Katarzyna Szymanek

The role of base excision repair in the development of primary open angle glaucoma in the Polish population.

Glaucoma is a leading cause of irreversible blindness in developing countries. Previous data have shown that progressive loss of human TM cells may be connected with chronic exposure to oxidative stress. This hypothesis may suggest a role of the base excision repair (BER) pathway of oxidative DNA damage in primary open angle glaucoma (POAG) patients. The aim of our study was to evaluate an association of BER gene polymorphism with a risk of POAG. Moreover, an association of clinical parameters was examined including cup disk ratio (c/d), rim area (RA) and retinal nerve fiber layer (RNFL) with glaucoma progression according to BER gene polymorphisms. Our research included 412 patients with POAG and 454 healthy controls. Gene polymorphisms were analyzed by PCR-RFLP. Heidelberg Retinal Tomography (HRT) clinical parameters were also analyzed. The 399 Arg/Gln genotype of the XRCC1 gene (OR 1.38; 95% CI 1.02-1.89 p = 0.03) was associated with an increased risk of POAG occurrence. It was indicated that the 399 Gln/Gln XRCC1 genotype might increase the risk of POAG progression according to the c/d ratio (OR 1.67; 95% CI 1.07-2.61 P = 0.02) clinical parameter. Moreover, the association of VF factor with 148 Asp/Glu of APE1 genotype distribution and POAG progression (OR 2.25; 95% CI 1.30-3.89) was also found. Additionally, the analysis of the 324 Gln/His MUTYH polymorphism gene distribution in the patient group according to RNFL factor showed that it might decrease the progression of POAG (OR 0.47; 95% CI 0.30-0.82 P = 0.005). We suggest that the 399 Arg/Gln polymorphism of the XRCC1 gene may serve as a predictive risk factor of POAG.

Altered Expression Levels of MMP1, MMP9, MMP12, TIMP1, and IL-1β as a Risk Factor for the Elevated IOP and Optic Nerve Head Damage in the Primary Open-Angle Glaucoma Patients

The aim of presented work was to analyze the impact of particular polymorphic changes in the promoter regions of the -1607 1G/2G MMP1, -1562 C/T MMP9, -82 A/G MMP12, -511 C/T IL-1β, and 372 T/C TIMP1 genes on their expression level in POAG patients. Blood and aqueous humor samples acquired from 50 patients with POAG and 50 control subjects were used for QPCR and protein levels analysis by ELISA. In vivo promoter activity assays were carried on HTM cells using dual luciferase assay. All studied subjects underwent ophthalmic examination, including BCVA, intraocular pressure, slit-lamp examination, gonioscopy, HRT, and OCT scans. Patients with POAG are characterized by an increased mRNA expression of MMP1, MMP9, MMP12, and IL-1β genes as compared to the control group (P < 0.001). Aqueous humor acquired from patients with POAG displayed increased protein expression of MMP1, MMP9, MMP12, and IL-1β compared to the control group (P < 0.001). Allele -1607 1G of MMP1 gene possesses only 42,91% of the -1607 2G allele transcriptional activity and allele -1562 C of MMP9 gene possesses only 21,86% of the -1562 T allele. Increased expression levels of metalloproteinases can be considered as a risk factor for the development of POAG.

7D.07: RETINAL ARTERIOLAR STRUCTURE IN PATIENTS WITH PHEOCHROMOCYTOMA.

Objective: The aim of the ongoing study is to analyze retinal arteriolar structure in patients with pheochromocytoma (Pheo), a form of secondary hypertension characterized by excessive catecholamine secretion as compared with well-matched patients with essential hypertension (EHT). Figure. No caption available. Design and method: We examined 24 (15f/9m) patients with Pheo (mean age 46.3 ± 11.8 years) and 24 (15f/9m) age, gender, body mass index, glycemic status and blood pressure levels matched patients (46.7 ± 12.0 years) with (EHT). In all patients evaluation of plasma free normetanephrine (NMN) and metanephrine (MN) concentrations by liquid chromatography with tandem mass spectrometry was performed. Diagnosis of pheochromocytoma was made based on increased free NMN and/or MN concentration and confirmed on pathological examination. Retinal arterioles morphology were assessed by Heidelberg Retina Flowmetry using scanning laser Doppler flowmetry (SLDF). Outer diameter (AD) and lumen diameter (LD) were measured with automatic full-field perfusion imaging analysis (AFFPIA). Wall-to-lumen ratio (WLR), wall thickness (WT) and wall cross-sectional area (WCSA) of retinal arterioles were calculated. Retinal capillary blood flow (RCF) was also assessed by SLDF. Results: Pheo and EHT groups were well matched for age, gender, BMI, office and ambulatory blood pressure levels as well as for glycemic status and number of hypertensive medication (p > 0.05 for all comparisons). Patients with pheochromocytoma were characterized by higher AD, LD, WT and WCSA as compared with EHT (Table). A tendency towards higher WLR in patients with pheochromocytoma as compared to EHT was also noted (Table). There was no difference in RCF between patients with pheochromocytoma and EHT. Conclusions: Patients with pheochromocytoma as compared with matched patients with essential hypertension are characterized by higher outer wall diameter, lumen diameter, higher wall thickness and higher wall cross-sectional area of retinal arterioles. This may indicate potential deleterious effect of high catecholamine levels on small caliber arterioles evaluated non-invasively by SLDF.

