ABSTRACT
Exosomes, nano-sized secreted extracellular vesicles (EVs), are actively studied for their diagnostic and therapeutic potential. In particular, exosomes secreted by dendritic cells (DCs) have been shown to carry MHC-peptide complexes allowing efficient activation of T lymphocytes, thus displaying potential as promoters of adaptive immune responses. DCs also secrete other types of EVs of different size, subcellular origin and protein composition, whose immune capacities have not been yet compared to those of exosomes. Here, we show that large EVs (lEVs) released by human DCs are as efficient as small EVs (sEVs), including exosomes, to induce CD4+ T-cell activation in vitro When released by immature DCs, however, lEVs and sEVs differ in their capacity to orient T helper (Th) cell responses, the former favouring secretion of Th2 cytokines, whereas the latter promote Th1 cytokine secretion (IFN-γ). Upon DC maturation, however, these functional differences are abolished, and all EVs become able to induce IFN-γ. Our results highlight the need to comprehensively compare the functionalities of EV subtypes in all patho/physiological systems where exosomes are claimed to perform critical roles.
Subject(s)
CD4-Positive T-Lymphocytes/drug effects , CD4-Positive T-Lymphocytes/immunology , Dendritic Cells/immunology , Dendritic Cells/metabolism , Extracellular Vesicles/metabolism , Lymphocyte Activation , HumansABSTRACT
We argue that the field of extracellular vesicle (EV) biology needs more transparent reporting to facilitate interpretation and replication of experiments. To achieve this, we describe EV-TRACK, a crowdsourcing knowledgebase (http://evtrack.org) that centralizes EV biology and methodology with the goal of stimulating authors, reviewers, editors and funders to put experimental guidelines into practice.
Subject(s)
Biomedical Research , Databases, Bibliographic , Extracellular Vesicles/physiology , InternationalityABSTRACT
The goal of this study was to explore quality of life in patients with advanced non-small-cell lung cancer (NSCLC) in an attempt to single out features that could help predict the possibility of non-completion of chemotherapy. The survey tool was the Quality of Life Questionnaire Core-30 (QLQ-C30) with the module Lung Cancer 13 (LC-13) developed by the European Organization for Research and Treatment of Cancer. The assessment of quality of life (QoL) was performed in 58 patients with advanced NSCLC before palliative chemotherapy and it was repeated in 43 patients who completed at least three cycles of chemotherapy. We found that the patients who failed to complete the chemotherapy course distinctly showed, in contradistinction to those who completed it, poor physical functioning in (67.6 ± 16.3 vs. 78.3 ± 21.3 points, respectively, p < 0.05) and the lack of appetite (27.1 ± 38.0 vs. 48.9 ± 37.5 points, respectively p < 0.05). At the end of palliative chemotherapy alopecia, sore throat, and constipation significantly worsened QoL, but global health status remained unchanged. In conclusion, poor physical functioning and loss of appetite seem to harbinger a risk of non-completion of chemotherapy in advanced NSCLC.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols , Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Quality of Life , Carcinoma, Non-Small-Cell Lung/drug therapy , Humans , Lung Neoplasms/drug therapy , Palliative Care/psychology , Palliative Care/standards , Surveys and QuestionnairesABSTRACT
Extracellular vesicles (EVs) have become the focus of rising interest because of their numerous functions in physiology and pathology. Cells release heterogeneous vesicles of different sizes and intracellular origins, including small EVs formed inside endosomal compartments (i.e., exosomes) and EVs of various sizes budding from the plasma membrane. Specific markers for the analysis and isolation of different EV populations are missing, imposing important limitations to understanding EV functions. Here, EVs from human dendritic cells were first separated by their sedimentation speed, and then either by their behavior upon upward floatation into iodixanol gradients or by immuno-isolation. Extensive quantitative proteomic analysis allowing comparison of the isolated populations showed that several classically used exosome markers, like major histocompatibility complex, flotillin, and heat-shock 70-kDa proteins, are similarly present in all EVs. We identified proteins specifically enriched in small EVs, and define a set of five protein categories displaying different relative abundance in distinct EV populations. We demonstrate the presence of exosomal and nonexosomal subpopulations within small EVs, and propose their differential separation by immuno-isolation using either CD63, CD81, or CD9. Our work thus provides guidelines to define subtypes of EVs for future functional studies.
