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Biomedical subjects

S Stefoni

Publications and source records attributed to S Stefoni.

At least 73 records · Page 4Linked to original sources

Comparison of hemodialysis versus hemoperfusion in the clearance of high-dose methotrexate in pigs.

To evaluate the different capabilities of hemo-perfusion (HP) and hemodialysis (HD) treatment in resolving the side effects during high-dose methotrexate (MTX) therapy, an experimental comparative study was done. Under general anesthesia, 12 female Large-White pigs, 35 +/- 5 kg body weight, underwent a bilateral renal vessel ligature to create an acute renal failure and avoid the physiologic elimination of MTX. An MTX i.v. infusion was performed using a dose of 12.5 g/m2 and for a specific time according to its pharmacokinetics. The animals were divided into 2 groups of 6 animals each and connected to the extracorporeal circuit; the first group was submitted to a 1-h HP with an anion exchange resin (Dow 1X-2, Dow Chemical) and the second group to 1-h HD with capillary polysulfone fibers (F6, Fresenius). Blood samples were taken at established intervals for hematological and biochemical evaluations. The results demonstrate a higher capability of HP treatment (P < 0.001) related to HD in MTX removal.

Animals↗

Structure of dialysis membranes and long-term clinical outcome.

The present comparative evaluation aims at establishing whether the basic structure of dialysis membrane is able to predict long-term clinical outcome. From a population of 1,256 patients on renal dialysis treatment, treated by the Institute of Nephrology and Dialysis of the St. Orsola University Hospital of Bologna from 1963 to 1993, 122 patients were retrospectively selected for the present study. Patients were divided into two different groups according to the kind of dialysis membrane used--cellulose-based (64 patients) and synthetic-based (58 patients) membranes. The parameters considered were: intradialytic biology, long-term biocompatibility, survival and morbidity, and cost/benefit. The results obtained demonstrate that cellulosic membranes can be said to cause a greater acute intradialytic biological response than synthetics, though not to a significant degree. There are, however, no significant differences in the biological changes from group to group. Nonsignificant differences were noted in long-term survival general morbidity. In terms of sheer cost, synthetic membrane treatment is anything up to 200% dearer than cellulosic.

Adult↗

Fibroblast proliferation over dialysis membrane: an experimental model for "tissue" biocompatibility evaluation.

The present study reports on a biological model based on fibroblast proliferation applied to 3 different types of flat-plate dialysis membrane, in order to ascertain whether the artificial materials currently used in hemodialysis cause in vitro cellular proliferation. The study plan we followed involved plate membrane isolation from non-used dialyzers and used dialyzers, observed through scanning electron microscopy (SEM) both before and after testing with human fibroblasts by means of cell culture. Fibroblast growth was assessed by phase contrast light microscopy examination and cytometric DNA content evaluation. Our investigations proved that the artificial materials we considered interact with fibroblast cultures. Noticeable proliferative response was observed both after contact with unused material and on mediation by the protein layer absorbed on the membrane surface at the end of dialysis sessions. In this last case fibroblast proliferative activity appeared higher than that observed with unused membranes, showing that the soluble molecules entrapped in the protein layer appeared able to exert a biological activity even in vitro tests.

Acrylic Resins↗

Immunologic changes in circulating leukocytes in the presence of atherosclerotic disease.

To assess the numeric and functional changes in circulating white blood cells in the presence of severe atherosclerosis, 25 subjects with marked, angiographically assessed, atherosclerotic lesions and 29 selected controls were studied. Of the differential leukocyte counts, only monocyte count was significantly higher in the atherosclerotic than in the control subjects (449.0 +/- 115.6 (1 S.D.) vs. 344.1 +/- 138.8/mmc; P = 0.0016). By flow-cytometry no significant differences concerning monocyte surface antigens were found, except a feeble decrease in beta 2-microglobulin in the atherosclerotic subjects. As to lymphocytes, an increase in the CD8 population (33.4 +/- 6.8 vs 28.6 +/- 6.5%; P = 0.0144) and decreases in class I HLA antigen (96.6 +/- 7.3 vs 99.4 +/- 0.7%; P = 0.0049), beta 2-microglobulin (97.9 +/- 2.1 vs 99.3 +/- 1.0%; P = 0.0055) and especially in vivo DNA synthesis (3.8 +/- 1.2 vs 5.3 +/- 2.1%; P = 0.0102) percent expressions were found in the atherosclerotic patients with respect to the controls. This study shows that circulating monocytes are increased in atherosclerotic disease, possibly due to their participation in the phagocytosis of lipids in the arterial wall, with no further immunologic involvement. Conversely, the replicative activity of T lymphocytes is decreased, which might be a consequence of or a factor predisposing to atherosclerosis.

