The Severity of Diabetic Retinopathy Corresponds with Corneal Nerve Alterations and Ocular Discomfort of the Patient.

Diabetic retinopathy (DR) remains the leading cause of blindness in the working-age population. Its progression causes gradual damage to corneal nerves, resulting in decreased corneal sensitivity (CS) and disruption of anterior-eye-surface homeostasis, which is clinically manifested by increased ocular discomfort and dry eye disease (DED). This study included 52 DR patients and 52 sex- and age-matched controls. Ocular Surface Disease Index (OSDI) survey, tear film-related parameters, CS, and in vivo corneal confocal microscopy (IVCM) of the subbasal plexus were performed. Furthermore, all patients underwent tear sampling for neurotrophin and cytokine analysis. OSDI scores were greater in DR patients than in controls (p = 0.00020). No differences in the Schirmer test score, noninvasive tear film-break-up time (NIBUT), tear meniscus or interferometry values, bulbar redness, severity of blepharitis or meibomian gland loss were found. In the DR group, both the CS (p < 0.001), and the scotopic pupil diameter (p = 0.00008) decreased. IVCM revealed reduced corneal nerve parameters in DR patients. The stage of DR was positively correlated with the OSDI (Rs = +0.51, 95% CI: + 0.35-+0.64, p < 0.001) and negatively correlated with IVCM corneal nerve parameters and scotopic pupillometry (Rs = -0.26, 95% CI: -0.44--0.06, p = 0.0097). We found negative correlations between the OSDI and IVCM corneal innervation parameters. The DR group showed lower tear film-brain-derived neurotrophic factor (BDNF) levels (p = 0.0001) and no differences in nerve growth factor (NGF)-ß, neurotrophin (NT)-4, vascular endothelial growth factor (VEGF), interleukin (IL)-1ß, IL-4, IL-5, IL-6, or IL-12 concentrations. Tumor necrosis factor (TNF)-α, IL-2, IL-8, IL-10, granulocyte macrophage colony-stimulating factor (GM-CSF), and interferon (IFN)-γ levels were decreased among patients with DR. Corneal innervation defects have a direct impact on patients' subjective feelings. The evolution of DR appears to be associated with corneal nerve alterations, emphasizing the importance of IVCM.

Associations of microRNAs, Angiogenesis-Regulating Factors and CFH Y402H Polymorphism-An Attempt to Search for Systemic Biomarkers in Age-Related Macular Degeneration.

Age-related macular degeneration (AMD) remains the leading cause of blindness in elderly people, but the pathophysiology of this disease is still largely unknown. We investigated the systemic expression of angiogenesis-regulating growth factors and selected miRNAs known to regulate angiogenesis in AMD patients. We also focused on possible correlations of their expression with the presence of CFH Y402H or ARMS A69S risk variants. A total of 354 AMD patients and 121 controls were enrolled in this study. The levels of angiogenesis-regulating factors were analyzed in plasma samples using Luminex technology. The expression of selected miRNAs was analyzed in peripheral blood plasma using real-time qPCR. The genetic analysis was performed with an Illumina NextSeq500 system. AMD was an independent factor associated with lower levels of angiogenin (ß = -0.29, p < 0.001), endostatin (ß = -0.18, p < 0.001), FGF-basic (ß = -0.18, p < 0.001), PlGF (ß = -0.24, p < 0.001), miRNA-21-3p (ß = -0.13, p = 0.01) and miRNA-155-5p (ß = -0.16, p = 0.002); and with higher levels of FGF-acidic (ß = 0.11, p = 0.03), miRNA-23a-3p (ß = 0.17, p < 0.001), miRNA-126-5p (ß = 0.13, p = 0.009), miRNA-16-5p (ß = 0.40, p < 0.001), miRNA-17-3p (ß = 0.13, p = 0.01), miRNA-17-5p (ß = 0.17, p < 0.001), miRNA-223-3p (ß = 0.15, p = 0.004), and miRNA-93 (ß = 0.11, p = 0.04). The expression of analyzed miRNA molecules significantly correlated with the levels of tested angiogenesis-regulating factors and clinical parameters in AMD patients, whereas such correlations were not observed in controls. We also found an association between the CFH Y402H polymorphism and miRNA profiles, whereby TT homozygotes showed evidently higher expression of miRNA-16-5p than CC homozygotes or TC heterozygotes (p = 0.0007). Our results suggest that the balance between systemic pro- and anti-angiogenic factors and miRNAs is vital in multifactorial AMD pathogenesis.

