Detection of multiple cytokines in the urine of patients with focal necrotising glomerulonephritis may predict short and long term outcome of renal function.

BACKGROUND: Detection of urinary cytokines in pauci-immune focal segmental necrotizing glomerulonephritis (FSNGN) may provide valuable information about disease pathogenesis and prognosis. METHODS: Epidermal growth factor (EGF), transforming growth factor (TGF-ß1) and vascular endothelial growth factor (VEGF) were measured by ELISA, and Interleukins, monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein (MIP1ß) by a multiplex cytokine assay, in 38 patients with FSNGN. Their levels were correlated with severity of histological findings and renal function outcome in short and long term. RESULTS: The percentage of crescents in renal biopsy had positive correlation with TGF-ß1 (p=0.004) and IL-15 urinary excretion (p=0.01), and negative correlation with EGF (p=0.01). Increased urinary excretion of IL-6, IL-15, VEGF and MIP-1ß was associated with poor renal function outcome, but increased levels of EGF, IL-2 and IL-9 predicted a favourable prognosis. In multiple regression analysis IL-6 and VEGF urinary levels were independent predictors of no-response at the acute phase (p=0.001 and p<0.0001, respectively), while, IL-6 was the only factor (p=0.03) predicted worse outcome at the end of follow-up (39.4±45 months). CONCLUSION: Increased urinary excretion of IL-6, IL-15, VEGF, TGF-ß1, MCP-1 and MIP-1ß and reduced EGF, IL-2, IL-9 may be associated with histological damage and influence response to treatment in pauci-immune FSNGN.

Kikuchi-Fujimoto disease and systemic lupus erythematosus: the EBV connection?

Kikuchi-Fujimoto disease (KFD) is a benign and self-limited disease of unknown etiology that affects mainly young women. It presents with localized lymphadenopathy, usually cervical, accompanied with fever, night sweats, and leucopenia. KFD has been rarely described in association with autoimmune disorders, mainly systemic lupus erythematosus (SLE). We report the case of a young patient presenting with KFD coinciding with SLE. The association of KFD and SLE is reviewed. Moreover, a possible pathogenetic role of Ebstein-Barr virus linking the two clinical entities is discussed.

The anticoagulant activity of enoxaparin sodium during on-line hemodiafiltration and conventional hemodialysis.

To study and compare the anticoagulant activity of enoxaparin sodium during on-line hemodiafiltration (OL-HDF) and conventional hemodialysis (C-HD). Enoxaparin was administered as an anticoagulant to 21 hemodialysis patients at the beginning of a single 4-hour OL-HDF session as an intravenous bolus dose of 80 IU/kg [DOSAGE ERROR CORRECTED] On-line hemodiafiltration was performed using a high-flux polyester polymer alloy dialyzer and a total of 18 L replacement fluid (session A). One week later, the study was repeated in the same patients during a single 4-hour session of C-HD using a low-flux polysulfone dialyzer (session B). Blood samples for the measurement of Hb, blood urea and nitrogen (BUN), activated partial thromboplastin time (APTT), and anti-Xa levels were taken before each study session and 5-minute postdialysis. In 13 more patients, the same study was performed during OL-HDF using a high-flux polysulfone dialyzer (session C). No differences were found between sessions A, B, and C when predialysis values for Hb, BUN, APTT, and anti-Xa were compared. The mean postdialysis APTT and anti-Xa values were 32.5+/-3.8 seconds and 0.19+/-0.11 IU/mL, respectively, in session A, 39.0+/-5.0 seconds and 0.71+/-0.17 IU/mL in session B, and 33.8+/-3.1 seconds and 0.35+/-17 IU/mL in session C (A vs. B, P<0.0001, for both parameters, A vs. C, P<0.003 for anti-XA, and B vs. C, P<0.005, for both parameters). The anticoagulant activity of enoxaparin sodium is decreased significantly during a 4-hour OL-HDF session compared with to a similar session of C-HD. The degree of the reduction seems to depend on the dialyzer's membrane.

Fibrotic mechanisms in idiopathic rapidly progressive glomerulonephritis: the role of TGF-beta1 and C5b-9.

