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5-Fluorouracil (5-FU) is now used in eye drops for the management of conjunctival malignant melanoma, intraepithelial neoplasia, and corneal and conjunctival squamous cell carcinoma. The previously used methods for 5-FU quantification in AqH were time-consuming and less sensitive. Herein, a highly perceptive bioanalytical UPLC–MS/MS method was developed for the quantitative determination of 5-FU in the aqueous humor (AqH) of rabbits using allopurinol as the internal standard (IS). The 5-FU and IS were well separated in an Acquity™ HILIC column. Acetonitrile and 10 mM of ammonium acetate at 95:5 (v/v) were isocratically pumped at a 0.3 mL/min flow rate with a total runtime of 2.5 min. AqH samples were processed with a liquid–liquid extraction method in ethyl acetate. The 5-FU and IS were identified in the negative mode with electrospray ionization. The parent to daughter ion transitions for the 5-FU and IS occurred at m/z 128.92→41.68 and 134.80→64.10, respectively, as quantified using the multiple reaction monitoring mode. The developed method was validated with the ICH-Harmonized Guideline for Bioanalytical Method Validation, and the parameters were within acceptable limits. The calibration curve was linear at the 10.5–2000 ng/mL concentration range, with a correlation coefficient (R2) of 0.9946, and the lower limit of detection was 3.55 ng/mL. The developed and validated method was rapid, sensitive, accurate and robustly able to quantify 5-FU in rabbit AqH. The method was effectively applied to establish the ocular pharmacokinetics of 5-FU following the topical instillation of 5-FU-containing preparations in rabbits.

Sónia Torres-Costa,1 António Benevides Melo,1,2 Sérgio Estrela-Silva,1,2 Fernando Falcão-Reis,1,2 João Barbosa-Breda1– 4 1Department of Ophthalmology, Centro Hospitalar e Universitário São João, Porto, Portugal; 2Department of Surgery and Physiology, Faculty of Medicine, University of Porto, Porto, Portugal; 3UnIC@RISE, Faculty of Medicine of the University of Porto, Porto, Portugal; 4Research Group Ophthalmology, Department of Neurosciences, KU Leuven, Leuven, BelgiumCorrespondence: Sónia Torres-CostaOphthalmology Department, Centro Hospitalar Universitário de São João, Alameda Professor Hernani Monteiro, Porto, 4200, Portugal, Tel +351 225 512 100, Fax +351 225 025 766, Email sonia.torres.costa@gmail.comPurpose: To evaluate whether previous clear-cornea phacoemulsification surgery affects the surgical outcomes of trabeculectomy in open-angle glaucoma (OAG).Methods: We performed a retrospective cohort study, which included 82 patients with OAG that underwent trabeculectomy between January 1, 2010, and December 31, 2017. The primary outcome was the probability of surgical failure. Failure was defined as IOP > 21 mmHg or reduced < 20% from baseline, IOP ≤ 5 mmHg in three consecutive visits, need for further glaucoma surgery, phthisis or loss of light perception vision due to glaucoma.Results: Eighty-two eyes (58 phakic and 24 pseudophakic) were included. Phakic group patients were younger than those in the pseudophakic group, 65.8 ± 11.7 vs 76.2 ± 7.9 years (p < 0.001). The most common type of glaucoma was primary OAG [59% (n = 34) phakic vs 63% (n = 15) pseudophakic], followed by exfoliative and pigmentary glaucomas. The mean preoperative IOP was not significantly different between groups nor was the number of preoperative hypotensive medications. The rate of surgical failure was not significantly different between groups at year 1 [17% (n = 10) phakic vs 29% (n = 7) pseudophakic; p = 0.361] nor at year 2 [28% (n = 16) phakic vs 46% (n = 11) pseudophakic; p = 0.110]. No significant differences were observed regarding the postoperative IOP or any secondary outcome measures at year 1 or 2.Conclusion: Previous clear-cornea phacoemulsification surgery does not lead to statistically significant differences in the rate of trabeculectomy failure. Despite not being significant, clinically relevant differences were observed between groups. Future studies with a larger sample and/or randomized are needed to clarify this association.Keywords: trabeculectomy, phacoemulsification, surgical failure, intraocular pressure

Purpose of review Mitomycin C (MMC) is an alkylating agent with extraordinary ability to crosslink DNA, preventing DNA synthesis. By this virtue, MMC is an important antitumor drug. In addition, MMC has become the gold standard medication for glaucoma filtration surgery (GFS). This eye surgery creates a passage for drainage of aqueous humor (AqH) out of the eye into the sub-Tenon's space with the aim of lowering the intraocular pressure. A major cause of failure of this operation is fibrosis and scarring in the sub-Tenon's space, which will restrict AqH outflow. Intraoperative application of MMC during GFS has increased GFS success rate, presumably mainly by reducing fibrosis after GFS. However, still 10% of glaucoma surgeries fail within the first year. Recent findings In this review, we evaluate risks and benefits of MMC as an adjuvant for GFS. In addition, we discuss possible improvements of its use by adjusting dose and method of administration. One way of improving GFS outcome is to prolong MMC delivery by using a drug delivery system.

Purpose of review Mitomycin C (MMC) is an alkylating agent with extraordinary ability to crosslink DNA, preventing DNA synthesis. By this virtue, MMC is an important antitumor drug. In addition, MMC has become the gold standard medication for glaucoma filtration surgery (GFS). This eye surgery creates a passage for drainage of aqueous humor (AqH) out of the eye into the sub-Tenon's space with the aim of lowering the intraocular pressure. A major cause of failure of this operation is fibrosis and scarring in the sub-Tenon's space, which will restrict AqH outflow. Intraoperative application of MMC during GFS has increased GFS success rate, presumably mainly by reducing fibrosis after GFS. However, still 10% of glaucoma surgeries fail within the first year. Recent findings In this review, we evaluate risks and benefits of MMC as an adjuvant for GFS. In addition, we discuss possible improvements of its use by adjusting dose and method of administration. Summary One way of improving GFS outcome is to prolong MMC delivery by using a drug delivery system.

Keratoconus (KC) is a corneal ectatic condition characterized by focal structural changes, resulting in progressive thinning, biomechanical weakening, and steeping of the cornea that can lead to worsening visual acuity due to irregular astigmatism and corneal scarring in more advanced cases. It is a relatively common ectatic disease of the cornea predominantly affecting the younger population. Despite its worldwide prevalence, its incidence is rather varied with a higher incidence among the Middle Eastern and South Asian population. Dysregulated corneal extracellular matrix remodeling underlies KC pathogenesis. However, a lack of absolute clarity regarding the factors that initiate and drive progression poses a significant challenge in its prevention and management. KC is a complex multifactorial disease as it is associated with a wide variety of etiological factors such as environmental stimuli/insults, oxidative stress, genetic predisposition, comorbidities, and eye rubbing. A series of studies using corneal tissues (epithelium, stroma), cultured corneal fibroblasts/keratocytes, tear fluid, aqueous humor, and blood from KC subjects has reported significant alterations in various biochemical factors such as extracellular matrix components, cellular homeostasis regulators, inflammatory factors, hormones, metabolic products, and chemical elements. It has become apparent that alterations in the biochemical mediators (related to various etiologies) could contribute to KC pathogenesis by altering the dynamics of extracellular matrix remodeling events such as collagen deposition, degradation, and cross-linking in the cornea. Determining key disease contributing biochemical mediators would aid in disease monitoring, prediction or abatement of disease progression, and development of targeted therapeutics to improve disease prognosis.

Keratoconus (KC) is a corneal ectatic condition characterized by focal structural changes, resulting in progressive thinning, biomechanical weakening, and steeping of the cornea that can lead to worsening visual acuity due to irregular astigmatism and corneal scarring in more advanced cases. It is a relatively common ectatic disease of the cornea predominantly affecting the younger population. Despite its worldwide prevalence, its incidence is rather varied with a higher incidence among the Middle Eastern and South Asian population. Dysregulated corneal extracellular matrix remodeling underlies KC pathogenesis. However, a lack of absolute clarity regarding the factors that initiate and drive progression poses a significant challenge in its prevention and management. KC is a complex multifactorial disease as it is associated with a wide variety of etiological factors such as environmental stimuli/insults, oxidative stress, genetic predisposition, comorbidities, and eye rubbing. A series of studies using corneal tissues (epithelium, stroma), cultured corneal fibroblasts/keratocytes, tear fluid, aqueous humor, and blood from KC subjects has reported significant alterations in various biochemical factors such as extracellular matrix components, cellular homeostasis regulators, inflammatory factors, hormones, metabolic products, and chemical elements. It has become apparent that alterations in the biochemical mediators (related to various etiologies) could contribute to KC pathogenesis by altering the dynamics of extracellular matrix remodeling events such as collagen deposition, degradation, and cross-linking in the cornea. Determining key disease contributing biochemical mediators would aid in disease monitoring, prediction or abatement of disease progression, and development of targeted therapeutics to improve disease prognosis.

