Carmelo Libetta

Oxidative stress and inflammation: Implications in uremia and hemodialysis

Oxidative response and inflammation constitute a major defense against infections, but if not properly regulated they could also lead to a number of deleterious effects. Patients affected by different stages of acute and chronic kidney disease, particularly patients on hemodialysis, present a marked activation of oxidative and inflammatory processes. This condition exposes these patients to an elevated risk of morbidity and mortality. This Review is up to date and it analyses the newest notions about pathophysiological mechanisms of oxidative stress and inflammation in patients with renal diseases, also considering the different strategies studied to counterbalance this high risk state.

Stimulation of Hepatocyte Growth Factor in Human Acute Renal Failure

Hepatocyte growth factor (HGF) is a potent mitogen for tubular cells. Experimental injury to the kidney is associated with HGF release both locally and by distant organs stimulated by circulating ‘injurins’. In this study, the serum HGF concentration was measured in patients with acute renal failure (ARF). Normal subjects and chronic renal failure patients either not on dialysis or on regular dialysis treatment served as controls. Human mesangial cells were incubated with sera from ARF patients and controls. The serum HGF concentration was strikingly increased in ARF patients (478 ± 68 ng/dl) and was normal in chronic renal failure patients not on dialysis (20 ± 3 ng/dl) and in those on regular dialysis treatment (25 ± 3 ng/dl). Serum of ARF patients strongly stimulated HGF release from mesangial cells (1,384 ± 55 ng/ml) in comparison with normal serum (67 ± 10 ng/ml). These results indicate that in ARF HGF participates in tubular repair both as an endocrine factor, released in the circulation, and as a paracrine substance, diffusing to the tubules from the mesangium.

Circulating Levels and Dietary Intake of the Advanced Glycation End-product Marker Carboxymethyl Lysine in Chronic Kidney Disease Patients on Conservative Predialysis Therapy: A Pilot Study

OBJECTIVE Advanced glycation end-products (AGEs) are proposed to influence inflammatory pathways and cardiovascular risk in chronic kidney disease (CKD). Dietary AGEs are believed to sustain circulating levels and toxicity in this condition. DESIGN AND PATIENTS We investigated this aspect in a cross-sectional pilot study measuring levels of the AGE marker carboxymethyl lysine (CML) and fluorescent AGEs in the blood of pre-dialysis patients with CKD and hemodialysis (HD) patients (n = 10 each), and in a group of matched healthy controls (Ctr). METHODS Plasma CML was measured by immuno-dot blot and fluorescent AGEs were determined by high-performance liquid chromatography (HPLC) analysis measuring the fluorescence of the cross-link pentosidine. The dietary intake of CML was assessed by dietary recall to trace total AGE intake in patients with CKD and the Ctr group. All the subjects included in the study were assessed for dietary intake while maintaining their usual diet. Main exclusion criteria for patients with CKD and HD were severe protein-caloric malnutrition and inflammation (measured by high sensitivity C-reactive protein and interleukin-6 levels). RESULTS Plasma CML, as well as free and protein-bound fluorescent AGEs, significantly increased in CKD and even more in HD patients than that of the Ctr group. In patients with CKD, the average dietary intake of CML was less than half than that of the Ctr group (6 vs. 13 MU/day) and the lowered protein intake adopted spontaneously by these patients appear to explain this finding. CONCLUSIONS The results show that the intake of CML does not affect circulating levels of this as well as of other AGEs, in well nourished predialysis CKD patients.

Severe Symptomatic Hyponatremia During Sibutramine Therapy: A Case Report

Sibutramine, a serotonin reuptake inhibitor, currently is used in treatment of obesity. The known side effects of sibutramine, ie, hypertension and tachycardia, depend on its adrenergic and serotoninergic effects. We describe a case of life-threatening hyponatremia associated with sibutramine use in an obese woman. We hypothesize that sibutramine, through its effect on neurotransmitters, may induce antidiuretic hormone secretion and lead to a syndrome of inappropriate antidiuretic hormone secretion. We advise careful monitoring of water-electrolytic balance during sibutramine therapy.