The macular function and structure in patients with diabetic macular edema before and after ranibizumab treatment.

PURPOSE: To evaluate macular function and structure in patients with diabetic macular edema prior to, as well as 3 and 6 months after intravitreal ranibizumab treatment. PATIENTS AND METHODS: Seventeen eyes of 17 patients with type 2 diabetes mellitus and diabetic macular edema (DME) were treated with intravitreal injections of 0.5 mg ranibizumab. Prior to the first injection, as well as after 3 and 6 months, the following examinations were performed: assessment of distance best-corrected visual acuity (log MAR), perception of metamorphopsia (M-Chart), slit lamp examination of the anterior and posterior segment of the eye (Volk 90D lens), evaluation of the retinal and choroidal circulation (fluorescein angiography), assessment of the structure and thickness of the macula (OCT), as well as evaluation of the macular function (PERG and mfERG). RESULTS: We observed that ranibizumab significantly improved visual acuity after 3 and 6 months from the beginning of the treatment, which was a consequence of reduced macular edema and vascular leakage. There was a statistically significant decrease in metamorphopsia frequency at month 3; however, at month 6 it was a statistically insignificant when compared to the baseline. The results of electrophysiological examinations revealed no improvement in ranibizumab-treated patients. CONCLUSION: Improvement of visual acuity and reduction in macular thickness were maintained up to the 6-month follow-up. The results of electrophysiological examinations revealed that ranibizumab injections tend to stabilize bioelectrical macular function of the outer, middle and inner retinal layers, which was impossible to recognize on the basis of visual acuity and OCT. Therefore, the electrophysiological examinations should be used as an additional objective tool for the evaluation of the anti-VEGF treatment effectiveness in DME.

Aqueous levels of VEGF correlate with retinal non-perfusion areas in patients with diabetic macular edema and macular edema secondary to central retinal vein occlusion.

AIM: To evaluate the association between the level of vascular endothelial growth factor in the aqueous humor and the size of capillary non-perfusion areas in patients with macular edema secondary to retinal vein occlusion and diabetic retinopathy. MATERIAL AND METHODS: The study group consisted of 24 patients (24 eyes) at the age of 55-78 years, with diffuse macular edema secondary to retinal vein occlusion and diabetic retinopathy. The control group consisted of 26 subjects aged 55-87 years who were admitted for scheduled cataract surgery. The VEGF aqueous humor levels, retinal thickness using optical coherence tomography, as well as the size of non-perfusion areas measured on fluorescein angiography images were evaluated in each enrolled subject. RESULTS: The vascular endothelial growth factor aqueous humor levels were found to be significantly higher in patients with macular edema as compared to controls (p = 0.0002). In the diabetic macular edema and retinal vein occlusion group, the con- centration of vascular endothelial growth factor in aqueous humor positively correlated with the extent of non-perfusion areas measured on fluorescein angiograms (Rs = + 0.45, p = 0.02;). Multivariate analysis of patients and controls performed using the general linear model, adjusted for age, sex, intraocular pressure and the presence of diabetes, revealed that macular edema was an independent factor associated with higher aqueous VEGF concentrations (ß = +0.74, p = 0.0012). CONCLUSIONS: Macular edema secondary to either retinal vein occlusion or diabetic retinopathy is associated with the increased levels of vascular endothelial growth factor in the aqueous humor. Therefore, the management of patients with macular edema secondary to retinal vein occlusion or diabetic retinopathy should aim at reducing the ocular vascular endothelial growth factor concentrations, especially in the presence of capillary non-perfusion areas.

[Increased expression of endothelin-1-- a novel diagnostic marker for early AMD detection?]. / Wzrost ekspresji endoteliny-1--nowy marker wczesnej diagnostyki AMD?

