Successive Acanthamoeba Corneal Isolates Identified in Poland Monitored in Terms of In Vitro Dynamics.

BACKGROUND: Amoebae of the genus Acanthamoeba cause a sight-threatening infection called Acanthamoeba keratitis. It is considered a rare disease in humans but poses an increasing threat to public health worldwide, including in Poland. We present successive isolates from serious keratitis preliminary examined in terms of the identification and monitoring of, among others, the in vitro dynamics of the detected strains. METHODS: Clinical and combined laboratory methods were applied; causative agents of the keratitis were identified at the cellular and molecular levels; isolates were cultivated in an axenic liquid medium and regularly monitored. RESULTS: In a phase-contrast microscope, Acanthamoeba sp. cysts and live trophozoites from corneal samples and in vitro cultures were assessed on the cellular level. Some isolates that were tested at the molecular level were found to correspond to A. mauritanensis, A. culbertsoni, A. castellanii, genotype T4. There was variability in the amoebic strain dynamics; high viability was expressed as trofozoites' long duration ability to intense multiply. CONCLUSIONS: Some strains from keratitis under diagnosis verification and dynamics assessment showed enough adaptive capability to grow in an axenic medium, allowing them to exhibit significant thermal tolerance. In vitro monitoring that was suitable for verifying in vivo examinations, in particular, was useful to detect the strong viability and pathogenic potential of successive Acanthamoeba strains with a long duration of high dynamics.

Pupil diameter during cataract surgery after intracameral injection of the first ready-to-use combination of mydriatics and anaesthetic at the beginning of surgery in patients with a preoperative pupil diameter <6 mm.

PURPOSE: We evaluated, in a real-life setting, the effect of Mydrane® (ready-to-use combination of tropicamide, phenylephrine hydrochloride and lidocaine, injected into the anterior chamber at the beginning of cataract surgery to induce mydriasis and intraocular anaesthesia) on the pupil diameter during cataract surgery in patients with a preoperative pupil diameter <6 mm after the use of topical mydriatics. METHODS: We collected and analysed the data of 59 consecutive patients whose pupils dilated to a diameter <6 mm after the administration of mydriatic eye drops during the preoperative visit and who received Mydrane® during cataract surgery. RESULTS: In the group of 59 patients with a preoperative pupil diameter <6 mm after topical mydriatics, cataract surgery was performed in 36 patients (61.0%) using only Mydrane® to obtain mydriasis, with no additional drug or medical device. The mean pupil diameters in this group (36 of 59) during the preoperative assessment after topical mydriatics and just before capsulorhexis when Mydrane® was injected during surgery were 5.1 ± 0.74 and 6.15 ± 1.14 mm. Additional drugs were used in 23 patients (39%). In this group, the mean pupil diameters after topical mydriatics and just before capsulorhexis using Mydrane® were 4.58 ± 1.06 and 5.6 ± 1.26 mm, respectively. CONCLUSION: In a real-life setting, the mean pupil diameter achieved during cataract surgery after the intracameral injection of Mydrane® in patients with a preoperative pupil diameter <6 mm was over 1 mm larger than the mean pupil diameter after topical mydriatics, despite the trauma caused by the operation.

Canaloplasty - Efficacy and Safety in an 18-Month Follow Up Period, and Analysis of Outcomes in Primary Open Angle Glaucoma Pigmentary Glaucoma and Pseudoexfoliative Glaucoma.

PURPOSE: Open-angle glaucoma (OAG), accounting for 90% of all glaucoma cases, is a progressive optic nerve neuropathy. It may lead to irreversible loss of visual field and complete blindness. When conservative treatment becomes insufficient to stop OAG progression, a surgical intervention is considered. Currently, canaloplasty procedure is being introduced instead of conventional trabeculectomy for invasive OAG treatment. The aim of the study is to asses safety and efficacy of canaloplasty. METHODS: This prospective study included 67 eyes that received 360° canaloplasty with placement of a tensioning suture. Primary OAG (n = 35), secondary OAG in pseudoexfoliative syndrome (n = 13), and pigmentary glaucoma (n = 19) patients were included. Control check-ups were conducted pre-operatively and in a 18-month follow-up time. Study endpoints involved reduction in IOP values and in the number of glaucoma medications after the intervention. RESULTS: The intervention led to a significant 38% reduction in IOP value from the preoperative baseline to 18 months after the intervention. The number of medications decreased significantly by 89%. At 18 months postoperative, 79% eyes did not require any glaucoma medications. The incidence of complications after canaloplasty was low, and none of the adverse effects were vision threatening. A surgically-induced astigmatism was the most frequent complication. Pigmentary glaucoma patients were the most beneficial subgroup, with 50% reduction in IOP, the highest success rate, and 98% reduction in the number of medications used. CONCLUSION: This study proved that canaloplasty is an efficient and safe procedure in OAG eyes.