BACKGROUND: Idiopathic IRPGN is a form of renal vasculitis in which a high chronicity index is present despite minimal impairment of renal function. The present study investigated the mechanisms underlining the relatively early appearance of fibrosis. METHODS: In all, 34 patients (17 males) with biopsy proven idiopathic RPGN were included. On light microscopy, the percentage and evolution stage of crescents, the presence of glomerular necrosis, the degree or severity of arteriosclerosis, as well as the extent of tubulointerstitial (TIN) infiltration, interstial fibrosis, and tubular atrophy were assessed. Monoclonal antibodies were used to identify infiltrating macrophages, HLA-DR (+), alpha-SMA (+), and PCNA (+) cells, the expression of the adhesion molecule ICAM-1, the growth factor TGF-beta1, and the terminal complement component C5b-9. RESULTS: The presence of glomerular necrosis correlated positively with the number of SMA (+) cells in TIN (p = 0.036). Glomerular TGF-beta1 expression had positive correlation with tubular C5b-9 expression. The tubulointerstitial TGF-beta1 expression correlated with tubular C5b-9 expression (p = 0.001) and TGF-beta1 expression (p = 0.009). Independent factors predicting the severity of renal function impairment were the CRP levels (p = 0.002) and the degree of arteriosclerosis (p = 0.01). CRP levels correlated with the severity of interstitial infiltration and fibrosis (p = 0.02), the expression of TGF-beta1 in the glomeruli (p = 0.009) and the interstitial space (p = 0.001), and the intensity of tubular ICAM-1 and C5b-9 expression (p = 0.023, p = 0.002, respectively). The severity of proteinuria showed a significant correlation with the expression of TGF-beta1 in the glomeruli (p = 0.033) and the tubulointerstitium (p = 0.019). CONCLUSIONS: The activation of interstitial fibroblasts seems to be an early phenomenon that is related to the extent of glomerular necrosis. Glomerular TGF-beta1 may induce tubular expression of C5b-9. Increased tubular C5b-9 expression may result in interstitial fibrosis through increased TGF-beta1 production.

Elevated hepatocyte growth factor levels at the beginning of high-flux hemodialysis are due to heparin administration.

BACKGROUND: It has been reported that hemodialysis (HD) stimulates hepatocyte growth factor (HGF) release, but it is not clear if this stimulation is due to HD itself or to heparin used during HD. To clarify this issue, we undertook the present study. METHODS: We studied 18 HD patients using high-flux dialyzers, during a single 4-hr hemodialysis session (session A). The dialyzers were pre-rinse with normal saline without heparin, and HD was started with zero ultrafiltration and without anticoagulation. Anticoagulation was administered as IV injection (80 IU/kg of LMWH enoxaparin sodium) 10 min after the beginning of HD. HD was continued for 10 more minutes and then as prescribed. HGF serum levels were measured before the beginning of the HD session (sample t0) as well as 10 and 20 minutes after the beginning of the session (samples t10 and t20). In six more patients (controls), the same study was repeated but without the administration of LMWH during the first 20 min of HD initiation (session B). RESULTS: In comparison with t0, t10 HGF serum levels changed significantly in neither session A nor in session B. However, at t20, HGF levels increased significantly in session A compared with t0 (increment 666.3 +/- 211.0%, p < 0.0001) and t10 (increment 894.2 +/- 506.0%, p < 0.0001), but not in session B. No differences were found between sessions A and B at samples t0 and t10 (p = NS). HGF serum levels at t20 in session A were found to be higher compared with corresponding levels in session B (p < 0.0001). CONCLUSION: Elevated HGF serum levels at the beginning of high-flux HD session are due to LMWH administration.

Predictors of outcome in idiopathic rapidly progressive glomerulonephritis (IRPGN).

