[Circumscribed choroidal hemangioma and non-pigmented choroidal melanoma: clinical, instrumental and molecular genetic features]. / Otgranichennaya gemangioma i bespigmentnaya melanoma khorioidei: kliniko-instrumental'nye i molekulyarno-geneticheskie osobennosti.

Circumscribed choroidal hemangioma (CCH) and early non-pigmented choroidal melanoma (CM) have similar clinical, ultrasound and morphometric features, which in some cases makes their differential diagnosis difficult. There are few studies in the literature devoted to a comparative analysis of the molecular genetic features of CCH and non-pigmented CM, and the results of those studies are contradictory. PURPOSE: This study attempts to develop a method of non-invasive molecular genetic differential diagnostics of CCH and non-pigmented CM. MATERIAL AND METHODS: Based on the results of clinical and instrumental examination methods, 60 patients (60 eyes) with CCH (n=30) and non-pigmented CM (n=30) were included in this prospective study. The control group consisted of 30 individuals without intraocular tumors. Mutations in the GNAQ/GNA11 genes were determined by real-time PCR using the analysis of genomic circulating tumor DNA isolated from peripheral blood plasma. The average follow-up period was 12.1±1.8 months. RESULTS: The study revealed a significant association of mutations in exons 4 and 5 of the GNAQ/GNA11 genes with the presence of non-pigmented CM (27/30; 90%). These mutations were not detected in the group of patients with CCH. Mutations in exons 4 and 5 of the GNAQ/GNA11 genes were also not detected in the control group of healthy individuals. CONCLUSION: This study proposes a method of non-invasive and low-cost differential diagnostics based on molecular genetic analysis and detection of mutations in exons 4 and 5 of the GNAQ and GNA11 genes, which are specific for CM (90%).

Association of BARD1 and BRIP1 Gene Polymorphisms with the Risk of Uveal Melanoma.

Considering the limited information about the role of hereditary predisposition to the development of uveal melanoma, we have performed an analysis of the frequencies of BARD1 (rs1048108, rs2229571, rs2070094) and BRIP1 (rs4986764) gene polymorphisms in patients with uveal melanoma and benign choroidal nevus in comparison with healthy volunteers (control). It has been found that the minor alleles of BRIP1 rs4986764 and BARD1 rs2070094 polymorphisms, as well as the homozygosity of T allele at the BARD1 rs1048108 locus are common genetic markers for the predisposition to uveal melanoma and benign choroidal nevus, while the homozygous genotype GG for the BARD1 rs2229571 polymorphism is a specific marker for the predisposition to uveal melanoma and progressive choroidal nevus. We have also found that the heterozygous genotype at BARD1 rs1048108 polymorphic locus is a specific marker for protection against uveal melanoma and progressive choroidal nevus. Thus, our results indicate the advisability of studying polymorphisms of the BARD1 gene (rs1048108, rs2229571, and rs2070094) and the BRIP1 gene (rs4986764) in patients with uveal melanoma and progressive choroidal nevus. The obtained findings can be used for forming risk groups, prevention of uveal melanoma, and differential diagnosis of intraocular neoplasms.

[Angioarchitecture of small retinoblastomas: the role of optical coherence tomography angiography]. / Angioarkhitektonika malykh retinoblastom: rol' opticheskoi kogerentnoi tomografii-angiografii.