Stable Mydriasis After Intracameral Injection of a Combination of Mydriatics and Anesthetic During Cataract Surgery: A Real-Life, Multicenter Study.

Purpose: To evaluate the effect of Mydrane (contains tropicamide, phenylephrine hydrochloride, and lidocaine hydrochloride) on time needed to induce mydriasis and mydriasis stability during cataract surgery. Methods: This was an observational, non-interventional, multicenter study of patients undergoing cataract surgery who received Mydrane for mydriasis and intraocular anesthesia. The study was conducted at seven ophthalmology departments at university hospitals in Poland. Patients admitted for cataract surgery within a 2-week period were asked to participate in the study. Patients whose pupils dilated to a diameter ≥6 mm after topical mydriatic administration during preoperative examinations were scheduled to receive Mydrane and included in the registry. No additional inclusion criteria were used. Patients' medical histories, examination results, and operative details were recorded. Pupil diameter was measured during surgery. Surgeons were asked to complete a Likert-based survey in parallel. Results: A total of 307 patients were enrolled. The mean pupil diameter was 7.0 ± 1.0 mm before capsulorhexis and 6.9 ± 1.2 mm before lens implementation. A pupil diameter ≥6 mm was achieved in 91.9% and 87.6% of patients before capsulorhexis and lens implantation, respectively. We asked 58 surgeons whether they agreed with the statement "Mydriasis was obtained in a short time after the administration of Mydrane"; the surgeons agreed with this statement after 92.2% (283/307) of surgeries. In addition, after 88.2% of surgeries, the surgeons agreed with the statement "Mydriasis was stable after the administration of Mydrane." Conclusions: Mydriasis was rapidly and stably obtained after Mydrane injection, as demonstrated by pupil diameter measurements during surgery and surgeons' feedback.

Retinal arterial remodeling in patients with pheochromocytoma or paraganglioma and its reversibility following surgical treatment.

OBJECTIVE: Structural abnormalities in resistance arteries are a hallmark of patients with hypertension. In hypertensive patients with pheochromocytoma or paraganglioma (PPGL), it is still a matter of debate whether structural vascular changes are because of elevated blood pressure (BP) or to toxic effects of elevated circulating catecholamines. Hence, the aim of our study was to assess whether catecholamine excess and/or elevated BP affect the structure of small retinal arteries in patients with catecholamine-producing tumors. METHODS: The study included 27 patients with PPGL and 27 hypertensive patients. All patients underwent biochemical tests for catecholamine excess, echocardiography and analyses of scanning-laser-Doppler-flowmetry (SLDF) both at baseline and 12 months following surgical resection of PPGL. RESULTS: Baseline retinal arterial diameter, arterial wall thickness and wall cross sectional area (WCSA) were higher in patients with PPGL as compared with subjects without PPGL (arterial diameter: 110 ±â€Š16.5 vs. 99.5 ±â€Š10.8 µm, wall thickness: 16.3 ±â€Š6.0 vs. 13.5 ±â€Š4.0 µm, WCSA: 4953.9 ±â€Š2472.8 vs. 3784.1 ±â€Š1446.3 µm, P < 0.05). Significant correlations were noted between wall thickness and WCSA and echocardiographic parameters assessing diastolic and systolic function of left ventricle. No correlations between retinal parameters, BP level and plasma concentrations of metanephrines were observed. In patients with PPGL, there were postoperative decreases in wall thickness (16.4 ±â€Š15.8 vs. 14.8 ±â€Š4.7 µm; P = 0.011) and WLR (0.42 ±â€Š0.13 vs. 0.37 ±â€Š0.10; P = 0.003) at 12 months after surgical removal of tumors. CONCLUSION: This is the first study to demonstrate that catecholamine excess is related to thickening of retinal arteries independent of BP and reversible after surgical cure. These data support a role of catecholamines in vascular remodeling in PPGL patients.

