[The Italian Registry for Therapeutic Apheresis: fifteen years of activity]. / Il registro italiano di aferesi terapeutica: quindici anni di attivita'

There is currently no consensus about the indications for therapeutic apheresis, also due to the lack of large clinical trials. A registry where all the data can be organized and analyzed therefore becomes a priority for all professionals involved in apheresis. The Apheresis Study Group of the Italian Society of Nephrology has instituted a registry for data collection. This report describes data collected from 1994 to 2008 by 68 units of different specialties in 15 Italian regions. Data about 29,260 treatments on 2,503 patients were recorded. Plasma exchange accounted for 43.4% of the procedures; 37% of these were performed by filtration. Plasma treatment was used in 48% of procedures, in particular with protein A immunoadsorption (11%), LDL-cholesterol apheresis by dextran sulfate adsorption (13%), and semiselective cascade or double filtration (12.5%). Cell apheresis, limited to photopheresis and leukocytapheresis, was used in 3.6% of cases, and whole blood treatment, with different techniques, in 5% of cases for the treatment of dyslipidemia, liver failure and sepsis. These procedures account for about 20% of the estimated therapeutic apheresis performed in Italy according to the national survey performed for the year 2005 by the Italian Apheresis and Cell Manipulation Society. The data collected are sufficiently informative to show a definite trend to use plasma/whole blood treatments as often as possible. The registry not only is a tool for consultation and information, but also a platform to plan and realize interdisciplinary and multicenter clinical trials.

Acute decompensation and absence of brain and kidney dysfunction predict long-term efficacy of plasma exchange in hyper-bilirubinemic cirrhotic patients awaiting liver transplantation.

Various artificial liver support systems are currently used in patients with decompensated chronic liver disease or acute liver failure as a bridge to recovery or to orthotopic liver transplantation (OLT). Between June 2004 and September 2006, 9 subjects were treated with plasma exchange (PE) for acute decompensation on chronic liver disease or chronic decompensation in end-stage liver disease. All of them were awaiting OLT or were listed at the moment of decompensation. Grade II to III hepatic encephalopathy (HE) was present in 4 patients, significant renal dysfunction in 3 patients, and ascites in 6 patients. Baseline serum total bilirubin was 35.1+/-11.2 mg/dL (mean value+/-SD). The patients underwent a mean of 12.1 2-hour exchanges over 1 to 8 weeks. The 3 who recovered were alive after a mean follow-up of 22.7+/-10.3 months. There were 3 patients who underwent transplantation and 3 who died due to liver failure during treatment. Only subjects with acute decompensation and without HE or significant renal dysfunction survived without OLT. PE did not significantly modify the grade of HE or the renal function. PE seemed to be a safe, long-term, effective therapeutic option for acute decompensation among subjects with chronic liver disease without brain or renal dysfunction.

Plasmaperfusion on triptophan columns can improve the clinical outcome of patients affected with myasthenia gravis.

UNLABELLED: Myasthenia Gravis (MG) is a neuromuscular disease often associated with thymic pathology due to neuromuscular transmission impairment by circulating antibodies directed against the cholinergic postsynaptic receptor on the neuromuscular junction (Anti-AchR-Ab). The treatment of MG includes cholinesterase inhibitors, steroids and thymectomy. Plasmapheresis can remove Anti-AchR-Ab but more recently plasma-perfusion (PP), a more specific apheresis for selective removal of noxious plasma components, has been developed. AIM OF THE STUDY: To study the effect of PP treatment, performed by using specific immunocolumns for Anti-AchR-Ab, on the clinical outcome of MG patients non-responder to steroid therapy or thymectomy. MATERIALS AND METHODS: We treated 8 patients suffering from severe MG by a cycle of 6 sessions of PP. We used columns containing triptophan as a specific ligand for Anti-AchR-Ab. In order to evaluate the effectiveness of treatment we used functional tests (muscular tests, respiratory function, electromyography) and laboratory tests (Anti-AchR-Ab; immunoglobulins, complement fractions, immunocomplexes). RESULTS: After one to three PP sessions, early clinical improvement in bulbar and respiratory symptoms were found in all patients and EMG showed improvement of neuromuscular transmission. Serum concentration of immunological markers decreased progressively and significantly during the treatment. Clinical improvements were progressive despite the tendency for Anti-AchR-Ab to reach initial values between one session and another. We observed no side effects due to the type of immunocolumns used. CONCLUSIONS: Triptophan columns appear to be able to remove large quantities of Anti-AchR-Ab and immunological markers from plasma. Our experience shows that PP performed using triptophan columns in patients suffering from severe MG provides good clinical results, improving patients' outcome, without any risk linked to the procedure.