Subject(s)
Antigens, CD/metabolism , Cell-Derived Microparticles/metabolism , HSP70 Heat-Shock Proteins/metabolism , Membrane Proteins/metabolism , Proteomics , Biomarkers/metabolism , HumansABSTRACT
Purpose: To assess the late complications after the I-125 brachytherapy of uveal melanoma. Material and methods: 129 patients (71 women 55%, and 58 men 45%) treated at the Department of Ophthalmology and Ocular Oncology of the University Hospital in Krakow between January 2005 and December 2009 were evaluated. I-125 isotope applicators were used for tumor irradiation. The mean follow-up was 42.5 months. Results: The following complications were observed: radiation cataract, secondary glaucoma, radiation retinopathy, radiation maculopathy, necrosis of sclera, retinal and vitreous hemorrhages and retinal detachment. Post-radiation cataract was the most common late complication (90.594.7%). Conclusions: Despite the undoubted effectiveness in the treatment of uveal melanomas, I-125 brachytherapy poses a high risk of complications. Tumor location in the proximity of critical visual structures negatively affected ultimate visual function. The tumor size and the dose of irradiation significantly affected the incidence of radiation-induced complications.
Subject(s)
Brachytherapy/adverse effects , Eye Diseases/etiology , Iodine Radioisotopes/adverse effects , Melanoma/radiotherapy , Radiation Injuries/etiology , Uveal Neoplasms/radiotherapy , Cataract/etiology , Female , Glaucoma/etiology , Humans , Iris/blood supply , Male , Middle Aged , Neovascularization, Pathologic/etiology , Poland , Retinal Diseases/etiology , Risk FactorsABSTRACT
Uveal melanoma can be typically diagnosed based on clinical presentation and the A and B mode ultrasound. In some atypical intraocular tumours or for prognostic purposes intraoperative biopsy may be performed. Uveal melanoma biopsy is not safe in 100% and can cause complications (vitreous hemorrhage, retinal detachment and endophthalmitis). Like all biopsies, a biopsy in uveal melanoma biopsy may show limited cellularity and can yield insufficient tissue specimen for histology, cytology and genetic testing. This is most likely in small tumours, below 3 mm in thickness. Another limitation of biopsy-based prognosis is the issue of intratumoural heterogeneity. As a biopsy allows for only a small sample to be removed from the tumour, it is possible to receive false negative results. The most devastating complication of uveal melanoma biopsy is the extraocular spread of the tumour. The study is a review of the current opinions and findings on the role of biopsy in uveal melanoma.
Subject(s)
Biopsy/adverse effects , Melanoma/diagnosis , Patient Safety , Uveal Neoplasms/diagnosis , HumansABSTRACT
Exosomes are extracellular vesicles (EVs) secreted upon fusion of endosomal multivesicular bodies (MVBs) with the plasma membrane. The mechanisms involved in their biogenesis have not yet been fully identified although they could be used to modulate exosome formation and therefore are a promising tool in understanding exosome functions. We have performed an RNA interference screen targeting 23 components of the endosomal sorting complex required for transport (ESCRT) machinery and associated proteins in MHC class II (MHC II)-expressing HeLa-CIITA cells. Silencing of HRS, STAM1 or TSG101 reduced the secretion of EV-associated CD63 and MHC II but each gene altered differently the size and/or protein composition of secreted EVs, as quantified by immuno-electron microscopy. By contrast, depletion of VPS4B augmented this secretion while not altering the features of EVs. For several other ESCRT subunits, it was not possible to draw any conclusions about their involvement in exosome biogenesis from the screen. Interestingly, silencing of ALIX increased MHC II exosomal secretion, as a result of an overall increase in intracellular MHC II protein and mRNA levels. In human dendritic cells (DCs), ALIX depletion also increased MHC II in the cells, but not in the released CD63-positive EVs. Such differences could be attributed to a greater heterogeneity in size, and higher MHC II and lower CD63 levels in vesicles recovered from DCs as compared with HeLa-CIITA. The results reveal a role for selected ESCRT components and accessory proteins in exosome secretion and composition by HeLa-CIITA. They also highlight biogenetic differences in vesicles secreted by a tumour cell line and primary DCs.