The relationship between HDAC6, CXCR3, and SIRT1 genes expression levels with progression of primary open-angle glaucoma

ABSTRACT Background: Primary open-angle glaucoma (POAG) belongs to neurodegenerative diseases. Its etiology is not fully understood. However, a lot of reports have indicated that many biochemical molecules are involved in the retinal ganglion cell damage. Therefore, the purpose of this study was to evaluate a relationship between HDAC6, CXCR3, and SIRT1 genes expression levels with the occurrence risk of POAG and its progression. Materials and methods: The study included 34 glaucoma patients and 32 subjects without glaucoma symptoms. RNA was isolated from peripheral blood lymphocytes. Level of mRNA expression was determined by real-time PCR method. Results: Our results have shown significant association of the HDAC6 and SIRT1 expression levels with progression of POAG according to rim area (RA) value, p = 0.041; p = 0.012. Moreover, the analysis of the CXCR3 expression level showed a correlation with progression of POAG based on RA and cup disc ratio (c/d) value, p = 0.006 and p = 0.012, respectively. Conclusions: The expression level of HDAC6, CXCR3, and SIRT1 genes may be involved in the progression of POAG.

Association of the expression level of the neurodegeneration-related proteins with the risk of development and progression of primary open-angle glaucoma

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[OP.5A.04] EVALUATION OF RETINAL MICROPERFUSION AND ARTERIOLAR STRUCTURE IN PATIENTS WITH FIBROMUSCULAR DYSPLASIA. THE PRELIMINARY DATA OF THE POLISH REGISTRY FOR FIBROMUSCULAR DYSPLASIA

Objective: To compare retinal microperfusion and arteriolal structure between patients with fibromuscular dysplasia (FMD) enrolled to the ARCADIA-POL registry and patients with essential hypertension (EHT). Design and method: From the first 84 patients with confirmed FMD in any vascular bed enrolled in the ARCADIA-POL registry in 2015 (instituted on the basis of as Polish-French collaboration), we analyzed data regarding retinal microperfusion and arteriolar structure available from 70 patients (51F, 19 M, mean age: 41.9 ± 14.8 years). All patients underwent detailed clinical evaluation including ABPM, biochemical evaluation and biobanking as well as duplex Doppler of carotid and abdominal arteries and whole body angio-CT. We also evaluated 42 age, gender, body mass index, glycemic status, blood pressure levels and number of medication (p > 0.05) matched patients (24 F, 18 M, mean age: 46.1 ± 10.5 years) with EHT. Retinal microperfusion (RCF) and retinal arterioles were assessed using scanning laser Doppler flowmetry (SLDF). The parameters of retinal morphology: outer diameter (AD), lumen diameter (LD), wall/lumen ratio (WLR), wall thickness (WT), and wall cross-sectional area (WCSA) were determined by automatic full-field perfusion imaging analysis (AFFPIA V.4.011). Results: When comparing 70 patients with FMD to the matched group of patients with EHT we observed no significant differences in parameters describing retinal morphology. In further analysis we included 23 patients with confirmed FMD in carotid and/or intracranial arteries (22F, 1 M, mean age 43.6 ± 15.2ys) and we compared them to 26 matched patients (p < 0.05) with EHT (23F, 3 M, mean age: 48.8 ± 10.1 years). Patients with FMD affecting carotid and/or intracranial arteries were characterized by lower WLR (0.34 ± 0.08 vs. 0.39 ± 0.08, p = 0.048) and tendency towards lower WT (12.9 ± 3.7 vs.14.5 ± 3.3 &mgr;m, p = 0.15) and lower WCSA (3691.6 ± 1359.6 vs. 4106.8 ± 1222.1 &mgr;m2, p = 0.29) as compared to patients with EHT. There was no differences in AD, LD, nor RCF between the groups. Conclusions: In our ongoing ARCADIA-POL study we observed significant changes in retinal arterioles structure in patients with confirmed FMD in carotid and/or intracranial arteries.

