Theonellasterone, a steroidal metabolite isolated from a Theonella sponge, protects peroxiredoxin-1 from oxidative stress reactions.

Peroxiredoxin-1, a key enzyme in the cellular detoxification pathway, has been identified through a chemoproteomic approach as the main partner of theonellasterone, a marine bioactive metabolite. A combination of chemical and biochemical assays disclosed its mechanism of action at the molecular level.

Cost-effectiveness of kidney transplantation from DCD in Italy.

INTRODUCTION: Kidney transplantation represents the best therapeutic option for patients with end-stage renal disease (ESRD), providing the best outcomes for survival, quality of life, and cost-effectiveness. To increase kidney donations, in 2007, the Italian IRCCS Policlinico San Matteo Foundation in Pavia designed and conducted Programma Alba, a protocol for organ donation after cardiac death (DCD). This study evaluated the costs and health outcomes of DCD transplantation and in all types of transplants compared with current clinical practice. PATIENTS AND METHODS: A Markov-based model was used to assess costs and health outcomes for new ESRD patients for 2008 to 2013. A health care founder perspective was used. Data sources were the Italian National Institute of Statistics and the Lombardy Registry of Dialysis and Transplantation. A microcosting analysis was performed to calculate costs related to clinical pathways for DCD. We assessed costs, survival, quality-adjusted survival, and cost-effectiveness. FINDINGS: Changing the actual practice pattern for new patients with ESRD and increasing the availability of kidneys from DCD to 10 extra transplants per year will induce an incremental cost per quality-adjusted life-year of €4255. Increases in transplantation to reach an extra 10% by transplant type would result in reduced costs and increased patient survival and quality of life compared with the current scenario. INTERPRETATION: Our data show that increasing DCD transplants would result in a cost-effective policy to expand the kidney donor pool compared with current ESRD treatment patterns. Italian policies should make an effort to increase transplant rates to optimize cost-effectiveness in ESRD service supply.

Non-heart-beating organ donation in Italy.

In 2007 the non-heart-beating organ donation (NHBD) "Programma Alba" (Sunrise Programme) started in Pavia, Italy. The initial plan was to cut down waiting list for kidney transplantation, while its final aim is to shorten organ transplantation waiting lists. When compared to European countries and the USA, the Italian NHBD program has taken longer to get established. Initially Italian physicians were not entirely aware of the NHBD organ viability for transplantation, furthermore ethical issues and the need to regulate medical requirements to Italian law slowed down the NHBD program. In particular, Italian legislation provides for death ascertainment after irreversible cardiac arrest, 20-minute flat electrocardiogram. This no-touch period is longer when compared to worldwide legislation, and organ viability has been a main concern for Italian transplant doctors over the years. However, recent data let up to 40-minute warm ischemia time to preserve organ viability; this has encouraged Pavia's group to establish the NHBD "Programma Alba". It was designed according to Italian legislation from death diagnosis to graft placement, from this perspective must the significant role of the Transplant coordinator be recognized. Since 2007 seven kidneys have been gathered from seven NHBD. Of these, six NHBD kidneys have been transplanted. Currently, four patients are out of dialysis. This report is a detailed description of NHBD "Programma Alba" and its preliminary results.

Toll-like receptor 4 expression is increased in circulating mononuclear cells of patients with immunoglobulin A nephropathy.

We investigated Toll-like receptors (TLR-3, -4 and -7) expression in circulating mononuclear cells of patients with immunoglobulin A nephropathy (IgAN), a disease with debated relationships with mucosal immunity. TLR-4 expression (detected by fluorescence activated cell sorter) and mRNA transcriptional levels (Taqman) were significantly higher in patients with IgAN than in healthy controls (P = 0.00200 and P = 0.0200). TLR-3 and TLR-7 were not modified significantly. In IgAN patients proteinuria was correlated significantly with TLR-4 expression (P = 0.0312). In a group of nephrotic syndromes, TLR-3, -4 and -7 expression was similar to healthy controls. A significant difference in TLR-4 expression and mRNA levels was found between very active IgAN patients (proteinuria > 1 g/1.73 m(2)/day in association with severe microscopic haematuria) and inactive patients (proteinuria < 0.5 g/1.73 m(2)/day, with absent or minimal haematuria). No correlation with levels of aberrantly glycosylated IgA1, age, renal biopsy features or therapy was found. This study shows for the first time an up-regulation of TLR-4 in circulating mononuclear cells of patients with IgAN, particularly in association with proteinuria and heavy microscopic haematuria.