BACKGROUND: Small vessel vasculitides are known to follow a devastating course towards end-stage renal disease, unless treated with immunosuppressive regiments. We investigated the value of clinical, histological and immunohistochemical parameters as predictors of outcome at diagnosis in patients with pauci immune necrotizing glomerulonephritis. METHODS: In 34 patients the percentage and evolution stage of crescents, the presence of glomerular necrosis, the degree or severity of arteriosclerosis, as well as the extent of tubulointerstitial infiltration, interstial fibrosis and tubular atrophy were assessed. Monoclonal antibodies were used to identify infiltrating macrophages, alpha-SMA(+) and PCNA(+) cells, the expression of integrins alpha3beta1 and LFA-1beta, the adhesion molecule ICAM-1, the growth factor TGF-beta1 and the terminal complement component C5b-9. RESULTS: 24 pts (70.6%) showed a complete or partial response to the treatment. The follow-up period was 20 +/- 22 months. At multivariate analysis, serum CRP (p = 0.024), the intensity of tubular expression of C5b-9 (p < 0.0001) as well as the extent of glomerular and tubular expression of alpha3beta1 integrin (p = 0.001 and 0.008 respectively) independently predicted the response to treatment. The response rate was better in ANCA(+) pts (p = 0.008). The extent of interstitial infiltrate (p < 0.0001), the severity of tubulointerstitial fibrosis (p < 0.0001) and the severity of tubular TGF-beta1 expression (p < 0.0001) were independent predictors of long term outcome of renal function. CONCLUSION: Patients with ANCA-associated renal vasculitis seem to respond better to the treatment. Acute phase reactants, such as CRP, implying a more intense parenchymal inflammatory reaction, as well as the intensity of the de novo expression of C5b-9 and the glomerular and tubular expression of alpha3beta1 integrin predict the response to therapy. The severity of TIN lesions and of the tubulo-interstitial TGF-beta1 and C5b-9 expression predict an unfavourable outcome.

HCO3 increment in arterial line can reveal significant vascular access recirculation in high-flux hemodialysis: a preliminary report.

We report a new and simple way that can reveal the presence of vascular access recirculation (VAR) in patients undergoing hemodialysis (HD). Acid-base and blood gas parameters (pH, pO(2), pCO(2), and HCO(3)) were measured in blood samples drawn from an arterial fistula needle before the initiation of HD and from arterial and venous lines simultaneously 5 min later, in 31 patients (group A). Vascular access recirculation was measured using the glucose infusion test (GIT) immediately after the withdrawal of the 5-min samples. The same study was repeated in 30 patients in whom HD lines were reversed (group B). A comparison with baseline (predialysis) values of an analysis of the arterial line in group A at 5 min revealed that pCO(2) increased by 1.14+/-2.5 mmHg and HCO(3) by 0.6+/-0.6 mM/L (p<0.02 and p<0.00001, respectively). The corresponding pO(2) and pH values did not show significant differences. Glucose infusion test at 5 min (GITa) was -0.058+/-0.03%. A comparison with baseline (predialysis) values of an analysis of the arterial line in group B at 5 min revealed that pCO(2) increased by 7.7+/-3.5 mmHg and HCO(3) by 2.9+/-1.0 mM/L (p<0.000001 in each case). The pH level was significantly lower in comparison with baseline values (p<0.00001), while pO(2) did not show a significant difference. Glucose infusion test at 5 min (GITb) was 12.0+/-6.1% (p<0.000001 in comparison with GITa values). Clinically significant VAR was defined as HCO(3) increment >1.8 mM/L, based on the receiver-operating characteristics curve, which showed a threshold value of HCO(3) increment >1.8 mmol/L as a predictor of GIT recirculation. Five minutes after the initiation of high-flux HD with a 0 ultrafiltration rate, there is a small increment in arterial HCO(3) values relative to predialysis values. Clinically significant VAR is present when this increment is higher than 1.8 mM/L.

The effect of two different doses comprising the simultaneous administration of intravenous B-complex vitamins and oral folic acid on serum homocysteine levels in hemodialysis patients.