Effect of 0.1% diclofenac ophthalmic solution on inflammatory response and macular thickness following phacoemulsification cataract surgery. / [Wplyw 0,1% kropli diklofenaku na odczyn zapalny w komorze przedniej oka i stan plamki po operacji fakoemulsyfikacji zacmy].

Purpose: To evaluate 0.1% diclofenac sodium as an adjunctive therapy with loteprednol etabonate on postoperative inflammation in the anterior chamber and on foveal and parafoveal retinal thickness. Material and methods: Eighty eyes eligible for phacoemulsification were enrolled in a randomized clinical trial. Patients in group I (N = 40) received anti-inflammatory treatment consisting of 0.1% diclofenac with 0.5% loteprednol; group II (N = 40) patients received 0.5% loteprednol alone. Best corrected visual acuity and intraocular pressure were measured, and laser flarephotometry was done. Foveal and parafoveal thickness were assessed by optical coherence tomography. Results: Median flare values decreased more rapidly in group I at 7 and 14 days (7.9 and 7.4 ph/ms, respectively) than in group II (13.7 and 11.8 ph/ms, respectively; p < 0.0001). Group II had significantly increased parafoveal thickness at 14 and 42 days (median 285.59 µm, p = 0.001 and 288.38 µm, p < 0.001, respectively). Parafoveal thickness differed significantly between groups at 14 and 42 days (p = 0.0085, p = 0.0004, respectively). Conclusions: Eyes treated with both diclofenac sodium and loteprednol etabonate showed less inflammatory response and were less likely to develop foveal and parafoveal thickening than those treated with steroid only.

The opinion of the Expert Group of the Polish Society of Ophthalmology on using nepafenac in the prevention of postoperative macular edema after cataract surgery in diabetic patients.

Nepafenac is an innovative non-steroidal anti-inflammatory drug used in ophthalmology for the prevention of macular edema after cataract surgery. Along with its anti-inflammatory effect, nepafenac has some unique properties which distinguish it from other non-steroidal anti-inflammatory drugs. It is a prodrug activated to amfenac after it penetrates through the corneal layers to the aqueous humour and the ciliary body. Having electrically neutral molecules of lipophilic properties, nepafenac does not accumulate in the cornea and does not cause its degeneration. Additionally, it quickly achieves higher concentrations in the aqueous humour as compared to other non-steroidal anti-inflammatory drugs. Nepafenac shows high selectivity and activity against COX-2 isoform, the key enzyme implicated in inducing inflammation, which is the main cause of macular edema. Furthermore, nepafenac has the unique scleral and suprachoroidal distribution pathways. Finally, its effect on the intraocular pressure is none to negligible. Nepafenac treatment should be initiated prior to cataract surgery and continued long enough to reduce the risk of late-onset macular edema. The Expert Group of the Polish Society of Ophthalmology consider using nepafenac in the prevention of post­operative macular edema in diabetic patients undergoing cataract surgery as expedient and reasonable. The proposed optimum pre- and postoperative treatment regimen can be modified for individualised therapy.

Transferrin receptor levels and polymorphism of its gene in age-related macular degeneration.

The aim of the present study was to investigate the association of age related macular degeneration (AMD) risk with some aspects of iron homeostasis: iron concentration in serum, level of soluble transferrin receptor (sTfR), and transferrin receptor (TFRC) genetic variability. Four hundred and ninety one AMD patients and 171 controls were enrolled in the study. Restriction fragment length polymorphism PCR was employed to genotype polymorphisms of the TFRC gene, and colorimetric assays were used to determine the level of iron and sTfR. Multiple logistic regression was applied for all genotype/allele-related analyses and the ANOVA test for iron and sTfR serum level comparison. We found that the genotypes and alleles of the c.-253G > A polymorphism of the TFRC gene were associated with AMD risk and this association was modulated by smoking status, AMD family history, living environment (rural/urban), body mass index and age. The levels of sTfR was higher in AMD patients than controls, whereas concentrations of iron did not differ in these two groups. No association was found between AMD occurrence and the p.Gly142Ser polymorphism of the TRFC gene. The results obtained suggest that transferrin receptor and variability of its gene may influence AMD risk.