[Regulatory T cells in kidney transplant recipients]. / Cellule T-regolatorie nel trapianto di rene.

Immunosuppressive drugs are essential for the prevention of acute transplant rejection but some may not promote long-term tolerance. Tolerance to self-antigens is ensured naturally by several mechanisms; one major mechanism depends on the activity of regulatory T lymphocytes (Treg), particularly CD4+CD25+ T cells. The transcription factor forkhead box protein 3 (Foxp3) has been identified as a molecular marker for Treg cells. The direct effects of immunosuppressive drugs on CD4+CD25+ cells are uncertain. In the clinical setting, basiliximab used in the induction phase of immunosuppression effectively reduced the number of acute rejection episodes. We studied the effects of the most widely used immunosuppressive induction regimens including cyclosporine, mycophenolate mofetil, steroids, and anti-CD25 monoclonal antibody (basiliximab) on the capacity to regulate human Treg in vivo. Twenty first cadaveric kidney transplant recipients (14 men, 6 women) were enrolled in the study. Blood samples were collected before kidney transplant and after one month. Blood sampling was done immediately before the administration of immunosuppressive therapy after an overnight fast. None of the transplant recipients presented laboratory or clinical signs of infection or acute rejection. The number and percentage of CD4+CD25+ and Foxp3+ T cells were determined by fluorescence activated cell sorter (FACS) analysis. Our results showed absence of both CD4+CD25+ and CD4+CD25+ Foxp3+ T cells one month after transplant. Peripheral CD4+CD25-Foxp3+ T cells significantly decreased after transplant but did not disappear. These preliminary data suggest that immunosuppressive induction therapy with basiliximab completely suppresses CD4+CD25+ regulatory cells and significantly reduces the total number of Foxp3+ lymphocytes.

Holoprosencephaly with neurogenic hypernatremia: a new case.

BACKGROUND: Holoprosencephaly (HPE) is the most common developmental defect of the forebrain and mid-face in humans. It is a disorder of neural induction in which a genetic programming defect results in noncleavage of the forebrain in the sagittal plane and variable hypoplasia of paramedian structures. It occurs in 5-12/10,000 live births. Clinically, there is a nearly continuous spectrum of malformations consistent with HPE. Endocrinopathies, such as diabetes insipidus, hypothyroidism, hypocorticism, and growth hormone deficiency, are frequently associated with HPE. Seizures may occur. CASE REPORT: We report a new case of semilobar-HPE complicated by neurogenic hypernatremia and no signs of dehydration in a child with microcephaly, spasticity, mental and psychomotor retardation, frontal bones hypoplasia, and mild facial dysmorphism.

Mycophenolate mofetil in primary glomerulopathies.

Mycophenolate mofetil is an immunosuppressive agent that blocks purine biosynthesis, inhibits T and B-lymphocyte and mesangial proliferation. Mycophenolate mofetil is not nephrotoxic like calcineurin inhibitors and is widely used in solid-organ transplantation. Recently, mycophenolate mofetil has been introduced in the treatment of autoimmune diseases and primary glomerulopathies. This review analyzes the literature currently available on the treatment of primary glomerulopathies with mycophenolate mofetil. Encouraging results have been obtained in minimal change nephropathy where it may help to reduce the use of steroids in these patients who are often very young. The results obtained in medium and high risk patients with focal segmental glomerulonephritis and idiopathic membranous nephropathy were less encouraging. Conflicting results have been reported on IgA nephropathy in controlled trials. None of these studies attained level A evidence, meaning that randomized control trials of sufficient statistical significance are necessary to estimate the real effectiveness of mycophenolate mofetil in primary glomerulopathies.