AIM: The relationship between ischemic vascular disease and age-related macular degeneration may indicate the role of vascular injury as the primary insult causing functional deficits in age-related macular degeneration. The vasoactive factors produced by endothelial cells include endothelin-1 (ET-1), which is one of the most potent vasoconstricting peptides. In this study we sought to explore the potential role of endothelial dysfunction in the pathogenesis of age-related macular degeneration by measuring the concentration of ET-1 in peripheral blood of individuals diagnosed with age-related macular degeneration and evaluating its intracellular expression in peripheral blood cells, on mRNA level. MATERIAL AND METHODS: Peripheral blood samples from 31 patients with diagnosed dry age-related macular degeneration and 46 patients with neovascular age-related macular degeneration were collected. Forty six age- and sex-matched volunteers without age-related macular degeneration were enrolled as a control group. ET-1 plasma levels were analyzed by ELISA and intracellular expression of ET-1 in peripheral blood cells was studied by using qRT-PCR. RESULTS: The expression of intracellular ET-1 was significantly elevated in peripheral blood cells of both dry and wet age-related macular degeneration patients compared with the control subjects. Immunofluorescence staining revealed that ET-1 was specifically expressed in the circulating endothelial cells. CONCLUSIONS: We assume that damaged endothelial cells may release a variety of vasoconstricting molecules, including ET-1, leading to derangement between the endothelium-derived relaxing and contracting factors. Local retinal ischemia consequently develops which may promote the development of retinal degeneration in patients with age-related macular degeneration,

[Idiopathic choroidal neovascularization--case report]. / Idiopatyczna neowaskularyzacja podsiatkówkowa--opis przypadku.

PURPOSE: To report a patient with peripapillary idiopathic choroidal neovascularization treated with an anti-VEGF (anti- vascular endothelial growth factor) agent (bevacizumab) observed during for 9 months. PATIENT AND METHODS: Twenty nine years old man was referred to the Department for diagnosis and treatment because of unilateral visual acuity decrease in the right eye (VA RE--0.1) and metamorphopsias. The routine ophthalmic examination revealed macular edema and peripapillaryedema with epiretinal and intraretinal hemorrhages. The optical coherence tomography, fluorescein angiography, as well as laboratory tests were performed in order to exclude uveitis. Due to the difficulties in the diagnosis indocyanine green angiography was also performed. Based on clinical symptoms and the findings of the additional diagnostic procedures, the patient was diagnosed with idiopathic choroidal neovascularization. The patient was qualified for anti-VEGF therapy and received three intravitreal injections of bevacizumab at the dose of 1.25 mg, at monthly intervals. RESULTS: Significant improvement of visual acuity (VA RE - 1.0) and regression of the peripapillary edema with hemorrhages were achieved after the third injection of 1.25 mg bevacizumab. At 6 months, peripapillary scarring was observed in the area involved by the primary lesions. CONCLUSIONS: Anti-VEGF therapy is the effective treatment of idiopathic choroidal neovascularization in the described case. The visual acuity improvement and rapid regression of posterior segment lesions after bevacizumab administration were observed.

PEDF and VEGF plasma level alterations in patients with dry form of age-related degeneration--a possible link to the development of the disease.

PURPOSE: The aim of the study is to explore the interaction between stimulators and inhibitors of angiogenesis by measuring pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF) plasma levels in patients with the wet and dry forms of age-related macular degeneration (AMD). MATERIAL AND METHODS: Forty-six subjects with the wet form, 31 with the dry form of AMD as well as 47 non-AMD healthy controls were enrolled in the study. Plasma concentrations of VEGF and PEDF were measured using ELISA test. RESULTS: A significant decrease in the PEDF plasma level in patients with the dry form of AMD was found. Multivariate analyses of patients and controls adjusted for age, sex, smoking, and concomitant vascular diseases as independent variables revealed that the dry form of AMD was the only independent factor associated with lower plasma PEDF levels (beta = -0.34; p = 0.026). On the contrary, in the wet AMD group, a strong positive correlation between VEGF and PEDF concentrations was observed (Rs = +0.63; p = 0.002), and significantly higher PEDF and VEGF plasma levels in patients with bilateral manifestations of the disease were also found. CONCLUSIONS: These findings suggest that different manifestations of AMD, i.e. the dry and wet forms, may be associated with various altered concentrations of counterbalancing stimulators and inhibitors of the angiogenesis process.

[Polypoidal choroidal vasculopathy with spontaneous regression of subfoveal changes--case report]. / Polipoidalna waskulopatia naczyniówkowa z samoistna regresja zmian poddolkowych--opis pacjenta.

PURPOSE: To report a patient with polypoidal choroidal vasculopathy (PCV) with spontaneous regression of subfoveal changes during follow-up. MATERIAL AND METHODS: The seventy six years old men was referred to the treatment of exudative type of age related macular degeneration (AMD) in the RE. The routine ophthalmological examination, the optical coherence tomography (OCT), fluorescein angiography (FA), and indocyanine green angiography (ICGA) were performed. RESULTS: Decreasing of visual acuity of the RE and abnormal result of the Amsler test, hemorrhagic and exudative changes near inferior-temporalis vascular arcade were observed. Intraretinal fluid in the OCT was noted. FA revealed parapapillaris changes suggesting CNV. ICGA showed the presence of branching vascular network extending from choroidal vasculature (BVN) and polypoidal and aneurysmal vascular terminal lesion (PL) localized under retinal pigment epithelium (RPE). CONCLUSIONS: Based on the results PCV was diagnosed and the patient was referred to laserotherapy. Due to the regression of the eye fundus changes during the period of observation, confirmed by control OCT and FA the treatment was not implemented.