Optical coherence tomography angiography (OCTA) is a non-invasive diagnostic method used in children and adults. Features of angioarchitecture of small retinoblastoma are not sufficiently covered. PURPOSE: The study investigated the angioarchitecture of small retinoblastomas using OCTA. MATERIAL AND METHODS: The study included 10 children with binocular retinoblastoma aged 2.7±0.5 months with small tumors of central localization (10 foci). The tumors were divided into 3 groups: group 1 (n=4) - tumor thickness 0.8±0.2 mm; group 2 (n=3) - 1.6±0.5 mm; group 3 (n=3) - 2.4±0.8 mm. OCTA was performed on Spectralis HRA+OCT (2460 scans in total). Vessels were identified in the superficial, deep and outer layers of the tumor on En Face images. Their average number was estimated by visualization of yellow pixels in the superficial layers on 10 sagittal sections. Statistical analysis was done using Microsoft Excel, Statistica 8.0. The Kruskal-Wallis H test was used for comparative analysis of independent variables with more than two samples. RESULTS: Retinal vessels with feeding anastomoses connecting them to multiple small tortuous tumor vessels in the superficial layers were identified in group 1. Number of yellow pixels - 16.5±0.5. In the deep layers - single chaotic vascular arcades. In flat small retinoblastomas the vascular component was not evaluated. In group 2 in the superficial layers of the tumor we found multiple geniculate vessels of large and small caliber anastomosing between themselves and the retinal vessels. Number of yellow pixels was 21±0.8. A few vessels were identified in the deep and outer layers. In group 3 we identified single convoluted vessels in the superficial layers with glow and quantity increasing in the deep layers. In the deep layers - emergence of a small number of vessels. The maximum number of multiple own tumor vessels was determined in the outer layers. Number of yellow pixels - 10±0.8. CONCLUSION: The obtained results confirm the possibility to preclinically identify the angioarchitecture of small retinoblastomas in order to determine the activity of tumor growth and serve as a marker of neoplasm regression in the future, after organ-preserving treatment.

[The incidence of infection in tumor and eye fluid system, and specific humoral immunity to herpes viruses in patients with uveal melanoma].

INTRODUCTION: Studies aimed at a direct research of human herpes viruses (HHVs) in the tumor material and eye media have not been carried out so far. Research goal to establish the frequency of detection HHVs DNA in the biomaterial of the eye and blood and to assess the specific humoral immunity to the causative agents of herpes virus infections in patients with uveal melanoma. MATERIALS AND METHODS: 38 patients with the uveal tract tumor were examined for the presence of DNA of HHV types 1 and 2 (HSV-1, 2), Cytomegalovirus (CMV), Varicella Zoster virus (VZV), EpsteinBarr virus (EBV) and herpes viruses 6 and 8 types (HHV-6, HHV-8) in tumor tissue, vitreous body, aqueous humour and blood plasma by real-time polymerase chain reaction; blood serum was studied by enzyme-linked immunosorbent assay (ELISA) for IgG and IgM antibodies to HHVs. RESULTS: EBV DNA was present in tumor tissue in 20.6% of cases, in vitreous body in 4.2%, in blood plasma in 2.7%, and was not found in aqueous humor. Ig G antibodies to HSV-1, 2 and CMV were detected in 97.3% of cases, VZV 94.6%, HHV-6 32.4%, antibodies to HHV-8 were not detected. 20 patients (55.6%) had reactivation of chronic HSV-1, 2 infection, and 14 (38.9%) patients had reactivation of CMV infection. Markers of chronic EBV infection were found in all patients, its atypical reactivation was observed in 2 cases (5.4%). CONCLUSION: Our findings suggest the possible participation of EBV in the oncogenesis of the uveal tract and emphasize the need for further in-depth study of this problem.

[Use of optical coherence tomography angiography in assessment in conjunctival vascular architecture in health and pathology]. / Vozmozhnosti opticheskoi kogerentnoi tomografii v angiorezhime v otsenke angioarkhitektoniki kon"yunktivy v norme i pri patologii.

