, TGF- β1 content showed uniform changes in the concentration of these growth factors at all observation times. Conclusion. It was shown for the first time that high IGF-II and low TGF- β1 concentrations revealed before the clinical manifestation of the disease can be viewed as prognostically unfavorable as far as subsequent development of severe ROP is concerned. Uniform changes of TGF- β1 and VEGF-A level in blood serum during ROP development may indicate to the synergistic participation of these growth factors in the pathological vasoproliferation. Further studies are required to determine the details of how this mechanism works // Russian Ophthalmological Journal, 2017; 1: 20-5. doi: 10.21516/2072-0076-2017-10-1-20-25

To study the role of HHV-6 in the development of eye diseases, 193 patients with various eye diseases were examined. Using polymerase chain reaction (PCR) tests DNA of HHV-6 (n = 146) in peripheral blood was detected, using immunoferment analysis markers of chronic (IgG antibodies, n = 142) and active (IgM antibodies, n = 74) HHV-6 infection were detected in blood serum. HHV-6 could be detected in patients with different ophthalmic pathology forms both in the chronic (66.9 %) and the active (43.2 %) form, and DNA of the virus could be found in the blood (20.5 %). A parallel simultaneous study of the blood (n = 40) revealed markers of chronic and/or active infection and DNA of the virus in patients of all nosological groups surveyed (uveitis, corneal ulcers of various etiologies, post-inflammatory cataracts, corneoscleral eye injuries and corneal graft disease). For the first time serological tests (IgG and IgM antibodies) were performed on patients with age-related macular degeneration, where 82.4 % of the patients turned out to be seropositive. The obtained data suggest that HHV-6, along with other viruses generally known to be ophthalmic, may be an etiologic or pathogenetic factor of eye diseases. In this regard, a comprehensive laboratory testing (serological diagnostics and PCR) for HHV-6 should become the object of further targeted study // Russian Ophthalmological Journal, 2016; 1: 73-77.

Purpose: to study the impact of human herpes viruses (HHV) on the development of infectious corneal ulcers (CU). Material and methods. 43 patients aged 21 to 77 with suspected corneal ulcers of bacterial etiology were examined. Based on the history and ophthalmological examination, the patients were divided into 3 groups: group 1 of 26 patients with herpetic ulcers (HU), group 2 of 13 patients with secondary bacterial ulcers (SBU), and group 3 of 4 patients with primary bacterial ulcers (PBU). Blood (43 samples) and corneal scrapings (29 samples) were examined by nested polymerase chain reaction (PCR) for the presence of DNA of various HHV, including herpes simplex virus (HSV) 1, 2, Epstein - Barr virus (EBV), HHV-6, and HHV-7. Results. HHV was detected in 41.6-53 % of the corneas and in 33.3-50 % of blood samples, depending on the patient group. Clinical patients with the DNA of HHV identified in their cornea had torpid CUs and showed no epithelization following the suppressed purulent process. The addition of antiherpetic drugs resulted in complete epithelialization of the cornea within 3-5 days. Conclusion.The frequent representation of HHV-6 and EBV DNA and less frequently detected HSV DNA in the corneal scrapings indicate the possible involvement of HHV in the etiopathogenesis of SBU. The effectiveness of complex antiherpetic drug therapy supports this conclusion. For citations: Neroev V.V., Slepova O.S., Kovaleva L.A., Krichevskaya G.I. Optimizing etiological diagnostics and improving the efficiency of treating centralized infectious corneal ulcers. Russian ophthalmological journal. 2017; 10 (3): 56-61. doi: 10.21516/2072-0076-2017-10-3-56-61 (in Russian).