Identification of adulteration in water buffalo mozzarella and in ewe cheese by using whey proteins as biomarkers and matrix-assisted laser desorption/ionization mass spectrometry.

A rapid and accurate method to identify bovine and ewe milk adulteration of fresh water buffalo mozzarella cheese by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) is described. The differentiation among mozzarella made from water buffalo milk and from mixtures of less expensive bovine and, more recently, ewe milk with water buffalo milk is achieved using whey proteins, alpha-lactalbumin and beta-lactoglobulins as molecular markers. It is worth noting that the method proposed here is, to our knowledge, the first strategy able to characterize possible fraudulent additions of ewe milk in samples of water buffalo milk devoted to the production of water buffalo mozzarella cheese. In addition, a linear relationship was found between the relative response of the molecular ion and the abundance of the analysed whey proteins. This demonstrates that this approach can be used to determine the amount of bovine or ovine milk added to water buffalo milk employed for mozzarella cheese production. Furthermore, this method also appears suitable for the analysis of ewe cheese. Hence these findings open the way to a new field for mass spectrometry in the evaluation of possible fraudulence in dairy industry production.

Sodium and fluid modulation in dialysis: new approach.

Hypotension during hemodialysis is still an unsolved problem. The treatment of patients with cardiovascular instability is efficaciously carried out with the use of 'profiled dialysis' (PD) with computerized modulation of ultrafiltration and conductivity. We tested a new profile model which involves progressive decrease of ultrafiltration associated with variable conductivity ('bell pattern'). We observed 8 stable long-term patients receiving hemodialysis (4 men and 4 women, mean age 63.5 years) for 4 h three times a week. Before our test, sodium balance had reached a steady state in all patients and remained stable during the entire observation period. The sodium balance was established by means of a simple pattern suggested by Ursino and coworkers. The patients were observed for two periods of 1 month each (protocols A and B). The intradialytic mean arterial pressure was studied, checking every hour of dialysis. Statistical analysis was done by ANOVA for repeated measures. We compared standard dialysis with constant ultrafiltration rate and conductivity (protocol A) with sessions performed involving a progressive decrease of ultrafiltration together with a variable conductivity of -0.2, +0.2, +0.6, +0.6, 0, -0.4, -0.4, and -0.4 mS/s (protocol B). We found a lower incidence of hypotension (p < 0.01) with better cardiovascular stability during and after treatment in 'profiled dialysis'.

Identification of adulteration in milk by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

The development is described of a rapid, simply and accurate analytical method aimed at evaluating both the presence of cow milk in either raw ewe and water buffalo milk samples employed in industrial processes and the addition of powdered milk to samples of fresh raw milk, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The presence of adulteration is defined by evaluating the protein patterns coming from the most abundant whey proteins, alpha-lactalbumin and beta-lactoglobulin, used as molecular markers. As no pretreatment of the milk samples is required and owing to the speed and ease of use of MALDI-MS the proposed analytical protocol can be used as a routine strategy for the identification of possible adulteration of the raw fresh milk samples that the dairy industry receives from producers every day.

Apheresis in primary focal segmental glomerulosclerosis of native and transplanted kidneys: a therapeutic protocol.

