Increased Granulocyte Heparanase Activity in Neutrophils from Patients with Lupus Nephritis and Idiopathic Membranous Nephropathy.

Heparanase is a ß-glucuronidase that cleaves sugar chains of heparan sulfate proteoglycans. It is believed that heparanase may be involved in the pathogenesis of proteinuria. The aim of this study was to assess the significance of heparanase in the pathogenesis of particular glomerulonephritis types. The evaluation of heparanase activity in serum, urine, and granulocytes and superoxide dismutase (SOD) activity in granulocytes of patients with lupus nephritis (n = 17), membranous nephropathy (n = 11), IgA nephropathy (n = 12), focal and segmental glomerulosclerosis (n = 18), mesangiocapillary glomerulonephritis (n = 12) and in 19 healthy volunteers were performed. The heparanase activity in granulocytes of patients with lupus nephritis and membranous nephropathy was higher than heparanase activity in granulocytes in the control group (p = 0.02 in both cases). This is the first observation of this phenomenon. There was no difference between SOD activity in granulocytes of patients with all assessed types of glomerulonephritis and the control group. A positive correlation between heparanase activity in urine and double-strain DNA antibodies (r = 0.51; p = 0.04), and reverse correlations between heparanase in urine and hemolytic activity of the complement (r = -0.57; p = 0.03) in the lupus nephritis group, and between heparanase activity in granulocytes and serum total protein level (r = -0.69; p = 0.02) in membranous nephropathy were observed. Increase in heparanase activity without changes in superoxide dismutase activity in the granulocytes from patients with lupus nephritis and membranous nephropathy was observed. It may be used as one of the markers of these disease activities.

Endostatin and vascular endothelial growth factor: potential regulators of endothelial progenitor cell number in chronic kidney disease.

INTRODUCTION: Cardiovascular mortality is significantly increased in patients with chronic kidney disease (CKD). The number of circulating endothelial progenitor cells (EPCs) may affect vascular regenerative potential and thus influence cardiovascular mortality. OBJECTIVES: The aim of the study was to assess the number of circulating EPCs and the factors that potentially affect these cells, including vascular endothelial growth factor (VEGF) and endostatin, in patients with CKD. PATIENTS AND METHODS: The study involved 139 patients divided into groups depending on the severity of renal impairment: 67 predialysis patients with CKD, 46 patients on hemodialysis (HD), and 26 patients on peritoneal dialysis (PD). Plasma levels of VEGF and endostatin were measured by enzyme-linked immunosorbent assays. The number of circulating EPCs, defined as CD34+VEGFR2+, was assessed in the whole blood using flow cytometry. RESULTS: There was a positive correlation between VEGF and CD34+VEGFR2+ and the glomerular filtration rate. Endostatin levels increased with renal impairment. The highest endostatin levels were observed in HD and PD patients. The number of EPCs was significantly lower in predialysis patients with CKD and in HD patients, while in PD patients it was nonsignificantly lower compared with the control group. CONCLUSIONS: In patients with CKD, a decrease in circulating EPCs may impair vascular regenerative potential and thus contribute to a higher cardiovascular risk. The effect of significantly increased endostatin levels on the endothelial function and progenitors in patients with CKD requires further investigation.

Plasma and urine leukocyte elastase-alpha1protease 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.

Vasculitis in systemic lupus erythematosus (SLE)--assessment of peripheral blood mononuclear cell activation and the degree of endothelial dysfunction: initial report.

