Jerzy Szaflik

Publications of Jerzy Szaflik

• Association between polymorphisms of the DNA base excision repair genes MUTYH and hOGG1 and age-related macular degeneration.

  Authors: Ewelina Synowiec, Janusz Blasiak, Malgorzata Zaras, Jerzy Szaflik, Jacek P Szaflik

  Experimental eye research. 03/2012;

  Age-Related Macular Degeneration (AMD) is an eye disease that results in progressive and irreversible loss of central vision and is considered as the primary cause of visual impairment, includingAge-Related Macular Degeneration (AMD) is an eye disease that results in progressive and irreversible loss of central vision and is considered as the primary cause of visual impairment, including blindness, in the elderly in industrialized countries. Oxidative stress has been implicated in the pathogenesis of AMD. The hOGG1 and the MUTYH genes play an important role in the repair of oxidatively damaged DNA in the base excision repair pathway. The DNA glycosylases encoded by the hOGG1 and MUTYH genes initiate this pathway by recognizing and removing 8-oxoguanine and adenine paired with 8-oxoguanine, respectively. Our study was designed to examine the association between the c.977 C > G polymorphism (rs1052133) of the hOGG1 gene and the c.972 G > C polymorphism (rs3219489) of the MUTYH gene and AMD as well as the modulation of this association by some clinical and lifestyle factors. Genotypes were determined in DNA from blood of 271 AMD patients, including 101 with wet and 170 with dry form of the disease and 105 sex- and age-matched individuals without AMD. We observed an association between AMD, dry and wet forms of AMD and the C/G genotype and the G allele of the c.977 C > G-hOGG1 polymorphism (p 0.006; 0.009; 0.021 and 0.004; 0.005; 0.016 respectively). On the other hand, the C/C genotype and the C allele reduced the risk of AMD as well as of its dry form or wet form (p 0.002; 0.003; 0.010 and 0.004; 0.005; 0.016, respectively). Therefore, the associations we detected were driven by the dry AMD. We observed some statistically significant association between the occurrence of AMD and its dry and wet forms and genotypes of the other polymorphism, the c.972 G > C-MUTYH polymorphism, but due to borderline character of all this association we do not consider them as medically relevant. Our findings suggest that the c.977 C > G-hOGG1 polymorphism may be associated with dry AMD. Further studies are needed to determine possible association between AMD and the c.972 G > C-MUTYH polymorphism.

• An association between environmental factors and the IVS4+44C>A polymorphism of the DMT1 gene in age-related macular degeneration.

  Authors: Daniel Wysokinski, Malgorzata Zaras, Mariola Dorecka, Maja Waszczyk, Jerzy Szaflik, Janusz Blasiak, Jacek P Szaflik

  Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie. 02/2012;

  BACKGROUND: Age-related macular degeneration (AMD) is an ocular disease affecting macula - the central part of the retina, resulting in the degeneration of photoreceptors and retinal epithelium andBACKGROUND: Age-related macular degeneration (AMD) is an ocular disease affecting macula - the central part of the retina, resulting in the degeneration of photoreceptors and retinal epithelium and causing severe central vision impairment. The pathophysiology of the disease is not completely known, but a significant role is attributed to genetic factors. The contribution of oxidative stress in AMD as a trigger of the degenerative process is well-established. Iron ions may act as a source of reactive oxygen species; therefore, maintaining iron homeostasis is important for redox balance in the organism. Diversity in iron homeostasis genes may counterpart in unbalanced redox state, and thus be involved in AMD pathophysiology. METHODS: In this work, we searched for an association between some single nucleotide polymorphisms in the divalent metal transporter 1 (DMT1) gene intronic IVS4+44C>A (rs224589) and 3'-UTR c.2044T>C (rs2285230) and environmental factors and AMD. Genotyping was performed using the PCR-RFLP method. DNA was obtained from 436 AMD patients and 168 controls. RESULTS: We did not find any association between the genotypes of the two polymorphisms and AMD occurrence. However, we observed that AMD patients living in a rural environment and having the CC genotype of the IVS4+44C>A polymorphism had an increased risk of AMD, while individuals with the CA genotype or the A allele had a decreased risk of the disease. Moreover, in male AMD patients the C allele increased the risk of the disease, while the AA genotype decreased it. CONCLUSIONS: These results suggest that the VS4+44C>A polymorphism of the DMT1 gene may interact with place of living and gender to modulate the risk of AMD.