PURPOSE: Investigation of the capabilities of anterior segment (AS) optical coherence tomography angiography (OCTA) in evaluation of conjunctival vascular architecture in healthy individuals and in various pathological conditions. MATERIAL AND METHODS: The study included 17 healthy volunteers (34 eyes; the control group) and 62 patients (68 eyes) with conjunctival lesions of various nature. All participants underwent AS-OCT and AS-OCTA with assessment of qualitative (vessels pattern, lumen, pathologic tortuosity) and quantitative parameters (vessel density (VD, %) in the lesion area). Mean VD (MVD) and local VD (LVD) were determined, as well as VD in perifocal tissues (PVD). RESULTS: OCTA scans in 8 conjunctival sectors showed mostly radial pattern of the vascular architecture, with vessel lumen remaining the same over their entire visible length. Larger-sized vessels in deeper conjunctival layers were discovered in most cases. The lowest VD value (33.3%) was registered in the superotemporal quadrant, and the highest (38.9%) - in the nasal. Tortuosity of the vessels with course disruption, uneven lumen over the length of the vessels and increase in VD were observed in the area of conjunctival lesions in all cases excluding congenital abnormalities, pingueculae and conjunctival melanocytic intraepithelial neoplasia. The malignant nature of the tumors was indicated by dense vessel distribution and difficulties for visualization of intravascular space, and confirmed by pathohistological analysis. An increase in the number of areas with a lace-like pattern was characteristic for melanomas, with mean VD of more than 50% in the most vascularized areas. CONCLUSION: AS-OCTA is an informative method for the visualization of vessels in healthy conjunctiva and in conjunctival pathology. Local VD in the lesion area should be measured when the vessels are unevenly distributed.

[The role of CXC and CC chemokines in patients with uveal melanoma]. / Rol' khemokinov klassov CXC i CC u bol'nykh s uveal'noi melanomoi.

Uveal melanoma is a malignant neoplasm with high metastatic potential; its pathogenesis is currently being studied. Chemokines play a key role not only in the inflammatory response, but also in enhancing angiogenesis, tumor invasiveness, increasing proliferative potential and metastasis. PURPOSE: To study the role of chemokines of classes CXC and CC in blood serum and tear fluid of patients with uveal melanoma. MATERIAL AND METHODS: The study included 118 people aged 53.7±12.2 years, among them 80 patients with uveal melanoma and 38 healthy donors. Group 1 included 32 patients with small tumors, group 2 (medium-sized tumors) - 26 patients; group 3 (large tumors) was comprised of 22 patients. Chemokines of classes CC (CCL2/MCP-1, CCL3/MIP-1α, CCL4/MIP-1ß, CCL5/RANTES, CCL11/Eotaxin) and CXC (CXCL1/GRO-α, CXCL8/IL-8, CXCL10/IP-10, CXCL12/SDF-1α) were determined by multiplex analysis of the blood serum and tear fluid. Statistical processing: Student's t-test, Fisher criteria, and Pierson's chi-squared test (χ2), differences were considered significant at p<0.05. RESULTS: Significantly increased level of chemokines with pro-inflammatory (CCL5/RANTES), proliferative (CXCL10/IP-10) and pro-angiogenic (CXCL12/SDF-1α) effects was found in the blood serum of patients with small-sized uveal melanoma in comparison with healthy donors. Concentration of all studied pro-inflammatory, proliferative, and pro-angiogenic chemokines in the lacrimal fluid was found to be significantly elevated in both the affected and the paired "healthy" eyes in all 3 groups of patients, with the maximum content seen in the large tumor group. CONCLUSION: The obtained data indicates that early local and systemic immune imbalance can be observed in uveal melanoma, and detection of chemokines can serve as a good reason for developing targeted therapy for small uveal melanoma.

Assessment of the Chemosensitivity of Uveal Melanoma Cells Ex Vivo.