Evaluation of treatment efficacy of intravitreal ranibizumab injections in patients with wet type of AMD.

PURPOSE: To evaluate foveal function, retinal circulation and foveal thickness before and after intravitreal ranibizumab injections in eyes with wet type of age-related macular degeneration (AMD). MATERIAL AND METHODS: The study group consisted of 21 eyes (20 patients) with choroidal neovascularisation (CNV) due to AMD. Inclusion criteria were based on fluorescein angiography (FA) and distance best corrected visual acuity (DBCVA)--log MAR scale. In each eye, 3 consecutive injections of ranibizumab every 4 weeks were administered and then individual course for re-injections according to DBCVA and optical coherence tomography (OCT) up to 12 months was applied. At baseline, 3, 6 and 12 months follow-up, the following tests were performed: DBCVA, multifocal electroretinogram (mfERG) and OCT. Additionally, FA was carried out before the treatment, 3 and 12 months from the baseline. RESULTS: At baseline, FA revealed mainly minimally occult choroidal neovascularisation--57% (12/21) of eyes. At 3 months choroidal neovascularisation diameter was stable; no leakage from active choroidal neovascularisation was seen in 76% (16/21) of eyes. After 12 months follow-up, increase in choroidal neovascularisation diameter was seen in 43% (9/21) of eyes and no leakage in 57% (12/21) of cases. The mean DBCVA significantly improved only after 3 months (p < 0.02). Significant decrease of mean foveal thickness was observed in each follow-ups (p < 0.01). The mfERG data from the macular region remained stable or improved slightly in some cases. CONCLUSIONS: In our series of patients with the wet type of AMD after intravitreal injections of ranibizumab in 12 months follow-up, the reduction of foveal thickness was noted while DBCVA and the bioelectrical function from the macular region measured by the mfERG remained stable.

[Combined photodynamic therapy and intravitreal injection of triamcinolone acetonide in patients with wet form of AMD. Introductory report]. / Zastosowanie terapii laczonej PDT z iniekcjami doszklistkowymi octanu triamcinolonu u pacjentów z wysiekowa postacia AMD. Doniesienie wstepne.

PURPOSE: To evaluate the efficacy of combined PDT and 4 mg intravitreal triamcinolone acetonide injection, performed 48-72 hours after PDT, in patients with wet form of AMD. MATERIAL AND METHODS: Nonrandomised, interventional case series, 13 eyes of 13 patients with subfoveal CNV due to AMD that did not respond to PDT monotherapy - 7 females, 6 males - at the age of 65-85 (average age 76.6 +/- 6.7 years); standard PDT was performed in all patients followed by a 4 mg intravitreal injection of triamcinolone acetonide given 48-72 hours after PDT. Follow up visits were scheduled 1 and 7 days after the injection and then every 3 months afterwards and included: BVCA (Snellen chart), IOP measurements, FA, OCT, slit lamp and eye fundus examination. Lesions with active CNV leakage in FA were retreated every 3 months. RESULTS: Average observation time was 10.8 +/- 3.5 months. Baseline visual acuity before PDT monotherapy was applied (Vo) was 0.17 +/- 0.12 (0.06-0.5), and after the therapy decreased to (V1) 0.14 +/- 0.13 (0.05-0.2). After combined PDT and Tc treatment BVCA increased to (V2) 0.21 +/- 0.13 (0.06-0.5), p<0,03. 76,9% of patients gained or maintained visual acuity after combined therapy in the observation time. In 70% of eyes no signs of active CNV was observed in AF and OCT after 1 session of combined PDT and Tc treatment. Only 4 patients required 1 repeated treatment session. CONCLUSIONS: 1. Combination of PDT and IVTA may be effective in patients with wet AMD with no response to PDT alone and significantly reduces the repeated treatment rate. 2. Intravitreal Tc injection performed 48-72 hours after PDT may improve the final functional effects in treated eyes as compared with PDT monotherapy. Our results need further investigation.