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

  Authors: Ewelina Synowiec, Magdalena Pogorzelska, Janusz Blasiak, Jerzy Szaflik, Jacek Pawel Szaflik

  Molecular biology reports. 02/2012;

  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 canIron 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.

• Influence of donor age, post-mortem time and cold storage on metabolic profile of human cornea.

  Authors: Tomasz Kryczka, Jacek P Szaflik, Jerzy Szaflik, Anna Midelfart

  Acta ophthalmologica. 11/2011;

  Purpose:  Limited knowledge exists about the influence of donor age and death-to-preservation interval (DPI) on the metabolic properties of the cornea. The aim of this study is to investigate thePurpose:  Limited knowledge exists about the influence of donor age and death-to-preservation interval (DPI) on the metabolic properties of the cornea. The aim of this study is to investigate the relationship between both factors and metabolite content of the cornea. Methods:  Corneas from 15 human donors (age: 41-78 years) were obtained within 16 hrs post-mortem and kept in cold storage for 8 days. The metabolic profiles of the samples were investigated using high-resolution, magic angle spinning (1) H nuclear magnetic resonance spectroscopy before and after 8 days of preservation. Results:  Twenty-two metabolites were detected and assigned in the corneal spectra. The significant metabolic differences before and after hypothermic storage were revealed between younger and older donors. DPI-related significant differences revealed before preservation of the corneas were not displayed after 8 days of cold storage. Conclusions:  Age of donor as well as post-mortem time influences the biochemical properties of the cornea. Cold storage decreases the metabolite differences between the tissues collected at different post-mortem time.

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

  Authors: Daniel Wysokinski, Ewelina Synowiec, Marta Chmielewska, Katarzyna Wozniak, Małgorzata Zaras, Anna Sklodowska, Janusz Blasiak, Jerzy Szaflik, Jacek Pawel Szaflik

  Medical science monitor : international medical journal of experimental and clinical research. 08/2011; 17(8):CR449-455.

  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.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. 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. 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. The results suggest that the c.544G>A polymorphism of the heme oxygenase-2 gene is not associated with AMD in this Polish subpopulation.

• Association of MMP1-1607 1G/2G and TIMP1 372 T/C gene polymorphisms with risk of primary open angle glaucoma in a Polish population.

  Authors: Ireneusz Majsterek, Lukasz Markiewicz, Karolina Przybylowska, Mira Gacek, Anna K Kurowska, Anna Kaminska, Jerzy Szaflik, Jacek P Szaflik

  Medical science monitor : international medical journal of experimental and clinical research. 07/2011; 17(7):CR417-21.