Mesenchymal stromal cells reset the scatter factor system and cytokine network in experimental kidney transplantation

BackgroundIn former studies we showed in a rat model of renal transplantation that Mesenchymal Stromal Cells (MSC) prevent acute rejection in an independent way of their endowing in the graft. In this study we investigated whether MSC operate by resetting cytokine network and Scatter Factor systems, i.e. Hepatocyte Growth Factor (HGF), Macrophage Stimulating Protein (MSP) and their receptors Met and RON, respectively.MethodsMSC were injected into the renal artery soon after reperfusion. Controls were grafted untreated and normal rats. Rats were sacrificed 7 days after grafting. Serum and renal tissue levels of IFN-γ, IL-1, IL-2, IL-4, IL-6, IL-10, MSP/RON, HGF/Met systems, Treg lymphocytes were investigated.ResultsIn grafted untreated rats IFN-γ increased in serum and renal tissue and IL-6 rose in serum. MSC prevented both the phenomena, increased IL-10 serum levels and Treg number in the graft. Furthermore MSC increased serum and tissue HGF levels, Met tubular expression and prevented the suppression of tubular MSP/RON expression.ConclusionsOur results demonstrate that MSC modify cytokine network to a tolerogenic setting, they suppress Th1 cells, inactivate monocytes/macrophage, recruit Tregs. In addition, MSC sustain the expression of the Scatter Factor systems expression, i.e. systems that are committed to defend survival and stimulate regeneration of tubular cells.

Hemodialysis related interleukin-2 receptor release by peripheral blood mononuclear cells.

To evaluate the effects of hemodialysis treatment on the spontaneous cell release of interleukin-2 receptor (IL-2R), we studied 19 hemodialyzed patients (HD), 9 non hemodialyzed patients with chronic uremia (UR, glomerular filtration rate: 8.4 +/- 1.8 ml/min), and 8 healthy control subjects (C). We measured the release of IL-2R in the supernatant of peripheral blood mononuclear cells (PBMC) cultured for 24 hrs in Iscoves medium as well as the plasma levels of IL-2R. A significant increase of IL-2R release was detected in the supernatant of PBMC harvested from HD patients (32.4 +/- 2.4 and 34.2 +/- 5.6 U/3 x 10(6) PBMC daily before and after HD, respectively) as compared with UR (16.6 +/- 5.2 U/3 x 10(6) PBMC daily) and C (21.4 +/- 3.8 U/3 x 10(6) PBMC daily). Similarly, IL-2R plasma levels were significantly higher in HD (378.5 +/- 164.6 U/ml) than in UR (189.5 +/- 89.3 U/ml) and C (11.2 +/- 2.68 U/ml). To summarize, the current study demonstrates: a) an enhancement of spontaneous IL-2R cell release in HD patients; b) an increase of sIL-2R plasma levels in UR patients possibly related to reduced metabolism and/or urinary excretion, because it was not associated with high IL-2R cell release; and c) a further increment of IL-2R systemic levels in HD likely secondary to the high cell release of IL-2R. Therefore, a chronic T cell activation with increased release of IL-2R secondary to the dialysis procedure is suggested.

Mesenchymal Stromal Cells Prevent Renal Fibrosis in a Rat Model of Unilateral Ureteral Obstruction by Suppressing the Renin-Angiotensin System via HuR

We studied Mesenchymal Stromal Cells (MSC) effects in experimental Unilateral Ureteral Obstruction (UUO), a fibrogenic renal disease. Rats were divided in 5 groups: sham, UUO, MSC treated-UUO, ACEi treated-UUO, MSC+ACEi treated- UUO. Data were collected at 1, 7, 21 days. UUO induced monocyte renal infiltration, tubular cell apoptosis, tubular atrophy, interstitial fibrosis and overexpression of TGFβ, Renin mRNA (RENmRNA), increase of Renin, Angiotensin II (AII) and aldosterone serum levels. Both lisinopril (ACEi) and MSC treatment prevented monocyte infiltration, reduced tubular cell apoptosis, renal fibrosis and TGFβ expression. Combined therapy provided a further suppression of monocyte infiltration and tubular injury. Lisinopril alone caused a rebound activation of Renin-Angiotensin System (RAS), while MSC suppressed RENmRNA and Renin synthesis and induced a decrease of AII and aldosterone serum levels. Furthermore, in in-vitro and in-vivo experiments, MSC inhibit Human antigen R (HuR) trascription, an enhancer of RENmRNA stability by IL10 release. In conclusion, we demonstrate that in UUO MSC prevent fibrosis, by decreasing HuR-dependent RENmRNA stability. Our findings give a clue to understand the molecular mechanism through which MSC may prevent fibrosis in a wide and heterogeneous number of diseases that share RAS activation as common upstream pathogenic mechanism.