The impact of vascular risk factors on the thickness and volume of the choroid in AMD patients.

Disturbances in choroidal microcirculation may lead to the onset and progression of age-related macular degeneration (AMD). We aimed to assess changes in the choroidal volume and thickness in the macular region in AMD eyes and to investigate whether coexisting vascular risk factors alter choroidal status. We enrolled 354 AMD patients (175 dry, 179 wet AMD) and 121 healthy controls. All participants underwent a complete ophthalmologic examination and assessment of choroidal thickness and volume. A multivariate analysis adjusted for age, sex, and smoking status revealed that wet AMD was an independent factor associated with higher average thickness of the central ring area (ATC) and average volume of the central ring area (AVC) and lower choroidal vascularity index (CVI) compared to controls (ß = + 0.18, p = 0.0007, ß = + 0.18, p = 0.0008, respectively) and to dry AMD (ß = + 0.17, p = 0.00003 for both ATC and AVC and ß = - 0.30 p < 0.0001 for CVI). ATC, AVC and average volume (AV) were lower in AMD patients with hypertension and ischaemic heart disease (IHD). The duration of hypertension was inversely correlated with ATC, AVC and AV (Rs = - 0.13, p < 0.05; Rs = - 0.12; p < 0.05, Rs = - 0.12; p < 0.05, respectively) while IHD duration negatively correlated with AV (Rs = - 0.15, p < 0.05). No such associations were observed in the control group. Our findings show that the choroidal vascular system in eyes with AMD is much more susceptible to damage in the presence than in the absence of systemic vascular disease.

Retinal Vessel Functionality Is Linked With ARMS2 A69S and CFH Y402H Polymorphisms and Choroidal Status in AMD Patients.

Purpose: We aimed to investigate the reactivity of retinal vessels to a flickering stimulus in patients with age-related macular degeneration (AMD) and healthy participants. We also assessed whether the parameters of retinal vessels are dependent on genetic predisposition. Methods: A total of 354 patients with AMD and 121 controls were recruited for the study. All participants underwent thorough ophthalmologic examination and static and dynamic retinal vessel analysis. AMD risk polymorphisms were genotyped in the CFH and ARMS2 genes. Results: We found no differences between the AMD group and controls in central retinal arteriolar equivalent (CRAE), central retinal venular equivalent (CRVE), arteriovenous ratio (AVR), dynamic analysis of arteries (DAAs), or dynamic analysis of veins (DAVs). Eyes with early AMD presented with significantly higher AVR values than eyes with late AMD. In the AMD group, DAA correlated positively with both choroidal thickness (Rs = 0.14, P = 0.00096) and choroidal volume (Rs = 0.23, P < 0.0001), and no such associations were observed in the controls. We found significantly lower DAA (1.47 ± 1.50) in TT homozygotes for the ARMS2 A69S polymorphism in comparison with GG homozygotes (2.38 ± 1.79) and patients with GG + GT genotypes (2.28 ± 1.84). We also observed less prominent DAV (3.24 ± 1.71) in patients with TC + CC genotypes in the CFH Y402H polymorphism compared with TT homozygotes (3.83 ± 1.68). Conclusions: Our findings suggest that retinal microcirculation appears to be associated with the genetic background, choroidal parameters, and clinical features of the patients with AMD.

Genetic factors associated with age-related macular degeneration: identification of a novel PRPH2 single nucleotide polymorphism associated with increased risk of the disease.

PURPOSE: Age-related macular degeneration (AMD) is associated with multiple environmental and genetic risk factors. Two main risk factors for AMD are variants in the CFH and ARMS2/HTRA1 genes. We investigated over 2000 variants in AMD patients and controls using high-throughput sequencing methods to search for variants associated with AMD. METHODS: A total of 296 AMD patients and 100 controls were enrolled in this study. Genetic analysis was performed with the Illumina NextSeq 500 system. RESULTS: Multivariate analysis of patients and controls, adjusted for age, sex and smoking status (pack-years), revealed that three SNPs were strong risk factors independently associated with AMD: CFH Y402H, ARMS A69S and PRPH2 c.582-67T>A (rs3818086). The TC genotype in CFH Y402H was associated with 1.90-fold higher odds, and the CC genotype was associated with 5.66-fold higher odds of AMD compared with the TT genotype. The GT genotype in ARMS A69S was associated with 2.40-fold higher odds, and the TT genotype was associated with 6.75-fold higher odds of disease compared with the GG genotype. In the case of rs3818086, the A allele could be considered a 'risk' allele, since AA + TA genotypes were associated with 2.33-fold higher odds of AMD compared with the TT genotype. CONCLUSIONS: Although PRPH2 mutations have been previously implicated in various forms of retinal degeneration, to the best of our knowledge, this study is the first to show that the rs3818086 variant increases the risk for AMD more than two times. Further studies on larger cohorts are required to elucidate how this variant affects protein structure.