Purpose/Aim: Many genes have been associated with primary open-angle glaucoma (POAG). Knowing exactly where they are expressed in the eye helps to unravel POAG pathology and to select optimal targets for intervention. We investigated whether RNA in situ hybridization (RNA-ISH) is a convenient technique to obtain detailed pan-ocular expression data of these genes. We tested this for four diverse candidate POAG genes, selected because of unclear ocular distribution (F5 and Dusp1) and relevance for potential new therapies (Tnf, Tgf beta r3). Optn, a POAG gene with well-known ocular expression pattern served as control. Methods: We made a list of candidate glaucoma genes reported in genetic studies. A table of their ocular expression at the tissue level was compiled using publicly available microarray data (the ocular tissue database). To add cellular detail we performed RNA-ISH for Optn, Tnf, Tgf beta r3, F5, and Dusp1 on eyes of healthy, 2-month-old, pigmented, and albino mice. Results: Expression of the Optn control matched with published immunohistochemistry data. Ocular expression of Tnf was generally low, with patches of higher Tnf expression, superficially in the corneal epithelium. F5 had a restricted expression pattern with high expression in the nonpigmented ciliary body epithelium and moderate expression in the peripapillary region. Tgf beta r3 and Dusp1 showed ubiquitous expression. Conclusions: RNA-ISH is a suitable technique to determine the ocular expression pattern of POAG genes, adding meaningful cellular detail to existing microarray expression data. For instance, the high expression of F5 in the nonpigmented ciliary body epithelium suggests a role of this gene in aqueous humor dynamics and intraocular pressure. In addition, the ubiquitous expression of Tgf beta r3 has implications for designing TGF-beta-related glaucoma therapies, with respect to side effects. Creating pan-ocular expression maps of POAG genes with RNA-ISH will help to identify POAG pathways in specific cell types and to select targets for drug development.

Background Uveitis is one of the most frequent ophthalmologic manifestations in rheumatology. Uveal inflammation can underlie a systemic inflammatory rheumatic disease (SIRD) in approximately 30% of cases with a significant burden on the quality of life since it represents a cause of blindness in up to 20% of cases in Western countries. Methods In this review, we provide a comprehensive overview of the pathophysiology of uveitis associated with SIRDs. According to our literature survey on the epidemiology of uveitis among SIRDs, spondyloarthritides, Behcet's disease and sarcoidosis get the major impact. Results In Behcet's uveitis, the key players are highly polarized Th1 and Th17 lymphocytes, natural killer T cells and gamma delta T cells. All contribute to a great destructive inflammatory environment with the most serious visual damage resulting from the involvement of the posterior segment of the eye. In contrast, spondyloarthritides-related uveitis derives from a complex interaction between genetic background and extra-ocular inflammatory mediators originating from enthesitis, arthritis, psoriatic lesions and microbiome pro-inflammatory alterations. In such conditions, the immune infiltration of CD4+ T cells, Th17 and natural killer cells along with pro-inflammatory cytokines, TNF-alpha among all, leads to intraocular inflammation. Lastly, granuloma formation represents the primary hallmark lesion in sarcoid uveitis. This suggests a profound link between the innate system that mainly recruits activated macrophages and adaptive system involving by Th1, Th17 and Th17.1 cells. Conclusions Awareness among rheumatologists of a potential severe ocular involvement generates new insights into targeted therapeutic approaches and personalized treatments for each patient.

The main risk factor for primary open-angle glaucoma (POAG) is increased intraocular pressure (IOP). It is of interest that about half of the patients have an IOP within the normal range (normal-tension glaucoma, NTG). Additionally, there is a group of patients with a high IOP but no glaucomatous neurodegeneration (ocular hypertension, OHT). Therefore, risk factors other than IOP are involved in the pathogenesis of glaucoma. Since the retina has a very high oxygen-demand, decreased autoregulation and a fluctuating oxygen supply to the retina have been linked to glaucomatous neurodegeneration. To assess the significance of these mechanisms, we have utilized a human experimental model, in which we stress participants with a fluctuating oxygen supply. Levels of oxidative stress molecules, antioxidants, and lipid mediators were measured in the plasma. Patients with NTG, OHT, and control subjects were found to have similar levels of oxidative stress markers. In contrast, patients with OHT had a higher level of total antioxidant capacity (TAC) and pro-homeostatic lipid mediators. Thus, we suggest that OHT patients manage fluctuating oxygen levels more efficiently and, thus, are less susceptible to glaucomatous neurodegenerations, due to enhanced systemic antioxidant protection.

Intraocular pressure (IOP) is maintained as a result of the balance between production of aqueous humour (AH) by the ciliary processes and hydrodynamic resistance to its outflow through the conventional outflow pathway comprising the trabecular meshwork (TM) and Schlemm’s canal (SC). Elevated IOP, which can be caused by increased resistance to AH outflow, is a major risk factor for open-angle glaucoma. Matrix metalloproteinases (MMPs) contribute to conventional aqueous outflow homeostasis in their capacity to remodel extracellular matrices, which has a direct impact on aqueous outflow resistance and IOP. We observed decreased MMP-3 activity in human glaucomatous AH compared to age-matched normotensive control AH. Treatment with glaucomatous AH resulted in significantly increased transendothelial resistance of SC endothelial and TM cell monolayers and reduced monolayer permeability when compared to control AH, or supplemented treatment with exogenous MMP-3. Intracameral inoculation of AAV-2/9 containing a CMV-driven MMP-3 gene (AAV-MMP-3) into wild type mice resulted in efficient transduction of corneal endothelium and an increase in aqueous concentration and activity of MMP-3. Most importantly, AAV-mediated expression of MMP-3 increased outflow facility and decreased IOP, and controlled expression using an inducible promoter activated by topical administration of doxycycline achieved the same effect. Ultrastructural analysis of MMP-3 treated matrices by transmission electron microscopy revealed remodelling and degradation of core extracellular matrix components. These results indicate that periodic induction, via use of an eye drop, of AAV-mediated secretion of MMP-3 into AH could have therapeutic potential for those cases of glaucoma that are sub-optimally responsive to conventional pressure-reducing medications.

AMAÇ: Trabekülektomiye giden glokom hastalarinda hümör aköz TGF-β2 seviyesinin cerrahi başari üzerine etkisini değerlendirmek.GEREÇ VE YÖNTEM: Çalışma, Karadeniz Teknik Üniversitesi Tıp Fakültesi Göz Hastalıkları Kliniği'nde Ocak 2016 ve Nisan 2017 tarihleri arasında prospektif olarak gerçekleştirildi. Çalışmaya trabekülektomi uygulanmış 18 hastanın 19 gözü dahil edildi. Göz içi basıncının, ilaçsız 16 mmHg veya altında olması tam başarı, ilaçla 16 mmHg veya altında olması nitelikli başarı, bunun dışındaki göz içi basıncı değerleri başarısız olarak kabul edildi. Başarının değerlendirilmesi için Mann-Whitney testi uygulandı. İstatistiksel olarak `p` değerinin

Objectives: The aim of this study was to evaluate intraocular pressure (IOP) and retinal nerve fiber layer (RNFL) changes in patients with carotid artery stenosis (CAS) after carotid artery stenting. Materials and Methods: This study was conducted as a cross-sectional, non-randomised clinical case series. Fifteen male patients (mean age: 63.6ae9.1) with CAS and more than 70% carotid artery narrowing were included. All of the patients were followed in the department of neurology and were operated in the interventional radiology division. Eighteen healthy male subjects (mean age: 63.7ae5.3) were included in the control group. All of the healthy subjects had a detailed ophthalmological examination and subjects with any chronic eye disease were excluded from the study. All of the participants had a detailed ophthalmological examination including tonometry using Goldmann applanation tonometry and RNFL analysis using optical coherence tomography (RTVue-100 5.1). Results: There were no ocular ischemic symptoms in any of the participants. The mean IOP value was 15.1ae2.1 mmHg in the control group and 16.6ae2.4 mmHg before stent implantation, 16.4ae2.2 mmHg at 1 week after implantation, 16.6ae2.5 mmHg at 1 month after implantation, and 16.7ae2.9 mmHg at 3 months after implantation in the CAS group. Mean RNFL thickness was 105ae6 mu m in the control group; in the CAS group, mean RNFL thickness values were 98ae27 mu m before stent implantation and 103ae11 mu m, 101ae10 mu m, and 101ae11 mu m at 1 week, 1 month, and 3 months after stenting. There were no significant differences between the CAS group and control group regarding IOP and RNFL thickness values (p> 0.05). IOP and RNFL thickness also did not show any statistically significant changes from preoperative measurements in 3 months postoperative follow-up in the CAS group (p> 0.05). Conclusion: IOP and RNFL thickness remained unchanged after carotid stent implantation in carotid artery stenosis patients with no signs of ocular ischemic syndrome.