Subject(s)
Endosomal Sorting Complexes Required for Transport/metabolism , Exosomes/metabolism , Calcium-Binding Proteins/genetics , Calcium-Binding Proteins/metabolism , Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolism , Dendritic Cells/metabolism , Endosomal Sorting Complexes Required for Transport/genetics , Gene Knockdown Techniques , HeLa Cells , Histocompatibility Antigens Class II/metabolism , Humans , Microscopy, Immunoelectron , Multivesicular Bodies/metabolism , RNA, Small Interfering/genetics , Tetraspanin 30/metabolismABSTRACT
Uveal melanoma is the most common primary intraocular neoplasm in adults. Its bilateral localization is extremely rare. The aim of the paper is analysis of the cases of bilateral uveal melanoma. Five bilateral uveal melanoma patients were diagnosed in the Department of Ophtalmology and Ocular Oncology beetwen 1980 and 2014. Both eyes of four patients were threated with brachytherapy. Final enulcleation of the one eye was performed in three patients. It was the primary treatment in one patient. The presence of uveal melanoma was confirmed by pathological examination in all cases after surgical removal of eyeball and in one after local resection of iris tumor. Metastatic lesions were diagnosed in lungs and liver in two patients. Three patients are still followed-up at our institution. The possibility of bilateral uveal melanoma should considered although it is extremely rare. bilateral uveal melanoma, brachytherapy, enucleation.
Subject(s)
Choroid Neoplasms/pathology , Choroid Neoplasms/therapy , Melanoma/pathology , Melanoma/therapy , Neoplasms, Multiple Primary/pathology , Neoplasms, Multiple Primary/therapy , Adult , Aged , Brachytherapy/methods , Eye Enucleation/methods , Female , Follow-Up Studies , Humans , Male , Middle Aged , Poland , Retina/pathology , Retrospective Studies , Treatment OutcomeABSTRACT
Tissues are spatially orchestrated ecosystems composed of heterogeneous cell populations and non-cellular elements. Tissue components' interactions shape the biological processes that govern homeostasis and disease, thus comprehensive insights into tissues' composition are crucial for understanding their biology. Recently, advancements in the spatial biology field enabled the in-depth analyses of tissue architecture at single-cell resolution, while preserving the structural context. The increasing number of biomarkers analyzed, together with whole tissue imaging, generate datasets approaching several hundreds of gigabytes in size, which are rich sources of valuable knowledge but require investments in infrastructure and resources for extracting quantitative information. The analysis of multiplex whole-tissue images requires extensive training and experience in data analysis. Here, we showcase how a set of open-source tools can allow semi-automated image data extraction to study the spatial composition of tissues with a focus on tumor microenvironment (TME). With the use of Lunaphore COMET platform, we interrogated lung cancer specimens where we examined the expression of 20 biomarkers. Subsequently, the tissue composition was interrogated using an in-house optimized nuclei detection algorithm followed by a newly developed image artifact exclusion approach. Thereafter, the data was processed using several publicly available tools, highlighting the compatibility of COMET-derived data with currently available image analysis frameworks. In summary, we showcased an innovative semi-automated workflow that highlights the ease of adoption of multiplex imaging to explore TME composition at single-cell resolution using a simple slide in, data out approach. Our workflow is easily transferrable to various cohorts of specimens to provide a toolset for spatial cellular dissection of the tissue composition.
Subject(s)
Ecosystem , Lung Neoplasms , Humans , Algorithms , Image Processing, Computer-Assisted , Biomarkers , Tumor MicroenvironmentABSTRACT
Although myeloid-derived immune cells can be dispersed throughout the tumor microenvironment (TME), anti-tumor effector cells are confined to the perivascular space. Here, we present a protocol to quantify immune cell distribution from tumor vasculature to its glioma microenvironment on sequential immunofluorescence multiplex images. We describe steps for sequential immunofluorescence multiplex staining, image generation, and storage. We then detail the procedures for tissue, vessel, and nuclei segmentation; cell phenotyping; data extraction; and training using RStudio and Spyder.
Subject(s)
Fluorescent Antibody Technique , Glioma , Tumor Microenvironment , Tumor Microenvironment/immunology , Glioma/immunology , Glioma/pathology , Glioma/blood supply , Humans , Fluorescent Antibody Technique/methods , Animals , Brain Neoplasms/immunology , Brain Neoplasms/pathology , Brain Neoplasms/blood supply , Image Processing, Computer-Assisted/methods , MiceABSTRACT
Intravital two-photon microscopy of the mouse brain requires visual access without affecting normal cognitive functions, which is crucial for longitudinal imaging studies that may last several months. In this protocol, we describe the surgical implantation of a metal-free cranial imaging window, which can be used to perform two-photon microscopy and magnetic resonance imaging in the same animal. This multimodal imaging platform enables the investigation of dynamic processes in the central nervous system at a cellular and macroscopic level. For complete details on the use and execution of this protocol in the context of brain cancer, please refer to Zomer et al.1.