Dietary and Lifestyle Factors Modulate the Activity of the Endogenous Antioxidant System in Patients with Age-Related Macular Degeneration: Correlations with Disease Severity.

Age-related macular degeneration (AMD) is a common cause of blindness in the elderly population, but the pathogenesis of this disease remains largely unknown. Since oxidative stress is suggested to play a major role in AMD, we aimed to assess the activity levels of components of the antioxidant system in patients with AMD. We also investigated whether lifestyle and dietary factors modulate the activity of these endogenous antioxidants and clinical parameters of disease severity. We recruited 330 patients with AMD (39 with early, 100 with intermediate and 191 with late form of AMD) and 121 controls in this study. At enrolment, patients' dietary habits and physical activity were assessed, and each study participant underwent a thorough ophthalmologic examination. The activity of several components of the antioxidant system were measured in red blood cells and platelets using both kinetic and spectrophotometric methods. Patients with AMD consumed much lower levels of fatty fish and eggs than the control group (p = 0.008 and p = 0.04, respectively). In the nAMD group, visual acuity (VA) correlated positively with green vegetable consumption (Rs = +0.24, p = 0.004) and omega-3-rich oil intake (Rs = +0.17, p = 0.03). In the AMD group, the total physical activity MET score correlated positively with VA (Rs = +0.17, p = 0.003) and correlated negatively with the severity of AMD (Rs = -0.14, p = 0.01). A multivariate analysis of patients and controls adjusted for age, sex, and smoking status (pack-years) revealed that AMD was an independent variable associated with a lower RBC catalase (ß = -0.37, p < 0.001) and higher PLT catalase (ß = +0.25, p < 0.001), RBC GPx (ß = +0.26, p < 0.001), PLT GPx (ß = +0.16, p = 0.001), RBC R-GSSG (ß = +0.13, p = 0.009), PLT R-GSSG (ß = +0.12, p = 0.02) and RBC GSH transferase (ß = +0.23, p < 0.001) activity. The activities of components of the antioxidant system were associated with disease severity and depended on dietary habits. The observed substantial increase in the activity of many critical endogenous antioxidants in patients with AMD further indicates that the required equilibrium in the antioxidant system is disturbed throughout the course of the disease. Our findings explicitly show that a diet rich in green vegetables, fish and omega-3-rich oils, supplemented by physical exercise, is beneficial for patients with AMD, as it might delay disease progression and help retain better visual function.

Elevated plasma levels of C3a complement compound in the exudative form of age-related macular degeneration.

AIM: Recent findings suggest that chronic inflammatory processes play a role in the progression of age-related macular degeneration (AMD). Here we asked whether the development of different forms of AMD is connected with the elevation of plasma C3a-desArg concentration. METHODS: We recruited 30 subjects with a clinical diagnosis of exudative AMD with newly diagnosed choroidal neovascularization (CNV), 30 subjects with dry AMD and 30 age- and sex-matched volunteers without AMD. The concentration of C3a-desArg complement compound was measured in the subjects' peripheral blood. We evaluated the association between the level of C3a-desArg and age, sex, smoking, atherosclerosis, and hypertension. RESULTS: We found that the levels of C3a-desArg were significantly elevated in patients with exudative AMD compared to the control group. The concentrations of C3a-desArg in patients with dry AMD were similar to those of controls. Additionally, patients and controls with documented atherosclerosis (AS) displayed significantly higher levels of C3a-desArg compared to subjects without AS. CONCLUSIONS: Our results suggest an association between systemic complement activation and the development of CNV. Moreover, we found an association of complement activation with atherosclerosis and confirmed the hypothesis that AMD can be a local manifestation of systemic disease.

The Interplay Between Systemic Inflammatory Factors and MicroRNAs in Age-Related Macular Degeneration.