The study was designed to create a primary cell culture of uveal melanoma and to evaluate its resistance to chemotherapy. Of the obtained 20 samples of uveal melanoma, the primary cultures with proliferation sufficient for MTT test were derived in only 7 cases. However, even these cultures were unable to survive more than 4 passages; the cells accumulated melanin and underwent apoptosis. Retinol palmitate and nepafenac produced no cytotoxic effect on uveal melanoma cells. Of 5 cultures treated with sodium valproate (Convulex), no pronounced cytotoxic effect was observed in one culture (UM4); in 2 cultures, 50% cells died in the presence of the lowest drug concentration of 1.88 mg/ml; and in 2 cultures, the same effect was achieved at drug concentrations 7-10 mg/ml. The cytotoxic effect of treosulfan was evaluated in only 4 cultures of uveal melanoma: the drug exhibited pronounced antitumor activity on all cultures, in 2 cases, it was effective at a concentration of 0.16 mg/ml. Gemcitabine in a concentration of 2.5 mg/ml produced a pronounced cytotoxic effect in 4 out of 7 cultures (death of 70-80% cells) and induced death of ~45% cells in the remaining 3 cultures. Mitoxantrone had ambiguous effect: in 2 of 5 cultures, the drug in high concentrations stimulated the growth of tumor cells, but in 3 cultures, the drug even in minimum concentrations induced death of 70-80% cells.

Development of In Vivo Model of Intraocular Retinoblastoma.

Here we describe creation of a model of intraocular retinoblastoma on 4 mature immunodeficient BALB/c nude mice of both sexes. A suspension of the primary retinoblastoma culture was injected into each of the 8 eyes of the test animals. The injections were performed under the control of an operating microscope using insulin syringes with a diameter of 29G transsclerally into the subretinal space in a volume of 0.3 ml at a rate of 10,000 cells in 25 µl physiological saline. The culture of Rb10 cells derived from a patient with retinoblastoma passed 12 passages at the time of the experiments. After 9-week follow-up, a clinical (ophthalmoscopy) and instrumental (ultrasound) study was performed with visualization of the tumor mass on the fundus. After enucleation, a morphological study was performed. The retinoblastoma tumor nodes were identified in two eyes. The proposed model can be used for further research and for testing new chemotherapeutic drugs and treatment regimens for retinoblastoma.

[Optical coherence tomography in diagnostics of small choroidal melanoma]. / Opticheskaia kogerentnaia tomografiia v diagnostike nachal'noi melanomy khorioidei.

PURPOSE: To determine signs of small choroidal melanoma with different pigmentation using enhanced depth imaging optical coherence tomography (EDI-OCT). MATERIAL AND METHODS: The study included 344 patients with small choroidal melanoma with different pigmentation examined using EDI-OCT: 1st group - pigmented melanoma (228 eyes), 2nd group - low pigmented (65 eyes), and 3rd group - amelanotic (51 eyes). RESULTS: In pigmented small choroidal melanomas - elevation of choroidal profile towards vitreous, compression of choriocapillaries with a narrow even 'belt' and a 'shadow' effect; thinning, defects in Bruch's membrane; thickening of the retina above the tumor, lobulated photoreceptors; intra- and subretinal exudate (diffuse, cystic edema, neuroepithelial detachment); defects and detachment of pigment epithelium with hyperreflective foci, disorganization of the pigment with the formation of hyperreflective foci at different retinal levels. In low-pigmented small choroidal melanomas - elevation of choroidal profile towards vitreous, visualized inner surface of the sclera, 'excavation' of the choroid, enlarged choriocapillaries, contour of tumor; thickening of the retina, accumulation of intra- and subretinal exudate (local neuroepithelial detachments); disorganization of the pigment in pigment epithelium with hyperreflective foci in the outer retinal layers. In amelanotic small choroidal melanomas - elevation of choroidal profile towards vitreous, visualized inner surface of the sclera, 'excavation' of the choroid; contouring of choriocapillaries, longitudinal hyperreflective bands in the tumoral stroma, smoothness of the Bruch`s membrane, structural losses of photoreceptors; thickening of the retina (neuroepithelial detachment, diffuse edema); uneven thickening of pigment epithelium. CONCLUSION: EDI-OCT can help identify microstructural changes in the choroid and adjacent retina in small choroidal melanomas with different degrees of pigmentation, suggesting at the early stages a more aggressive course of the tumoral process affecting the prognosis of the disease. In addition, identification of the microstructure and degree of pigmentation of initial choroidal melanomas is necessary for planning an organ-preserving treatment.