The effect of sirolimus- or cyclosporine-based immunosuppression effects on T-cell subsets in vivo.

While sirolimus (SRL) is thought to be a non-nephrotoxic agent, cyclosporine A (CsA) toxicity is a serious problem in kidney transplantation. We compared the effects of the two drugs on T-helper (Th) subsets in kidney transplant patients. We examined 24 first cadaver kidney recipients equally randomized to receive SRL/mycophenolate mofetil (MMF)/methylprednisolone (MP), or cyclosporine with either MMF or MP. The Th1 and Th2 subsets in peripheral blood were separated based on their production of interferon-gamma (INFgamma) or interleukin (IL)-4/IL-5. The lymphocytes were stimulated with phytohemoagglutinin or with allogenic CD3-depeted and irradiated antigen-presenting cells. Furthermore, the conversion potential of Th0 to Th1 was determined by measuring IL-12 and IL-18 levels after lipopolysaccharide challenge. When peripheral blood lymphocytes taken from SRL-treated patients were stimulated by phytohemoagglutinin, there were significantly lower INFgamma-producing cells compared with the lymphocytes taken from patients treated with CsA. The number of IL-4/IL-5-producing cells did not differ among the patient groups. Release of IL-12 but not IL-18 from peripheral lymphocytes following treatment with lipopolysaccharide was significantly lower in the SRL-treated patients. These results show that compared with CsA, SRL caused a significant decrease in the Th1 lymphocyte subset associated with a significant reduction of IL-12 release.

Influence of methylprednisolone on plasma homocysteine levels in cadaveric renal transplant recipients.

The aim of the present study was to investigate plasma homocysteine levels in renal transplant recipients in the course of steroid-based or steroid-free immunosuppression. Data from 32 patients were retrospectively analyzed according to the steroid immunosuppressive regimen. The 20 recipients on methylprednisolone (MP) plus cyclosporine (CyA) or tacrolimus (TRL) (n = 20) showed similar creatinine levels when compared with those on calcineurin inhibitors plus mycophenolate mofetil (MMF; n = 12), (1.6 +/- 1.5 vs 1.6 +/- 0.4 mg/dL; P = NS) but significantly higher total plasma homocysteine (tHcy) levels (28.5 +/- 12.5 vs 16.3 +/- 5.5 micromol/L; P < .05). No differences of tHcy levels have been observed when patients were analyzed according to CyA- or TRL-based immunosuppression regardless of MP or MMF associations. Our data suggest that recipients, particularly those on steroid-based immunosuppression, should receive homocysteine-lowering treatment early after transplantation.

[Changes is Th1/Th2 balance in hemodialysis patients: effect of different membranes]. / Alterazione del bilancio Th1/Th2 nei pazienti in trattamento emodialitico: effetto di differenti membrane.

BACKGROUND: Hemodialysis (HD) patients present an immunodeficiency that is mainly related to the defect of cell-mediated immunity. We have previously shown the polarisation of T-helper cells toward the phenotype in HD treatment with cuprophan membrane. In the present study, we have examined the effect of a Vitamin E-coated dialyser (Excebrane, VE) on the activity of the two Th subsets. METHODS: We studied 8 healthy controls and 10 patients on RDT for at least 6 months with cellulose membrane (AC), then switched to HD-VE. Blood was collected from HD patients while on treatment with AC, and after 1 year of treatment with VE. CD4+ cells were isolated from peripheral blood mononuclear cells (PBMC) by negative selection, treating PBMC with a cocktail of anti-CD8, -CD16, -CD19, -CD36 and -CD56 antibodies labelled with magnetic beads, and passing them through a magnetic field. The collected Th cells were cultured for 48 h with and without phytohemagglutinin (PHA). IFNgamma and IL-4 were measured in the supernatant using the ELISA assay. RESULTS: The constitutive release of IL-4 by CD4+ cells was abated significantly by treatment with VE. IFNgamma released by mitogen-stimulated CD4+ recovered with VE. CONCLUSIONS: This study demonstrates that treatment with vitamin E-coated dialyser improves the defect of PBMC function associated with cellulose membrane dialyser consisting of altered spontaneous and mitogen-stimulated cytokine release. The effects of vitamin E-coated filter, in particular the recovery of reactive IFNgamma production by Th1 cells and the restriction of spontaneous IL-4 release by Th2 cells may have clinical importance.