We aimed to explore the expression of systemic inflammatory factors and selected intracellular miRNAs that regulate inflammatory signaling pathways potentially involved in age-related macular degeneration (AMD) pathogenesis. A total of 179 patients with wet AMD, 175 with dry AMD and 121 controls were enrolled in the study. Soluble inflammatory factors were analyzed in plasma samples using Luminex technology. Expression of selected miRNAs was analyzed in isolated nucleated peripheral blood cells (PBNCs) using real-time qPCR. Wet AMD was an independent factor associated with higher concentrations of IL-6 (ß = +0.24, p = 0.0004), GM-CSF (ß = +0.31, p < 0.001), IFN-γ (ß = +0.58, p < 0.001), higher expression of miRNA-23a-3p (ß = +0.60, p < 0.0001), miRNA-30b (ß = +0.32, p < 0.0001), miRNA-191-5p (ß = +0.28, p < 0.0001) and lower concentration of IL-1ß (ß = -0.25, p = 0.0003), IL-5 (ß = -0.45, p < 0.001), IL-10 (ß = -0.45, p < 0.001), IL-12 (ß = -0.35, p < 0.001), lower expression of miRNA-16-5p (ß = -0.31, p < 0.0001), miRNA-17-3p (ß = -0.18, p = 0.01), miRNA-150-5p (ß = -0.18, p = 0.01) and miRNA-155-5p (ß = -0.47, p < 0.0001). Multivariate analysis revealed that dry AMD was an independent factor associated with higher concentration of GM-CSF (ß = +0.34, p < 0.001), IL-6 (ß = +0.13, p = 0.05), higher expression of miRNA-23a-3p (ß = +0.60, p < 0.0001), miRNA-126-3p (ß = +0.23, p = 0.0005), miRNA-126-5p (ß = +0.16, p = 0.01), miRNA 146a (ß = +0.14, p = 0.03), and mRNA191-5p (ß = +0.15, p = 0.03) and lower concentrations of TNF-α (ß = +0.24, p = 0.0004), IL-1ß (ß = -0.39, p < 0.001), IL-2 (ß = -0.20, p = 0.003), IL-5 (ß = -0.54, p < 0.001), IL-10 (ß = -0.56, p < 0.001), IL-12 (ß = -0.51, p < 0.001), lower expression of miRNA-16-5p (ß = -0.23, p = 0.0004), miRNA-17-3p (ß = -0.20, p = 0.003) and miRNA-17-5p (ß = -0.19, p = 0.004). Negative correlations between visual acuity and WBC, lymphocyte count, TNF-α, IL-1 ß, IL-2, IL-4, IL-6, IL-10 concentrations and miRNA-191-5p, as well as positive correlations between visual acuity and miRNA-126-3p, -126-5p, and -155-5p PBNCs expression were found in AMD patients. No such correlations were found in the control group. Our results may suggest the role of both intra- and extracellular mechanisms implicated in inflammatory response regulation in multifactorial AMD pathogenesis.

[Frequency and incidence of myopia among medical students]. / Wystepowanie krótkowzrocznosci szkolnej u studentów medycyny.

PURPOSE: To estimate the frequency and dynamic changes in refractive errors in population of students of medical (MF) and dental (DF) faculties of Pommeranian Medical Academy in Szczecin. MATERIAL AND METHODS: Medical records from ophthalmic examinations performed in 176 medical students (117 females and 59 males) during the 2nd and 4th course of studies were analyzed (2-years long observation). RESULTS: Refractive error was found in 42% of MF students and 39.5% of DF students on the 2nd course and in 51% and 47% on the 4th course respectively. Myopia was found in 75% of ametropic eyes. In 2 years observation time, there was a statistically significant progression of refracive error in 50% of myopic students (from -2.9 +/- 2.3 D to -3.4 +/- 1.9D in MF and from -2.0 +/- 1.5 to -2.4 +/- 1.5D in DF). Myopia developed in 13% of initially emmetropic students.

Different populations of circulating endothelial cells in patients with age-related macular degeneration: a novel insight into pathogenesis.