Amaç: Retinopatisi olan ve olmayan diabetik hastalarda aköz hümör ve serum BDNF düzeylerini belirlemek ve bu faktör düzeyinin hastaların RNFL ve GHK kalınlıklarıyla ilişkisini değerlendirmektir.Gereç ve Yöntem: Çalışma kapsamında retinopatisi olmayan DM'li, Non-PDR'li ve PDR'li hastaların kanları ve katarakt ekstraksiyonu sırasında aköz hümörleri toplandı ve BDNF düzeyleri ELİSA yöntemi ile değerlendirildi. Tüm gruplarda OCT ile RNFL ve GHK kalınlıkları ölçüldü.Bulgular: Ortanca serum BDNF düzeyleri kontrol grubunda 485.90 ng/ml (129.25-919.01), retinopatisi olmayan DM'li grupta 367.13 ng/ml (91.72-459.16), Non-PDR'li grupta 276.55 ng/ml (200.47-641.91), PDR'li grupta 188.69 ng/ml (102.93-591.79) olarak değerlendirildi. Ortanca aköz hümör BDNF düzeyleri kontrol grubunda 0.81 ng/ml (0.39-1.04), retinopatisi olmayan DM'li grupta 0.62 ng/ml (0.17-0.77), Non-PDR'li grupta 0.61 ng/ml (0.07-0.71), PDR'li grupta 0.54 ng/ml (0.07-0.70) olarak ölçüldü (sırasıyla p=0.002 ve p=0.0016, Kruskal Wallis). Bütün hasta gruplarında aköz hümör BDNF düzeyleri RNFL ve GHK düzeyiyle orta düzeyde pozitif korelasyon gösterirken, serum BDNF düzeyleri bu parametreler ile korele değildi.Sonuç: Beyinden türetilmiş nörotrofik faktör, retinopatisi olan ve olmayan diabetik hastaların serum ve aköz hümörlerinde azalmaktadır ve bu azalma RNFL ve GHK tabakalarındaki incelmeyle ilişkilidir. Purpose: To determine aqueous humor and serum BDNF levels in diabetic patients with and without retinopathy, and to assess the relationship between these factors and RNFL and GCC in the patients.Materials and Methods: BDNF levels in patients' blood and aqueous humor collected during cataract extraction were evaluated with ELISA in diabetic patients without retinopaty, with non-proliferative retinopathy and with proliferative retinopathy. In all groups, RNFL and GCC thicknesses were measured with OCT.Results: Median serum BDNF levels were found to be 485.90 ng / ml (129.25-919.01) in the control group, 367.13 ng / ml (91.72-459.16) in the DM group without retinopathy, 276.55 ng / ml (200.47-641.91) in the non-PDR group and 188.69 ng / ml (102.93-591.79) in the PDR group. The median aqueous humor BDNF levels were 0.81 ng / ml (0.39-1.04) in the control group, 0.62 ng / ml (0.17-0.77) in the DM group without retinopathy, 0.61 ng / ml (0.07-0.71) in the non-PDR group, 0.54 ng / ml (0.07-0.70) in the PDR group (p = 0.002 and p = 0.0016, Kruskal Wallis, respectively). While aqueous humor BDNF levels showed moderate positive correlation with RNFL and GCC levels in all patient groups, serum BDNF levels did not correlate with these parameters.Conclusion: BDNF is reduced in serum and aqueous humor of diabetic patients with and without retinopathy, and this decrease is related to the thinning of the RNFL and GCC layers. 77

Background: The aim of is to determine the association of vitamins with glaucoma by performing a systematic review and meta-analyses. Methods: Studies on the relation of vitamins and glaucoma published up to December 2017 were identified in the PubMed and Embase database. Data on vitamins (method of assessment), glaucoma (type and method of assessment), study characteristics and quality were recorded. In case of multiple studies for one nutrient a meta-analysis was performed. Results: A total of 629 articles were identified of which 36 were included in the systematic review. The meta-analysis included five of them (940 open-angle glaucoma (OAG) cases and 123,697 controls in total) and resulted in an odds ratio [95% confidence interval] (OR [95% CI]) of 0.58 [0.37-0.91] for dietary vitamin A, though heterogeneity was high (I-2 = 51%). After omitting studies that contributed significantly to the heterogeneity, the pooled OR [95% CI] was 0.45 [0.30-0.68] for dietary vitamin A on OAG (I-2 = 0%). For vitamin B1, C and E no significant association with OAG was found (OR [95% CI]: 0.84 [0.47-1.51]; 0.68 [0.38-1.22]; 0.95 [0.75-1.19]; respectively). However, after addressing heterogeneity, vitamin C showed a protective effect as well. Especially, foods high in these vitamins (e.g., dark green vegetables) were protective for OAG. Conclusions: Dietary intake of vitamin A and C showed a beneficial association with OAG; however, findings on blood levels of vitamins do not show a clear relation with OAG.

Purpose: To determine the intraocular and serum vascular endothelial growth factor (VEGF) levels in patients with acute retinal necrosis (ARN) and compare those with VEGF levels found in patients with ocular toxoplasmosis (OT).Methods: Paired intraocular fluid and serum samples of 17 patients with ARN and of 16 patients with OT were analyzed by ELISA for VEGF levels, and the clinical records were reviewed.Results: The mean intraocular VEGF levels in patients with ARN were higher than in patients with OT (P = 0.005), whereas the serum levels did not differ. Intraocular VEGF levels exceeded the serum levels in 8 of 17 patients (47%) with ARN compared with 1 of 16 patients (6%) with OT (P = 0.009). The group with high intraocular VEGF was associated with a more extensive retinitis and lower visual acuity at 3-month follow-up (P Conclusion: Intraocular VEGF levels were elevated in patients with ARN compared with OT patients. Our results suggest strong intraocular VEGF production in ARN, which might be of importance for the treatment of these patients. RETINA 30:1734-1738, 2010

Bu çalışmanın amacı psödoeksfoliasyon sendromunun gelişimine dahil olan olası mekanizmaları aydınlatmaktır.PES'li hastaların aköz hümör içeriğinin proteomiks analizi gerçekleştirildi ve PES olmayan katarakt hastalarının aköz hümör örnekleri ile karşılaştırıldı.15 PES'li hasta ve benzer demografik özelliklerde PES olmayan 15 kişiden oluşan kontrol grubu olmak üzere katarakt ameliyatı esnasında aköz hümör örnekleri alındı.Her örnek için protein konsantrasyon ölçümü yapıldıktan sonra havuzlama metodu uygulandı. Örnekler temizlenerek IPG striplerine yüklendi. Standart protokol kullanılarak odaklama ve ikinci boyutta ayrım gerçekleştirildi.Jeller Sypro ruby boyası ile boyandı ve PDQest Advance yazılımı ile analiz edildi.300 protein spotu tespit edildi ve bu spotlardan 10 tanesinin farklı ekspresyon özellikleri gösterdiği MALDI TOF/TOF ile tanımlandı. Analiz edilen proteinlerden özellikle iki tanesinin, APOA4 ve transtiretin, PES grubunda yüksek olarak eksprese olduğu görüldü.Bu sonuçler PES'in moleküler mekanizmasını anlamada yol gösterici olabilir ve biyomarker olarak kullanılabilir. The purpose of this study was to identify the possible mechanisms involved in the development of pseudoexfoliation syndrome. A proteomic analysis of the aqueous humor composition in the eyes of patients with PEX was performed and compared with that in the eyes of patients with cataract (non-PEX; controls). During cataract surgery ,aqueous humor from 15 patients with PEX and 15 patients with cataract (non-PEX) was collected. After measuring protein concentrations of each samples, pools were formed and cleaned up before loading onto IPG strips (11 cm, pH3-10). Focusing and second dimension separation were carried using standard protocols. Gels were stained with Spro Ruby and analyzed with PDQuest Advance software. 300 proteins spot were detected and 10 spots displaying differences in their expression were identified using MALDI TOF/TOF. Two of these proteins, namely ApoA4 and transthyretin, were highly over expressed in the PEX group.They may be significant in understanding molecular mechanism of PEX and may hold values as biomarkers. 68

Amaç: Bu çalısmada diyabetik ve non-diyabetik katarakt hastalarında hümöraközde total oksidatif stres (TOS), total antioksidan kapasite (TAK) seviyeleri karsılaştırılarak, diyabetik retinopatinin ilerlemesinde oksidatif stresin rolü değerlendirildi.Gereç ve Yöntem: Prospektif olarak planlanan bu çalısmada, olgular Ağustos 2014-Aralık 2014 tarihleri arasında katarakt cerrahisi tedavisi planlanan diyabetik ve diyabetik olmayan hastalar(toplam n:86) arasından seçildi. Katarakt tanısı dısında ek sistemik hastalığı olmayan hastalar 1. Grup (n:31), diyabeti olup retinopatisi olmayan hastalar 2. Grup (n:19), non-proliferatif diyabetik retinopatili hastalar 3. Grup (n:17) ve poliferatif diyabetik retinopatili hastalar 4. Grup (n:19) olarak ayrıldı. Hastalardan katarakt cerrahisi öncesi hümör aköz örneği alındı. Örneklerdeki TAS, TOS düzeyleri kolorimetrik yöntemle belirlendi ve gruplar arasında karşılaştırma yapıldı.Bulgular: 4 grubun hümör aköz sonuçları karsılastırıldıgında, total antioksidan seviye (TAS) düzeylerinin kontrol grubundan diyabetik retinopati evresine doğru ilerledikçe azaldığı, kontrol grubu (1.Grup) ile DRP grubu (Grup 3 ve 4) arasında anlamlı fark olduğu bulundu (sırasıyla p:0.042 ve p:0.02). Total oksidatif stres (TOS) düzeylerinde ise, 1. Grupta TOS düzeylerinin tüm diyabetik gruplara (Grup 2, 3 ve 4) göre anlamlı olarak daha düşük olduğu saptandı (p