BACKGROUND: Inflammatory-immune changes in the vascular endothelium are one of the main factors initiating vessel wall damage. Enhanced expression of endothelial adhesion molecules and their receptors on the surface of circulating leukocytes seems to play an important role in the pathogenesis of vasculitis. Increasing evidence indicates endothelial cell activation/damage in SLE. In patients with SLE complicated by vasculitis, enhanced expression of integrin activation markers on the surface of peripheral blood mononuclear cells (PBMCs) has been reported. It seems relevant to assess the mechanisms of inflammatory response involving PBMCs and endothelial cells at particular stages of SLE microangiopathy. AIM: The main aim was to assess the surface expressions of the integrin adhesion molecules VLA-4 (CD49d) and LFA-1 (CD11a) on PBMCs as well as the number of circulating endothelial cells (CECs) in patients with SLE and complications related to inflammatory microangiopathy and to determine whether these parameters vary depending on disease activity. PATIENTS: Twenty-nine women with SLE (mean age: 38.72+/-10.23 years) were divided into subgroup I: those with severe disease activity according to the modified disease activity index SLEDAI, characterized by the presence of inflammatory microangiopathy-related complications such as systemic central nervous system affection and/or vasculitis and/or nephritis (15 women, mean age: 38.33+/-11.02 years), and subgroup II: patients with mild or moderate disease activity according to SLEDAI and without vascular complications (14 women, mean age: 39.14+/-9.72 years). METHODS: Expressions of VLA-4 and LFA-1 on the surface of peripheral blood lymphocytes and monocytes were assessed by flow cytometry using monoclonal antibodies. CECs (a marker of endothelial damage) were isolated from peripheral blood with anti-CD146(S-Endo 1)-coated immunomagnetic Dynabeads. Tests for the lupus anticoagulant, antinuclear antibody, anti-dsDNA, and anticardiolipin antibody were performed in every study subject by ELISA. Erythrocyte sedimentation rate and serum levels of fibrinogen, C-reactive protein, the complement components C3 and C4, urea, creatinine, and uric acid were determined by standard methods. Peripheral blood counts and a general urinalysis were also performed. RESULTS: The mean CEC count was significantly higher in SLE patients than in the control group (15.29+/-12.10 vs. 3.08+/-1.46 cells/ml, p<0.001). CEC counts was notably elevated in patient subgroup II compared with the control group (9.14+/-5.16 vs. 3.08+/-1.46 cells/ml, p<0.05) and in subgroup I compared with subgroup II (21.03+/-13.96 vs. 9.14+/-5.19 cell/ml, p<0.05). In patients with severe SLE flares, CEC count visibly correlated with disease activity assessed by SLEDAI score (R=0.92, p<0.001). The expressions of VLA-4 and LFA-1 on peripheral blood lymphocytes in both patient subgroups were significantly higher than in the control group (subgroup I vs. controls: 1.70+/-1.56 vs. 0.39+/-0.26%, p<0.05, and 1.97+/-2.60 vs. 0.67+/-0.83%, p<0.05; subgroup II vs. controls: 1.71+/-1.04 vs. 0.39+/-0.26%, p<0.001, and 3.32+/-2.48 vs. 0.67+/-0.83%, p<0.05, for VLA-4 and LFA-1, respectively). There was no significant difference between the two subgroups of patients (1.70+/-1.56 vs. 1.71+/-1.04%, p>0.05, and 1.97+/-2.60 vs. 3.32+/-2.48%, p>0.05, respectively). Similarly, the surface expression of LFA-1 on circulating monocytes in patients in both subgroups was notably enhanced over that of the control group (91.44+/-16.00 vs. 84.95+/-19.86%, p<0.05, and 90.11+/-10.34 vs. 84.95+/-19.86%, p<0.05, in subgroups I and II respectively) and was comparable in both subgroups of patients (91.44+/-16.00 vs. 90.11+/-10.33%, p>0.05). The surface expression of VLA-4 on peripheral blood monocytes was considerably higher in patients with severe disease activity than in the control group and in patients with less active disease (77.10+/-13.56 vs. 64.90+/-19.13%, p<0.05, and 77.10+/-13.56 vs. 63.40+/-20.95%, p<0.05, respectively). However, there was no significant difference between patients with mild or moderate disease activity and the control group (63.40+/-20.95 vs. 64.90+/-19.13%, p>0.05). CONCLUSIONS: 1) The number of CECs increases in the course of SLE and correlates with disease activity, indicating progressive endothelial damage.2) The expressions of VLA-4 and LFA-1 on the surface of peripheral blood lymphocytes as well as that of LFA-1 on circulating monocytes are enhanced in SLE patients regardless of disease activity. 3) The expression of VLA-4 on the surface of circulating monocytes is enhanced only in patients with severe disease activity, characterized by the presence of complications connected with inflammatory microangiopathy, which may indicate that the upregulation of VLA-4 expression in monocytes plays a leading role in the pathogenesis of vasculitis in SLE.

