Choosing new adsorbents for endogenous ultrapure infusion fluid: performances, safety and flow distribution.

Adsorption may notably contribute to the removal of uremic toxins and to the efficiency of hemodialysis. We examined different uncoated stationary matrixes, charcoals and synthetic resins to establish their adsorptive capacities in relation to low (urea, creatinine) and high molecular weight (beta2-microglobulin, myoglobin) compounds in in vitro conditions (steady state and flow-through) using isotonic solutions or uremic ultrafiltrate. Trace metal, particle release analyses and scanning electron microscopy of different adsorbents were performed. Dynamic flow-distribution studies were made using 99Technetium and analysing the different regions of interest by single head gamma-camera. We show that adsorbents may differ greatly as to their adsorptive capacity depending on flow rate, nature, and total mass of the compounds to be removed from the ultrafiltrate. These studies suggest a methodological approach for screening stationary matrixes for possible application in hemodialysis.

Hemodiafiltration with online regeneration of the ultrafiltrate.

The concept of regeneration of dialysis fluids and of ultrafiltrate in particular has been recently revisited. Hemodiafiltration with online regeneration of the ultrafiltrate allows the concomitant infusion of sodium, calcium, and bicarbonate. Here, we studied the adsorptive characteristics of an integrated two-step sorbent system relative to different solutes present in the ultrafiltrate: sodium, calcium, phosphate, bicarbonate, uric acid, creatinine, and beta2-microglobulin. In vitro studies were performed in order to differentiate the relative roles for each sorbent (mineral-activated charcoal or hydrophobic resin) in adsorbing a given solute. Ex vivo studies were performed in order to evaluate the presence of cytokines (interleukin-1 beta and tumor necrosis factor-alpha), of cytokine (interleukin-1 beta and tumor necrosis factor-alpha)-inducing activities, and of the cytokine release in response to exogenous bacterial lipopolysaccharide by normal whole blood incubated with ultrafiltrate samples obtained at 15, 120, and 240 minutes after the start of treatment. The results of the present studies show the presence of immunomodulatory substances in the ultrafiltrate and the significant (P < 0.01) increase in the lipopolysaccharide-induced release of both interleukin-1 beta and tumor necrosis factor-alpha. The biological relevance of the ultrafiltrate and the possible relevance of the online, endogenous reinfusion are discussed.

Retention of limulus amoebocyte lysate reactive bacterial products by polysulfone dialyzers is affected by the type of disinfectant.

To reduce the level of contamination by bacterial products, ultrafiltration systems have been introduced and validated for their capacity to block the passage of bacterial components reactive to the limulus amoebocyte lysate (LAL) test. In this study, the absorptive capacity of polysulfone membranes undergoing disinfection cycles with free chlorine and peracetic acid were evaluated at various concentrations and contact times. The results of this study implicate a relevant physicochemical derangement of the polysulfone membranes treated with sodium hypochlorite but not with peracetic acid, diluted peracetic acid (Dialox) or Amuchina. The implications for the practical use of ultrafilters are discussed.

In vitro and in vivo biocompatibility of substituted cellulose and synthetic membranes.