Amaç: Serum ve aköz apolipoprotein (Apo) A1, Apo B değerleri ve Apo B/A1 oranı ile diyabetik retinopati şiddeti arasındaki ilişkiyi değerlendirmekGereç ve Yöntem: 60 tip 2 diabetes mellituslu hasta ve 61 kontrol hastası çalışmaya alındı. Serum ve aköz Apo A1, Apo B değerleri ve Apo B/A1 oranları karşılaştırıldı. Yaş, diyabet süresi, hipertansiyon (HT), açlık kan şekeri, HbA1c, serum lipid profili, makula kalınlık ölçümleri ile diyabetik retinopati arasındaki ilişki değerlendirildi.Bulgular: Çalışmaya katılan hastaların ortalama diyabet süresi 14.12±7.30 yıldı. Diyabetik hasta grubu ve kontrol grubu arasında serum Apo A1, Apo B değerleri, serum Apo B/A1 oranı ve aköz Apo A1 değerleri arasında fark yoktu (P>0.05). Aköz sıvıda Apo B saptanması kontrol grubunda %4.9; diyabetik hasta grubunda %61.7 idi. Aközde Apo B saptanan hastalarda diyabetik retinopati şiddeti daha fazlaydı (P0.05). Apo B level was measured as 4.9% in control group and 61.7% of diabetic patients in aqueous samples. Diabetic patients with Apo B in the aqueous had more severe diabetic retinopathy (P

The purpose of this study was to evaluate in vitro cytotoxicity and antifibrotic effects of mitomycin C on normal and scarred human vocal fold fibroblasts.Fibroblasts were subjected to mitomycin C treatment at 0.2, 0.5, or 1 mg/mL, or serum control. Cytotoxicity, immunocytochemistry, and Western blot for collagen I/III were performed at days 0, 1, 3, and 5.Significant decreases in live cells were measured for mitomycin C-treated cells on days 3 and 5 for all doses. Extracellular staining of collagen I/III was observed in mitomycin C-treated cells across all doses and times. Extracellular staining suggests apoptosis with necrosis, compromising the integrity of cell membranes and release of cytosolic proteins into the extracellular environment. Western blot indicates inhibition of collagen at all doses except 0.2 mg/mL at day 1.A total of 0.2 mg/mL mitomycin C may provide initial and transient stimulation of collagen for necessary repair to damaged tissue without the long-term risk of fibrosis.

Amaç: Koruyucu olarak polyquad içeren antiglokomatöz damlalar ile koruyucu olarak benzalkonium chloride (BAC) içeren antiglokomatöz damlaların kornea epiteli üzerine ve gözyaşı üretimi üzerine etkilerini değerlendirmek ve bu etkileri karşılaştırmak. Gereç ve yöntem: Koruyucu olarak polyquad içeren antiglokomatöz damla tedavisi başlanan grupta (Pol grubu) 21 primer açık açılı glokom veya oküler hipertansiyonlu hastanın 40 gözü, koruyucu olarak benzalkonium chloride içeren antiglokomatöz damla tedavisi başlanan grupta (BAC grubu) 20 primer açık açılı glokom veya oküler hipertansiyonlu hastanın 37 gözü bulunmaktaydı. Hastalar 4 ile 7 ay boyunca takip edilmişlerdir. Çalışmada 20 sağlıklı gönüllünün 40 gözü de kontrol grubunu oluşturmuştur. Epitel dansitesini değerlendirmede, İn vivo konfokal mikroskop (Heidelberg Retina Tomografisi/Rostock Kornea Modülü), gözyaşı üretimini değerlendirmede ise Schirmer 1 testi kullanılmıştır.Bulgular: Takibin sonunda başlangıçtakine göre kornea yüzeyel epitel dansitelerinde azalma gerçekleşmiş ve bu azalma hem Pol grubunda hem de BAC grubunda istatistik olarak anlamlı bulunmuştur (Her iki grupta p

Amaç: Tavşanlarda Korneal Kollajen Cross-linking öncesinde, standart, transepitelyal ve korneal epitel bütünlüğünü bozmadan iyontoforez yardımı ile riboflavin uygulaması sonrasında kornea ve humör aközde riboflavin konsantrasyonlarının incelenmesi.Metod: Çalışmada 12 erişkin Yeni Zelanda tavşanının 24 gözü kullanılmıştır. Her grupta 6 göz olacak şekilde 4 grup oluşturulmuştur. Standart yöntemde epitel debridmanı sonrası %0.1 riboflavin , transepitelyal yöntemde 1.5 dakika arayla 3 kez BAC, EDTA, Tris, HPMC, % 0.25 riboflavin ardından 4 kez % 0.25 riboflavin, iyontoforez yardımı ile korneal epitel bütünlüğünü bozan maddeler kullanılmadan 10 dk boyunca 1mA elektrik akımı kullanılarak %0.1 ve %0.25 riboflavin uygulanmıştır. İşlem sonrası alınan humör aköz ve korneadaki riboflavin konsantrasyonları Ultra Performans Sıvı Kromatografisi (UPLC-ESI-MS/MS) kullanılarak ölçülmüştür.Sonuç: Standart yöntemde korneal ve humör aközdeki riboflavin konsantrasyonları diğer gruplardan yüksek saptandı. İyontoforez yardımı ile %0.25 riboflavin uygulanan gözlerdeki korneal riboflavin konsantrasyonu hem transepitelyal yöntemden (p=0.009), hem de iyontoforez yardımı ile %0.1 riboflavin uygulanan gözlerden (p=0.026) anlamlı düzeyde yüksek ölçüldü. Transepitelyal ve iyontoforez yardımı ile %0.1 riboflavin uygulanan gözlerde korneal riboflavin konsantrasyonu açısından fark saptanmadı (p=0.937)Yorum: Korneal epitel bütünlüğünü bozmadan iyontoforez yardımı ile %0.25 riboflavin uygulaması sonrasında, standart uygulamadan daha düşük, transepitelyal uygulamadan daha yüksek düzeylerde intrastromal ve aköz riboflavin konsantrasyonları ölçülmüştür. Riboflavin konsantrasyonunu çok hassas bir şekilde ölçümlediğimiz bu çalışmada, iontoforez yönteminin epitel bütünlüğünü bozan maddeler kullanılmadan da riboflavin molekülünün korneaya geçişini arttırdığı izlenmiştir. Purpose: To examine riboflavin concentrations in corneas and aqueous humor with standard and transepithelial methods and iontophoresis without disrupting integrity of corneal epithelium in rabbits before corneal collagen cross-linking.Methods: Twenty-four eyes of 12 adult New Zealand rabbits were used. They were assigned into four groups, each including six eyes. Group 1 was exposed to the standard method and given riboflavin 0.1% following epithelial debridement. Group 2 was exposed to the transepithelial method and given BAC, EDTA, TRIS, HPMC and riboflavin 0.25% three times at 1.5-minute intervals followed by riboflavin 0.25%. Group 3 was given riboflavin 0.1% by using 1mA electric current for ten minutes with the help of iontophoresis without using substances disrupting integrity of corneal epithelium. Group 4 received the same treatment as Group 3 except that it was given riboflavin 0.25%. Following these treatments, riboflavin concentrations in aqueous humor and corneas were measured with ultra-performance liquid chromatography (UPLC-ESI-MS/MS).Results: Riboflavin concentrations in the cornea and aqueous humor were higher in Group 1 than in the other groups. They were significantly higher in Group 4 than in Group 2 (p=0.009), and Group 3 (p=0.026). There was not a significant difference in corneal riboflavin concentrations between Group 2 and Group 3 (p=0.937)Conclusion: Intrastromal and aqueous riboflavin concentrations after administration of riboflavin 0.25% through iontophoresis without disrupting integrity of the corneal epithelium were lower than those after the standard method, but higher than those after the transepithelial method. In the present study, in which riboflavin concentrations were measured with a very sensitive method, iontophoresis was observed to increase transmission of riboflavin molecules into the cornea without using substances disrupting the epithelial integrity. 80