  Primary open angle glaucoma (POAG) is considered to be a leading cause of irreversible blindness worldwide. Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) have been extensively studiedPrimary open angle glaucoma (POAG) is considered to be a leading cause of irreversible blindness worldwide. Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) have been extensively studied as POAG risk factors. Recently, several single-nucleotide polymorphisms (SNPs) for MMPs and TIMPs encoding genes have been reported in POAG patients. The aim of this study was to investigate association of the -1607 1G/2G MMP1 and 372 T/C TIMP1 gene polymorphisms with risk of POAG in a Polish population. In the present case-control study we examined a group of 449 unrelated Caucasian subjects consisting of 196 POAG patients (66 males and 130 females; mean age 70 ± 14) and 253 controls (72 males and 181 females; mean age 67 ± 16). The MMP1-1607 1G/2G and TIMP1 372 T/C gene polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The odds ratios (ORs) and 95% confidence intervals (CIs) for each genotype and allele were calculated. We found a statistically significant increase of the 2G/2G genotype (OR 1.73; 95% CI 1.05-2.86; p=0.019) as well as the 2G allele frequency (OR 1.34; 95% CI 1.03-1.75; p=0.017) of MMP1 in POAG patients in comparison to healthy controls. There were no differences in the genotype and allele distributions and odds ratios of the TIMP1 polymorphism between patients and controls group. We also did not find any association of TIMP1 with MMP1 gene-gene interaction and risk of POAG occurrence. In conclusion, we suggest that the -1607 1G/2G polymorphism of MMP1 gene may be considered as an important risk factor associated with primary open angle glaucoma in a Polish population. However, further in vivo study is needed to evaluate biological importance of MMPs polymorphisms as a risk factor of POAG.

• An association between polymorphism of the heme oxygenase-1 and -2 genes and age-related macular degeneration.

  Authors: Ewelina Synowiec, Jerzy Szaflik, Marta Chmielewska, Katarzyna Wozniak, Anna Sklodowska, Maja Waszczyk, Mariola Dorecka, Janusz Blasiak, Jacek Pawel Szaflik

  Molecular biology reports. 06/2011; 39(3):2081-7.

  Iron may be implicated in the generation of oxidative stress by the catalyzing the Haber-Weiss or Fenton reaction. On the other hand, oxidative stress has been implicated in the pathogenesis ofIron may be implicated in the generation of oxidative stress by the catalyzing the Haber-Weiss or Fenton reaction. On the other hand, oxidative stress has been implicated in the pathogenesis of age-related macular degeneration (AMD) and heme oxygenase-1 (HO-1), encoded by the HMOX1 gene and heme oxygenase-2 (HO-2), encoded by the HMOX2 gene are important markers of iron-related oxidative stress and its consequences. Therefore, variability of the HMOX1 and HMOX2 genes might be implicated in the pathogenesis of AMD through the modulation of the cellular reaction to oxidative stress. In the present work, we investigated the association between AMD and a G → C transversion at the 19 position in the HMOX1 gene (the 19G>C-HMOX1 polymorphism, rs2071747) and a A → G transition at the -42 + 1444 position in the HMOX2 gene (the -42 + 1444A>G-HMOX2 polymorphism, rs2270363) and its modulation by some environmental factors. 279 patients with AMD and 105 controls were recruited in this study and the polymorphisms were typed by restriction fragment length polymorphism and allele-specific polymerase chain reaction (PCR). We observed an association between the occurrence of dry AMD and the G/A genotype of the -42 + 1444A>G-HMOX2 polymorphism (odds ratio (OR) 2.72), whereas the G/G genotype reduced the risk of dry AMD (OR 0.41). The G/C genotype and the C allele of the 19 G>C-HMOX1 polymorphism and the G/G genotype and the G allele of the -42 + 1444A>G-HMOX2 polymorphism were associated with progression of AMD from dry to wet form (OR 4.83, 5.20, 2.55, 1.69, respectively). On the other hand, the G/G genotype and the G allele of the 19 G>C-HMOX1 polymorphism and the A/G genotype and the A allele of the -42 + 1444A>G-HMOX2 polymorphism protected against AMD progression (OR 0.19, 0.19, 0.34, 0.59, respectively). Therefore, the 19G>C-HMOX1 and the -42 + 1444A>G-HMOX2 polymorphisms may be associated with the occurrence and progression of AMD.

• [Pharmacological treatment of recurrent idiopathic anterior uveitis].

  Authors: Jerzy Szaflik, Artur Bachta, Maciej Kulig, Mateusz Tłustochowicz, Andrzej Stankiewicz, Witold Tłustochowicz

  Klinika oczna. 01/2011; 113(4-6):111-6.