[Ocular manifestations in hereditary diseases with defects in DNA repair]. / Zaburzenia narzadu wzroku w przebiegu chorób genetycznych zwiazanych z naprawa DNA.

DNA repair is involved in maintaining the stability of the genome and accurate sending of genetic information. DNA repair pathways remove many DNA damages induced by endo- and exogenous factors. There are several DNA repair pathways in human cells, including base or nucleotide excision system, homologous recombination system and non-homologous end joining. Mutation in DNA repair genes may results in rare genetic disorders, including Xeroderma pigmentosum, Cockayne syndrom, trichothiodystrophy, Nijmegen syndrome, ataxia teleangiectasia, Werner syndrome, Bloom syndrome, Rothmund-Thomson syndrome. These diseases may be associated with various visual disturbances. In this work we review we focus on human genetic diseases linked with mutations in DNA repair genes associated with visual impairment.

Polymorphisms of the apoptosis-related FAS and FAS ligand genes in keratoconus and Fuchs endothelial corneal dystrophy.

Keratoconus (KC) is a non-inflammatory eye disease characterized by progressive corneal thinning and asymmetrical conical protrusion of the cornea. Fuchs endothelial corneal dystrophy (FECD) is a degenerative, slowly progressive disease of the corneal endothelium that is characterized by alteration in corneal endothelial cell morphology and progressive loss of these cells. They are unrelated eye diseases that may ultimately lead to vision loss. Their pathogenesis is largely unknown, although impaired apoptosis has been suggested to be responsible for both diseases. Therefore, we studied the frequency of the c.-671A>G polymorphism of the apoptosis-related FAS gene and the c.-844T>C polymorphism of the FAS ligand (FASLG) gene in patients with FECD (221 individuals) or KC (264) and controls (300). Each polymorphism is located within the putative cis-acting element of the respective promoter. Risk of KC or FECD was estimated with unconditional multiple logistic regression with adjustment for various factors, including age, sex, allergies, and family history. The T/T genotype and the T allele of the c.-844T>C polymorphism were associated with increased occurrence of KC, while the C allele was associated with decreased KC occurrence. The G allele of the c.-671A>G polymorphism was associated with increased occurrence of FECD, while the A allele was associated with decreased FECD occurrence. The C/C-A/A combined genotype was associated with reduced risk of FECD, whereas the T/T-G/A combined genotype increased risk of KC. In conclusion, variability in the expression of the FAS and FASLG genes may be involved in the pathogenesis of KC and FECD.

Association between the 25129A > C polymorphism of the nuclear respiratory factor 2 gene and age-related macular degeneration.

PURPOSE: Oxidative stress belongs to the main factors of pathogenesis of age-related macular degeneration, characterized by the damage to the retinal pigment epithelium cells and photoreceptors. Retinal pigment epithelium cells are rich in mitochondria, producing large amount of reactive oxygen species, which are by-products of the activity of the respiratory chain. The distribution in the activity of the chain may be evoked by the release of cytochrome C from the mitochondrion to the cytoplasm. This process may activated by nuclear respiratory factor 2, Nrf2, which is encoded by a highly polymorphic gene. In this study we examined the association between age-related macular degeneration risk and the 25129A > C polymorphism of the gene encoding nuclear respiratory factor 2 (rs12594956). MATERIAL AND METHODS: Genotypes were determined in DNA from peripheral blood lymphocytes of 281 patients with age-related macular degeneration (181 with wet form of the disease and 101 with its dry form), and 105 controls by PCR-restriction fragment length polymorphism. RESULTS: A weak association (OR 1.96; p = 0.023) between the C/C genotype of the 25129A > C polymorphism and the occurrence of age-related macular degeneration was found. A stronger association was observed between dry age-related macular degeneration occurrence and the C/C genotype of the polymorphism (OR 2.23; p = 0.018). The A/C genotype decreased the risk of age-related macular degeneration and its dry form (OR 0.51; p = 0.023 and 0.44; p = 0.018, respectively). Potential risk factors such as age, gender, smoking habit, living environment (rural or urban) and family status of age-related macular degeneration increased the risk of AMD associated with the C/C genotype (OR 2.52; p = 0.012). CONCLUSIONS: The 25129A > C polymorphism of the NRF2 gene may be associated with age-related macular degeneration. mitochondria, reactive oxygen species, gene polymorphism, NRF2 gene, age-related macular degeneration - AMD.