Difüzyon ağırlıklı MR görüntüleme (DAMRG) su moleküllerinin difüzyon hareketineduyarlı bir görüntüleme yöntemidir. DAMRG günümüzde en sık akut serebral iskemininerken dönem tanısında kullanılmaktadır. Son yıllarda DAMRG bazı orbital patolojilerindeğerlendirilmesinde de kullanılmaya başlanmıştır.Klinik olarak çeşitli sebeplerle bölümümüzde beyin diffüzyon MRG tetkiki yapılan 40Diabetes Mellitus hastası ve oküler, serebral patoloji saptanmamış 40 kontrol grubu hastasıolmak üzere toplam 80 beyin diffüzyon MRG üzerinden retrospektif olarak yürüttüğümüz buçalışmada DM? i olan ve olmayanlar arasında yaş grupları, cinsiyet ve göz tarafı değişkeninebağlı olarak oküler ADC değerleri arasında olabilecek farklılıkları araştırmayı amaçladık.DM grubunun yaş ortalaması 54.58 ±10.13 yıl, kontrol grubunun yaş ortalaması 53.68 ± 5.38yıl olarak saptandı. DM grubunda 24 (% 60) erkek, 16 (% 40) kadın olgu vardı. Kontrolgrubunda 20 (% 50) erkek, 20 (% 50) kadın olgu vardı.40 kişilik DM grubundan elde edilen 80 gözün orbital ADC değerleri ile tüm orbitadanalınan ADC değerleri, kontrol grubundan istatistiksel olarak anlamlı düzeyde düşüktü. DM?i olan grup ve DM? i olmayan kontrol grubunda sağ göz ve sol gözden alınan ADC değerleriarasında istatistiksel olarak anlamlı fark saptanmadı. DM grubundaki olgulardeğerlendirildiğinde; 50 yaş ve altındaki bireylere ait ADC değerleri, 50 yaş üstü bireylereait ADC değerlerinden istatistiksel olarak anlamlı düzeyde yüksekti. Kontrol grubundakiolgular değerlendirildiğinde; 50 yaş ve altındaki olguların ADC değerleri ile 50 yaş üstüolguların değerleri arasında istatistiksel olarak anlamlı düzeyde fark yoktu. DM? i olan grupve DM? i olmayan kontrol grubunda kadınlar ve erkeklerden alınan ADC değerleri arasındaistatistiksel olarak anlamlı fark saptanmadı.Sonuç olarak Diabet hastalarında vitreus humor değişikliklerinin değerlendirilmesindedifüzyon MRG kullanılabilir. Diffusion-weighted magnetic resonance imaging (DW-MRI) is an imaging methodsensitive to the movement of the diffusion of water molecules. DW-MRI is mostwidely used in the diagnosis of early acute cerebral ischemia. In recent years, DWMRIhas been used in the assessment of some of the orbital pathology.We aimed to investigate possible differences in ocular ADC values in patients withand without DM. We also searched for the effect of age and gender on the ocularADC values. Brain DW-MRI scans of 40 patients with diabetes mellitus, and 40patients without diabetus mellitus with normal MRI were included in to thisretrospective study.Mean age 54.58 ± 10.13 years DM group and the control group mean age was 53.68± 5:38 years. DM group, 24 (60%) men and 16 (40%) were female patients. Thecontrol group consisted of 20 (50%) males and 20 (50%) female patients.Mean orbital ADC value of the DM group (80 eyes of 40 patients) was significantlylower than the control group.There was no statistically significant difference between the mean ADC values fromthe right and the left eyes in the group with DM and the control group.When the DM group patients were evaluated; ADC values of under the age of 50 wasstatistically significantly higher than ADC values of over the age of 50.6When the control group patients were evaluated; there was no significant differencebetween ADC values of patients under 50 years of age and over 50 years of age .There was no statistically significant difference between the ADC values of womenand men in both the DM group patients and the control group patients without DM.As a result, the assessment of changes in vitreous humor, diffusion-weighted MRIcan be used in patients with diabetes. 73

Purpose: The ciliary body (CB) of the human eye consists of the non-pigmented (NPE) and pigmented (PE) neuro-epithelia. We investigated the gene expression of NPE and PE, to shed light on the molecular mechanisms underlying the most important functions of the CB. We also developed molecular signatures for the NPE and PE and studied possible new clues for glaucoma.Methods: We isolated NPE and PE cells from seven healthy human donor eyes using laser dissection microscopy. Next, we performed RNA isolation, amplification, labeling and hybridization against 44xk Agilent microarrays. For microarray conformations, we used a literature study, RT-PCRs, and immunohistochemical stainings. We analyzed the gene expression data with R and with the knowledge database Ingenuity.Results: The gene expression profiles and functional annotations of the NPE and PE were highly similar. We found that the most important functionalities of the NPE and PE were related to developmental processes, neural nature of the tissue, endocrine and metabolic signaling, and immunological functions. In total 1576 genes differed statistically significantly between NPE and PE. From these genes, at least 3 were cell-specific for the NPE and 143 for the PE. Finally, we observed high expression in the (N)PE of 35 genes previously implicated in molecular mechanisms related to glaucoma.Conclusion: Our gene expression analysis suggested that the NPE and PE of the CB were quite similar. Nonetheless, cell-type specific differences were found. The molecular machineries of the human NPE and PE are involved in a range of neuro-endocrinological, developmental and immunological functions, and perhaps glaucoma.

Bu çalışmada; serum ve aköz hümörde iskemi ile modifiye olmuş albümin (IMA), total thiol, totol antioksidan kapasite (TAK) ve total oksidatif sters (TOS) düzeylerine bakılarak diabetik retinopati (DRP) gelişiminde iskemi ve oksidatif stresin rolünü belirlemek amaçlanmıştır.Prospektif olarak planan bu çalışmaya, olgular 01.01.2012 ile 30.07.2012 tarihleri arasında, C. Ü. Tıp Fak. Göz Hast. A. D. Polikliniğine, puslu görme şikayeti ile başvuran ve katarakt tanısı alıp, cerrahi tedavi önerilen DRP' si olan ve diyabetik olmayan hastalar arasından seçildi. Katarakt operasyonu yapılacak olan; DRP' si olan diabetes mellitus (DM) hastaları çalışma grubu olarak, DM'u olmayan katarakt cerrahisi uygulanan hastalar kontrol grubu olarak çalışmaya dahil edildi. Her bir grup ayrı ayrı 22 hasta olacak şekilde toplam 44 hasta çalışmaya dahil edildi.Sonuç olarak, çalışmamızda DRP'li grupta ortalama serum IMA düzeyini istatistiksel olarak anlamlı yüksek, total thiol düzeyini istatistiksel olarak anlamlı düzeyde düşük bulduk. TAK ve TOS düzeyleri açısından istatistiksel olarak anlamlı bir fark yoktu. Çalışmanın ikinci basamağı olan aköz hümörde ise TOS düzeyini istatistiksel olarak anlamlı yüksek bukduk. IMA hiç tespit edemedik, total thiol ve TAK açısından ise istatistiksel olarak anlamlı bir fark saptamadık. Bu bulgular; DRP patogenezinde oksidatif stres ve iskeminin rol aldığı düşüncesini desteklemektedir. Ancak daha geniş vaka serilerinde ve retinopatinin eşlik etmediği diyabetik hasta gruplarınında eklendiği kapsamlı çalışmalara ihtiyaç duyulmaktadır.Anahtar Kelimeler: Diyabetik retinopati, iskemi ile modifiye olmuş albümin, total thiol, total anti oksidan kapasite, total oksidatif stres, aköz hümör, oksidatif stres. In this study Ischemia modified albumin (IMA), total thiol, total antioxidant capacity (TAC) and total oxidative stress (TOS) levels were examined in human serum and aquose humor to investigate the role of ischemia and oxidative stress in developement of diabetic retinopathy.This prospective study includes patients diagnosed with or without diabetic retinopathy who applied to C. U . Medical Faculty Ophthalmology Department Policlinic with hazy vision who are diagnosed with cataract and adviced to undergo cataract extraction. Patients who are diagnosed with cataracts and diabetic retinopathy are identified as study group,other patients who does not have diabetes but cataracts are identified as control group.In each group there were 22 patients which equals 44 patients total.In conclusion, we observed that patients diagnosed with DRP has mean levels of IMA were higher but total thiol levels were lower. However, TAC and TOS levels weren?t statistically significant. In second part of the study where aquos humor is examined TOS levels were higher, IMA could not ve identified and total thiol and TAC weren?t statistically significant. These findings proves that oxidative stres and ischemia take part in pathogenesis of DRP. However, further studies which includes larger case series and patients with diabetes who does not have diabetic retinopathy are needed.Key Words: Diabetic retinopathy, ischemia modified albumin, total thiol, total antioxidant capacity, total oxidative stres, aquose humor, oxidative stress. 44

Open-angle glaucoma (glaucoma) is a major eye disorder characterized by optic disc pathology. Recent genome-wide association studies identified new loci associated with clinically relevant optic disc parameters, such as the optic disc area and vertical cup-disc ratio (VCDR). We examined to what extent these loci are involved in glaucoma. The loci studied include ATOH7, CDC7/TGFBR3 and SALL1 for optic disc area, and CDKN2B, SIX1, SCYL1/LTBP3, CHEK2, ATOH7 and DCLK1 for VCDR. We performed a meta-analysis using data from six independent studies including: the Rotterdam Study (n = 5736), Genetic Research in Isolated Populations combined with Erasmus Rucphen Family study (n = 1750), Amsterdam Glaucoma Study (n = 296) and cohorts from Erlangen and Tubingen (n = 1363), Southampton (n = 702) and deCODE (n = 36 151) resulting in a total of 3161 glaucoma cases and 42 837 controls. Of the eight loci, we found significant evidence (P = 1.41 x 10(-8)) for the association of CDKN2B with glaucoma [odds ratio (OR) for those homozygous for the risk allele: 0.76; 95% confidence interval (CI): 0.70-0.84], for the role of ATOH7 (OR: 1.28; 95% CI: 1.12-1.47) and for SIX1 (OR: 1.20; 95% CI: 1.10-1.31) when adjusting for the number of tested loci. Furthermore, there was a borderline significant association of CDC7/TGFBR3 and SALL1 (both P = 0.04) with glaucoma. In conclusion, we found consistent evidence for three common variants (CDKN2B, ATOH7 and SIX1) significantly associated with glaucoma. These findings may shed new light on the pathophysiological protein pathways leading to glaucoma, and point to pathways involved in the growth and development of the optic nerve.