Studying immunological aspects of pathogenesis of retinopathy of prematurity (ROP) helps develop new approaches to prevention, rational diagnosis and increase of the effectiveness of treatment of the disease. Purpose: to analyze the level of growth factors in blood serum of premature infants belonging to the group of ROP risk at the pre-clinical stage and find out how this level is related to further development and course of the disease. Material and methods. 85 prematurely born infants of the ROP risk group were examined. The average gestational age at birth was 27.7 ± 2.2 weeks, the average birth weight - 1086.1 ± 266.1 g. The examination included dynamic ophthalmoscopy and determination of the content of VEGF-A in blood serum (using a flow cytometer BD FACS Canto II) and TGF- β1, IGF-I, IGF-II (by enzyme-linked immunosorbent assay (ELISA) using a c-test system Bender MedSystems, Austria). Results. During the first ophthalmoscopic examination, i.e. before the clinical manifestation of the disease, blood serum of children who later developed ROP that subsequently required laser coagulation of avascular retinal areas, was found to have relatively higher concentrations of VEGF-A (above 1300 pg/ml) and IGF-II (above 140 pg/ml) and lower IGF-I concentrations (below 24 pg/ml) and TGF- β1 (below 8000 pg/ml) as compared with the groups of safe children. A dynamic study of VEGF-A content and, simultaneously, TGF- β1 content showed uniform changes in the concentration of these growth factors at all observation times. Conclusion. It was shown for the first time that high IGF-II and low TGF- β1 concentrations revealed before the clinical manifestation of the disease can be viewed as prognostically unfavorable as far as subsequent development of severe ROP is concerned. Uniform changes of TGF- β1 and VEGF-A level in blood serum during ROP development may indicate to the synergistic participation of these growth factors in the pathological vasoproliferation. Further studies are required to determine the details of how this mechanism works // Russian Ophthalmological Journal, 2017; 1: 20-5. doi: 10.21516/2072-0076-2017-10-1-20-25

Results of comprehensive ELISA tests of blood serum for the presence of IgM-, IgA-, and IgG-antibodies to herpes simplex virus types 1 and 2, cytomegalovirus, Epstein-Barr virus, human herpesvirus 6, human herpes virus 8 type, Chlamydia trachomatis in 38 patients with uveal melanoma are presented. The polymerase chain reaction was used to detect DNA of these pathogens in tumor biopsies, vitreous body of 10 enucleated eyes, as well as in plasma IgG-antibodies to HHV 6 were revealed in 50% of patients; IgG-antibodies to HHV 8, in 5.3% of patients. Among the 16 patients with uveal melanoma at advanced stages, 6 patients had antibodies indicative of EBV reactivation (1.2-3.3). Chlamydia trachomatis genome was detected in both biopsies; in one of them, in conjunction with EBV and CMV DNA . Tissue samples from the identified infectious agents were related only to the spindle-cell histologic type AB of uveal melanoma. In plasma, genomes of pathogens were not determined. The results indicate the presence of infectious agents in patients with uveal melanoma and require further study of the pathogenetic role of infections in the pathogenesis of uveal melanoma.

Biopsy samples of 32 patients with uveal melanoma (10), RBL (11), orbital lymphoma (7), lacrimal gland adenomas (2), rhabdomyosarcoma of conjunctiva (2) were tested by real time polymerase chain reaction (PCR) to detect the DNA of HSV 1, HSV 2, EBV, CMV, HHV-6, HHV-8, Toxoplasma gondii, Mycoplasma hominis, Ureaplasma spp. Totally, 14 of 32 (43.75 %) tumor samples revealed DNA herpesviruses, Chlamydia trachomatis, mycoplasma hominis or ureaplasma spp. Positive PCR results were revealed in orbital lymphoma almost 2 times more frequently than in RBL (6/7 - 85.7 % and 5/11 - 45.5 %; p = 0.15), and 4 times more frequently than in uveal melanoma (6/7 - 85.7 % and 2/10 - 20%; р = 0.015). The spectrum of pathogens in different tumors varied. Only genomes of lymphotropic HHV (EBV, HHV-6, HHV-8) with predominance of EBV are found in orbital lymphoma. In RBL, we could identify DNA of HHV 6, EBV, CMV, and ureaplasma. Infectious DNA were detected the least frequently (2 out of 10 patients) in biopsies of uveal melanoma: in both cases, Chlamydia trachomatis genome was found, in one of them it combined with EBV and CMV DNA. Plasma investigation revealed positive PCR results significantly less frequently than in biopsies (1/28 and 14/32; p = 0.002). Further studies are needed to assess the role of persistent pathogens, primarily human herpesviruses, Chlamydia and ureaplasma in the pathogenesis of orbit and eye tumors // Russian Ophthalmological Journal, 2016; 2: 42-46 . doi: 10.21516/2072-0076-2016-9-2-42-46 .