  Recurrent anterior uveitis (AU) is an autoimmune disease that in 20% of patients leads to severe visual loss or blindness, thereby a severe disability and significant quality of life impairment. ThatRecurrent anterior uveitis (AU) is an autoimmune disease that in 20% of patients leads to severe visual loss or blindness, thereby a severe disability and significant quality of life impairment. That is why, it is important to conduct a long term treatment after an acute phase of the disease, to keep the remission as long as possible. Up to 50% of patients with recurrent AU have no additional symptoms of systemic disease. This situation, classified as idiopathic AU (IAU), causes a lot of problems when a therapeutic strategy has to be chosen. Nevertheless, IAU is considered to be a first symptom of spondylarthropathy (SpA), even several dozen years prior to other typical manifestations of SpA. There have been published few papers concerning only small groups of patients with IAU, treated with disease modifying anti-rheumatic drugs. The authors present the recommendations for systemic treatment of IAU, based on their own clinical experience, and systematic review of the literature.

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

  Authors: Janusz Blasiak, Jerzy Szaflik, Jacek Pawel Szaflik

  Frontiers in bioscience : a journal and virtual library. 01/2011; 16:1551-9.

  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, alteredReactive 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.

• Evaluation of oxidative stress markers in pathogenesis of primary open-angle glaucoma.

  Authors: Ireneusz Majsterek, Katarzyna Malinowska, Malgorzata Stanczyk, Michal Kowalski, Jan Blaszczyk, Anna K Kurowska, Anna Kaminska, Jerzy Szaflik, Jacek P Szaflik

  Experimental and molecular pathology. 01/2011; 90(2):231-7.

  Primary open-angle glaucoma (POAG) is the leading cause of blindness in the industrial countries. It is reported that oxidative stress might be an important risk factor in the pathogenesis of POAG.Primary open-angle glaucoma (POAG) is the leading cause of blindness in the industrial countries. It is reported that oxidative stress might be an important risk factor in the pathogenesis of POAG. Forty subjects including 20 patients with open-angle glaucoma (9 men and 12 women, mean age 61.8±12.1yr) and 20 controls without glaucoma symptoms (9 men and 12 women, mean age 58.1±17.7yr) were enrolled in our study. The main aim of the work was to evaluate oxidative stress markers in the pathogenesis of open-angle glaucoma. In our work the activity of antioxidant enzymes: catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPX) as well as the total antioxidant status (TAS) was estimated. An alkaline comet assay was used to measure DNA damage of strand breaks (SB), oxidized purines as glicosylo-formamido-glicosylase (Fpg) sites, and oxidized pirmidines as endonuclease III (Nth) sites. We measured endogenous as well as exogenous DNA damage after 10μM hydrogen peroxide treatment (H(2)O(2)). We did not observe any statistical changes of DNA strand break lesion in examined POAG patients according to healthy subjects (P>0.05). However, either endogenous (P<0.01) or exogenous (P<0.001) levels of oxidative DNA damage in POAG patients were found to be statistically higher than controls. A significant decrease of antioxidant enzymes: CAT (P<0.001), SOD (P<0.05), and GPX (P<0.001) and a non-statistical decrease of TAS status (P>0.05) in glaucoma patients according to controls were also indicated. In conclusion our data revealed that oxidative stress had a pathogenic role in primary open-angle glaucoma. Therefore, we suggested that the modulation of a pro-oxidant/antioxidant status might be a relevant target for glaucoma prevention and therapy.

• 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.

  Authors: Daniel Wysokinski, Jerzy Szaflik, Anna Sklodowska, Urszula Kolodziejska, Mariola Dorecka, Dorota Romaniuk, Katarzyna Wozniak, Janusz Blasiak, Jacek Pawel Szaflik

  The Tohoku journal of experimental medicine. 01/2011; 223(4):253-61.

  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. OxidativeAge-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.

• Comparison of the biometric measurements obtained using noncontact optical biometers LenStar LS 900 and IOL Master V.5.

