Association between Monocyte Chemoattractant Protein-1 -2518 (A/G) Single Nucleotide Polymorphism and Chronic Periodontitis in End-stage Renal Disease Patients - A Case-control Study.

Monocyte chemoattractant protein 1 (MCP-1) plays an important role in the development of periodontitis. The purpose of this investigation was to evaluate association of the  MCP-1 gene polymorphism with chronic periodontitis in patients with end-stage renal disease (ESRD). One hundred fifty ESRD patients with chronic periodontitis (CP), 100 without CP and 190 healthy controls were included in this study. Genomic DNA from all participants was genotyped for the -2518 (A/G) polymorphism by a polymerase chain reaction - restriction fragment length polymorphism (PCR--RFLP) assay. Significant differences were observed in the genotype and allele frequencies between patients with ESRD and CP and controls.  The G allele frequency was significantly higher in patients than in control subjects, with odds ratio 1.77 (95 % CI 1.2-2.5), p = 0.0014. For the GG genotype the OR was 3.63 (95 % CI 1.5-8.76), p = 0.041.  No significant differences in the polymorphism distribution were observed between ESRD patients  without CP and control subjects. Comparison of the MCP-1 gene polymorphism distribution in ESRD patients with various primary diseases leading to ESRD did not show any significant differences. The mean MCP-1 serum levels were compared between subgroups. They were significantly higher in ESRD patients with CP (582 ± 112 pg/ml) than in patients without CP (309 ± 103 pg/ml) and controls (265 ± 85 pg/ml). Our results suggest that the MCP-1-2518 A/G  polymorphism might be a novel risk factor for developing chronic periodontitis in patients with ESRD.

Interleukin-1ß Gene (IL1B) Polymorphism and Risk of Developing Diabetic Nephropathy.

Objective: Cytokines play an important role in the pathogenesis of type 2 diabetes (T2DM) and its complications. The aim of the study was to evaluate an association of the -511 (C/T) polymorphism in the IL1B gene with diabetic nephropathy (DN). Methods: The study population included 860 patients with T2DM (506 with diabetic nephropathy and 354 without nephropathy) as well as 505 healthy individuals. Genomic DNA was genotyped for the IL1B -511 (C/T) polymorphism using PCR-RFLP technique. Results: The IL1B -511 C/T polymorphism was genotyped in 860 T2DM patients with or without DN and 505 healthy individuals. The average age of patients was 65.3 years in DN+ and 62.2 years in DN- subgroups. The genotype distribution did not differ significantly between patients and controls. Only a tendency to a slight increase of T allele frequency was observed in patient group. Genotype and allele frequencies of -511 C/T polymorphism were compared in patients with DN and those without it. The minor allele (T) and homozygote TT frequencies were significantly different between subgroups. The T allele was more frequent in DN+ patients, with odds ratio 1.45 (95% CI 1.2-1.8), p = 0.0003. The TT genotype frequency was also higher in DN+, with OR 1.76 (96% CI 1.1-2.7), p = 0.01. Conclusion: In a studied population the -511 C/T polymorphism in the IL1B gene is associated with diabetic nephropathy in dialyzed T2DM patients. Further studies are required to confirm the clinical significance of this finding.

IL4 gene VNTR polymorphism in chronic periodontitis in end-stage renal disease patients.

OBJECTIVE: Interleukin-4 gene polymorphisms were found to be associated with periodontitis. The purpose of this case-control study was to evaluate association of IL4 VNTR polymorphism with periodontitis in patients with end-stage renal disease (ESRD). SUBJECTS AND METHODS: We examined 180 ESRD patients with chronic periodontitis, 82 without CP and 180 healthy controls. Genomic DNA from all subjects was genotyped for the IL4 VNTR polymorphism by polymerase chain reaction (PCR). RESULTS: Genotype distribution in all groups followed Hardy-Weinberg equilibrium. Significant differences in genotype and allele frequencies were observed between groups. The patient group had higher frequency of P1 allele than controls, with odds ratio for P1 allele 1.6 (95% CI 1.1-2.3) and P1P1 genotype 2.73 (95% CI 1.06-7.5). There were no differences in polymorphism distribution between ESRD patients without CP and controls. Periodontal disease was more severe in older patients (≥50 years). Similarly, patients with T2DM had more severe manifestation of CP than patients without diabetes (p = 0.01 for plaque index, p = 0.004 for bleeding index and p = 0.03 for gingival index). CONCLUSION: Our findings suggest that VNTR polymorphism in IL4 gene might be a risk factor for chronic periodontitis in patients with ESRD.