In this work, the results of a comprehensive laboratory examination of 37 children with retinoblastoma were described. The presence of Igm-, IgA, - IgG- antibodies to the herpes simplex virus types 1 and 2, cytomegalovirus (СMV), epstein-Barr virus (eBV), human herpes virus (HHV) type 6, Toxoplasma gondii, mycoplasma hominis and ureaplasma urealyticum in the serum was tested using ELISA. In the polymerase chain reaction the DNA of these pathogens were detected in the blood plasma of 18 patients and tumor biopsy specimens from 10 eyes. The results showed that children with RB were predominantly infected by the herpesviruses, among which prevailed CMV. in 4 of 5 enucleated eyes the DNA of herpesvirus [CMV (2 eyes), EBV (1 eye), HHV 6 (1 eye)] and ureaplasma urealyticum (1 eye) were also present in tumor tissue. Nucleic acid of infectious microorganisms were considerably more often detected in the tumor tissue than in plasma (5 of 10, 1 of 18, respectively; p = 0.023), suggesting thereby the presence of the virus in the eye and its adverse role in the pathogenesis of the RB.

To study the role of HHV-6 in the development of eye diseases, 193 patients with various eye diseases were examined. Using polymerase chain reaction (PCR) tests DNA of HHV-6 (n = 146) in peripheral blood was detected, using immunoferment analysis markers of chronic (IgG antibodies, n = 142) and active (IgM antibodies, n = 74) HHV-6 infection were detected in blood serum. HHV-6 could be detected in patients with different ophthalmic pathology forms both in the chronic (66.9 %) and the active (43.2 %) form, and DNA of the virus could be found in the blood (20.5 %). A parallel simultaneous study of the blood (n = 40) revealed markers of chronic and/or active infection and DNA of the virus in patients of all nosological groups surveyed (uveitis, corneal ulcers of various etiologies, post-inflammatory cataracts, corneoscleral eye injuries and corneal graft disease). For the first time serological tests (IgG and IgM antibodies) were performed on patients with age-related macular degeneration, where 82.4 % of the patients turned out to be seropositive. The obtained data suggest that HHV-6, along with other viruses generally known to be ophthalmic, may be an etiologic or pathogenetic factor of eye diseases. In this regard, a comprehensive laboratory testing (serological diagnostics and PCR) for HHV-6 should become the object of further targeted study // Russian Ophthalmological Journal, 2016; 1: 73-77.

Results of comprehensive ELISA tests of blood serum for the presence of IgM-, IgA-, and IgG-antibodies to herpes simplex virus types 1 and 2, cytomegalovirus, Epstein-Barr virus, human herpesvirus 6, human herpes virus 8 type, Chlamydia trachomatis in 38 patients with uveal melanoma are presented. The polymerase chain reaction was used to detect DNA of these pathogens in tumor biopsies, vitreous body of 10 enucleated eyes, as well as in plasma IgG-antibodies to HHV 6 were revealed in 50% of patients; IgG-antibodies to HHV 8, in 5.3% of patients. Among the 16 patients with uveal melanoma at advanced stages, 6 patients had antibodies indicative of EBV reactivation (1.2-3.3). Chlamydia trachomatis genome was detected in both biopsies; in one of them, in conjunction with EBV and CMV DNA . Tissue samples from the identified infectious agents were related only to the spindle-cell histologic type AB of uveal melanoma. In plasma, genomes of pathogens were not determined. The results indicate the presence of infectious agents in patients with uveal melanoma and require further study of the pathogenetic role of infections in the pathogenesis of uveal melanoma.