  Authors: Wojciech Kołodziejczyk, Tomasz Gałecki, Monika Łazicka-Gałecka, Jerzy Szaflik

  Klinika oczna. 01/2011; 113(1-3):47-51.

  Evaluation of the measurement accuracy using a new optical biometer LenStar LS 900 (Haag Strait) and comparison according to the IOL Master V.5 (Zeiss) device. In a prospective clinical studyEvaluation of the measurement accuracy using a new optical biometer LenStar LS 900 (Haag Strait) and comparison according to the IOL Master V.5 (Zeiss) device. In a prospective clinical study biometric measurements along with artificial lens power calculation using the LenStar LS 900 device were performed. A total number of 106 patients qualified for a cataract extraction procedure were included in the study. Measurements along with lens power calculation were repeated using the IOL Master V.5. device. Results were elaborated using Pearson's correlation and Bland-Altman plot. Measurements were performed in 204 eyes of 106 patients. Mean values and ranges of biometry results were as follows: 23.46 +/- 2.81 mm [20.79-29.80] for IOL Master, and 23.47 +/- 2.83 mm [20.79-29.86] for LenStar LS 900. In keratometry for flat meridian (K1) the following data was obtained: mean 43.24 +/- 3.22 D [range 38.27-47.94] for IOL Master and 42.44 +/- 3.15 D [38.27-47.94] for LenStar LS 900. For steep meridian (K2) the data obtained were 44.14 +/- 3.40 D; [39.29-49.13] and 43.27 +/- 3.34 D [38.61-48.4] respectively. The obtained calculation results for each eye were as follows: mean 21.23 +/- 8.07, [range 3.25-28.99] for Hagis formula, 21.14 +/- 6.90; [4.83-27.6] for SRK II, 21.04 +/- 7.78; [3.05-28.05] for SRK/T, 21.09 +/- 8.13; [2.43-28.61] for Holladay using IOL Master and 21.41 +/- 8.23; [2.99-29.15] for Haigis; 21.24 +/- 7.00; [4.6-27.71] for SRK II, 21.13 +/- 7.90; [2.76-28.18] for SRK/T; 21.09 +/- 8.13; [2.16-28.76] for Holladay using LenStar LS 900. The LenStar LS 900 device enables to perform accurate and repetitive biometric measurements and implant power calculations. Implant calculation results obtained using the LenStar LS 900 device are comparable to those achieved using the IOL Master V.5 device, which has been commonly accepted as standard for over a decade. The use of both devices is limited by significant lens opacification and posterior capsule calcification. In such cases, additional ultrasound biometry should be performed. Keratometry results obtained using both devices should not be used alternatively because of the different measurement methods and different refraction indexes. The LenStar LS 900 device comparing to the IOL Master additionally enables pachymetry, macular retinal thickness, lens thickens and pupil diameter measurement. Accuracy of those measurements should be studied in the future.

• Prevention of vision loss after cataract surgery in diabetic macular edema with intravitreal bevacizumab: a pilot study.

  Authors: Jacek P Szaflik, Jerzy Szaflik

  Retina (Philadelphia, Pa.). 09/2010; 30(8):1328-9; author reply 1329.

• Reactive oxygen species promote localized DNA damage in glaucoma-iris tissues of elderly patients vulnerable to diabetic injury.

  Authors: Jacek P Szaflik, Pawel Rusin, Anna Zaleska-Zmijewska, Michał Kowalski, Ireneusz Majsterek, Jerzy Szaflik

  Mutation research. 02/2010; 697(1-2):19-23.