[Effectiveness of oral supplements of folates, vitamins B12 and B6 in hemodialysed patients. Preliminary results after 5 months of follow-up]. / Efficacia dei supplementi orali di folti, vitamine B12 e B6 in pazienti emodializzati. Risultati preliminari dopo 5 mesi di follow-up.

BACKGROUND: Elevated serum levels of homocysteine have increasingly been associated as a risk factor of cardiovascular disease. Recent reports demonstrated that supplements of folates, vitamin B12 (B12) and vitamin B6 (B6) are effective in correcting serum Hcy levels in hemodialysed patients. AIM: to assess the effectiveness of oral supplements of folates, B12 and B6, in order to reduce serum Hcy levels in our cohort of hemodialysed patients. METHODS: Sixty-one hemodialysed patients have been enrolled in the study (age 68+/-13 years; hemodialysis 62+/-42 months). Oral supplements of calcium folinate (30 mg 3 times a week), B12 (500 mg 3 times a week) and B6 (200 mg 3 times a week) were administered at the end of each hemodialysis session. Serum levels of Hcy, folic acid and B12 were tested at the beginning of the study and at 2 month intervals. RESULTS: After 5 months of follow-up, serum levels of Hcy were normalised in 19% of our patients and in total 70% of them showed a reduction >8% when compared with the basal Hcy levels. No side effects related to folates, B12 or B6 supplementation were observed. CONCLUSIONS: Oral supplements of folates, B12 and B6 are a safe and effective treatment of hyperhomocysteinemia in hemodialysed patients.

Sardinia: a battlefield approach to type I diabetes epidemiology. Sardinia-IDDM Study Groups.

Sardinia and Finland have the highest incidence of insulin-dependent diabetes mellitus (IDDM) in the world. Therefore, both regions represent ideal observatories for investigating the environmental, genetic and immunological factors which have led to this dramatic increase. We have concentrated our efforts on Sardinia. Among several projects, there is the mapping of the island for hot and cold spots for overt IDDM. In order to map the island for pre-IDDM, we have collected and bled around 10,000 school children (age 6-14 years) and we are now in the process of enrolling around 30,000 new-born babies. We report here our initial results, which show that progression to IDDM is accompanied in both cohorts by the presence of a combination of islet-cell antibodies with either glutamic acid decarboxylase or IA-2 antibodies or both. This approach should lead to the design of reliable models of IDDM prediction in the general population, which will benefit an early insulin treatment and, hopefully, an effective prevention of the disease.

The "Sardinia-IDDM study": an attempt to unravel the cause of insulin-dependent diabetes mellitus in one of the countries with the highest incidence of the disease in the world.

Sardinia and Finland have the highest incidence of IDDM in the world. Thus, both regions represent ideal observatories for investigating the environmental, genetic and immunological factors, which have led to this dramatic increase. We have concentrated our efforts in Sardinia. Among several projects, there is the mapping of the Island for hot and cold spots for overt IDDM. In order to map the Island for pre-IDDM, we have collected and bled around 10,000 school children (age 6-14 years) and we are now in the process to enroll around 30,000 newborn. We report here our initial results, which show that progression to IDDM is accompanied in both cohorts by the presence of a combination of ICA with either GAD and IA-2 antibodies or both. This approach should lead to design reliable models of IDDM prediction in the general population, which will benefit an early insulin treatment and, hopefully, an effective prevention of the disease.