Costimulatory Pathways in Kidney Transplantation: Pathogenetic Role, Clinical Significance and New Therapeutic Opportunities

Costimulatory pathways play a key role in immunity, providing the second signal required for a full activation of adaptive immune response. Different costimulatory families (CD28, TNF-related, adhesion and TIM molecules), characterized by structural and functional analogies, have been described. Costimulatory molecules modulate T cell activation, B cell function, Ig production, cytokine release and many other processes, including atherosclerosis. Patients suffering from renal diseases present significant alterations of the costimulatory pathways, which might make them particularly liable to infections. These alterations are further pronounced in patients undergoing kidney transplantation. In these patients, different costimulatory patterns have been related to distinct clinical features. The importance that costimulation has gained during the last years has led to development of several pharmacological approaches to modulate this critical step in the immune activation. Different drugs, mainly monoclonal antibodies targeting various costimulatory molecules (i.e. anti-CD80, CTLA-4 fusion proteins, anti-CD154, anti-CD40, etc.) were designed and tested in both experimental and clinical studies. The results of these studies highlighted some criticisms, but also some promising findings and now costimulatory blockade is considered a suitable strategy, with belatacept (a CTLA-4 fusion protein) being approved as the first costimulatory blocker for use in renal transplantation. In this review, we summarize the current knowledge on costimulatory pathways in the setting of kidney transplantation. We describe the principal costimulatory molecule families, their role and clinical significance in patients undergoing renal transplantation and the new therapeutic approaches that have been developed to modulate the costimulatory pathways.

Standard Hemodiafiltration Improves Diuretic Responsiveness in Advanced Congestive Heart Failure

who were responsive to diuretics were treated with intravenous boluses of furosemide. Renal function was similar in the two groups ( table 1 ). Group A patients received HDF using an AN-69 membrane (M100, Hospal, Italy) with a surface area of 1 m 2 ; the mean time duration of a single HDF session was 220 8 49 min for 4 8 1.6 sessions. Group B patients received furosemide as intravenous boluses of 160 8 Elevated cytokine levels have been associated with an increased risk of congestive heart failure (CHF) even in people without history of myocardial infarction [1, 2] . This has prompted new therapeutic approaches from continuous ultrafi ltration to intravenous immunoglobulins [3, 4] , aimed at modulating circulating levels of proinfl ammatory cytokines and at improving diuretic responsiveness. We tested the hypothesis that an effective removal of proinfl ammatory cytokines in patients with advanced CHF unresponsive to diuretic treatment was associated with diuresis restoration and with a signifi cant reduction of B-type natriuretic peptide (BNP) circulating levels. We prospectively enrolled 10 patients with decompensated CHF (NYHA class III–IV). Data are presented as mean 8 standard deviation. Five patients aged 57 8 12 years who were unresponsive to diuretic treatment (group A) underwent a short course of hemodiafi ltration (HDF) whereas 5 patients (group B) aged 49 8 13 years (p = nonsignifi cant vs. group A) Received: August 15, 2005 Accepted: October 5, 2005 Published online: December 15, 2005

Progressive Hemodialysis: Is It The Future?

Progressive hemodialysis is based on the simple idea of adjusting its dose according to residual renal function (RRF). The progressive, infrequent paradigm is slowly gaining a foothold among nephrologists, despite a lot of skepticism in the scientific world. Given the importance of RRF preservation in conservative therapy, it seems a contradiction to ignore the contribution of RRF when patients initiate hemodialysis (HD), especially when it is routinely considered with peritoneal dialysis. While a three‐times‐weekly HD regimen is broadly considered the standard starting regimen for new patients, twice‐weekly HD has been used in selected patients and is currently a common practice in South‐East Asia. Small studies indicate that a once‐weekly HD regimen may be a viable starting option as well. Progressive hemodialysis still requires validation, yet it is promising. We share the belief that a randomized clinical trial to investigate progressive hemodialysis is much needed, but we also strongly recommend including a once‐weekly HD starting dose as part of any such investigation.