Oxidative DNA damage and total antioxidant status in serum of patients with esophageal squamous cell carcinoma.

BACKGROUND/AIMS: Oxidative stress is connected with activation of somatic mutations and rates of cell proliferation existing in cancer tissue. High level of reactive oxygen species is a mutagenic factor for DNA damage. Antioxidants are the components of the cellular defense mechanism against reactive oxygen molecules. The aim of our study was to analyze DNA peroxidation products' concentration and total antioxidant level in serum of the patients with esophageal squamous cell carcinoma before and after esophagectomy. We examined these parameters as markers of cancer development. METHODOLOGY: We tested 18 patients (2 woman and 16 men, mean age 59.4 years) with esophageal squamous cell cancer before and after esophagectomy and 12 healthy people as a control group. Concentrations of 8-OHdG and enzymatic antioxidants level were analyzed in serum. Data were statistically analyzed by Mann-Whitney test. RESULTS: We observed statistically significant higher concentrations of 8-OHdG and significant lower levels of enzymatic antioxidants in the patients with cancer in comparison to the control group. After esophagectomy we observed normalization of these parameters. In four patients the level of total antioxidants was low and 8-OHdG concentration was high during the whole time of treatment. These patients had disease progression. CONCLUSIONS: Estimation of serum 8-OHdG concentration and total antioxidant status may be helpful for monitoring cancer therapy in patients with esophageal squamous cell cancer.

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 Bowman's 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.

[Toll-like receptors (TLR) in the pathogenesis of kidney diseases]. / Receptory Toll-podobne (TLR) w patogenezie chorób nerek.

In the present work general characteristics and occurrence of TLR receptors have been presented. The participation of TLR receptors in kidney pathology in experimental models in the course of urinary system infection, acute renal failure and interstitial fibrosis has been discussed. In addition, the importance of TLRs in various forms of glomerular nephritis and in haemodialytic patients as well as in postrenal-transplant patients has been shown. It is believed that in lipopolysaccharide-induced renal failure in the course of infections caused by Gram negative bacteria TLR4 plays a fundamental role. In the event of damage of renal tubular epithelial cells by mechanical, toxic, or ischemic factors activation of TLRs induces inflammatory processes leading to acute renal failure. In the course of progressive fibrosis of renal interstitial tissue TLR 2 and 4 receptors are stimulated, which results in the fact that immunological and structural cells of renal tissue release chemokines and cytokines, which causes increased inflow of leucocytes and intensification of interstitial nephritis and progressive fibrosis. The study on experimental models on mice MLR (mixed lymphocyte reaction) with genetically conditioned lupus-like disease showed that, CpG-DNA stimulation as a TLR 9 specific agonist intensifies inflammatory symptoms in mice. Similarly in apoferritin induced glomerulopathy (model of immune complex disease) CpG-DNA nucleotide increased glomerulopathy symptoms. It has been proved that activation of mechanisms of inherent immunity through TLR4 receptors affects the frequency and intensity of acute rejections in human organ transplantations. Incidence of acute kidney and lung [transplant rejections was significantly lower in recipients with mutated variants of Toll-like receptor 4 (TLR-4 Asp 299Gly and TLR-4-Tyr399-IIe).

[Influence of tobacco smoking and periodontitis on selected factors of oxidative stress]. / Wplyw nikotynizmu i zapalen przyzebia na wybrane parametry stresu oksydacyjnego.