BACKGROUND: Several regimens using different doses of folic acid (FA) alone or supplemented with B-complex vitamins (BCVs) have been tested for their ability to reduce total homocysteine (tHcy) serum levels in hemodialysis (HD) patients. In the present study, we assessed the effect of two different doses comprising the simultaneous administration of intravenous (IV) BCVs and an oral FA supplementation on serum tHCy levels in HD patients. PATIENTS-METHODS: In a cohort of 49 patients (31 male, 18 female) undergoing chronic HD treatment for a mean of 40.0+/-40.7 months, serum concentrations of tHcy, folate and vitamin-B12 (vB12) were determined at the end of three sequential periods as follows: 20 weeks without any BCV and/or FA supplementation (period A), 20 weeks with a dose comprising the simultaneous administration of IV BCVs and an oral supplementation of 5 mg of FA once a week (period B), and 20 weeks with a dose comprising the simultaneous administration of IV BCVs and an oral supplementation of 5 mg of FA thrice a week (period C). An IV dose of BCVs consisting of a 5 mL solution containing vitamin B1 (250 mg), vitamin B6 (250 mg) and vitamin B12 (1.5 mg) was administered at the end of hemodialysis. RESULTS: Mean serum tHcy levels were significantly higher at the end of period A relative to levels at the end of periods B and C (35.8+/-23 micromol/L vs. 22.0+/-17.6 and 15.0+/-4.5 micromol/L, respectively; p<0.000001). Mean serum folate levels and mean serum vB12 levels were significantly lower at the end of period A relative to levels at the end of periods B and C (p<0.000001). Mean serum tHcy levels were lowest at the end of period C (p<0.000001 in comparison to periods A and B), and 26 of the 49 HD patients (67.3%) possessed tHcy levels below 16 micromol/L. CONCLUSIONS: In HD patients, high doses consisting of the simultaneous administration of IV BCVs and an oral FA supplementation resulted in the efficient reduction of serum tHcy levels.

pO2 and pCO2 increment in post-dialyzer blood: the role of dialysate.

Blood returning from a dialyzer during hemodialysis has a higher pO2 and pCO2 content than blood entering the dialyzer, and this has been attributed to the dialysate. The present study investigates this phenomenon. Acid-base and blood-gas parameters (pH, pO2, pCO2 and HCO3) were measured in three groups of stable chronic hemodialysis patients (A, B, and C) undergoing high-flux hemodialysis. In group A (n = 15), "arterial" (a) and "venous" (v) samples were withdrawn simultaneously before dialysis (samples A0), 5 min after circulation of the blood with the dialysate in the by-pass mode (samples A5), and 5 min after high-flux hemodialysis at a zero ultrafiltration rate (samples A10). In group B (n = 11) (a) and (v) samples were withdrawn simultaneously before dialysis (samples B0), 5 min after isolated-ultrafiltration with closed dialysate ports ("isolated-closed" ultrafiltration) (samples B5), and 5 min after high-flux hemodialysis at a zero ultrafiltration rate (samples B10). In group C (n = 14), after an initial arterial blood sample withdrawal before hemodialysis (sample C0), high-flux hemodialysis at a zero ultrafiltration rate was initiated. Five minutes later, blood and dialysate samples were withdrawn simultaneously from the hemodialysis lines (samples C5). In all cases blood and dialysate (bicarbonate) flow rates were set at 0.300 and 0.700 L/min, respectively. FLX-18 hemodialyzers (membrane PEPA 1.8 m2) were used in this study. Analysis of variance revealed significant changes only in venous samples. A comparison of arterial and venous samples revealed no differences between groups A and B before the initiation of dialysis (A0a vs. A0v and B0a vs. B0v, P = NS). The pO2 content was higher in A5v samples than in A5a samples (83.5 +/- 11.2 vs. 88.8 +/- 14.0 mm Hg, P < 0.02), while the level of HCO(3) was higher in A5a samples than in A5v samples (20.8 +/- 2.0 vs. 20.4 +/- 1.8 mEq/L, P < 0.05). A10a samples possessed a higher pH and lower levels of pO2, pCO2, and HCO3 in comparison to A10v samples (P < 0.001 for all). Mean pO2 and pCO2 values in A5v and A10v samples increased by 6.3% and 12.1% and by 1.29% and 52% in comparison to corresponding values of A5a and A10a samples, respectively. The pO2 level was the only parameter that differed significantly between B5a and B5v samples (B5a = 84.6 +/- 10.1 vs. B5v = 98.0 +/- 12.6 mm Hg, P < 0.005). B10a samples possessed a higher pH and lower levels of pCO2, pO2, and HCO3 in comparison to B10v samples (P < 0.0005 for all comparisons). Mean pO2 and pCO2 values in B5v and B10v samples increased by 16.2% and 16.3% and by -0.29% and 64.8% in comparison to corresponding values of B5a and B10a samples, respectively. C5a samples possessed a higher pH and lower levels of pCO2, pO2, and HCO3 in comparison to C5v samples (P < 0.001 for all). Mean pO2 and pCO2 values in C5v samples were, respectively, 16.0% and 65.0% higher than corresponding values of C5a samples. These results indicate that blood returning from the dialyzer after 5 min of high-flux hemodialysis has a higher pO2 and pCO2 than blood entering the dialyzer, and that this difference is due to O2 and CO2 transfer from the dialysate space into the blood.