[Mast cells in the microenvironment of uveal melanoma]. / Tuchnye kletki v mikrookruzhenii uveal'noi melanomy.

OBJECTIVE: To study the morphological features of the microenvironment of a tumor nodule in the eyes with uveal melanoma, focusing on mast cells. MATERIAL AND METHODS: A total of 43 enucleated eyes with uveal melanoma (260 histological specimens) were examined. The patients' age averaged 54±2.7 years. The morphopathological types of tumors were as follows: epithelioid-cell (n=9; 20.9%), spindle-cell type AB (n=15; 34.9%), and mixed-cell (n=19; 44.2%). The tumor prominence was 4.7±1.3 mm; the base diameter was 13.5±3.3 mm. Statistical methods, such as Microsoft Excel, Statistica 10.1, Spearman's rank correlation coefficient (rs), were applied. RESULTS: Mast cells in the microenvironment of uveal melanoma were present in 18 (41.9%) of the 43 eyes. There was a significantly higher correlation between the mixed-cell type of tumor and the accumulation of mast cells (rs=0.636). The correlation coefficient (rs) of the number of mast cells with the degree of tumor pigmentation in terms of densely and weakly pigmented forms was 0.571 and 0.717, respectively. Tumor invasion through the sclera was detected in 7 (16.3%) eyes with mast cells (rs=0.395). Tumor growth in the emissarium in the presence of mast cells was determined in 8 (18.6%) cases (rs=0.469). Comparison established a correlation between the number of mast cells and the sections of tumor blood vessels (rs=0.21). Granulated cells were noted in 15 (34.9%) cases; degranulated ones were seen in 3 (7%) cases of the 43 examined eyes. with an ejection of granules around the tumor cells, which may be evidence of their interaction. Granules were ascertained to be released around the tumor cells, which may be suggestive of their interaction. CONCLUSION: The study of the mast cell population as one of the components of the tumor microenvironment can be used to elaborate novel approaches for the targeted treatment of uveal melanoma, in particular for its impact on tumor angiogenesis, by using mast cell inhibitors. Therefore, it is relevant and promising to conduct further investigation of mast cells in uveal melanoma.

[Unilateral multifocal uveal melanoma (a clinical and genetic study)]. / Monolateral'naia mul'tifokal'naia uveal'naia melanoma (kliniko-geneticheskoe issledovanie).

The study presents clinical and genetic analysis of a case of unilateral multifocal uveal choroidal melanoma in a patient of 67 years. Results of ophthalmoscopy, echography, fluorescent angiography, optical coherence tomography are described. Molecular genetic testing of peripheral blood samples was performed, including detection of the occurrences of CC genotype in C3435T polymorphism of the gene ABCB1/MDR1 associated with unfavorable vital prognosis. Analysis of the genes GNAQ and GNA11 revealed two mutually exclusive mutations in the genes GNAQG183A and GNAQA209C showing genetic heterogeneity of the two tumor lesions. Organ preservation treatment of unilateral multifocal uveal melanoma was proven possible with brachytherapy method. Uveal melanoma with multicentric growth is of interest to ophthalmologists because it requires differential diagnostics from a variety of diseases including metastases in the choroid, such as metastases of uveal melanoma and skin melanoma, as well as other intraocular neoplasms.

[Melanoma-associated vitelliform retinopathy (a clinical case study)]. / Melanoma-assotsiirovannaia vitelliformnaia retinopatiia (klinicheskoe nabliudenie).

