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.

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.

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.

Effectiveness of single daily intraperitoneal administration of aztreonam and cefuroxime in the treatment of peritonitis in continuous ambulatory peritoneal dialysis (CAPD).

To discover if the management of peritonitis in continuous ambulatory peritoneal dialysis (CAPD) may be effectively simplified by single daily i.p. administration of aztreonam (A) and cefuroxime (C), 10 adult CAPD patients (pts) with peritonitis were trained to start the following treatment procedure: a) sterile collection of dialysate effluent for cultures; b) 4 rapid in-and-out exchanges with antibiotic free dialysate; c) addition of 2 g C and 2 g A to a 2-L exchange for 6-h dwell time (the same dosage was repeated once a day in the overnight exchange); d) routine CAPD exchanges. Concentrations of C and A were measured in dialysate and serum of the patients 2, 12, 18, and 21-23 h after the i.p. administration: C remained within therapeutic range in all samples, while serum and dialysate A levels fell below such range 16 h after the i.p. administration. Seventeen peritonitis episodes occurred during the observation period (12 months): initial dialysate cultures grew Staph. aureus in 6 episodes, Staph epidermidis in 6, Pseudomonas aer. in 2, Streptococcus faecalis in 1, Citrobacter in 1, and Candida in 1. All patients but 1 (with Candida-positive culture) responded to this treatment with no relapse in 2 months. We conclude that once a day i.p. administration of 2 g aztreonam plus 2 g of cefuroxime is an effective and simple way of treating CAPD pts with gram-positive and gram-negative peritonitis.

[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.

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.

[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.

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.

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.

Outcome of alcoholism treatment among 5578 patients in an urban comprehensive hospital-based program: application of a computerized data system.

The Elmhurst Alcoholism Program data system relates fraction of time patients are in remission to length of time they are know, and permits a systematic search for groups of variables which predict greater abstinence rates. Our program (inpatient detoxification ward, multidisciplinary outpatient clinic, and halfway house) sets abstinence as a goal. After initial data collection (social and demographic variables, income, education, residence, drinking and drug history, legal problems, and prior treatment), we collect data at 30 days after admission and at 2- or 3-month intervals thereafter from all patients who remain in contact. Forty-five percent of all patients are lost before 30 days. Only 32% of those still under observation at 1 yr and 27% of those still under observation at 2 yr claim continual abstinence. From among many combinations of variables examined for predictive power, a group of five was chosen as best. Patients who at their first follow-up interview reported attending AA meetings and taking Antabuse and who in addition had a job at admission, were older, and were admitted through the clinic rather than through inpatient detoxification had a greater fraction of time abstinent. Nevertheless, the best cluster of predictor variables did not account for more than 25% of the variation in rates of abstinence. Many social variables are not independent, and some may merely be indicators of duration or severity of alcoholism. Contemporary alcoholism treatment is, at best, of limited effectiveness.

Cardiovascular impairment, dialysis strategy and tolerance in elderly and young patients on maintenance haemodialysis.

Cardiovascular impairment is considered a major cause of reduced tolerance to haemodialysis in elderly patients and is thought to require a soft dialysis strategy in these patients. To verify these observations we compared dialysis strategy and tolerance of patients over 65 years old to those of patients under 45 years, since atherosclerotic/involutional changes of vasculature should be quite different in these groups. Seventy-seven elderly and 57 young patients (age 72 +/- 5 and 35 +/- 6 years respectively, mean and SD) were selected from a population of 292 patients undergoing regular dialysis treatment exclusively on the basis of the age criterion. Cardiovascular impairment was found to be greater (as expected) in older than in younger patients, as documented by a larger proportion of diabetic patients (19% vs 3%), a greater incidence of cardiovascular manifestations both before the beginning of dialysis (27% vs 10% of patients affected) and after the beginning of dialysis (35% vs 16%), a greater incidence of ECG abnormalities (78% vs 56%), a longer exposure to hypertension (9.2 vs 3.5 years), and a greater utilisation of digitalis (36% vs 5%) and nitrates (19% vs 2%). Dialysis strategy in elderly patients differed from that in young patients only for a moderately lower blood flow rate (243 vs 279, 244 vs 279, ml/min, males and females respectively). Dialysis sessions were shorter in older than in younger males (11.8 vs 12.4 h/week). The incidence of intradialytic hypotension did not differ significantly between the two groups (44% vs 32%), although the elderly showed a greater pre-to-post-dialysis systolic arterial pressure reduction (21 vs 15 mmHg).(ABSTRACT TRUNCATED AT 250 WORDS)

Mesangial expression of intercellular adhesion molecule-1 in primary glomerulosclerosis.

Frozen sections of renal biopsy specimens from eight patients with primary focal segmental glomerulosclerosis (FSGS) and 10 patients with membranous nephropathy (MN) were stained in immuno-peroxidase with the intercellular adhesion molecule-1 (ICAM-1) monoclonal antibody (MoAb), CL203.4. ICAM-1 was expressed by mesangial cells in six patients with FSGS. On the other hand, ICAM-1 was not detected in mesangial cells in patients with MN or in the non-affected portion of tumoral kidneys used to control normal renal expression of ICAM-1. De novo mesangial expression of ICAM-1 in FSGS suggests that sclerosis results from an inflammatory process, possibly associated with local release of cytokines.