Circulating levels of ICAM-1, VCAM-1, and MCP-1 are increased in haemodialysis patients: association with inflammation, dyslipidaemia, and vascular events.

BACKGROUND: Increased levels of circulating adhesion molecules and chemokines have been reported in haemodialysis (HD) patients but the influence of the HD membranes on their secretion, as well as their pathophysiological implications, remains largely unknown. METHODS: Circulating levels of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and monocyte chemoattractant protein-1 (MCP-1) were measured by immunosorbent assay (ELISA) in 81 HD patients (45 male, mean age 57+/-13 years) and 35 normal subjects. All patients had been stabilized on renal replacement therapy for >3 months and were free of active infection. Thirty-three patients (40.7%) were routinely dialysed with modified cellulose membranes and 48 patients (59.3%) were dialysed with polysulfone membranes. Blood samples were taken directly from the arteriovenous fistula immediately before and at the end of a routine HD session. RESULTS: Pre-dialysis levels were significantly elevated in HD patients compared with controls (ICAM-1 515+/-177 vs 238+/-664 ng/ml, P<0.0001; VCAM-1 2107+/-648 vs 1012+/-115 ng/ml, P<0.0001; MCP-1 427+/-148 vs 125+/-42 pg/ml, P<0.0001). The HD session resulted in a significant increase in the levels of all three molecules measured (515+/-177 vs 679+/-187 ng/ml, P<0.0001; 2107+/-648 vs 2662+/-800 ng/ml, P<0.0001; 427+/-148 vs 567+/-153 pg/ml, P<0.0001, respectively). There was no difference in pre- or post-dialysis levels of the above molecules between patients routinely dialysed with either modified cellulose or polysulfone membranes. MCP-1 levels had a positive correlation with ICAM-1 levels (r=0.41, P<0.0005). VCAM-1 levels had a negative correlation with HDL levels (r=-0.30, P<0.01) and were significantly elevated in patients with HDL <35 mg/dl compared with patients with HDL > or = 35 mg/dl (2300+/-606 vs 1890+/-633 ng/ml, P<0.005). Log-transformed exact C-reactive protein (CRP) values were significantly correlated with ICAM-1 and VCAM-1 levels (r=0.41, P<0.005 and r=0.43, P<0.005, respectively). In addition, compared with patients with normal CRP values, patients with elevated CRP had significantly increased levels of ICAM-1 (466+/-166 vs 580+/-172 ng/ml, P<0.005). Patients with cardiovascular, cerebrovascular, or peripheral vascular diseases had significantly increased serum CRP and ICAM-1 levels compared with patients with no evidence of vascular disease (19.2+/-12.9 vs 7.9+/-11.8 mg/l, P<0.001 and 608+/-189 vs 474+/-155 ng/ml, P<0.005 respectively). CONCLUSIONS: Serum levels of ICAM-1, VCAM-1, and MCP-1 are increased in HD patients and probably result from either inadequate clearance or enhanced synthesis and release. HD session resulted in a significant increase of the above molecule levels but the exact mechanism(s) responsible for these alterations are yet to be fully elucidated. Increased levels of adhesion molecules are associated with inflammation, dyslipidaemia, and cardiovascular events. However, the potential link between these processes and its clinical significance warrants further investigation.