UNLABELLED: The aim of the study was to estimate the influence of tobacco smoking and the clinical type of periodontitis on the chosen parameters of oxidative stress (superoxide anion generation on the ground of cytochrome c reduction in the whole blood, the concentration of autoantibodies for oxidized low-density lipoprotein in venous and gingival blood, the level of 8-hydroxy-2-deoxyguanosine (8-OHdG) as a marker of oxidative DNA damage in venous and gingival blood and the total antioxidant capacity in venous and gingival blood). MATERIAL AND METHODS: The study included 50 periodontitis patients (20 patients with generalized aggressive periodontitis - GAP and 30 patients with chronic periodontitis--CP), aged 28-55 (the mean value 40.3). There were 28 females and 22 males. The control group consisted of 25 volunteers in good general health, aged 2-50 (the mean value 37.8). There were 15 women in this group. In clinical examination ofperiodontium following indexes were included: plaque index (Silness and Löe), aproximal plaque index (Lange et al.), bleeding upon probing (Saxer and Mühlemann), pockets depth, the number of teeth. RESULTS: Significant influence of periodontal diagnosis on the level of 8-OHdG in gingival blood and total antioxidant capacity and also all analyzed clinical parameters was revealed. Significant total influence ofperiodontal diagnosis with tobacco smoking on total antioxidant capacity was also seen in venous blood (p = 0.004). CONCLUSIONS: 1) periodontal status impacted in significant way on the level of 8-OHdG in gingival blood and total antioxidant capacity in venous blood, 2) tobacco smoking and chronic periodontitis presence were connected with significantly decreasing levels of total antioxidant capacity in venous blood; this fact may be under importance in connection between periodontitis and atherosclerosis, 3) tobacco smoking was a significant factor in the progression ofperiodontitis (higher levels of plaque indexes, deeper periodontal pockets in nicotine addicted patients).

[IL-1beta and PGE2 production in whole blood and gingival fluid in women with periodontitis and preterm low birth weight]. / Stezenie IL-beta i PGE2 w krwi pelnej i plynie dziaslowym u kobiet z zapaleniem przyzebia oraz niska masa urodzeniowa noworodka wynikajaca z przedwczesnego zakonczenia ciazy.

DESIGN: The aim of the investigation was to evaluate of IL-1beta and PGE2 concentrations in gingival fluid, whole blood as well as IL-1beta, PGE2 production after Escherichia coli lipopolysaccharide stimulation in whole blood in women with preterm low birth weight (PLBW), as compared to the control group. MATERIAL AND METHODS: A case-control study of 88 postpartum women aged 17 to 39 was performed. The case group consisted of 52 women with PLBW and the control group consisted of 36 women giving birth in time. Concentration of inflammatory mediators in gingival fluid, blood serum and IL-1beta, PGE2 production in whole blood after bacterial lipopolysaccharide stimulation were determined by means of immunoenzymatic method. RESULTS AND CONCLUSIONS: The levels of IL-1beta and PGE2 in gingival fluid were significantly higher in all PLBW mothers (also PLBW primiparous) than in the control group. In addition in the primiparous with PLBW group significantly higher PGE2 concentration in blood serum was found compared to the primiparous controls. There were no significant differences between women with PLBW and the controls together with a significantly higher production of IL-1beta and PGE2 in whole blood after LPS stimulation in women with periodontitis and gingivitis compared to subjects with healthy periodontium. Such findings suggest that inflammatory mediator synthesis is mainly result of specific cells exposition to bacterial products. Therefore it seems that more frequent occurrence of the phenotype of hyperactive cells that synthesise these mediators is not responsible for PLBW.

[Antiphospholipid antibodies in the transplanted kidney pathology]. / Przeciwciala antyfosfolipidowe w patologii przeszczepionej nerki.

The purpose of the paper was to estimate clinical problems of the antiphospholipid antibodies (PAF), the mechanisms of their development and function in general thrombotic complications. Special attention has been paid to thrombotic complications in the transplanted kidney. In the group of patients with early renal allograft failure PAF are more frequent than in patients with functioning grafts. The presence of antiphospholipid antibodies in patients prior to the transplantation may indicate the patients' susceptibility to thrombotic complications and failure of the transplanted kidney function. In the case of less intensive coagulation the lifetime of the functioning kidney may be considerably shortened. So far, no satisfactory anticoagulation therapy has been developed. Patients were treated with warfarin, heparin as well as fractionated heparin and intravenous immunoglobulin, aspirin and prednisone. The anticoagulation therapy should be applied in compliance with the laboratory markers of coagulation and with special emphasis put on the general health status of a patient.