Diabet, insülin sekresyon yetersizliği ya da dokularda insülin direnci artması sonucu gelişen metabolik bir hastalıktır. Diabet ve diabetin komplikasyonlarının oluşumunda eser element metabolizması önemli bir yer tutmaktadır.Çeşitli çalışmalarda diabette farklı doku, kan ve idrar örneklerinde pek çok elementin seviyelerinin değiştiği belirtilmektedir. Diabete bağlı katarakt ve glokom oluşumunda lenslerdeki element değişimleri incelenmesine rağmen, plazma ve intraoküler sıvıda (aköz hümör sıvı) element konsantrasyonlarının diabetik ve diabetik olmayan durumlarda karşılaştırmalı olarak değerlendiren araştırma oldukça azdır. Bu nedenle diabetik tavşan modelinde tavşanların kan, plazma ve intraoküler (aköz hümör) sıvılarında Zn, Cu ve Cr eser element seviyelerini araştırmayı amaçladık.Çalışmamızda Yeni Zellanda ırkı erkek tavşanlar kullanıldı. Tavşanlar kontrol (n=8) ve alloksan uygulanarak oluşturulan diabet (n=8) grubu olarak ikiye ayrıldı. Deney süresi sonunda (6 hafta) tavşanlardan alınan kan, plazma ve aköz hümör sıvılarının osmotik basınçları osmometre ile plazma ve aköz hümör sıvı Zn, Cu ve Cr elementlerinin konsantrasyonu ICP-OES cihazı ile ölçüldü.Diabet grubunun kan, plazma ve aköz hümör sıvı osmotik basınç değerlerinin kontrollere göre anlamlı (sırası ile p

Glokom, uygunsuz göz içi basıncı ve oküler kan dolaşım bozukluğu sonucu, optik sinir başında atrofi ile sonlanan bir grup hastalığa denir. Glokoma eşlik eden katarakt, hastanın glokomdan kaynaklanan görme azlığını daha da azaltarak yaşam kalitesini düşürdüğü gibi glokomun takibini de zorlaştırmaktadır. Katarakta bağlı görme azlığı, glokomdan kaynaklananın aksine cerrahi ile geri döndürülebilmektedir. Katarakt için günümüzde en uygun ameliyat yöntemi fakoemülsifikasyon cerrahisidir.Bu çalışmaya kliniğimizde katarakt cerrahisi uygulanan 45 glokomlu (hasta grubu) ve 21 glokomu olmayan (kontrol grubu) hasta dahil edilmiştir. Hastaların hepsine fakoemulsifikasyon cerrahisi ve intraoküler lens implantasyonu uygulanmıştır. Hiçbir hastada komplikasyon görülmemiştir. Çalışmada gruplar arasında ameliyat öncesi ve ameliyat sonrası ilk 6 ayda göz içi basıncı seviyesi, görme keskinliği, ön kamara derinliği, ön kamara açısı, kullanılan antiglokomatöz ilaç sayısı ile cup/disk oranlarının karşılaştırılması planlanmıştır.Çalışmamızda yapılan istatistiksel analizler sonucunda görme keskinliği hem glokomlu hastalarda hem de kontrol grubunda ameliyat sonrasında istatistiksel olarak anlamlı artış gösterirken (p

Amaç: Ülkemizde sık rastlanan psödoeksfoliasyon sendromunda (PES) etiopatogenez halen tartışmalı bir konudur. Bu çalışmada PES bulunan ve PES bulunmayan bireyler arasında hümör aközde (HA) ve serumda matriks metalloproteinaz doku inhibitörleri 1 (TIMP-1), matriks metalloproteinaz 9 (MMP-9), bağ dokusu büyüme faktörü (CTGF) ve vasküler endotelyal büyüme faktörü (VEGF) düzeylerinin farklı olup olmadığını ve dolayısıyla bu maddelerin regülasyonunun gelecekte PES tedavisinde potansiyel rolü olup olamayacağını incelemeyi amaçladık.Gereç ve Yöntem: Çalışmaya katarakt ameliyatı geçiren toplam 72 hastanın 72 gözü (PES grubu n:30, katarakt haricinde okuler patolojisi bulunmayan kontrol grubu n:42) dahil edildi. Pseudoeksfoliatif glokomu (PEG) olan PES hastaları ?PEG +? alt grubu, glokom saptanmayan PES hastaları ise ?PEG -? alt grubu olarak belirlendi. Hastalardan operasyon gününde koldan 5 ml kan örneği, operasyon başlangıcında ön kamaradan 100-200 ?l HA örneği alındı. Kan örneklerinin santrifüjü sonrası elde edilen serum örnekleri ile HA örnekleri derin dondurucuda analiz edilinceye kadar muhafaza edildi. HA örneklerinde TIMP-1 ve MMP-9 düzeyleri, serum örneklerinde TIMP-1, MMP-9, CTGF ve VEGF düzeyleri ELİSA kitleri kullanılarak belirlendi.Bulgular: Ortalama HA MMP-9 ve TIMP seviyeleri PES grubunda sırasıyla 3,54±5,09 ng/ml ve 31,2±37,9 ng/ml, kontrol grubunda sırasıyla 4,16±6,11 ng/ml ve 21,1±16,08 ng/ml olarak saptandı (p>0.05). Serum MMP-9 ve TIMP seviyeleri PES grubunda sırasıyla 283,7±348,6 ng/ml ve 122,4±150,3 ng/ml, kontrol grubunda sırasıyla 427,4±435,7 ng/ml ve 97,5±57,9 ng/ml olarak saptandı (p>0.05). Serum örneklerinde ortalama VEGF ve CTGF düzeyleri PES grubunda sırasıyla 333,08±241,73 ng/ml ve 570,95±1222,13 ng/ml, kontrol grubunda sırasıyla 381,73±226,2 ng/ml ve 449,48±459,64 ng/ml olarak saptandı (p>0.05). HA' de ortalama TIMP-1 seviyesi PEG+ alt grubunda PEG- alt grubuna göre belirgin derecede yüksek (p=0.012), HA ortalama MMP-9 seviyesi ise PEG+ alt grubunda PEG- alt grubundan daha düşük olarak belirlendi (p>0.05). Ortalama serum TIMP-1 seviyesi PEG- alt grubunda PEG+ alt grubuna göre daha yüksek, ortalama serum MMP-9 seviyesi PEG+ alt grubunda PEG- alt grubuna göre daha düşük olarak saptandı (p>0.005). Ortalama serum CTGF düzeyi PEG+ alt grubunda PEG- alt grubundan daha yüksek iken (p>0.005), ortalama serum VEGF düzeyi iki alt grup arasında çok yakın düzeydeydi (p>0.005).Sonuç: PES bulunan olgularda gerek HA gerekse serumda MMP , TIMP ve CTGF düzeylerinde PES bulunmayan olgulardan farklılıklar mevcuttur. Bu durum ekstrasellüler matriks homeostazının ekstrasellüler matriksin birikimi yönünde bozulduğunu desteklemektedir. PEG mevcut olgularda MMP-9, TIMP ve CTGF düzeylerinin glokomu olmayan PES olgularından farklı bulunması PEG' in PES tablosunun daha ileri bir formu olduğunu düşündürmektedir. Gelecekte MMP-9, TIMP ve CTGF arasındaki ilişkileri düzenleyecek koruyucu tedavilerin geliştirilmesiyle PES' in gelişiminin engellenebileceği veya yavaşlatılabileceği öngörülebilir.Anahtar kelimeler: psödoeksfoliasyon sendromu, psödoeksfoliasyon glokomu, hümör aköz, ekstrasellüler matriks, MMP-9, TIMP-1, CTGF. Purpose: Etiopathogenesis of pseudoexfoliation syndrome (PES) which is commonly seen in our contry is stil controversial. In this study we aimed to determine if aqueous humor (AH) and serum levels of tissue inhibitor of matrix metalloproteinase 1 (TIMP-1), matrix metalloproteinase 9 (MMP-9), connective tissue growth factor (CTGF) and vascular endothelial growth factor (VEGF) differred between patients with PES and without PES, thus whether regulation of these substances might have a potential role in the treatment of PES.Materials and Methods: Seventy-two eyes of 72 patients (PES group n:30, control group without any other ocular pathology except cataract n:42) undergoing cataract surgery were included in the study. Patients with pseudoexfoliative glaucoma (PEG) were further considered as PEG+ subgroup and patients without glaucoma as PEG- subgroup. Blood specimens (5 ml) were collected on the day of cataract surgery from the forearm and AH samples from the anterior chamber (100 ?l ) during cataract surgery. Serum specimens obtained after centrifugation of blood samples and AH samples were stored in deep freezer until analysis. TIMP-1 and MMP-9 levels were determined in AH samples and TIMP-1, MMP-9, CTGF and VEGF levels were determined in serum samples using specific enzyme immunoassays (ELISA).Results: The mean MMP-9 and TIMP-1 AH levels were 3.54 ± 5.09 ng / ml and 31.2 ± 37.9 ng / ml in the PES group, and 21.1 ± 16.08 ng / ml and 31, 2 ± 37.9 ng / ml in the control group, respectively (p>0.05) . The mean serum levels of MMP-9 and TIMP-1 were 283.7 ± 348.6 ng / ml and 122.4 ± 150.3 in the PES group, and 427.4 ± 435.7 ng / ml and 97.5 ± 57.9 ng / ml in the control group, respectively (p>0.05). The mean VEGF and CTGF levels in serum samples were 333,08±241,73 ng/ml and 570,95±1222,13 ng/ml in the PES group, and 381,73±226,2 ng/ml and 449,48± 459,64 ng/ml, in the control group respectively (p>0.05). HA TIMP levels were significantly higher in PEG+ subgroup compared to PEG- subgroup (p=0.012), while HA MMP-9 levels were lower in PEG+ subgroup compared to PEG- subgroup (p>0.05). Mean serum TIMP-1 level was higher in PEG - subgroup compared to PEG+ subgroup , and mean serum MMP-9 level was lower in PEG+ subgroup compared to PEG- subgroup (p>0.005). Mean serum CTGF levels were higher in PEG+ subgroup compared to PEG- subgroup (p>0.005), while mean serum VEGF levels were very close in both subgroups (p>0.005).Conclusion: MMP-9, TIMP-1 and CTGF levels differ in PES patients from patients without PES in both AH and serum. This suppports that extracellular homeostasis is disturbed resulting in accumulation of extracellular matrix. The findings that patients with PEG had different levels of MMP-9, TIMP-1 and CTGF compared to PES patients without glaucoma make think that PEG is a more severe form of PES. It might be foreseen that development of preventive treatments in the future by regulation of interactions between MMP-9, TIMP-1 and CTGF will inhibit or slow down the progression of PES.Key Words: pseudoexfoliation syndrome, aqueous humor, extracellular matrix, MMP-9, TIMP-1, CTGF 70