[PP.12.17] EVALUATION OF SELECTED PARAMETERS OF NEURO-HORMONAL ACTIVITY AND RETINAL AND INTRARENAL PERFUSION IN PATIENTS WITH POLYCYTHEMIA VERA

Objective: Polycythemia vera (PV) is a myeloproliferative neoplasm primarily characterized by erythrocytosis. There is limited data on frequency and pathogenic mechanism of HT in patients with PV. Therefore the aim of the study is to present a selected clinical, neuro-humoral and hemodynamic characteristics of HT in patients with PV. Design and method: In the on-going study we included 20 consecutive patients (13F, 7 M, mean age 62 ± 9 years) with PV diagnosed (according to the WHO criteria) in one clinical center and 10 age, gender, body mass index, blood pressure (BP) and number of medication (p > 0.05) matched patients with essential HT (EHT). All patients underwent detailed clinical investigation which included among others: office and ambulatory BP measurements (ABPM), hormonal evaluations (renin, aldosterone and plasma metanephrines) and Doppler duplex ultrasound examination of renal arteries (resistive index [RI] was evaluated). Retinal microperfusion (RCF) and retinal arterioles were assessed using scanning laser Doppler flowmetry (SLDF). The parameters of retinal morphology were determined by automatic full-field perfusion imaging analysis (AFFPIA V.4.011). Muscle sympathetic nervous activity (MSNA) was evaluated by microneurography. Results: Hypertension on office or ABPM was present in 15 patients with PV (75%). Patients with PV as compared with EHT patients were characterized by less pronounced BP fall at night. There were: a trend towards lower serum aldosterone concentration and significantly lower plasma free epinephrine levels (25.9 ± 8.5 vs 36.8 ± 12.7 pg/mL; p = 0.048) in patients with PV. Patients with PV were characterized by lower 24 h heart rate on ABPM (63 ± 7 vs 75 ± 11 beats/min; p = 0.006) and lower MSNA as compared with EHT patients. RCF was lower in patients with PV (245 ± 51 vs 306 ± 49; p = 0.032) and it correlated significantly with red blood cell count in this group (r = -0.44; p = 0.012). There were no differences in parameters of retinal morphology between the groups. There was a significant difference in renal RI between PV and EHT groups (0.67 ± 0.06 vs 0.61 ± 0.07; p = 0.033). Conclusions: Patients with PV were characterized by lower indices of sympathetic nervous system activity and a tendency towards lower serum aldosterone concentrations. Alterations of retinal and renal perfusion in PV were also observed.

7D.11: Retinal arteriolar structure in patients with primary aldosteronism

Objective: Since retinal arteriolar structure has not been evaluated in patients with secondary hypertension we addressed a question if retinal arteriolar structure in patients with primary aldosteronism (PA) is altered as compared with well-matched patients with essential hypertension (EHT). Figure. No caption available. Design and method: We examined 30 (18m/12f) patients with PA (54.1 ± 9.5 years) and 30 (18m/12f) age, gender, body mass index, glycemic status, blood pressure levels and number of medication (p > 0.05) matched patients (55.8 ± 8.4 years) with EHT. All patients with PA underwent adrenal venous sampling to differentiate between aldosterone producing adenoma (APA) and bilateral adrenal hyperplasia (BAH). Retinal microperfusion (RCF) and retinal arterioles’ morphology were assessed by Heidelberg Retina Flowmetry using scanning laser Doppler flowmetry (SLDF). The parameters: outer diameter (AD) and lumen diameter (LD) were determined by automatic full-field perfusion imaging analysis (AFFPIA). Wall/lumen ratio (WLR), wall thickness (WT), and wall cross-sectional area (WCSA) were calculated. Results: Patients with PA were characterized by higher AD, WT, WLR and WCSA as compared with EHT (Table). There was no significant difference in LD, as well as in RCF between the groups (Table). Parameters describing retinal arterioles’ morphology were not correlated to office and ambulatory blood pressure both in the PA group and in the EHT group. There was no significant difference in parameters describing retinal morphology between BAH and APA groups. Conclusions: Patients with primary aldosteronism as compared to matched hypertensive controls are characterized by higher outer wall diameter, wall thickness, wall-to-lumen ratio and wall cross sectional area of retinal arterioles reflecting hypertrophic vascular remodeling. This may indicate the detrimental effect of excessive aldosterone on small retinal arterioles evaluated non-invasively by SLDF method.