The membrane with polylactide and hyaluronic fibers for skin substitute.

PURPOSE: Skin substitutes are heterogeneous group of scaffolds (natural or synthetic) and cells. We hypothesize that nanofibers with layer composition made of polylactide (PLA) and sodium hyaluronate (HA) obtained using electrospinning method are a good matrix for cell adhesion and proliferation. METHODS: Optimal conditions of electrospinning of PLA and HA nanofibers to create layered compositions (PLA membrane covered with HA nonwovens) were determined by modifying parameters such as the appropriate amount of solvents, polymer concentration, mixing temperature and electrospinning process conditions. By changing the parameters, it was possible to control the diameter and properties of both polymer fibers. The spinning solution were characterized by surface tension and rheology. A scanning electron microscope (SEM) was used to determine the morphology and fiber diameters: PLA and HA. Structure of the PLA/HA nonwoven was analyzed using spectroscopy (FTIR/ATR). Biocompatibility of the nonwoven with fibroblasts (ECM producers) was assessed in the in vitro conditions. RESULTS: The results showed that stable conditions for the formation of submicron PLA fibers were obtained using a 13% wt. solution of the polymer, dissolved in a 3:1 mixture of DCM:DMF at 45 °C. The hyaluronic fibers were prepared from a 12% wt. solution of the polymer dissolved in a 2:1 mixture of ammonia water and ethyl alcohol. All materials were biocompatible but to a different degree. CONCLUSIONS: The proposed laminate scaffold was characterized by a hydrophobic-hydrophilic domain surface with a maintained fiber size of both layers. The material positively underwent biocompatibility testing in contact with fibroblasts.

Interleukin-18 gene polymorphism and risk of CVD in older patients with type 2 diabetes mellitus.

OBJECTIVE: Interleukin-18 (IL-18), a proinflammatory cytokine, plays a key role in the acute and chronic inflammatory processes. It is associated with risk of developing cardiovascular disease (CVD). The aim of this study was to evaluate association between G(-137)C polymorphism (rs187238) in the IL-18 gene and risk of diabetes and CVD in type 2 diabetes patients. METHODS: We examined 1548 T2DM patients and 590 controls. All subjects were genotyped for the G(-137)C promoter region polymorphism by polymerase chain reaction (PCR-SSP). RESULTS: Genotype distribution of the G(-137)C polymorphism showed no significant difference between T2DM patients and controls (p=0.115). An association with CVD was analyzed in two age groups: ⩾65 and <65years. In patients younger than 65years there was a tendency to association of CC genotype with CAD (OR 1.87, 95% CI 1.0-3, p=0.051). In contrast, in subjects aged 65 or older, the C allele and CC genotype showed the significant association with the presence of CVD, with the OR 1.99, p=0.001 and OR 5.31, p=0.006, respectively. The C allele carriers showed the higher prevalence of CVD compared to non-carriers (61% vs. 39%, p<0.0001). CONCLUSION: Older T2DM patients carrying the C allele of IL-18 G(-137)C polymorphism have a significantly increased risk of CVD.

The effect of Toll-like receptor 4 gene polymorphism on vascular complications in type 2 diabetes patients.