ÖZETBu çalışmada diyabetik ve non-diyabetik katarakt hastalarında hümör aközde ve eş zamanlı venöz kan serumunda, total oksidatif stres, total antioksidan kapasite, paraoksonaz, arilesteraz, lipid peroksidaz seviyelerini karşılaştırarak, diyabetik retinopatinin ilerlemesinde oksidatif stresin rolü değerlendirildi.Prospektif olarak planlanan bu çalışmaya, olgular 10.03.2010 ile 30.08.2010 tarihleri arasında, C.Ü. Tıp Fak. Göz Hast. A.D. Polikliniğine, puslu görme şikayeti ile başvuran ve katarakt tanısı alıp, cerrahi tedavi önerilen diyabetik ve diyabetik olmayan hastalar arasından seçildi. Diyabeti olup retinopatisi olmayan hastalar 1. grup, diyabetik retinopatili hastalar 2. grup ve katarakt tanısı dışında ek sistemik hastalığı olmayan hastalar 3. grup olarak ayrıldı. Her bir grup ayrı ayrı 26 hasta olmakla birlikte toplam 78 hasta çalışmaya dahil edildi.Her 3 grubun eş zamanlı venöz kan serumu sonuçları, total oksidatif stres, total antioksidan kapasite, paraoksonaz ve HbA1c düzeyleri istatistiksel olarak önemli bulunmuşken, lipid peroksidaz ve arilesteraz açısından önemsiz bulunmuştur. 3 grubun hümör aköz sonuçları karşılaştırıldığında ise, total oksidatif stres, total antioksidan kapasite, paraoksonaz, lipid peroksidaz ve arilesteraz düzeyleri istatistiksel olarak önemsiz bulunmuştur.Çalışmamızda, eşzamanlı venöz kan serum sonuçlarının, benzer çalışmaların sonuçlarıyla karşılaştırıldığında; diyabette ve diyabetik retinopatide, TAK, TOS ve PON1 düzeyinde benzer sonuçlar elde ettik. Hümör aköz sonuçların ise, gerek istatistiksel olarak önemsiz çıkması, gerekse literatürde yapılmış başka çalışma olmaması nedeniyle farklı çalışmalara ihtiyaç duyulduğunu düşünüyoruz. ABSTRACTIn this study, the role of oxidative stress in the progression of diabetic retinopathy were evaluated by comparing the levels of total oxidative stress, total antioxidant capacity, paraoxonase, arylesterase and lipid peroxidase in aqueous humor and simultaneous venous blood serum in diabetic and non-diabetic cataract patients.In this prospective study, cases were selected from among the patients that referred Institutional Faculty of Medicine Hospital Ophthalmology Department between 10.03.2010-30.08.2010 with complaints of hazy vision and took the diagnosis of cataract and recommended surgical treatment, diabetic and nondiabetic patients. Patients were divided as first group who have diabetes patients without retinopathy, as second group who have diabetic retinopathy and as third group who have not additional systemic disease other than the diagnosis of cataract. Total of 78 patients enrolled in the study as well as there were 26 patients in each group separately,The results of simultaneous venous blood serum in all 3 groups, total oxidative stress, total antioxidant capacity, paraoxonase and HbA1c levels were found to be statistically significant, while lipid peroxidase and arylesterase was not significant. When the results of aqueous humor of the 3 groups were compared, the levels of total oxidative stress, total antioxidant capacity, paraoxonase, lipid peroxidase and arylesterase were found to be statistically insignificant.In our study, when simultaneous venous blood serum results were compared with results of similar studies, we found similar results at the level of TAC, TOS and PON1 in diabetes and diabetic retinopathy. We believe that the different studies are needed because the results of aqueous humor are statistically insignificant and there is no study made in literature.Key Words: diabetic retinopathy, aqueous humor, oxidative stress, paraoxonase, arylesterase. 47

The eye structure and function highly depend on the stability of the intraocular pressure (IOP). Glaucoma is an ophthalmic disease that can potentially lead to irreversible blindness. Glaucoma results from optic nerve damages due to persistent elevated IOP. Aqueous humor production, drainage and resistance to egress are the key elements that determine the IOP. After travelling from the posterior to the anterior chamber, the aqueous humor passes through the trabecular meshwork to collect into Schlemm's canal, where collector channels eventually drain into the venous system. The highest resistance to egress is located at the outer portion of the trabecular meshwork and inner wall of Schlemm's canal. In this work we intended to simulate the basic geometry and physiological parameters that determine the aqueous humor dynamics and the IOP values. A 3-D modeling of the human eye was made based on histology. Great attention was given to details in the geometry of the trabeculum, Schlemm's canal and the collector channels. Using Matlab program, the pressure distribution in the eye was computed. Flow was computed using Poisseuille's law. The 3-D modeling was meshed by ICEM Ansys before being implemented using Fluent 6.3. 2-D and 3-D meshes were obtained. The results were showing the pressure distribution from the posterior and anterior chambers, the trabeculum, Schlemm's canal and the collector channels. Most of the resistance to aqueous egress occurred at the level of the trabeculum and Schlemm's canal where the pressure drop was the greatest. Refinements in the mesh density would greatly enhance the quality of the modeling. In conclusion this simulation of the outflow pathway in the human eye provided interesting data about the pressure drop and the localization of the main resistance to aqueous egress.

Aim: To determine intraocular and plasma levels of cellular fibronectin (cFN) in patients with uveitis or diabetes mellitus (DM) and to assess the association with disease activity, macular oedema, and vascular leakage on fluorescein angiography. In addition, to examine whether cFN is locally produced in the eye.Methods: Intraocular and plasma levels of cFN were determined by ELISA in 39 patients with uveitis ( 23 active, 16 non-active), in 11 patients with DM ( eight with and three without diabetic retinopathy) and in 17 control patients. The influence of diabetic retinopathy, inflammatory activity, vascular leakage, and macular oedema ( MO) on intraocular and plasma cFN levels was studied. Local production of cFN was determined by calculating absolute and relative intraocular to plasma ratios. Aqueous and vitreous levels of cFN were compared.Results: No differences in plasma cFN levels were found between patients with uveitis, DM, or controls. Intraocular cFN levels were significantly raised in patients with uveitis and DM, specifically in those with active disease ( active uveitis and active diabetic retinopathy versus controls: p = 0.001 and 0.002 respectively). Further, intraocular cFN levels were significantly elevated in patients with macular oedema or vascular leakage, irrespectively of whether associated with uveitis or DM ( p = 0.001 and 0.002). Intraocular cFN levels were consistently higher in the vitreous than the aqueous. Intraocular production of cFN was documented by elevated absolute and relative intraocular to plasma ratios in nine out of 11 patients tested.Conclusions: Elevated intraocular cFN levels were found in uveitis and DM, especially in those with active processes, intraocular vascular damage, and MO. These results suggest that locally produced cFN levels reflect intraocular vascular damage.