Melanoma-associated vitelliform retinopathy is a manifestation of paraneoplastic syndrome in skin melanoma. Paraneoplastic syndrome, while not being a tumor or a metastatic disease, is regarded as a tumor-associated disease related to extraocular localization of neoplasm. In this clinical case, the diagnosis of melanoma-associated vitelliform retinopathy was based on a combination of clinical, angiographic, autofluorescence and morphometric signs of bilateral lesion. Analysis of the case showed that in common oncological diseases and complaints of visual impairment, examination of eye fundus is mandatory in order to timely diagnose the changes associated with tumor lesion. Detection of bilateral lesions with oval grey-yellow multiple foci at the level of retinal pigment epithelium may indicate melanoma-associated vitelliform retinopathy that requires diagnostic search for skin melanoma. A complex of instrumental studies including fluorescent angiography, optical coherence tomography and autofluorescence with feature identification allowed establishing the correct diagnosis in the particular case.

[Role of optical coherence tomography angiography in diagnostics of early choroidal melanoma and circumscribed choroidal hemangioma]. / Opticheskaia kogerentnaia tomografiia-angiografiia v diagnostike nachal'noi melanomy i otgranichennoi gemangiomy khorioidei.

Small choroidal melanoma is a malignant tumor that is prone to early metastasis, its amelanotic form is often similar to circumscribed choroidal hemangioma. The main attribute for tumor identification is its vascularization, which is the target of various examination methods. Optical coherence tomography angiography (OCTA) has not been previously used in complex diagnostics of early choroidal melanoma and circumscribed choroidal hemangioma for detection of tumor vessels and the nature of their branching, as well as for vessel caliber comparison. Purpose to examine vascularization of early uveal melanoma and circumscribed choroidal hemangioma by optical coherence tomography angiography. MATERIAL AND METHODS: The study included 23 patients with early choroidal melanoma (13 subjects) and circumscribed choroidal hemangioma (10 subjects) that were examined by optical coherence tomography angiography. According to ultrasound investigation, mean tumor prominence was 1.1±0.3 mm, mean base diameter - 8.1±0.6 mm. RESULTS: Optical coherence tomography angiography in 13 patients with small choroidal melanoma revealed presence of a neovascular component localized under retinal pigment epithelium (RPE) that had marginal avascular zone corresponding to the tumor slope. The loop-like shape of tumor vessels with numerous twists and interweaving was noted under retinal vessels. A tree-shaped neovascular component with large-caliber vessels in the form of a tree trunk with multiple branches extending from it was seen under RPE in 4 cases with circumscribed choroidal hemangioma; diffuse vascularization in the form of numerous tiny tortuous vascular branches was seen in 6 patients. CONCLUSION: Optical coherence tomography angiography allows detection of tumor`s own vessels with characteristics of their vascularization in early choroidal melanoma and circumscribed choroidal hemangioma. Increasing the frequency of detection of tumor`s own vessels will make possible early differential diagnostics of a malignant or benign tumor and will help establish adequate conserving therapy.

Retinoblastoma Cell Culturing and Evaluation of Their Drug Resistance.

We obtained primary culture of retinoblastoma cells and evaluated the resistance of cultured ells to chemotherapy. The study included 19 patients aged 6-64 months (mean 27.9±17.4 months); of these, 6 (31.6%) patients with bilateral retinoblastoma and 13 (68.4%) patients with unilateral form. In 18 (94.7%) patients, group E retinoblastoma was diagnosed. Enucleation was performed in all patients; in 94.7% cases, low-differentiated retinoblastoma was identified. Samples of the tumor tissue were taken to derive a cell culture and to study drug resistance and metabolic activity of cells (MTT test). In 4 cases, adhesion primary cultures of retinoblastoma were derived. Cytological verification of the obtained cultures was performed. The primary cultures were derived from 4 of 6 bilateral tumors and from none of 13 unilateral tumors (p=0.003). There were no statistically significant correlations with patient age (p=0.33) and the presence of calcifications in the tumor (p=0.26). MTT test revealed no differences in the sensitivity of cell cultures to irinotecan and ifosfamide. Pronounced differences in the resistance of cell cultures were observed for oxaliplatin and ascorbic acid. MTT test with evaluation of drug resistance can be used both in clinical practice for adjusting chemotherapy regimen and in development of new approaches to the treatment of retinoblastoma with assessment of in vivo tumor cell resistance in animal models.