OBJECTIVE: The aim of our study was to assess the association between the TLR4 Asp299Gly polymorphism and vascular complications in patients with type 2 diabetes. METHODS: We examined 1090 patients with T2DM and 716 healthy controls. All subjects were genotyped for the Asp299Gly polymorphism by polymerase chain reaction (PCR) and restriction analysis. RESULTS: The genotype frequencies of the Asp299Gly polymorphism were similar in T2DM patients and controls (p=0.512 and 0.311, respectively). The polymorphism was analyzed in subgroups of patients with macro- and microvascular complications. The distribution of genotypes was significantly different between patients with CVD and those without CVD. A significant increase of G allele frequency was observed in CVD+ patients, with odds ratio 2.06 (1.27-3.34), p=0.0035. The same effect was found when patients with diabetic retinopathy were compared with those without it (OR for G allele 2.12, 95% CI 1.43-3.12, p=0.0002). There were no statistically significant differences in genotype distribution between patients with diabetic nephropathy or neuropathy and those without these complications. CONCLUSIONS: The results of our study demonstrated that the G allele of the Asp299Gly polymorphism of the TLR4 gene is associated with increased risk of cardiovascular disease and diabetic retinopathy in type 2 diabetes patients.

Semi-automated kinetic perimetry provides additional information to static automated perimetry in the assessment of the remaining visual field in end-stage glaucoma.

PURPOSE: To test the hypothesis whether semi-automated kinetic perimetry (SKP) provides additional information to static automated perimetry (SAP) in the assessment of the remaining visual field in end-stage glaucoma, as defined by disc appearance (cup-to-disc ratio worse than 0.9) and SAP criteria (MD worse than 20 dB). METHODS: Fifty eyes of 44 patients presenting with end-stage glaucoma were examined first with SAP within the central 30° using stimulus size III, followed by SKP within 90° using test targets III4e and V4e. RESULTS: Overall, SKP provided additional information over SAP in more than half (54%) of the cases. In 16 instances (32%), SKP revealed visual field island beyond 30° that was undetected by SAP. In eight cases (16%), SKP showed both a central island and peripheral island of visual field. In three cases (6%) altitudinal scotomatous loss was found using SKP, but not in SAP. In 23 cases (46%) the central visual field island was defined both with SAP and SKP. The mean examination duration was 4 min for SAP and 9 min for SKP. CONCLUSIONS: In clinical practice, SKP with III4e and V4e test targets provides more information than 30° SAP regarding the remaining peripheral VF in patients with end-stage glaucoma; however, a longer test time is required for SKP.

Relationship between the area of isopters and Vigabatrin dosage during two years of observation.

BACKGROUND: The aim of the study was to evaluate the relationship between the area of isopters obtained using semi-automated kinetic perimetry (SKP) and Vigabatrin dosage in epilepsy patients with pretreatment baseline examination during 2-years of the follow-up. METHODS: 29 epilepsy patients were included into the study, but 15 individuals were excluded due to cognitive impairment, intracranial pathologies or eye diseases. Finally, 14 patients were examined with SKP before VGB treatment and after 6, 12, 18, and 24 months. Reaction time (RT)-corrected areas of three isopters (III4e, I4e and I2e) were measured for each of five examinations and compared intra-individually during 2-years period. Additionally, six epilepsy patients on other antiepileptic drugs were examined five times with SKP as a control. RESULTS: There was a significant decrease of I2e, I4e and III4e isopters' area during the follow-up of two years. Correlation was found between the I2e isopter's area and both cumulative dose and mean daily dose of VGB. With increasing RT, there was decreasing of all isopters' area in patients receiving VGB. In epilepsy patients who were not receiving VGB, there were no significance differences in isopters' area during follow-up. CONCLUSION: There was attenuation of area of III4e, I4e and I2e isopters obtained with SKP during a period of 2 years. RT, the cumulative dose and the mean daily dose of VGB influenced isopters' area obtained with SKP.

Polymorphism in microRNA-196a2 contributes to the risk of cardiovascular disease in type 2 diabetes patients.