The article presents results of multiplex cytokine assays in blood serum and lacrimal fluid at the initial and intermediate stages of age-related macular degeneration (AMD). Some features of local and systemic disturbances in the cytokine profile were detected in these patients. It was revealed that the initial stage of AMD was associated with elevated IL-17 levels in lacrimal fluid, along with imbalance between the local increase and systemic decrease of TGF-β1 amounts. Intermediate-stage AMD was associated with increased levels of the most cytokines assayed (except of TGF-β1) in blood serum and lacrimal fluid, thus suggesting stimulation of both pro-inflammatory and angiogenic responses, like as activation of anti-inflammatory and anti-infective factors.

Purpose . Analysis of refractive errors’ frequency in children with retinoblastoma. Patients and Methods . A retrospective study included 57 children with retinoblastoma (31 male and 26 female) who received treatment in 2013–2017. The criteria for patient selection was the presence of mono- or bilateral retinoblastoma, as well as autorefractometry of both eyes before treatment. In all cases, the study was performed with cycloplegia. The parameters studied included gender, age at the time of the initial examination, visual acuity, spherical and cylindrical component of refraction, the axis of the cylinder. Results . Bilateral retinoblastoma was determined in 34 children (59.6 %), the average age of the patients was 18.0 ± 2.6 months. Monolateral retinoblastoma was detected in 23 patients (40.4 %), the average age at the time of check-up was 26.2 ± 3.1 months. In total, anisometropia of more than 2.0 diopters was detected in 14 patients (24.6 %), of which with monolateral RB in 6 patients (26.1 %) and in bilateral patients in 8 (23.5 %). In 26 cases (45.6 %) anisometropia was determined from 1.0 to 2.0 diopters, of which 41.1 % (n = 14) with bilateral lesion and 52.1 % (n = 12) with monolateral. The frequency of hyperopia over 3.0 diopters in the total cohort was 25.2 %, myopia 3.3 %. Significant differences (p = 0.025) were obtained by comparing the frequency of astigmatism determination of more than 1.0 D (39.6 % in the total cohort versus 13.0 % for healthy eyes). When analyzing the frequency of ametropia depending on age, a trend was observed (0.05 < p < 0.1) to reduce the frequency of hypermetropia and increase the frequency of astigmatism. Additionally, an analysis of the astigmatism’s frequency depending on the tumor localization was made. Patients with central or paracentral localization of the tumor lesion amounted to 69.4 % (n = 25), and from the peripheral — 30.6 % (n = 11) (p = 0.002). Conclusions . In this paper, we analyzed the frequency and nature of refractive errors in children with monolateral and bilateral retinoblastoma. The identified risk factors may affect the development of refractive amblyopia in children with retinoblastoma in more adulthood, which requires the development of an individual approach to the correction of refractive disorders taking into account the localization of tumor foci, the nature of the lesion and the individual characteristics of patients.

To study the role of the HHV-6 type in the development of eye diseases PCR tests of blood (152), cornea biopsies (61), and intraocular fluids (11) for HHV-6 and other viruses of the herpes group (HSV type 1 and 2, CMV, EBV) were conducted. It was found that the HHV-6, along with other representatives of the Herpesviridae, can be detected in patients with different clinical forms of ophthalmopathology (174 patients were surveyed). Viral DNA was detected in blood, cornea, and in the anterior chamber fluid. The obtained data allow that the HHV-6 to be suggested as a possible cause of the ophthalmic herpes along with the other viruses of this group. It makes finding the virus DNA an essential step towards setting the etiologic diagnosis of the ophthalmological patients.