Acute Disease↗

Hemoperfusion in chronic uremia.

Hemoperfusion is a blood purification technique involving direct contact between blood and adsorbent substances (sorbents). There are three basic kinds of sorbent: activated charcoal, immunoadsorbents, resins. Following our previous experience on charcoal hemoperfusion, a new coated anionic exchange resin for blood purification specifically designed to remove phosphates was experimentally employed in animals. 3 pigs, in which uremia had been surgically induced, underwent 6 extracorporeal hemoperfusion sessions (2 per pig) with a cartridge containing 100 gr of resin. The phosphate clearance proved satisfactory, values being 120 ml/min after 10' and around 80 ml/min after 2 hours. The biocompatibility of the resin and of the coating membrane was satisfactory. The negligible variation in pH and plasma bicarbonate during all sessions confirmed the low absorption by the tested resin of other blood anions competing with phosphate.

Animals↗

Strategies in renal failure and the impact of lipids.

The main points to note in terms of strategies in renal failure and the impact of lipids are: 1) Timing and typing of dyslipidemia; 2) Occurrence of dyslipidemia in the course of strategies (conservative, dialysis and transplantation); 3) How the strategies can handle the impact of lipids. Analysis of point 1 confirms what a complex profile uremic dyslipidemia presents, involving the type, class, composition and enzyme systems involved in lipid metabolism. In conservative and dialysis, type IV (triglycerides) predominates; in transplantation, type II (cholesterol). Examination of point 2 shows the non obligatory relationship between dyslipidemia and the various strategies of treatment. Lipid abnormalities, type IV or II, occur in 50-60% of patients. Uremic factors for dyslipidemia include: 1) enhanced hepatic stimulation or altered removal in conservative strategies; 2) the same causes plus "specific" promotors in dialysis (dialysis fluid, plasticizer leaching; bioincompatibility, etc.); 3) steroid therapy and other "accessories" in transplantation. A genetic predisposition is very likely present in all patients. Point 3, finally, analyzes the various "supplements" that each strategy requires to cope with the lipid impact. Generic rules (ranging from doing nothing, to diet, drugs, etc.) are of value in all strategies when dyslipidemia occurs. More specific rules include: a) Conservative strategies: appropriate dietetic optimization and modulation (protein-lipid-carbohydrate ratio in terms of calories); b) Dialysis: timing treatment and improving biocompatibility; c) Transplantation: reducing steroids as much as possible.

Humans↗

Value of panel reactive antibodies (PRA) as a guide to the treatment of hyperimmunized patients in renal transplantation.

Patient presensitization represents a considerable problem in candidacy for renal transplantation. While it is well known that hyperimmunized patients--panel reactive antibody (PRA) higher than 60%--create difficulties in donor matching and have a worse outcome than non-hyperimmunized patients, less information is available on patients with an intermediate degree of sensitization (30-60%). In order to evaluate how graft outcome relates to such degrees of sensitization, 241 consecutive transplanted patients were divided into two groups on the basis of their previous year's PRA peak: group A, PRA 0-29%; group B, PRA 30-60%. Group A showed a significantly better survival both in the first year (90% vs 79%, P < 0.05) and in the third year (82% vs 64%, P < 0.01). However, detailed analysis of group B demonstrated that some parameters may significantly influence graft outcome: (1) better compatibility on locus DR; (2) a primary kidney transplant; (3) a dialysis duration of less than 6 months; and (4) the prophylactic use of antilymphocyte globulin (ALG).