Association of the 399Arg/Gln XRCC1, the 194 Arg/Trp XRCC1, the 326Ser/Cys OGG1, and the 324Gln/His MUTYH gene polymorphisms with clinical parameters and the risk for development of primary open-angle glaucoma.

BACKGROUND: Numerous data have shown that progressive loss of human trabecular meshwork (TM) cells may be connected with oxidative stress. This hypothesis may suggest an association of base excision repair with the risk of primary open angle glaucoma development. PURPOSE: The aim of this study was to evaluate the role of the 399Arg/Gln XRCC1, the 194 Arg/Trp XRCC1, the 326SerCys OGG1, and the 324Gln/His MUTYH gene polymorphisms with clinical parameters and the risk for development of POAG. METHODS: Our research included 170 patients with POAG and 193 healthy controls. Gene polymorphisms were investigated by PCR-RFLP. The Heidelberg Retinal Tomography (HRT) clinical parameters were also analyzed. RESULTS: The 399Arg/Gln genotype of the XRCC1 gene was associated with an increased risk for POAG (OR 2.50; 95% CI, 1.54-4.07, P=0.0002). The 399Gln/Gln XRCC1 genotype may increase the risk for POAG progression according to clinical parameters such as cup/disk ratio (c/d) (OR 1.93; 95% CI, 1-3.73, P=0.04) and Rim area (RA factor) (OR 3.88; 95% CI, 1.01-14.82, P=0.04). Moreover, an association was found of retinal nerve-fiber layer (RNFL factor) with the 399Arg/Gln XRCC1 genotype distribution and POAG progression (OR 2.46; 95% CI, 1.06-5.68, P=0.03). In contrast, analysis of the 324Gln/His MUTYH gene polymorphism distribution in the patient group according to RA factor showed that it may reduce the progression of POAG (OR 0.14; 95% CI, 0.02-0.89, P=0.05). Our current study demonstrates an association between the 326Ser/Cys OGG1 gene polymorphism and the 326Cys allele of the OGG1 gene, and progression of POAG. In addition, the presence of the 326His allele of the MUTYH gene may increase the risk for POAG progression, according to the VF parameter (OR 2.57; 95% CI, 1.47-4.57, P=0.0001). CONCLUSION: We suggest that the 399Arg/Gln genotype and the 399Gln allele of the XRCC1 gene may be risk factors for POAG development. Moreover, we postulate that the 399 Arg/Gln XRCC1, the 326 Ser/Cys OGG1 and the 324 Gln/His MUTYH genes polymorphisms may be associated with progression of POAG.

Genetic polymorphism of the iron-regulatory protein-1 and -2 genes in age-related macular degeneration.

Iron can be involved in the pathogenesis of AMD through the oxidative stress because it may catalyze the Haber-Weiss and Fenton reactions converting hydrogen peroxide to free radicals, which can induce cellular damage. We hypothesized that genetic polymorphism in genes related to iron metabolism may predispose individuals to the development of AMD and therefore we checked for an association between the g.32373708 G>A polymorphism (rs867469) of the IRP1 gene and the g.49520870 G>A (rs17483548) polymorphism of the IRP2 gene and AMD risk as well as the modulation of this association by some environmental and life-style factors. Genotypes were determined in DNA from blood of 269 AMD patients and 116 controls by the allele-specific oligonucleotide-restriction fragment length polymorphism and the polymerase chain reaction-restriction fragment length polymorphism. An association between AMD, dry and wet forms of AMD and the G/G genotype of the g.32373708 G>A-IRP1 polymorphism was found (OR 3.40, 4.15, and 2.75). On the other hand, the G/A genotype reduced the risk of AMD as well as its dry or wet form (OR 0.23, 0.21, 0.26). Moreover, the G allele of the g.49520870 G>A-IRP2 polymorphism increased the risk of the dry form of the disease (OR 1.51) and the A/A genotype and the A allele decreased such risk (OR 0.43 and 0.66). Our data suggest that the g.32373708 G>A-IRP1 and g.49520870 G>A-IRP2 polymorphisms may be associated with increased risk for AMD.