Association between polymorphic markers in candidate genes and the risk of manifestationof endocrine ophthalmopathy in patients with Graves' disease.

AIM: To analyze the association between the polymorphic markers in CTLA4, TNF, IL10 and IL16 genes and the risk of manifestation of endocrine ophthalmopathy (EO) in patients with Graves' disease (GD). MATERIALS AND METHODS: Case-control study included 248 patients with GD. Using polymerase chain reaction we studied the distribution of alleles and genotypes of polymorphic markers such as A60G (rs3087243) in CTLA4 gene, G(-308)A (rs1800629) in TNF gene, G(-1082)A (rs1800896) in IL10 gene, T3249C (rs4778641) in IL16 gene among 141 patients with Graves' disease and EO and 107 patients with GD without EO. RESULTS: The frequencies of A alleles and the AA genotypes were significantly increased and the frequencies of G alleles and the GG genotype polymorphic markers rs3087243 of CTLA4 gene and rs1800896 of IL10 gene, as well as the GG genotype polymorphic marker rs1800629 of TNF gene were reduced in patients with GD and EO. The polymorphism in CTLA4 gene was also associated with the activity and the severity of EO. The comparative analysis of the allele and genotype frequency distribution of polymorphic markers of IL16 gene did not show the significant difference. CONCLUSION: The risk of manifestation and the development of EO in patients with Graves' disease can be caused by not only environmental, but also genetic risk factors.

[Specific immunoglobulins G and M in blood serum in retinoblastoma and 'pseudoretinoblastoma']. / Spetsificheskie immunoglobuliny G i M v syvorotke krovi pri retinoblastome i 'psevdoretinoblastomakh'.

Perinatal inflammatory retinal diseases and intrauterine retinal maldevelopments are mistaken for retinoblastoma as often as in 8-16% of cases. AIM: To analyze the infectious status in children with retinoblastoma and pseudoretinoblastoma at different ages. MATERIAL AND METHODS: A total of 47 retinoblastoma suspects aged 4-69 months were enrolled. Pseudoretinoblastoma (inflammatory retinal diseases and intrauterine maldevelopments of the retina) was detected in 14 children (group 1), retinoblastoma - in 33 children (group 2). In each group, two subgroups were identified: 'a' - children under 12 months of age (1a - 5 patients, 2a - 10 patients) and 'b'- children over 12 months of age (1b - 9 patients, 2b - 23 patients). Their blood sera were examined for antibodies to herpes simplex virus types 1 and 2, cytomegalovirus, Epstein-Barr virus, toxoplasma, toxocara, chlamydia, and mycoplasma (enzyme-linked immunosorbent assay). RESULTS: According to serological screening, all patients from group 1a (children under 12 months of age with pseudoretinoblastoma), in contrast to other groups, were infected perinatally with cytomegalovirus infection. All 47 patients were seronegative to toxoplasma. Toxocara infection was identified in children over 12 months of age: in 3 out of 9 patients with pseudoretinoblastoma and in 2 out of 23 patients with retinoblastoma (p>0.05). Markers of Epstein-Barr viral activity were detected only in 3 retinoblastoma children over 12 months of age. CONCLUSION: The results suggest that cytomegalovirus infection plays the leading role in the development of perinatal eye pathology, which, in infants, is clinically similar to retinoblastoma. In children over 12 months of age we found no serological signs that could be regarded as specific of either retinoblastoma, or pseudoretinoblastoma. The only thing worth paying attention to is the activation of Epstein-Barr virus infection in children over 12 months of age with retinoblastoma.