  Glaucoma is typically an insidious-onset disease with serious visual consequences that has been positively linked to diabetes mellitus (DM). Glaucoma is more often present in the elderly. ImportantGlaucoma is typically an insidious-onset disease with serious visual consequences that has been positively linked to diabetes mellitus (DM). Glaucoma is more often present in the elderly. Important prognostic factors of glaucoma may be oxidative stress resulting from the toxic effects of glucose, and diabetes-associated vascular complications. Fifty-five patients and control subjects aged 71.0+/-10.1 yrs were enrolled in this study. Iris-tissue samples from DM type-2 patients, primary open-angle glaucoma-positive and -negative DM patients, and from healthy subjects were examined by use of the alkaline comet assay. We measured the DNA damage as numbers of strand breaks (SBs), oxidized purines as glycosyl-formamido-glycosylase (Fpg)-susceptible sites, and oxidized pyrimidines as endonuclease III (Nth)-susceptible sites. It was found that the level of oxidative damage in iris tissue was statistically higher in DM and glaucoma patients than that in healthy controls (oxidized purines: 38.0% and 34.7% vs 15.4%; oxidized pyrimidines: 43.3% and 39.0% vs 23.3%; P<0.001). Interestingly, we found strongly elevated levels of oxidized purines and pyrimidines in glaucomatous patients who also had DM, in comparison with healthy controls (oxidized purines: 55.7% vs 15.4%; oxidized pyrimidines: 61.8% vs 23.3%; P<0.001). Our observations suggest that the generation of reactive oxygen species may promote localized DNA damage in glaucoma-iris tissues of elderly patients vulnerable to diabetic injury.

• Demodex infections in general Polish population, in patients suffering from blepharitis, and among people who work with microscopes.

  Authors: Aleksandra Garbacewicz, Monika Udziela, Barbara Grytner-Ziecina, Jacek P Szaflik, Jerzy Szaflik

  Klinika oczna. 01/2010; 112(10-12):307-10.

  In this study we examined 264 people to estimate the prevalence of Demodex infection. The subjects were divided into 4 groups. First two groups consisted of young people (mean age 22 years) andIn this study we examined 264 people to estimate the prevalence of Demodex infection. The subjects were divided into 4 groups. First two groups consisted of young people (mean age 22 years) and seniors (mean age 66 years) respectively. The third group included people who work with microscopes (mean age 44 years). Forth group consisted of patients with diagnosed blepharitis (mean age 65 years). From every individual 3-4 lashes were epilated from the eyelids of both eyes and examined under the microscope for Demodex mites. The statistical analysis based on logistic regression was used to estimate the probability of infection. Age was independent variable significant for this model (p < 0.001). Significant relation between age and probability of Demodex infection was showed. The lowest number of infected individuals was observed in the group of young subjects (only 5%), the highest in the blepharitis patients (74% infected). Among microscope users 30% were infected, in the group of seniors 34% were infected.

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

  Authors: Katarzyna Janik-Papis, Malgorzata Zaras, Anna Krzyzanowska, Katarzyna Wozniak, Janusz Blasiak, Jerzy Szaflik, Jacek P Szaflik

  Experimental and molecular pathology. 09/2009;

  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 vascularThe 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.

• DNA damage and repair in age-related macular degeneration.

  Authors: Jacek P Szaflik, Katarzyna Janik-Papis, Ewelina Synowiec, Dominika Ksiazek, Magdalena Zaras, Katarzyna Wozniak, Jerzy Szaflik, Janusz Blasiak

  Mutation research. 07/2009;