[Blood and tissue eosinophilia in patient with angioedema caused by inhibitor of angiotensin converting enzyme (ACE)--case report and a view of literature]. / Eozynofilia obwodowa i tkankowa u pacjenta z obrzekiem naczynioruchowym wywolanym inhibitorem enzymu konwertujacego angiotensyne (ACE)--opis przypadku i przeglad pismiennictwa.

A case of angioedema caused by enalapril, undiagnosed for 5 years was presented. Enhanced blood and tissue eosinophilia shown in nasal smear was observed. In addition increased activity of coagulation system was shown manifested by enhance of concentration of Hageman factor and cardiolipin antibodies IgM and IgA isotype. The role of coagulation, complement and fibrinolysis systems in pathogenesis of ACE-inhibitors induced angioedema was discussed. The influence of bradykinin on activity of eosinophils was analyzed.

[Endothelin, endothelial cells and von Willebrand factor in peripheral blood of asthmatic patients]. / Endotelina, czynnik von Willebranda i komórki sródblonka we krwi obwodowej u chorych na astme.

As recent studies prove, endothelial cells and particularly one of their products, endothelin, are not only active agents taking part in sustaining homeostasis in the peripheral blood system, but are also involved in immunological responses and angiogenesis. This is especially true about endothelin, which is a powerful spasmogen and comitogen, in particular for the smooth muscles of the bronchi. Endothelial cells play an important role in angiogenesis of the lungs, observed in asthmatic patients. The marker of the condition of endothelium is von Willebrand factor (vWF). The aim of this research was to study the three elements (endothelial cells, endothelin, vWF) in peripheral blood of asthmatic patients. The study group consisted of 27 asthmatic patients (moderate and severe asthma) and 15 healthy individuals. Immunological method employing specific monoclonal antibodies was used to assess endothelial cells; immunoenzymatic method was used to assess endothelin and vWF. The concentration of endothelin in the blood of asthmatic patients was much higher than in the control group (1.02 +/- 2.13 vs 0.45 +/- 0.14), but the difference was not statistically significant. The numbers of circulating endothelial cells were significant higher in asthmatics than in healthy individuals (1.3 +/- 0.7 vs 0.48 +/- 0.3) and the concentrations of vWF were the same in both groups (105.3 +/- 18.7 vs 102.8 +/- 32.2). Additionally, positive correlation was found between endothelin and endothelial cells, endothelin and vWF, as well as endothelial cells and vWF. The results suggest that endothelium and endothelin play an active part in the process of remodeling in asthma and may serve as a tool in the prognosis of remodeling.

Analysis of collagen IV and fibronectin in blood and urine in evaluation of nephrotic fibrosis in children with chronic glomerulonephritis.

BACKGROUND: Changes in the concentration of extracellular matrix components in body fluids may reflect the degree of kidney fibrosis. The aim of our study was to evaluate fibronectin (FN) and type IV collagen (COL IV) concentrations in blood serum and urine, and to refer these values to immunoreactivity in renal biopsy material. MATERIAL/METHODS: Our research involved 37 children with glomerulonephritis (GN) aged 4-15 and 18 healthy children. The patients were divided into 3 groups according to the histopathological diagnosis of GN: I - minimal change GN (MCGN, n=17), II - lupus nephritis (LN, n=7), III - other types of GN (n=13). RESULTS: Significantly higher COL IV concentrations in serum and urine were noted in the patient group in comparison to the controls, but no differences in FN concentrations. The highest values of serum and urine COL IV were observed in the LN patients. Positive correlation was found between serum and urine COL IV and its renal content in the LN group. CONCLUSIONS: The results indicate that serum and urine COL IV rises in children with GN, above all in the course of LN. This also reflects changes in renal COL IV content.