PURPOSE: Circulating endothelial cells (CECs) and endothelial progenitor cells (EPCs) may serve as novel markers of endothelial dysfunction. The presence and clinical implications of CECs and the expression of endothelin (ET)-1, one of the most potent vasoconstrictors, have not been evaluated in patients with the neovascular form of age-related macular degeneration (AMD). This study was conducted to determine the different populations of endothelial cells (ECs) in the peripheral blood of AMD patients and to correlate these findings with the expression of ET-1 and the cytokines and growth factors responsible for EC migration and function. METHODS: Peripheral blood samples were collected from 29 patients with diagnosed neovascular AMD and from 38 healthy control subjects. CD133(-)CD144(+) CECs and CD34(+)CD133(+)CD144(+) EPCs were counted and analyzed by flow cytometry. The intracellular expression of ET-1 in peripheral blood nuclear cells (PBNCs) was studied by using qRT-PCR, Western blot, and immunocytofluorescence assays, and ET-1, IGF-1, VEGF, SDF-1, and HGF plasma concentrations were measured in enzyme-linked immunosorbent assays. RESULTS: Increased CECs and EPCs were found in the AMD patients compared with the counts in healthy individuals. The expression of intracellular ET-1 was significantly elevated in PBNCs from the AMD patients compared with the control subjects. In addition a significantly higher plasma concentration of IGF-1 was observed, but a lower SDF-1 level in the group of AMD patients. CONCLUSIONS: These findings suggest that circulating endothelial cells, together with high ET-1 content, may contribute to the development of AMD. Further prospective investigations on the mechanism involved may be relevant to the potential treatment of this disease.

[Evaluation of visual function among patients with exudative AMD treated with an intravitreal injection of triamcinolone acetonide]. / Ocena funkcji wzroku u pacjentów z wysiekowa postacia zwyrodnienia plamki zwiazanego z wiekiem leczonych doszklistkowym podaniem acetonidu triamcinolonu.

AIM: The aim of our study was to evaluate the visual acuity, visual field, retinal bioelectrical function and fluorescein angiography (FA) results of patients with exudative AMD treated with an intravitreal injection of triamcinolone acetonide (IVTA). MATERIAL AND METHODS: Visual acuity (Snellen chart), Humphrey automatic static perimetry (HASP) 30-2 W-W, mfERG were performed before, one month and three months after a single intravitreal injection of about 20 mg triamcinolone acetonide in 17 eyes of 17 patients (that did not meet the criteria for PDT). Fluorescein angiography was evaluated before and 3 months after treatment. RESULTS: There was no significant improvement in visual acuity after a month and three months after treatment when comparing to the initial examination. Although a slight improvement of 1-2 lines on Snellen chart was noted in individual cases. No significant changes were observed in static perimetry results after 3 months follow-up. In mfERG, there was a significant decrease in both, response density and P1 wave amplitude in fifth peripheral ring. In the rest of the rings, in comparison to the initial examination significant changes were not observed. In 76% of the analyzed eyes stabilization of CNV activity was noted in FA. In majority of eyes, the active CNV diameter did not change or was reduced and transformed into a scar. CONCLUSIONS: Three months after an intravitreal injection oftriamcinolone acetonide, no significant improvement of visual functions was noted in the observed group of patients with exudative AMD. A decrease of leakage in FA and a lack of a significant decrease in visual function may lead to a conclusion that IVTA treatment may slow down the natural course of the disease.

[In vitro level of L-ascorbic acid radical in lenses of patients with senile or diabetic cataract--preliminary study]. / Poziom rodnika askorbylowego in vitro w soczewkach z zacma starcza i cukrzycowa--badania wstepne.

INTRODUCTION: To estimate the in vitro level of L-ascorbic acid radical in lenses of patients with senile or diabetic cataract using EPR (Electron Paramagnetic Resonance) spectroscopy. MATERIAL AND METHODS: 24 human cataractous nuclei obtained during extracapsular removal were used. 5 of them (21%) were from diabetic patients. The analysis was carried out at the Department of Medical Physics of the Pomeranian Medical University using EPR spectroscopy. The in vitro level of L-ascorbic acid radical was calculated as the number of unpaired spins in the lens calculated in units x 10(16)/gram (spin/g), which is proportional to the in vivo level of vitamin C in lenses. RESULTS: The average L-ascorbic acid radical level in lenses of diabetic patients amounted 0.53 x 10(16) spin/g +/- 0.22 x 10(16) spin/g, and was lower than in group of non-diabetic patients in which it ranged 0.87 x 10(16) spin/g +/- 0.31 x 10(16) spin/g (p = 0.036). There was no significant correlation between L-ascorbic acid radical level and sex, age or visual acuity in these two groups of patients. CONCLUSIONS: (1) EPR spectroscopy can be used to determine the in vitro level of L-ascorbic acid radical in human lenses. (2) The in vitro L-ascorbic acid radical level in cataract lenses of patients with diabetes was lower than in patients without diabetes. (3). Lower in vitro level of L-ascorbic acid radical in lenses of patients with diabetes means lower in vivo level of vitamin C, what suggests an increased intensity of free radical reactions in the group of patients with diabetes than in the group without diabetes.