Wacław Kopeć

Total antioxidant status and 8-hydroxy-2'-deoxyguanosine levels in gingival and peripheral blood of periodontitis patients.

Introduction:The aim of this study was to determine 8-OHdG concentration as a biomarker of oxidant-induced DNA damage and to assess total antioxidant status (TAS) in gingival and peripheral blood during periodontal lesion.Materials and Methods:The study included 56 untreated periodontitis patients (26 with aggressive periodontitis, and 30 with chronic periodontitis (CP). The control group consisted of 25 healthy volunteers without pathological changes in the periodontium. Competitive ELISA was used to measure 8-OHdG. A colorimetric method based on the reduction of ABTSo+ radical cation generation was used to measure TAS.Results:Significantly higher 8-OHdG concentrations were detected in the gingival blood in both groups of patients with periodontitis than in the control group. Subjects with CP had significantly decreased TAS levels in the gingival blood compared with the control group. A significantly decreased TAS level in the peripheral blood in both patient groups compared with the control group was found. Significant positive correlation between TAS levels in venous and gingival blood in all the periodontitis patients and in the CP group was observed.Conclusions:The oxidative burst in periodontitis may lead to significant local damage to nucleic acids. The significantly decreased TAS level in the gingival blood of CP patients compared with the healthy subjects suggests the possibility of a significant decrease in local antioxidant system capacity during the course of periodontitis. The decreased TAS level in the peripheral blood in the group of all patients with periodontitis may be one of the pathogenic mechanisms underlying the links between periodontal disease and several systemic diseases for which periodontitis is regarded as a independent risk factor.

Imbalance of Metallaproteinase/Tissue Inhibitors of Metalloproteinase System in Renal Transplant Recipients With Chronic Allograft Injury

INTRODUCTION Nowadays, renal allografts continue to be lost at the rate of 2% to 4% per year beyond the first year after transplantation due to chronic allograft injury. Excessive accumulation of extracellular matrix results from overproduction and/or defective degradation by proteolytic enzymes, among which metalloproteinases (MMPs) play a major role. The aim of this study was to assess the role of MMPs in renal transplant recipients (RTR) in the context of allograft injury or proteinuria. MATERIALS AND METHODS Plasma and urine MMP-2 and MMP-9 and tissue inhibitors of metalloproteinases (TIMPs) were assessed by enzyme-linked immunoassay in 150 RTR including 66% males with an overall mean age of 49.2±11.5 years. The subjects were examined at a mean of 73.4±41.2 months (range=12-240) after kidney transplantation. Thirty-seven healthy volunteers including 54% male with an overall mean age of 48.4±14.1 years served as a control group. RESULTS Renal transplant recipients displayed significantly decreased plasma MMP-2 activity compared with healthy controls (P<.000) probably due to increased inhibitory plasma (p) TIMP-2 activity (P=.0029), and lower plasma MMP-2:TIMP-2 index (P<.0001). Plasma MMP-9 and pTIMP-1 activities were twofold increased in RTR compared with controls (P=.0015 and P<.000) but with a nearly stable plasma MMP-9:TIMP-1 index (P=NS). There was no difference between RTR and controls according to urine (u) MMP-2 activity, but uMMP-9 was increased in RTR compared with healthy controls (P=.0032). Urine MMP-9 potential was probably diminished by increased uTIMPs (uTIMP-2, P=.017; uTIMP-1, P=.000), which contributed to graft impairment or proteinuria. CONCLUSION Our study revealed profibrotic MMP/TIMP constellations in RTR that show an imbalance in plasma MMP-2 and MMP-9 with increased plasma and urinary TIMPs. The net proteolytic potential of increased plasma and urinary MMP-9 may be diminished significantly by enhanced plasma and urine TIMP activities.

Plasma and urine leukocyte elastase–α1protease inhibitor complex as a marker of early and long-term kidney graft function