ABSTRACT
Purpose: The presentation of results of an ophthalmic plaque displacement as a brachytherapy treatment method of large diffuse uveal melanomas. Material and methods: This was a retrospective analysis of treatment results of 9 patients with large diffuse uveal melanomas using ophthalmic plaque displacement. Patients were treated with this method in our center between 2012 and 2021 (last follow-up visit in 2023). To achieve appropriate radiation dose distribution for large tumors with a base greater than 18 mm, brachytherapy (106Ru in 7 patients and 125I in 2 patients) with applicator displacement was used as primary treatment. Median follow-up was 2.9 years, and for patients with positive primary treatment results, it was 1.7 months. Median time to local relapse was 2.3 years. Results: In 5 patients, a positive result of local treatment was obtained, out of whom, one patient underwent enucleation due to complications. In the next 4 cases, local recurrence developed. In all tumors, the use of applicator displacement method caused that planning target volume (PTV) was effectively covered with treatment isodose. Conclusions: Brachytherapy with ocular applicator displacement allows for the treatment of tumors with base measurements larger than 18 mm. The application of this method may be considered as an alternative for eye enucleation in particular cases of large diffuse tumors, such as a neoplasm of the eye with vison, or when a patient does not consent to enucleation.
ABSTRACT
Non-small-cell lung cancer (NSCLC) represents 85% of new cases of lung cancer. Over the past two decades, treatment of patients with NSCLC has evolved from the empiric use of chemotherapy to more advanced targeted therapy dedicated to patients with an epidermal growth factor receptor (EGFR) mutation. The multinational REFLECT study analyzed treatment patterns, outcomes, and testing practices among patients with EGFR-mutated advanced NSCLC receiving first-line EGFR tyrosine kinase inhibitor (TKI) therapy across Europe and Israel. The aim of this study is to describe the Polish population of patients from the REFLECT study, focusing on treatment patterns and T790M mutation testing practice. A descriptive, retrospective, non-interventional, medical record-based analysis was performed on the Polish population of patients with locally advanced or metastatic NSCLC with EGFR mutations from the REFLECT study (NCT04031898). A medical chart review with data collection was conducted from May to December 2019.The study involved 110 patients. Afatinib was used as the first-line EGFR-TKI therapy in 45 (40.9%) patients, erlotinib in 41 (37.3%), and gefitinib in 24 (21.8%) patients. The first-line EGFR-TKI therapy was discontinued in 90 (81.8%) patients. The median progression-free survival (PFS) on first-line EGFR-TKI therapy was 12.9 months (95% CI 10.3-15.4). A total of 54 patients started second-line therapy, of whom osimertinib was administered to 31 (57.4%). Among 85 patients progressing on first-line EGFR-TKI therapy, 58 (68.2%) were tested for the T790M mutation. Positive results for the T790M mutation were obtained from 31 (53.4%) tested patients, all of whom received osimertinib in the next lines of therapy. The median overall survival (OS) from the start of first-line EGFR-TKI therapy was 26.2 months (95% CI 18.0-29.7). Among patients with brain metastases, the median OS from the first diagnosis of brain metastases was 15.5 months (95% CI 9.9-18.0). The results of the Polish population from the REFLECT study highlight the need for effective treatment of patients with advanced EGFR-mutated NSCLC. Nearly one-third of patients with disease progression after first-line EGFR-TKI therapy were not tested for the T790M mutation and did not have the opportunity to receive effective treatment. The presence of brain metastases was a negative prognostic factor.
ABSTRACT
Tissues are complex environments where different cell types are in constant interaction with each other and with non-cellular components. Preserving the spatial context during proteomics analyses of tissue samples has become an important objective for different applications, one of the most important being the investigation of the tumor microenvironment. Here, we describe a multiplexed protein biomarker detection method on the COMET instrument, coined sequential ImmunoFluorescence (seqIF). The fully automated method uses successive applications of antibody incubation and elution, and in-situ imaging enabled by an integrated microscope and a microfluidic chip that provides optimized optical access to the sample. We show seqIF data on different sample types such as tumor and healthy tissue, including 40-plex on a single tissue section that is obtained in less than 24 h, using off-the-shelf antibodies. We also present extensive characterization of the developed method, including elution efficiency, epitope stability, repeatability and reproducibility, signal uniformity, and dynamic range, in addition to marker and panel optimization strategies. The streamlined workflow using off-the-shelf antibodies, data quality enabling downstream analysis, and ease of reaching hyperplex levels make seqIF suitable for immune-oncology research and other disciplines requiring spatial analysis, paving the way for its adoption in clinical settings.