[Iridal lymphoma (clinical case)]. / Limfoma raduzhki (klinicheskoe nablyudenie).

This is a case report of rare iridal involvement in intraocular lymphoma confirmed by the full range of diagnostic measures, including ultrasound biomicroscopy, optical coherence tomography of the anterior segment of the eye, iridectomy with biopsy and further cytological and histopathological examination of the obtained material.

[Pathological and molecular genetic characteristics in patients with extrabulbar growth of uveal melanoma]. / Patomorfologicheskie i molekulyarno-geneticheskie osobennosti u bol'nykh pri ekstrabul'barnom roste uveal'noi melanomy.

AIM: to analyze the association of extrabulbar tumor growth with pathological and molecular genetic changes in patients with uveal melanoma (UM). SUBJECTS AND METHODS: A total of 134 UM patients aged 22 to 84 years were examined and treated. The mean height of the tumor was 9.2±2.9 mm; the diameter of its base was 15.3±3.5 mm. Enucleation of the affected eye was performed in 97.8% of cases. Spindle-cell (n=61 (45.6%)), mixed cell (n=46 (34.3%)), and epithelioid cell (n=27 (20.1%)) tumors were identified according to their histological structure. Polymerase chain reaction-restriction fragment length polymorphism analysis was used to determine full and partial monosomy of chromosome 3, deletion of the short arm of chromosome 1, and RASSF1A gene methylation (n=134). The patients were divided into two groups: 1) those with extrabulbar growth (EG) (n=12) and 2) those without EG (n=122). RESULTS: There was a topographic association between the tumor invasion zone and the largest area of exit of the scleral vessels, along which the tumor invaded: the anterior and posterior segments of the eyeball. The specific features of the invasion pattern of UM were shown: there was its broader invasion in the posterior segment and thinner growing tissue interlayers in the anterior segment. Two UM types stopping the process of UM invasion through the scleral fibrous tunic of the eye were established: 1) that with nodule formation and 2) that with tumor cell dissemination within the episclera. The cellular composition of growing tumor tissue in the episclera was ascertained to differ from the main UM focus in the choroid towards its more atypization. The rate was shown to be significantly lower (20% versus 47.9% for the relatively favorable spindle cell type of UM) in the EG group. The frequency of full or partial chromosome 3 monosomy was significantly higher in the extrabulbar tumor growth group (80% versus 50.4%). CONCLUSION: The morphological features of the EG of UM were defined. The use of a statistically significant sample of patients with UM confirmed the favorable course of the tumor in its spindle cell type and the negative role of chromosome 3 monosomy, as well as the relationship to extrabulbar tumor growth.

[. Role of molecular and genetic factors in survival from uveal melanoma]. / Vyzhivaemost' pri uveal'noi melanome: rol' molekulyarno-geneticheskikh faktorov.

AIM: to analyze survival rates in uveal melanoma (UM) patients and establish correlations with chromosome 3 monosomy, chromosome 1p deletion, and RASSF1A methylation. MATERIAL AND METHODS: Methylation-specific PCR analysis was performed in 104 patients with histologically verified UM. RESULTS: A statistically significant correlation has been found between chromosome 3 monosomy, on the one hand, and mixed/epithelioid cell melanomas and ciliary body involvement, on the other. As for chromosome 1p deletion, it has demonstrated association with extrabulbar tumor growth. We have also calculated 5-year survival and mortality rates in «large¼ UMs and their relationship with chromosome 3 monosomy, chromosome 1p deletion, and RASSF1A methylation. CONCLUSION: Chromosome 3 monosomy is associated with lower survival rates, while RASSF1A methylation - with a better prognosis. A combination of molecular and genetic changes (particularly, chromosome 3 monosomy and chromosome 1p deletion) also leads to reduced survival in UM patients.

[Viral infections and retinoblastoma in children].

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.