  Age-related macular degeneration (AMD) is a retinal degenerative disease that is the main cause of vision loss in individuals over the age of 55 in the Western world. Clinically relevant AMD resultsAge-related macular degeneration (AMD) is a retinal degenerative disease that is the main cause of vision loss in individuals over the age of 55 in the Western world. Clinically relevant AMD results from damage to the retinal pigment epithelial (RPE) cells thought to be mainly caused by oxidative stress. The stress also affects the DNA of RPE cells, which promotes genome instability in these cells. These effects may coincide with the decrease in the efficacy of DNA repair with age. Therefore individuals with DNA repair impaired more than average for a given age may be more susceptible to AMD if oxidative stress affects their RPE cells. This may be helpful in AMD risk assessment. In the present work we determined the level of basal (measured in the alkaline comet assay) endogenous and endogenous oxidative DNA damage, the susceptibility to exogenous mutagens and the efficacy of DNA repair in lymphocytes of 100 AMD patients and 110 age-matched individuals without visual disturbances. The cells taken from AMD patients displayed a higher extent of basal endogenous DNA damage without differences between patients of dry and wet forms of the disease. DNA double-strand breaks did not contribute to the observed DNA damage as checked by the neutral comet assay and pulsed field gel electrophoresis. The extent of oxidative modification to DNA bases was grater in AMD patients than in the controls, as probed by DNA repair enzymes NTH1 and Fpg. Lymphocytes from AMD patients displayed a higher sensitivity to hydrogen peroxide and UV radiation and repaired lesions induced by these factors less effectively than the cells from the control individuals. We postulate that the impaired efficacy of DNA repair may combine with enhanced sensitivity of RPE cells to blue and UV lights, contributing to the pathogenesis of AMD.

• [Best's disease--family case report]

  Authors: Sława Kwiecien, Magdalena Ulińska, Robert Sulak, Jerzy Szaflik

  Klinika oczna. 02/2009; 111(1-3):62-5.

  Friedrich Best described in 1905 for the first time a genetic disease which is subject to autosomal dominant inheritance. Yellow material is assembled under pigment epithelium, especially in theFriedrich Best described in 1905 for the first time a genetic disease which is subject to autosomal dominant inheritance. Yellow material is assembled under pigment epithelium, especially in the macula. Afterwards pigment epithelium and choroid become involved in the process and visual acuity decreases. In the last phase the fibrotic scar appears in the macula with deterioration of visual acuity. In some late cases choroidal neovascularization develops too. We present a two-generation-family which suffers from Best's disease. Our patients: mother, her daughter and her son demonstrate changes in different phases of development. In mother's case we found fibrotic scar and atrophy of pigment epithelium, which is typical of the late phase of the disease. Daughter complained of decrease in visual acuity, the subretinal neovascularisation was found in the macula and photodynamic therapy was performed. Visual acuity improved from 0.5 to 0.8 and has been stable for 11 months. Son did not report any symptoms. Some yellow material was found under the macula in both eyes without decrease of visual acuity. Described case shows importance of the whole family members' examination in case of Best's disease. Early diagnosis of this serious disease makes possibility of proper treatment.

• [Pseudoexfoliation syndrome (PEX)--a systemic disorder]

  Authors: Anna K Kurowska, Anna Kamińska, Justyna Izdebska, Jacek P Szaflik, Jerzy Szaflik

  Klinika oczna. 02/2009; 111(4-6):160-4.

  Pseudoexfoliation syndrome is an age-related elastosis. The current pathogenetic concept describes PEX as an elastic microfibrillopathy involving TGF-betal, oxidative stress and impaired cellularPseudoexfoliation syndrome is an age-related elastosis. The current pathogenetic concept describes PEX as an elastic microfibrillopathy involving TGF-betal, oxidative stress and impaired cellular protection mechanisms as a key pathogenetic factors. Extracellular PEX material is produced, aggregated and accumulated in the anterior segment of the eye. It is also present in the skin, lungs, liver and other visceral organs. These findings suggest that pseudoexfoliation syndrome is a systemic disorder. It is also associated with coronary artery disease, hypertension, Alzheimer's disease and other cardio-vascular problems. The pseudoexfoliation syndrome has been found to be associated with higher mean intraocular pressure, secondary open-angle glaucoma, mature and nuclear cataract. Ocular depositions of pseudoexfoliation material can lead to many intraocular complications. That's why special attention is required before, during and after surgery. z

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

  Authors: Jacek P Szaflik, Janusz Błasiak, Anna Krzyanowska, Małgorzata Zaraś, Katarzyna Janik-Papis, Anna I Borucka, Katarzyna Woźniak, Jerzy Szaflik

  Klinika oczna. 02/2009; 111(4-6):125-7.

  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 macularPURPOSE: 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.