AIMS: To investigate the effect of the microRNA-196a2 gene polymorphism (rs11614913) on risk of cardiovascular disease in type 2 diabetes patients. METHODS: We examined 920 patients with diabetes and 834 healthy controls. All subjects were genotyped for the miRNA-196a2 SNP by polymerase chain reaction (PCR) and restriction analysis. RESULTS: The genotype distribution among controls and patients was in Hardy-Weinberg equilibrium (p=0.227 and 0.308, respectively). The frequency of the T allele was lower in patients than in controls (p=0.044). The odds ratio 0.66 (95% CI 0.54-0.79) suggests an association of the T allele with decreased risk of T2DM. For the main purpose of the study, T2DM patients were stratified into patients with CVD and those without it. The T allele and TT genotype were significantly more frequent in patients with CVD compared to those without CVD (p=0.013, p<0.001, respectively). The odds ratio for the T allele in the CVD+subgroup vs. CVD- was 1.76 (1.35-2.30), p<0.0001, mostly due to the overrepresentation of TT homozygotes. The highest risk of development of CVD was observed in the additive model for TT homozygotes (OR 3.33, 95% CI 2.05-5.42, p<0.0001). CONCLUSION: Our findings suggest that miRNA-196a2 T/C polymorphism (rs11614913) is associated with an increased risk of CVD in type 2 diabetes patients. This provides further insights on pathogenesis of cardiovascular disease in type 2 diabetes patients.

M-charts as a tool for quantifying metamorphopsia in age-related macular degeneration treated with the bevacizumab injections.

BACKGROUND: This article is aimed to assess quantitatively metamorphopsia using M-charts in patients suffering from wet age-related macular degeneration (AMD) treated with the intravitreal bevacizumab injections and to compare the results with traditional Amsler grid and ocular coherence tomography (OCT). METHODS: Thirty-six patients diagnosed with wet AMD were examined one day before and one month after the intraocular injection of bevacizumab. Horizontal and vertical metamorphopsia scores using M-charts, distance visual acuity, Amsler test and OCT were performed at each visit. Additionally, 23 healthy subjects were examined as a control group. RESULTS: The rate of metamorphopsia detection was 89% with M-charts and 69% with Amsler test. The horizontal metamorphopsia score improved in 22 patients, the vertical metamorphopsia score improved in 16 patients, the Amsler grid results improved in 6 patients, visual acuity improved in 17 patients. There was no correlation between the degree of metamorphopsia and the visual acuity or the central retinal thickness (CRT). The specificity of both the M-charts and Amsler grid was 100%. CONCLUSIONS: The rate of metamorphopsia detection in wet AMD patients was better with M-charts than with Amsler grid. M-charts may be used in the assessment of efficacy of treatment with intravitreal bevacizumab injections as another outcome measure, moreover they can be used even at home for the self-assessment. M-charts provide additional information concerning the visual function, independent of the visual acuity, CRT and morphological changes in OCT.

Presence and distribution of L-kynurenine aminotransferases immunoreactivity in human cataractous lenses.

PURPOSE: To investigate the presence and distribution of l-kynurenine aminotransferases immunoreactivity in human and animal lenses during cataract formation. METHODS: Immunohistochemistry was conducted using polyclonal antibodies against KAT I, KAT II and KAT III on sections of 26 anterior capsules from patients undergoing surgical treatment of anterior subcapsular cataract (ASC) and 22 cataractous lenses from human eyes enucleated because of choroidal malignant melanoma. Additionally, the eyes of 11-month-old DBA/2J mice (6 eyes) were investigated (with KAT I and II). Ten clear human lenses and four BL6 mice lenses were used as controls. Spatial immunoreactivity patterns of enzymes were compared with Periodic Acid - Schiff (PAS)-stained sections. RESULTS: Immunohistochemical analysis revealed presence of KAT I, KAT II and KAT III in extracellular structures of all studied types of cataract in human eyes showing specific pattern of the stain. In cortical cataract, immunoreactivity was observed on cortical lens fibres. In nuclear cataract, KAT II revealed stronger and diffused staining than KAT I. Additionally, both KAT showed more pronounced staining at the edge of small clefts. In normal human lenses, KAT I, II and III, immunoreactivity was not observed. Presence of KAT I and KAT II in the intercellular substance of DBA/2J mice cataract was observed. In BL6 mice lenses without cataract, only weak KAT I and KAT II staining was observed. CONCLUSIONS: Presence of l-kynurenine aminotransferases in extracellular matrix (ECM) during human cataract formation suggests that products of l-kynurenine pathway might be involved in mechanisms of cataractogenesis.