BACKGROUND: Patients with focal segmental glomerulosclerosis (FSGS) develop nephrotic syndrome and terminal renal failure in most cases. FSGS reappears in 15-50% of transplanted kidneys and frequently causes the graft loss. Sera from patients with FSGS of native or transplanted kidneys contain some proteinuric or permeability factors (PF) which can be removed by means of plasma exchange (PE) or protein A Immunoadsorption (IA). METHODS: We suggest a therapeutic protocol, for patients with biopsy proven FSGS of native or transplanted kidneys, resistant to steroid and immunosuppressive therapy, based on the association of PE or IA to conventional drug therapy. Daily proteinuria, renal function, serum albumin and circulating level of proteinuric factors (permeability test) will be monitored at regular time intervals during the apheresis cycle, which will be intensive at the beginning (8-10 sessions in 4 weeks) and very gradually discontinued. Results. We will consider satisfactory remission the reduction of proteinuria below 1 g/day, improvement of renal function, normalization of serum albumin level (> 3.5 g/dl). Partial remission will be considered: proteinuria below 3 g/day, stable renal function, serum albumin level between 3 and 3.5 g/dl. Permeability test, if positive at baseline examination, should be negative after apheresis. CONCLUSIONS: The primary endpoint of our protocol is: lasting remission (satisfactory or partial) after the apheresis suspension. Secondary endpoints are: maintained remission with continuing apheresis sessions, correlation between permeability activity and disease activity, identification of responders and non responders patients on the basis of positive permeability test.

Low levels of hepatitis C virus RNA in blood of infected patients under maintenance haemodialysis with high-biocompatibility, high-permeability filters.

BACKGROUND: Patients undergoing maintenance haemodialysis are often infected with hepatitis C virus, yet the clinical course of liver disease is usually mild. We investigated whether the hepatitis C virus viral load is influenced by the haemodialytic procedure and the type of dialyser. METHODS: Hepatitis C virus-RNA levels were measured using a quantitative polymerase chain reaction assay in predialysis blood from 51 hepatitis C virus-infected patients dialysed with different membranes. Paired pre- and post-dialysis measurements were also obtained in 18 patients. RESULTS: Patients dialysed using cuprammonium-regenerated cellulose membranes with low (cuprofan) or intermediate (cellulose acetate or diacetate) biocompatibility had higher pre-dialysis hepatitis C virus-RNA levels (p<0.05] compared to those dialysed with synthetic high-biocompatibility, high-permeability polymeric membranes (polyacrylonitrile, polysulfone, polymethylmethacrylate or polycarbonate). Hepatitis C virus-RNA levels were unrelated to the duration of haemodialysis and the presence of abnormal transaminases. A significant reduction (p=0. 04) of serum hepatitis C virus-RNA levels was observed after a single haemodialysis, particularly in patients with high pre-dialysis viral load. Patients with low pre-dialysis hepatitis C virus-RNA levels (<2. 5 x 10(3) copies/ml) exhibited only minimal changes following the procedure. Four patients with medium-high basal viral load switched from a low-biocompatibility/low-permeability to a high-biocompatibility/high permeability filter had a marked reduction of viraemia after three weeks, in one case followed by a new increase after return to the original filter. CONCLUSIONS: Haemodialysis with high-biocompatibility/high-permeability filters in hepatitis C virus-infected patients is associated with low blood levels of hepatitis C virus-RNA. This finding may be of clinical relevance, especially in patients listed for kidney transplantation.

Semi-selective immunoadsorption treatment in myasthenia gravis.

We treated some patients with severe generalized myasthenia gravis (MG) with semi-selective adsorption by plasma perfusion(PP) on IM-T350 column. We studied efficiency and undesired effects of the employed material to verify the safety and effectiveness of methodology. Thus a treatment protocol has been prepared: 6 PP per treatment cycle; 3-7 days between PP sessions; plasma to treat per session 1,2 lt; only one adsorbent column for every cycle. Our experience, on 13 cycles of treatment, showed that PP in autoimmune MG has provided good clinical results without any improvement in genetic MG.

Myasthenia gravis (MG) treatment with immunoadsorbent columns.