Adolescent↗

Flow cytometry evaluation of urinary sediment in renal transplantation.

The value of exfoliative urinary cytology for the diagnosis of different pathological conditions in renal transplantation is widely recognized. The method, however, has not yet gained full acceptance, mainly because identification of the different cells is not always possible by means of standard staining techniques. In view of its characteristics, flow cytometry (FC) seems to represent a consistently reliable, rapid and innovative approach for differentialing the various cells present in the urinary sediment and assessing their number. This study gives the examination result of 223 urinary specimens from 127 transplanted patients selected according to pathology. Sediment cells, collected from fresh urine samples, were washed, treated with a lysing solution, resuspended in saline solution and directly analysed in a FACSCAN cytometer. Morphological evaluation showed: a small number of cells in patients with stable renal function; a larger number of cells, with predominance of lymphocytes, during acute rejection episodes; an absolute predominance of neutrophils during bacterial infection; large-sized cellular debris in cases of post-transplant tubular necrosis; and small cell debris in cases of cyclosporine cytotoxicity. Lymphocyte surface-marker evaluation made it possible to differentiate lymphocyte populations observed during acute rejection episodes (cytotoxic T-cell, CD8 and HLA class II and NK cells) from those detected during bacterial infection (T-cell CD4 positive). These results suggest that urinary FC may be a reliable diagnostic tool in clinical renal transplantation.

Antigens, CD↗

Application of flow cytometry in clinical renal transplantation.

Flow cytometry (FC) may be considered as a fundamental technique in studying cell biology and pathology. It combines the quantitative character of biochemical methods with the multiparametric capacities of microscope analysis in a high-precision process for rapid analysis of individual cell characteristics. Three original FC techniques routinely applied in the field of renal transplantation are reported in the present study. They concern the donor-recipient cross-match test, the morphological analysis of urinary sediment and the modulation of the density of various membrane antigens on the lymphocyte surface. A common factor underlies all these methods: they aim to provide the physician with a reliable diagnostic tool in clinical renal transplantation.

Antigens, Surface↗

Epidemiology of hepatitis C in a population of hemodialysis patients.

A search for antibodies against hepatitis C virus (HCV) was performed in 185 patients on chronic hemodialysis by means of 1st and 2nd generation ELISA tests. Immunoblot assays were performed on positive sera. This study shows a 38% prevalence of HCV-positive patients in our dialysis population according to the 2nd generation ELISA test which shows a higher specificity and sensitivity when compared to the 1st generation one (38 vs. 20%). A correlation was found between the prevalence of HCV-positive patients and how long they had been on dialysis and how many blood transfusions they had received.

Cross Infection↗

Phosphate removal by resin hemoperfusion efficacy and biocompatibility of a new exchange resin.

A new coated anionic exchange resin for blood purification specifically designed to remove phosphates was experimentally employed in animals. 3 pigs, in which uremia had been surgically induced, underwent 6 extracorporeal hemoperfusion sessions (2 per pig) with a cartridge containing 100 gr of resin. The phosphate clearance proved satisfactory, values being 120 ml/min after 10' and around 80 ml/min after 2 hours. The biocompatibility of the resin and of the coating membrane was satisfactory. The negligible variation in pH and plasma bicarbonate during all sessions confirmed the low absorption by the tested resin of other blood anions competing with phosphate.

Animals↗

Hemoperfusion with a new anion exchange resin corrects the metabolic alkalosis in pyloric stenosis: an experimental demonstration.

An experimental model of hypertrophic pyloric stenosis was made by suture of the pyloric wall and gastrostomy in 10 rabbits under general anesthesia. Blood sampling indicated severe alkalosis and hypochloremia 3h 30 min after surgery. To correct the derangement, we tested an ion exchange resin (Dowex SAR), coated with a methacrylic hydrogel. A cartridge containing 18 g of this resin was inserted in an extracorporeal circuit. This chloride charged resin achieved uptake of HCO3- ions, and elution of Cl- ions. The electrolytic balance was fully restored after 10 min of treatment.

Alkalosis↗