Effects of hypercholesterolemia of renal hemodynamics: study in patients with nephrotic syndrome.

Experimental and clinical studies have demonstrated a positive relationship between hyperlipidemia and rate of progression of renal disease, suggesting that lipids can induce or aggravate glomerular injury mainly by interacting with mesangial cells. Nevertheless, recently has been demonstrated that increased cholesterol levels can also induce endothelial cell dysfunction. Thus, since endothelium is known to play a major role in modulating the vascular tone, we have tested the possibility that hypercholesterolemia impairs the renal hemodynamics in patients with active nephrotic syndrome and elevated serum cholesterol levels. In this single-blind, nonrandom study, 12 patients were treated with pravastatin (group T, treated, n = 12) and 8 with placebo (group C, controls, n = 8). The controls were studied after the pravastatin group had been completed. Before starting the treatment the patients underwent basal determinations including routine laboratory investigations and PAH and inulin clearances. The same determinations were repeated after 48 h, and 6 and 12 weeks from the beginning of the treatment. The study at 48 h was performed to see if pravastatin had a direct, cholesterol-independent effect on renal function. The following basal results were reported (mean +/- SEM; group T vs. group C): serum cholesterol (mmol/l) 9.7 +/- 0.4 vs. 9.1 +/- 0.3 (NS); proteinuria (g/24 h): 6.2 +/- 0.2 vs. 7.0 +/- 0.7 (NS); PAH clearance (ml/min): 353 +/- 21 vs. 385 +/- 31 (NS); inulin clearance (ml/min): 62.5 +/- 7.7 vs. 67 +/- 9.3 (NS). After 48 h, no changes were observed in both groups. Subsequently, in group T, the following percentage changes of basal levels were observed: serum cholesterol -21.4 +/- 3.2% at 6 weeks (p < 0.05) and -34.9 +/- 3.2% at 12 weeks (p < 0.01); inulin clearance +3 +/- 3.7% at 6 weeks (NS) and +9.3 +/- 2.9% at 12 weeks (p < 0.05); PAH clearance +7 +/- 3.1% at 6 weeks (p < 0.05) and +21.2 +/- 5.5% at 12 weeks (p < 0.01). By contrast, no significant changes of these parameters occurred in group C at any time, so that the percent changes of baseline values of CPAH were significantly greater in group T (at 6 weeks: p < 0.05; at 12 weeks p < 0.005). These results indicate that the reduction of cholesterol is associated with a significant increase in renal plasma flow, thus, suggesting that hypercholesterolemia may actually impair the renal hemodynamics. We speculate that this effect may contribute to increase the risk of ischemic acute renal failure in nephrotic patients and, along with changes induced in the mesangium by other mechanisms, to contribute to the progression of renal disease.

Serial morphometric analysis of sclerotic lesions in primary "focal" segmental glomerulosclerosis.