We performed ten plasma perfusion (PP) treatments on eight patients affected by Myasthenia Gravis (MG) with high serum levels of autoantibodies against acetylcholine receptors (anti AChR-AB), and one PP treatment on a patient with MG of probable genetic origin and without specific antibodies. All patients (Osserman group III-IV) had undergone thymectomy and immunosuppressive therapy. Each patient received a treatment cycle of six PP sessions. Clinical conditions were assessed before and after the treatment with evaluation of muscular strength, ventilatory function, and electromyographic testing. Immunologic markers were tested before and after each PP. The patient without specific antibodies showed no improvement with PP and was excluded from our study. All the other patients showed continued improvement, with increased muscle strength and improved respiratory function. Four patients (follow-up 16-24 months) still maintain the clinical improvement; two, unexpectedly relapsed 11-12 months after PP, received a new treatment successfully (follow-up 9-10 month).

Computerised monitoring of sodium and fluid during haemodialysis.

Four stable chronic haemodialysis patients were studied during two periods of 3 weeks: at first a 'standard' haemodialysis (S/HD) was used, followed by 'computer-modulated' haemodialysis (CM/HD). We used a monitor Hospal Monitral-S with Hospal DPM (Display Programmer Module) that allows the programming of 'weight loss rate' and 'dialysate osmolality'. During standard haemodialysis fluid was removed at a constant rate of 1 kg/h with constant sodium dialysate of 143 mmol/l. During computer-modulated haemodialysis the dialysate sodium concentration and the fluid removal of 4 kg per session was modulated by display programmer module.

Plasma perfusion in myasthenia gravis.

Seven patients with myasthenia gravis (MG) unresponsive to thymectomy and steroid treatment (Osserman group III) underwent plasma perfusion (PP). All patients showed palpebral ptosis, diplopia, dysphonia, dysphagia, and muscle weakness; five of them had impaired ventilatory function. Separated plasma was perfused onto a column to adsorb anti-AChR-Ab. Each patient received a treatment cycle of six PP sessions. Clinical conditions were assessed before and after the treatment, with evaluation of muscular strength, ventilatory function, and electromyographic testing (RSS). Immunologic markers were tested before and after each PP. Clinical improvement in bulbar symptoms and respiratory function was noted in all patients after one to three PP. Limb muscle strength began to improve later. Serum concentration decreased (mean % +/- SD) after each PP:anti-AChR-Ab 36.47 +/- 17.43; IgA 20.44 +/- 11.26; IgG 21.24 +/- 32.56; IgM 23.22 +/- 11.40; C3 36.78 +/- 10.15; C4 42.69 +/- 14.82. In five of seven patients the improvement continues (follow-up 1 to 10 months). In one patient it lasted only 1 month, and in another a relapse occurred after 10 months of benefit, but was successfully reversed by retreatment.

Extracorporeal blood recirculation and beta-2-microglobulin plasma levels during hemodialysis.

Beta-2-microglobulin (beta 2m) plasma levels during hemodialysis have recently been considered markers of membrane biocompatibility. The aim of this study was to assess if generation of beta 2m from blood cells in contact with the membrane could account for changes in beta 2m plasma concentration during hemodialysis. The role of heparin was studied as well by comparing conventional with heparin-free dialysis. beta 2m plasma levels were measured in six patients during 12 Cuprophan (Cu) and 12 Eval (E) non-consecutive hemodialysis sessions without ultrafiltration. E membranes were compared with and without heparin. beta 2m plasma levels significantly increased with Cu but not with E. A total of 200 ml of blood from four other patients was recirculated ex vivo before Cu hemodialysis in Cu or E and then reinfused into the patient. A drop in beta 2m concentration was observed in the recirculating blood, recovery of beta 2m from the washed filter was higher with E, and reinfusion of recirculated blood to the patient was not associated with significant increases in beta 2m plasma levels. Hemodialysis with a new Cu filter, following reinfusion, did not produce significant increase in plasma beta 2m concentration. In conclusion, 1) beta 2m plasma levels change during Cu, but are not associated with hemoconcentration from ultrafiltration nor with local release from the blood leaving the filter, 2) beta 2m binding to the membrane may account for decreased levels in recirculated blood, 3) E membranes both with and without heparin do not increase beta 2m plasma levels and reinfusion of recirculated blood blunts the beta 2m changes observed with Cu.