Subject(s)
Antibodies , Proteomics , Proteomics/methods , Reproducibility of Results , Fluorescent Antibody Technique , BiomarkersABSTRACT
Tumors evolve in a dynamic communication with their native tissue environment and recruited immune cells. The diverse components of the tumor microenvironment (TME) can critically regulate tumor progression and therapeutic response. In turn, anticancer treatments may alter the composition and functions of the TME. To investigate this continuous dialog in the context of primary brain cancers, we developed a multimodal longitudinal imaging strategy. We combined macroscopical magnetic resonance imaging with subcellular resolution two-photon intravital microscopy, and leveraged the power of single-cell analysis tools to gain insights into the ongoing interactions between different components of the TME and cancer cells. Our experiments revealed that the migratory behavior of tumor-associated macrophages is different in genetically distinct glioblastomas, and in response to macrophage-targeted therapy. These results underscore the importance of studying cancer longitudinally in an in vivo setting, to reveal complex and dynamic alterations in the TME during disease progression and therapeutic intervention.
ABSTRACT
UNLABELLED: Uveal metastases are the most common intraocular malignant tumors. Most patients who develop metastatic carcinoma to the choroid are managed by radiation or chemotherapy. Since TTT is an optional treatment for choroidal melanomas and hemangiomas, we ought to determine whether TIT is suitable for treatment of solitary choroidal metastasis at the posterior pole. PURPOSE: To evaluate effectivenes of TTT treatment for intraocular metastases. MATERIAL AND METHODS: 45 patients (59 eyes) with intraocular metastases were treated in the Ophthalmological Department of Jagiellonian University in Kraków. There were 30 women and 15 man, at the age 31-84 years (av. 57.5). The primary tumor was a breast cancer in 22 women, lung cancer in 5 men and 2 women, kidney (3), colon (2), uterus (1), larynx (1), testicle (1), esophagus (1). 10 patients had also metastases in other organs. TTT was performed in all treated eyes, in 11 combined with 106Ru brachytherapy (BT). Chemotherapy as adjuvant treatment was performed in 18 patients. The results of treatment were evaluated in mean 14.5 months (1-61) follow-up. RESULTS: After TTT, tumor shrinking was observed in most treated tumors (in 37 eyes, 62.7%), inhibition of tumor growth in 5 (8.4%), and in 4 cases (6.7%) progression of tumor growth. In 3 cases (5.1%) with flat scar and intraocular tumor shrinking, extrascleral extension located close to the tumor base appeared after treatment (TTT in 2 eyes, TTT combined with BT in one case). CONCLUSIONS: TTT is an effective treatment method for small choroidal metastases located in the posterior pole. TTT combined with 106Ru brachytherapy can be useful treatment in medium sized choroidal metastases. Efficient results encourage further application of thermotherapy in the treatment of intraocular metastases. It allows for the conservative treatment of the eyeball and also useful visual acuity. Extraocular extension appearing after TT needs further study.
Subject(s)
Carcinoma/secondary , Carcinoma/therapy , Choroid Neoplasms/secondary , Choroid Neoplasms/therapy , Hyperthermia, Induced/methods , Neoplasms/pathology , Pupil , Adult , Aged , Aged, 80 and over , Brachytherapy/methods , Combined Modality Therapy , Female , Fluorescein Angiography/methods , Follow-Up Studies , Humans , Male , Middle Aged , Neoplasm Staging , Neoplasms/therapyABSTRACT
BACKGROUND/AIM: This study analysed the prevalence of the characteristics evaluated in dermatoscopy for melanocytic infiltrations of the conjunctiva with various degrees of malignancy. PATIENTS AND METHODS: A total of 160 conjunctival pigmented lesions were studied. Each lesion was scored using dermatoscopic patterns and the characteristics of malignancy described by Kittler. Also, the Authors' own clues were added to the evaluation. RESULTS: In melanomas, the following characteristics were identified: asymmetry of the pattern and colour, larger average number of colours, the presence of grey colour, structureless area, polymorphic vessels and feeder vessels. A pattern of black dots and a black colour was typical of malignant lesions and pre-cancerous (premalignant) lesions - primary acquired melanosis (PAM) with atypia. Cysts were observed only in the group of naevi. CONCLUSION: The patterns evaluated with dermatoscopy are present in pigmented lesions of the conjunctiva. There are, however, some characteristics which allow differentiation between melanoma and pigmented naevus and melanosis and also between PAM.