Lack of association between the c.544G>A polymorphism of the heme oxygenase-2 gene and age-related macular degeneration.

BACKGROUND: Age-related macular degeneration (AMD) is a primary cause of blindness among the elderly in developed countries. The nature of AMD is complex and includes both environmental and hereditary factors. Oxidative stress is thought to be essential in AMD pathogenesis. Iron is suggested to be implicated in the pathogenesis of AMD through the catalysis of the production of reactive oxygen species, which can damage the retina. Heme oxygenase-2 is capable of degradation of heme producing free iron ions, thus, diversity in heme oxygenase-2 gene may contribute to AMD. In the present work we analyzed the association between the c.544G>A polymorphism of the heme oxygenase-2 gene (HMOX2) (rs1051308) and AMD. MATERIAL/METHODS: This study enrolled 276 AMD patients and 105 sex- and age-matched controls. Genotyping of the polymorphism was performed with restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) on DNA isolated from peripheral blood. RESULTS: We did not find any association between the genotypes of the c.544G>A polymorphism and the occurrence of AMD. This lack of association was independent of potential AMD risk factors: tobacco smoking, sex and age. Moreover, we did not find any association between AMD and smoking in our study population. CONCLUSIONS: The results suggest that the c.544G>A polymorphism of the heme oxygenase-2 gene is not associated with AMD in this Polish subpopulation.

The A Allele of the -576G>A polymorphism of the transferrin gene is associated with the increased risk of age-related macular degeneration in smokers.

Age-related macular degeneration (AMD) is the leading cause of blindness among the elderly in developed countries, and its pathogenesis is underlined by genetic and environmental factors. Oxidative stress is a major environmental risk factor of AMD; namely, AMD is associated with the increased level of reactive oxygen species, which may be produced in reactions catalyzed by iron present in the retina. Therefore, variability of the genes of iron metabolism may be important in the AMD risk. In the present study, we analyzed the association between AMD and the -576G>A polymorphism of the transferrin gene or the 1892C>T polymorphism of the transferrin receptor 2 (TFR2) gene in 278 patients with AMD and 105 controls. The former polymorphism is located in the promoter region of the transferrin gene and may affect the level of its transcription, while the latter is a synonymous mutation in the exon 16, which may affect the efficiency of translation of TFR2 mRNA. Transferrin and TFR2 are important in iron homeostasis. The A allele of the -576A>G polymorphism was significantly associated with the increased risk of AMD in tobacco smokers, whereas the 1892C>T polymorphism did not influence the risk of AMD related to smoking. Moreover, each polymorphism does not influence the risk of AMD associated with age, sex or the family history of the disease. In conclusion, the A allele of the -576A>G polymorphism of the transferrin gene may increase the risk of AMD in smokers.

Implications of altered iron homeostasis for age-related macular degeneration.

Reactive oxygen species (ROS) may contribute to the pathogenesis of age-related macular degeneration (AMD) and they can be produced in the Fenton reaction catalyzed by Fe3+ ions. Therefore, altered homeostasis of iron in the retina may be the source of ROS and its damage resulting in clinically detectable AMD symptoms. The results of some post mortem research indicate a higher concentration of iron in AMD retinas in comparison with non-affected retinas, although those results do not determine whether iron overload is the reason or a consequence of AMD. However, the results of some other research suggest that iron may contribute to the pathogenesis of AMD. Those include increasing of macular iron level with age, involvement of iron in the pathogenesis of some degenerative diseases linked with AMD, upregulation of transferrin in AMD, developing of AMD-like syndromes in mice deficient in ceruloplasmin and hephaestin, association between polymorphism of the iron homeostasis genes and AMD and others. Better understanding of the role of altered iron homeostasis may be useful in prevention and curing of AMD.

OCT and in vivo confocal microscopy of a pigmented corneal tumor-like lesion.