BACKGROUND Neutrophils are mediators of ischaemia/reperfusion (I/R) injury following kidney transplantation (kTx). Leukocyte elastase (LE) complex with alpha(1)protease inhibitor (LE-alpha(1)PI) is a marker of neutrophil degranulation. The aim of this study was to evaluate LE-alpha(1)PI as a marker of I/R kidney damage and to search for correlations between leukocyte activation and post-transplant complications. METHODS Plasma and urine LE-alpha(1)PI were estimated in 55 deceased-donor kidney graft recipients on postoperative days (POD) 1, 3 and 7, as well as in the late post-transplant period. RESULTS The plasma LE-alpha(1)PI level peaked on POD 1 after kTx, and the urine LE-alpha(1)PI peaked on POD 3. On POD 1 and POD 3, the urine LE-alpha(1)PI levels were higher in delayed graft function (DGF) patients than in patients with immediate graft function (IGF: P < 0.001 and P < 0.003, respectively). Urine LE-alpha(1)PI excretion on POD 1 was significantly higher in patients with longer cold ischaemia time (CIT) than in patients with shorter CIT, P < 0.002. Multivariate regression model revealed two factors influencing the occurrence of early acute rejection-urine LE-alpha(1)PI complex on POD 3 and human leukocyte antigen (HLA) mismatches. There was a significant association between the plasma LE-alpha(1)PI on POD 3 and serum creatinine level 6 and 12 months after kTx (r(2) 0.24; P < 0.005 and 0.19; P < 0.005, respectively). CONCLUSIONS This study is the first presentation of a simple, non-invasive measurement of neutrophil activation after kTx. It also demonstrates a strong correlation between the early post-transplant LE-alpha(1)PI complex level and kidney graft function.

Elastase deposits in the kidney and urinary elastase excretion in patients with glomerulonephritis – evidence for neutrophil involvement in renal injury

Objective. Elastase is a key proteolytic enzyme released during polymorphonuclear leukocyte degranulation. There are abundant data of elastase involvement in the development of injury in experimental models of glomerulonephritis (GN), but scant direct evidence of its involvement in human primary GN. The aims of this study were to determine the immunolocalization of elastase deposits in kidney biopsy specimens from patients with primary idiopathic GN, to attempt to correlate the distribution and intensity of deposits with urinary elastase excretion, and to determine clinical markers of renal injury in several types of primary idiopathic GN. Material and methods. The immunohistochemical localization and intensity of elastase deposits in kidney biopsies, the urinary excretion of leukocyte elastase, and proteinuria and serum creatinine levels were evaluated in 23 patients with primary GN and the associations between these factors were sought. Results. Patients with crescentic proliferative GN had the highest intensity of elastase deposits. In this group of patients, elastase was present in the glomerular endothelium, as well as in the tubular epithelium and interstitium. Patients with a high intensity of elastase deposits within the glomerular endothelium and Bowmans capsule had significantly higher urinary excretion of elastase. Patients with interstitial, mesangial and perivascular elastase deposits had significantly higher serum creatinine than those without. Patients with elastase deposits in the glomerular endothelium and in the interstitium had insignificantly higher proteinuria than those without. Conclusion. Our data provide morphological evidence of leukocyte elastase involvement in renal injury occurring in the course of primary idiopathic GN, in particular in the proliferative types.

Serum levels of proANP and albumin are independent predictors of mortality in the high-risk patients (elderly and diabetics) treated by haemodialysis (HD) and continuous peritoneal dialysis in 4-year prospective observation

1.73 m, the mean time to reach CKD stage 5 with dual RAS blockade would be 106 years. Finally, 8502 patients were exposed to dual RAS blockade in the ONTARGET Study, for a median follow-up of 56 months. During this 39 676 patient-year follow-up, the claimed significant increased risk of renal failure requiring renal replacement therapy was due to an excess of 15 cases of acute renal failure, without any difference in the incidence of chronic dialysis [2]. This excess risk of <4 events for 1000 patients treated per year is in fact a rather good tolerance profile for patients with a strong RAS blockade. Nevertheless, we should be aware that treatment strategies aiming at further decreasing glomerular pressure and proteinuria, with uptitration of either RAS blockade or diuretic dosage, requires cautious monitoring to prevent pre-renal failure. The second comment suggests that RAS blockade in combination with diuretics may increase urine volume and subsequently increase fluid intake via drinking, contributing to microalbuminuria reduction [6]. Indeed, the suppression of vasopressin (AVP) by increased water ingestion reduces proteinuria, glomerulosclerosis and tubulointerstitial fibrosis in 5/6 nephrectomized rats [7]. The AVP receptor antagonists also decrease proteinuria in animal models via haemodynamic and non-haemodynamic effects of AVP blockade, but without increasing urinary output because of AVP-resistant downregulation of aquaporin-2 and aquaporin-3 in CKD [8]. Indeed, a defective urine concentrating capacity is a manifestation of CKD. This may explain a post hoc analysis of the Modification of Diet in Renal Disease (MDRD) Study that found an association between high urine volumes and rates of GFR decline, suggesting that high fluid intakes make CKD progression worse [9]. Therefore, the safety and efficacy of increased water intake have yet to be confirmed in a prospective randomized controlled study. Nevertheless, loop diuretics increase diuresis only during the first 2–3 days following treatment institution, until a new equilibrium is attained [10], and may not increase water intake in the long term.