The possibility of missing the diagnosis of focal segmental glomerulosclerosis (FSGS) has been primarily attributed to the focal distribution of the sclerotic lesions, but this assumption has not been verified by any serial morphometric analysis of renal biopsy specimens. The aim of this study is to assess the size and the distribution of sclerotic lesions in primary FSGS and to establish the minimum number of glomeruli and sections necessary for the diagnosis. Fourteen biopsies from adult nephrotic patients with primary FSGS were carefully selected from a group of 41 biopsies, to minimize the possibility of finding and misinterpreting nonspecific glomerular scars, and were serially cut to obtain 1485 consecutive 2 microns-thick sections that, after PAS staining, showed 182 glomeruli. Fifty-seven glomeruli were "complete", i.e., they emerged after the first section and disappeared before the last section. The percentage of glomeruli with sclerotic lesions was 31.5% in the starting section, 71.8% after the observation of all serial sections, and 81.7% when only the complete glomeruli were considered. The morphometric analysis on complete glomeruli revealed that the volume of the sclerotic lesions averaged just 12.5% +/- 2.2 SE of the entire glomerular volume, and the statistical analysis revealed that the minimum number of glomeruli needed in the starting section to exclude sclerotic lesions is eight (P < 0.01) or nine (P < 0.001). If fewer glomeruli are seen, it is necessary to cut 2 microns-thick serial sections, but to examine just one of every 11 (P < 0.001), the number of sections to examine being proportional to the number of glomeruli found. In conclusion, this study shows that the distribution of sclerotic lesions in primary FSGS is not focal, but diffuse; however, because of the small size of the sclerotic lesions, the probability of missing the diagnosis is statistically relevant when fewer than eight glomeruli are found in the starting section, unless a serial morphological analysis, even on a reduced number of sections, is made.

Acute renal and immunological effects of low-dose cyclosporin in humans.

BACKGROUND: In recent years a reduction of oral cyclosporin A (CsA) dose has been adopted to minimize its adverse renal effects. To date, however, little is known about the intrinsic renal and immunological effects of low-dose CsA. METHODS: Four oral doses of the drug (2, 3, 4 and 5 mg/kg body wt) and placebo (P) were randomly administered in two half-doses to seven healthy subjects. Studies were performed during water diuresis 4 h after administration of the 2nd half-dose, i.e. when the biological activity of the drug is considered maximal. Renal function was evaluated after all doses. In the same subjects, the levels of interleukin-2 (IL-2) and IL-2 receptor (IL-2R), that are the main immunological targets of CsA, were measured in the supernatant of peripheral blood mononuclear cells cultured with phytohaemagglutinin (PHA) after P, 3 and 5 mg/kg of the drug. RESULTS: CsA induced a dose-dependent and proportional decrease of GFR and RPF associated with increasing renal vascular resistances (RVR) in presence of unmodified blood pressure. Similarly, Na+ urinary excretion decreased in a dose-dependent manner due to both GFR reduction and to an higher tubular reabsorption mainly localized at the level of the proximal nephron. All these changes were significant only after 4 and 5 mg/kg. A significant suppression of PHA-stimulated IL-2 and IL-2R cell release was observed following 5 mg/kg only. CONCLUSIONS: This study suggests that nephrotoxic and immunosuppressive effects of low-dose CsA are strictly linked.

Functional versus structural changes in the pathophysiology of acute ischemic renal failure in aging rats.

The aim of this study was to gain further insight into the greater susceptibility to acute ischemic renal failure (ARF, 30 min of renal arteries clamping) of old rats (O, 18 months) as against young rats (Y, 3 months). All the rats ate a hypoproteic diet (14% of casein) to avoid age-related glomerulosclerosis in O. Basal renal dynamics was similar in O and Y (Groups CON). One day after ARF, the decrease in GFR was more severe in O than in Y (-82% and -57% vs. respective CON, P < 0.05), due to a greater rise of RVR in O (+258%) than in Y (+104%). The histological renal damage after ischemia was comparable in the two groups with ARF. Five days after ARF, the recovery of renal function was characterized by a slower rise of GFR in O than in Y. In two further groups, two different scavengers of oxygen-free radicals, dimethylthiourea (DMTU) and superoxide dismutase (SOD), were administered at the time of arterial occlusion. DMTU had protective effects in Y but not in O (delta GFR was -28% and -72%, respectively); in contrast, SOD was more effective in O (delta GFR = -58%) than in Y rats (delta GFR = -40%). To test the hypothesis that such a difference was related to the capacity of SOD to increase the levels of nitric oxide (NO), four more groups of Y and O rats were pretreated with L-arginine (ARG), precursor of NO, in tap water (1.5%). No difference in renal dynamics was detected in basal conditions.(ABSTRACT TRUNCATED AT 250 WORDS)