A 43-year-old woman presented with a pigmented flat tumor situated at the posterior surface of the cornea nasally in her left eye. Anterior-segment optical coherence tomography revealed that the lesion was similar to the iris leaf, was limited to the cornea, and did not communicate with the iridocorneal angle. In vivo scanning slit confocal microscopy imaged dense hyperreflective tissue behind the endothelium and bright spots dispersed on the adjacent endothelial surface. Multiple hyporeflective formations resembling cell nuclei were visualized within the hyperreflective mass and the cell borders were distinguished. The diagnosis of pigmented nevus or retrocorneal membrane was suspected. The authors conclude that anterior-segment optical coherence tomography and in vivo scanning slit confocal microscopy are useful in assessing the microstructure and penetration of pigmented corneal lesions.

Association between vascular endothelial growth factor gene polymorphisms and age-related macular degeneration in a Polish population.

The pathogenesis of age-related macular degeneration (AMD) is thought to be determined by an array of environmental and genetic factors. The association of increased expression of vascular endothelial growth factor (VEGF) with AMD, especially the wet form of AMD, was reported in several studies. The VEGF gene is highly polymorphic and some of its polymorphisms may affect its expression. In our work, we searched for an association between the -460C> (rs833061) and -634G>C (rs2010963) polymorphisms of the VEGF gene and the occurrence of AMD and its dry and wet forms. We have chosen these polymorphisms because they were shown to be significant in other studies and we previously showed their association with diabetic retinopathy. A total of 401 individuals were enrolled in this study: 136 controls, and 88 patients with dry and 177 with wet AMD. The polymorphisms were determined with DNA from peripheral blood lymphocytes by allele-specific and restriction fragment length polymorphism polymerase chain reaction. The significance of the polymorphisms was assessed by multiple logistic regression, producing odds ratios (ORs) and 95% confidence intervals (CIs). We observed a weak association (OR 2.90) between AMD occurrence and the C/T genotype of the -460C>T polymorphism. An association (OR 3.77) between the C/T genotype of the -460C>T polymorphism and the occurrence of dry AMD was observed. The T/T genotype considerably lowered the risk of dry AMD (OR 0.19). Dry AMD was associated with the C/C genotype of the -634G>C polymorphism (OR 3.68). Another weak association (OR 2.63) was found between the C/T genotype of the -460C>T polymorphism and the occurrence of wet AMD. The occurrence of AMD was correlated with the presence of the combined C/T-G/G genotype of both polymorphisms (OR 2.41), whereas the T/T-G/G and T/T-G/C genotypes exerted a protective effect against the disease (OR 0.22 and 0.48, respectively). The presence of the C/T-G/G and C/T-C/C combined genotypes increased the risk of dry AMD (OR 2.08 and 3.77, respectively), whereas the presence of the T/T-G/G and T/T-G/C genotypes decreased the risk (OR 0.15 and 0.28, respectively). In the wet form of AMD, the combined genotype C/T-G/G slightly favored the disease (OR 2.61) and the T/T-G/G genotype had a protective effect (OR 0.25). Analysis of haplotypes of both polymorphisms yielded similar results for AMD in general as well as for the dry and wet forms of the disease: the CG haplotype favored both forms of AMD, whereas the TG haplotype protected against both forms of AMD. The results obtained indicate that the -460C>T and -634G>C polymorphisms of the VEGF gene may be associated with the dry and wet forms of AMD in a Polish population.

Distribution of the C-460T polymorphism of the vascular endothelial growth factor gene in age-related macular degeneration.

PURPOSE: To assess the association between genotypes and alleles of the C-460T polymorphism of the vascular endothelial growth factor (VEGF) gene and the risk of wet form of age-related macular degeneration (AMD). MATERIALS AND METHODS: 100 patients with clinically diagnosed wet form of AMD and 104 healthy individuals were enrolled in this study. The patients were diagnosed by optical coherence tomography, fluorescein angiography and indocyanin green angiography. The allele-specific polymerase chain reaction was used to determine the genotypes of the C-460T polymorphism of the VEGF gene. The odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using a logistic regression model to assess the assoctiation betweeen genotypes of the C-460T polymorphism and AMD occurrence. RESULTS: A difference was observed in the genotype distributions between patients and controls. An association (OR 3.04, 95% CI 1.65-5.60) was found between wet form of AMD and the C/T genotype. On the other hand, the T/T genotype displayed the protective effect against the disease. CONCLUSION: The C-460T polymorphism of the endothelial growth factor can be considered as a potential marker for the wet form of age-related macular degeneration.