Subject(s)
Conjunctival Neoplasms/diagnostic imaging , Melanoma/diagnostic imaging , Nevus, Pigmented/diagnostic imaging , Skin Neoplasms/diagnostic imaging , Adolescent , Adult , Aged , Aged, 80 and over , Algorithms , Child , Conjunctival Neoplasms/pathology , Dermoscopy , Female , Humans , Male , Melanoma/pathology , Middle Aged , Nevus, Pigmented/pathology , Skin Neoplasms/pathology , Young AdultABSTRACT
Not required for Clinical Vignette.
Subject(s)
Thymoma , Thymus Neoplasms , Choroid/diagnostic imaging , Humans , Thymoma/diagnostic imaging , Thymus Neoplasms/diagnostic imaging , Tomography, Optical Coherence , UltrasonographyABSTRACT
PURPOSE: A retrospective evaluation of effectiveness of brachytherapy or enucleation in treatment of rare form of uveal melanoma: ring melanoma. MATERIAL AND METHODS: Analysis comprised 49 patients treated from 2000 to 2019 for ring melanoma involving ciliary body. In 15 patients, primary treatment consisted of brachytherapy (106Ru or 125I), whereas in 34 patients, eyeballs were enucleated. The evaluation concerned differences between analyzed groups relating to the clinical and histopathological features as well as overall survival, cancer-specific overall survival, and disease-free survival. RESULTS: No significant differences between the analyzed groups were found with regards to clinical and histopathological features, apart from intra-ocular pressure (increased in the enucleation group). Kaplan-Meier analysis did not reveal any significant differences between the group treated with enucleation and the group undergoing brachytherapy, both with regards to overall survival (p = 0.325) and cancer-specific overall survival (p = 0.477). A significant difference was observed in disease-free survival (p = 0.009), which was significantly shorter in the group undergoing brachytherapy. In the analysis of parameters of the applied brachytherapy, no significant differences between patients with and without local recurrence were found. Mean observation period was 350.8 weeks (range, 24-996 weeks, SD = 231.6). A local recurrence occurred in 11 (22.4%) patients, including 3 (6.1%) in enucleation and 8 (16.3%) after brachytherapy groups. Metastasis developed in 11 (22.4%) cases after a mean follow-up of 133 weeks (33.25 months), range 3-655 weeks. CONCLUSIONS: Preliminary observations may suggest that brachytherapy in this rare form of uveal melanoma, such as ring melanoma involving the ciliary body, may be taken into consideration as a useful alternative to enucleation. However, the confirmation of such an approach requires a larger patients' group to be gathered, and also a longer follow-up period. This is especially important in patients with a good baseline visual acuity in the affected eye, or when the neoplasm is present in the remaining eye with vision.
ABSTRACT
BACKGROUND/AIM: This study aimed to develop an algorithm allowing the differentiation between conjunctival melanoma and other melanocytic infiltrations of the conjunctiva, on the basis of a dermatoscopic examination. PATIENTS AND METHODS: A total of 160 conjunctival pigmented lesions were studied (40 melanomas and 120 non-melanoma conjunctival infiltrations). The clinical characteristics of the tumours were assessed with the use of dermatoscopic characteristics as described by Kittler, and with taking into consideration the typical characteristics of conjunctival lesions. RESULTS: On the basis of modified dermatoscopic criteria, an algorithm was generated consisting of an assessment of the presence of 9 suspicious characteristics, e.g. more than two colours, colour asymmetry, pattern asymmetry, vascular polymorphism, presence of short vessels, linear vascular pattern, the presence of a peripheral structureless area, the presence of a grey structureless area and black dots in any part of the lesion. The presence of any of these characteristics scores 1 point. If a melanocytic lesion scores ≥3 points, the probability of diagnosing melanoma is on the level of p>0.001. CONCLUSION: The use of the proposed algorithm, based on modified dermatoscopic characteristics, may be a valuable tool for the diagnosis of conjunctival melanoma.