Einleitung: Die Wundheilung kornealer Endothelzellen erfolgt hauptsächlich durch Migration benachbarter Zellen. Die Endothelzellen sind in vivo ständig in Kontakt mit Kammerwasser (KW). TGF-ß2 und Ascorbin- säure (AS) sind in hoher Konzentration im KW enthalten. Der Einfluß von humanem KW, TGF-ß2 und AS auf Proliferation und Migration boviner kornealer Endothelzellen (BCEC) in Kultur wurde untersucht. Methoden: Proliferation: BCEC der 1.Passage wurden zu 1,5x104 Zellen/Well ausgesät. Mit frischem Medium und Zusatz von humanem KW (10% und 100%), TGF-ß2 (0,1; 1 und 10 ng/ml) und AS (50, 100 und 200 µg/ml) wurden die Kulturen für 72 oder 96 h inkubiert und anschließend gezählt. Migration: Konfluente Zellkulturen der 1.Passage wurden verwendet. Mit einem modifizierten Trepan (Durchmesser 5,5 mm) wurde eine zentrale, zirkuläre "Wunde" gesetzt. Die Kulturen wurden mit frischem Medium, das humanes KW, TGF-ß2 und AS in den o. g. Konzentrationen enthielt, für 72 oder 96 h inkubiert. Zur Auswertung wurden die in den Wundbereich migrierten Zellen an 5 verschiedenen Stellen, ausgehend vom Wundrand, gezählt. Ergebnisse: Die Proliferation der BCEC wurde durch humanes KW (unverdünntes KW: nahezu 100%; 10%iges KW: 30%) und Ascorbinsäure (ca. 90% bei AS 200 µg/ml) gehemmt. TGF-ß2 stimulierte die Proliferation bis zu einem 3fachen Wert der Kontrollen. Die Migration wurde durch humanes KW (unverdünntes KW: nahezu 100%; 10%iges KW: 30%) und TGF-ß2 (ca. 50% bei allen Konzentrationen) gehemmt. AS hatte einen stimulierenden Einfluß (20-50%) auf die Migration. Schlußfolgerung: Noch ist unklar, welche Substanzen im KW für den proliferationshemmenden Einfluß auf korneale Endothelzellen verantwortlich sind. Sowohl AS als auch TGF-ß2 zeigten einen gegensätzlichen Effekt auf Proliferation und Migration kultivierter BCEC. Die Bedeutung dieser Beobachtungen für die Wundheilung humaner kornealer Endothelzellen in vivo ist Gegenstand weiterer Untersuchungen. Purpose: Corneal endothelial cells are non-proliferating, wound healing primarily occurs by migration of adjacent cells. Endothelial cells in vivo are constantly exposed to aqueous humor (AH). Elevated concentrations of TGF-ß2 and ascorbic acid (AA) are present in aqueous humor. The influence of human AH, TGF- ß2 and AA on proliferation and migration of bovine corneal endothelial cells in vitro was investigated. Methods: Proliferation assays: BCEC at first passage were seeded at 1.5x104 cells/well. Fresh medium containing human AH (10% and 100%), TGF-ß2 (0,1;1 and 10 ng/ml) and AA (50, 100 and 200 µg/ml) was added to the cells. 72 or 96 hours later the cells were counted. Migration assays: BCEC at first passage were grown to confluency. A central, circular "wound" was made with an especially designed trephine (diameter 5.5 mm). The cultures were incubated with fresh medium containing human AH, TGF-ß2 and AA in the same concentrations for 72 or 96 hours. The cells were then counted in five randomly chosen sections from the wound edge. Results: Proliferation of BCE cells was inhibited by human AH (pure AH: nearly 100%, 10% AH: 30%) and AA (about 90% at 200 µg/ml). TGF-ß2 stimulated the proliferation up to 3 fold compared to the controls. Migration was inhibited by human AH (pure AH: nearly 100%, 10% AH: 30%) and TGF-ß2 (about 50% at all concentrations). AA had a stimulatory effect (20-50%) on migration. Conclusion: Presently, it remains unknown which substances in AH are responsible for the inhibiting effect on corneal endothelial proliferation. Both TGF-ß2 and AA showed to have a differential effect on proliferation and migration of cultured BCE cells. The significance of these observations for wound healing of human corneal endothelial cells in vivo has to be investigated.

53 6. ÖZET Süperoksit dismutaz, katalaz ve glutatyon peroksidaz gibi antioksidan enzimler reaktif oksijen türlerini yıkarak hücreleri oksidatif strese karşı korumada önemli bir rol oynarlar. Glutatyon peroksidaz organik hidroperoksitlerin ve de hidrojen peroksitin indirgenmesi reaksiyonunu substrat olarak redükte glutaryonu kullanarak katalizler. Selenyum insanlarda esansiyel bir elementtir. Glutatyon peroksidaz ve iyodotronin deiyodinazm bir bileşenidir. Selenyum organizmanın antioksidan strese karşı savunmasında yardım eder. Uzun süre selenyum eksikliğinde bütün vücut sıvılarında glutatyon peroksidaz aktivitesinin azaldığı gösterilmiştir. Çalışmamızda excimer laser ile yüzeyel (50 um ablasyon) ve derin ablasyon (100 pm ablasyon) uygulanan tavşan gözlerinde kornea glutatyon peroksidaz aktivitesi ile kornea dokusuna komşu ve bu dokunun beslenmesinde en büyük role sahip hümör aközde de selenyum düzeylerine bakılmıştır. Kontrol grubu ile karşılaştırıldığında 100 um ablasyonlu gözlerin GPx aktivitesi kontrollere göre istatistiksel olarak anlamlı şekilde düşük bulunmuştur (p0,05). Sonuç olarak yüksek doz laser sonrasında tavşan kornea dokusu antioksidan enzimlerinden GPx'in aktivitesi azalırken, hümör aköz selenyum düzeyleri değişmemektedir. 65

VI. ÖZET Son zamanlarda serbest oksijen radikalleri ve keratosit ölümü ile artan yara iyileşmesi cevabı PRK sonrası oluşan haze ve regresyondan sorumlu tutulmaktadır. NO'in toksik metaboliti olan peroksinitrit benzer şekilde oksidatif doku hasan ve hücre ölümüne yol açmaktadır. Ancak PRK sonrası NO seviyesinde değişiklik olup olmadığı bilinmemektedir. Çalışmamızda 6 gruba ayrılan tavşan deneklere epitel kazınması, epitel kazınması + yüzeyel PRK ve epitel kazınması + derin PRK yapıldı. Epitel kazınması ve epitel kazınması + yüzeyel PRK yapılan deneklerden 4 saat sonra alınan aköz hümör örneklerinde nitrit/nitrat düzeyleri çalışıldı. Epitel kazınması, epitel kazınması + yüzeyel PRK ve epitel kazınması + derin PRK yapılan deneklerden 24 saat sonra alınan aköz hümör örneklerinde nitrit/nitrat düzeyleri ile kornea dokularında SOD ve Gpx aktivheleri çalışıldı. Elde edilen değerler cerrahi yapılmayan kontrol grubu ile karşılaştırıldı. Nitrit/nitrat düzeyi 4. saatte belirgin olarak azalırken (p0,05). Yüzeyel ve derin PRK gruplarından 24 saat sonra alınan örnekler karşılaştırıldığında, derin PRK grubunda nitrit/nitrat belirgin olarak düşük bulundu (p0,05). Cerrahi gruplarda Gpx aktivitesi azalırken (p0,05). Ancak derin PRK yapılan grupta, yüzeyel PRK grubuna göre Gpx aktivitesi daha düşüktü (p

ÖZET Penetran Keratoplasti uygulanan olguların ön kamara sıvılarında proenflamatuvar sitokinlerden İnterlökin-6 (IL-6) ve İnterlökin-8'in (IL-8) düzeyini belirlemek amacıyla Çukurova Üniversitesi Tıp Fakültesi (ÇÜTF) Göz Hastalıkları Anabilim Dalı Kornea Biriminde Eylül 1997-Mayıs 1998 tarihleri arasında penetran keratoplasti uygulanan, yaşları 18-81 (ortalama 49,5±20,73) arasındaki 32 olgunun 32 gözü, kontrol grubu olarak katarakt cerrahisi uygulanan yaşları 48-74 (ortalama 59,6+30,28) arasında olan 16 olgunun 16 gözünden peroperatif aköz humör örneği alındı. Aköz humör IL-6 ve IL-8 düzeyi mikroelisa yöntemi ile saptandı. İstatiksel araştırmalar Kruskall Wallis ve varyans korelasyon analiz testi kullanılarak yapıldı. Vaskülarize lökom ve regreft uygulanan olguların oluşturduğu yüksek risk grubuna ait aköz humör IL-6 ve IL-8 düzeyleri, keratokonus, büllöz keratopati ve lökom simpleks nedeni ile keratoplasti uygulanan düşük risk gruplarına göre yüksek bulundu. (p

35 ÖZET Sağ göz arka kamaraya a.chymotrypsin enjeksiyonu ile unilateral deneysel glokom oluşturulan 21 tavşanın aköz sıvısında karbonik anhidraz aktivitesi araştırıldı. Aköz sıvıdaki karbonik anhidraz aktivitesi, sağ- glokomatöz gözlerde, sol-normal gözlerden istatistiki olarak anlamlı derecede fazla idi (p

50 ÖZET Çağımızın önemli görmezlik nedenlerinden olan kataraktın oluşumunda eser elementlerin rolünü araştırmak amacıyla 48 senil kataraktlı hastanın serumu, bunlardan 42' sinin lensi ve 10' unun hümör aközünde ve kontrollerde çinko, bakır ve selenyum düzeyleri atomik absorpsiyon spektrofotometresi ile ölçülmüştür. Kataraktlı hastaların serum bakır değerleri kontrollere göre yüksek, selenyum değerleri ise düşük bulunmuştur (p

Isobutylmethylxanthine (IBMX), a phosphodiesterase/adenosine receptor inhibitor, was combined with norepinephrine (nE), epinephrine (Epi) and isoproterenol, respectively, to evaluate their effect on intraocular pressure (IOP). Application of topical IBMX alone had no measurable effect on IOP. When IBMX was combined with nE or Epi the ocular hypotension in rabbits and beagles increased. The Emax for nE alone was 2.9 +/- 0.4 mmHg, for Epi alone 7.3 +/- 0.5 mmHg and for isoproterenol alone 5.1 +/- 0.3 mmHg. The EC50 was 0.2 +/- 0.05% (nE), 0.05 +/- 0.01% (Epi) and 0.003 +/- 0.001% for isoproterenol. When given in combination with 1% IBMX the Emax for nE was 7.4 +/- 1.7 mmHg, for Epi 9.0 +/- 0.8 mmHg and for isoproterenol 6.1 +/- 0.3 mmHg. The corresponding values for EC50 were 0.07 +/- 0.03% (nE), 0.02 +/- 0.006% (Epi) and 0.002 +/- 0.001% for isoproterenol. Combining 1% IBMX with 0.1% Epi increased the aqueous humour cyclic AMP-levels at 1, 3 and 5 hr in rabbits. The results of this study demonstrate that a phosphodiesterase/adenosine receptor inhibitor such as IBMX enhances the reduction in IOP induced by adrenergic agonists.

80