AIM:to study the role of infectious agents in the development and progression of AMD by means of serodiagnostic assay for a complex of ophthalmotropic and opportunistic infections.MATERIAL AND METHODS:The study enrolled 61 patients (23 men, 38 women) aged 43-68 years and diagnosed in accordance with AREDS classification of AMD. Serodiagnostic tests were performed for 11 infections with the purpose of identifying antigen-specific antibodies in the serum.RESULTS:Activation markers of one or several infections were detected in most of the patients with AMD.CONCLUSION:Herpes viruses (HSV1 and CMV) along with some opportunistic pathogens are important factors in the pathogenesis of AMD, thereby, specific treatment should be performed in these patients.

PURPOSE: Comparative analysis of the biomechanical parameters of the fibrous tunic of the eyeball and extracellular matrix (ECM) characteristics in patients with suspected POAG, early-stage POAG and healthy elderly people, as well as control groups of various ages. METHODS: The study included 60 people aged 21 to 74 years. The first group consisted of 16 healthy young adults (n=16; mean age 26.3±5 years; “young" control group), the second group included 17 healthy elderly subjects (n=17; 69.2±4 years “age" control group) with no signs of ophthalmological pathology. The third group consisted of 17 people with suspected POAG (n=17; mean age 68.2±6 years; with IOP above 21 mm Hg for three consecutive measurements (average central corneal thickness) and absence of visual field defects on the threshold perimetry test). The fourth group consisted of 10 patients with early-stage POAG (n=10; mean age 67.6±5 years; lesion of one or both eyes). Therapy was discontinued 2 weeks prior to the study. Ophthalmic examination included both the standard (visometry, tonometry, biomicroscopy, gonioscopy, automatic static perimetry) and additional methods (laser scanning tomography). Investigation of the fibrous tunic biomechanical parameters was conducted using the Ocular Response Analyzer (ORA) («Reichert», USA). All subjects underwent lab tests. The content of the regulation of extracellular matrix factors, MMP-9 and TIMP-4, in the serum and tear fluid was determined by enzyme linked immunosorbent assay (ELISA; diagnostic test systems Bender MedSystems® («eBiosciences», Austria; «R & D Diagnostics Inc.», US). We investigated 60 tear fluid samples and 60 serum samples. Statistical processing was made in the SPSS program 17.0 (SPSS, Inc., Chicago, IL). Two independent groups were compared with the use of nonparametric U-test Mann-Whitney test. Correlation analysis was performed using the Spearman test (r). Differences were considered statistically significant at p <0.05. RESULTS: The results showed that elderly age, suspected POAG, and the early-stage of POAG were associated with a set of changes in the cornea-scleral capsule and more or less expressed changes in the ECM regulation system both on local and systemic levels. Thus, the data obtained from the bi-directional applanation tonometry showed a reduction of viscoelastic properties of fibrous tunic associated with age and the development of the early-stage of POAG: the correlation between the age and corneal hysteresis (CH) (significant decrease with age), CH and changes in the early-stage of POAG (similar dynamics). In the study of MMP-9 and TIMP 4 content a significant increase in TIMP-4 level in tear fluid was detected in the 2-nd group compared to the 1-st (p=0.0001). This tendency to a progressive increase of tear fluid TIMP 4 was observed in patients with suspected of POAG and early-stage of POAG. Pair correlations analysis revealed a statistically significant strong inverse relationship between CH and TIMP-4 level in the tear fluid (r=-0,510; p=0,01). It was found that physiological aging was associated with a significant rise of TIMP-4 on local level (p=0.0001), whereas patients suspected of POAG had an increased MMP-9 production on systemic level. CH and CH/corneal resistance factor (CRF) decreased with age, and in initial POAG. We also revealed a significant correlation between the CH and TIMP-4 levels in tear fluid (r=-0,510; p=0,01), which suggests the presence of age associated cornea-scleral capsule restructuring resulting in an increase of its stiffness. Significant increase of the systemic level of MMP-9 may play a key role in the launch of pathological changes in the cornea-scleral capsule of the eye and be one of the major factors leading to the development of POAG.