Regenerated cellulosic membranes are held as bioincompatible due to their high complement - and leukopenia - inducing properties. Adherence of polymorphonuclear neutrophils and monocyte purified from normal human blood to the three membranes were evaluated in an in vitro recirculation circuit in the presence or absence of fresh, autologous plasma after recirculation in an in vitro circuit using minimodules with each of the three membranes. In in vivo studies, 9 patients were treated with conventional haemodialysis for 2 weeks with each membrane and 1 week for wash-out using haemodialysers with the following surface: 1.95 m2 for benzyl-cellulose, 1.8 m2 for acetate-cellulose and low-flux polysulfone. Measurement of leukopenia, plasma C3a des Arg and elastase-alpha1 proteinase inhibitor complex levels as well as urea, creatinine, phosphate and uric acid clearances was performed. Plasma-free neutrophils adhered maximally to acetate-cellulose (65% remaining in the circulation), while there was no significant difference between low-flux polysulfone and benzyl-cellulose (80% circulating neutrophils, at 15 min, p<0.001 vs acetate cellulose). In the presence of fresh plasma, as source of complement, the differences between acetate cellulose vs polysulfone and benzyl-cellulose were even more evident, suggesting the role of complement-activated products in neutrophil adherence. A similar trend was observed for monocyte adherence with the three membranes in the absence or presence of plasma. In vivo studies showed that the nadir of leukopenia was at 15 and 30 min with acetate-cellulose (79%) and benzyl-cellulose (50%) (p<0.05 acetate- vs benzyl-cellulose) and at 15 min with polysulfone (24%) (p<0.01 vs acetate- and benzyl-cellulose). Plasma C3a des Arg levels arose to 2037 +/- 120 ng/ml, 1216 + 434 ng/ml and 46 +/- 55 ng/ml with acetate-, benzyl-cellulose and polysulfone, respectively. No pre- vs post-dialysis increase in the intracellular content of TNF-alpha was detected with any of three membranes. Clearance values of urea, creatinine and uric acid were superimposable for all the three membranes. However, benzyl cellulose had a significantly higher clearance for phosphorus (normalized for surface area) (p<0.01 vs acetate-cellulose, 0.001 vs polysulfone). These results implicate that synthetic modification of the cellulose polymer as for the benzyl-cellulose significantly reduces the in vitro adherence, delays the in vivo activation of "classic" biocompatibility parameters and notably improves the removal of inorganic phosphorus.

[Struma cordis: first case in Mexico]. / Struma cordis: primer caso en México.

The Intracardiac ectopic thyroid is an extremely rare condition and there is no previous report on this subject in Mexico. This is the case of a 33 years old woman, with normal thyroid function. She was found to have an intracardiac tumor in the interventricular septum. The intraoperative biopsy showed typical thyroid follicules; tumor removal left a septal defect that was closed with a dacron patch suture. Two years follow-up showed normal echocardiographic images, good clinical status an normal thyroid functioning. A brief review of the literature is included.

Mechanisms and kinetics of the synthesis and release of platelet-activating factor (PAF) by polyacrylonitrile membranes.

Platelet-activating factor is a recognized mediator of anaphylaxis and bioincompatibility. Here, the mechanisms and the kinetics of the production of platelet-activating factor were studied in vivo during high-flux hemodialysis and in vitro in a recirculation model with polyacrylonitrile membranes, the AN-69 and the more recent SPAN, where the Na-metallilsulfonate group is partially substituted with the less polar methacrylate group. In in vivo studies, eleven patients were studied in cross over. Patients were randomly allocated to the AN-69 (5 patients) and to the SPAN membrane (6 patients) for two weeks. Measurements were made in the second week of use. After completion of the second week, the patients were switched to the other membrane for a further two weeks. Samples for leukocyte and platelet counts, PAF in whole blood or bound to platelets, the C3a des Arg and the C5b-C9 membrane attack complex as well as samples for clearances of urea, creatinine and phosphates were taken at different time intervals during treatment. PAF was detected by biological assay after methanol extraction of whole blood or of platelet pellets obtained by sequential centrifugation. C3a des Arg and the C5b-C9 fraction were detected by commercially available immunoassays. Results were analyzed by Minitab statistical package. PAF was detectable only during treatment with AN-69 but not with SPAN 1 min after start of the extracorporeal circulation in both whole blood (4.5 +/- 2.7 ng/ml) and on platelet surface (4.1 +/- 1.2 ng/ml). No statistical significant differences were observed between AN-69 and SPAN with regard to leukocyte and platelet counts, plasma C3a des Arg and C5b-C9 levels. The structure modification did not alter functional performances as indicated by the lack of statistically significant differences in clearance values between the two membranes. In in vitro experiments performed with normal washed and whole blood recirculated in a closed circuit demonstrated the presence of a plasma-dependent, complement-independent mechanisms responsible for the triggering of PAF synthesis and release with AN-69 but not SPAN membrane. PAF was extractable from the inner and outer side of both polyacrylonitrile membranes (AN-69: inner, 4.9 +/- 0.5 ng/ml; outer, 0.1 +/- 0.05 ng/ml; SPAN: inner, 5.5 +/- 0.6 ng/ml, outer: 3.3 +/- 0.7 ng/ml, SPAN vs. p < 0.001), suggesting that absorption may be relevant with both membranes.

Production of platelet activating factor by human neutrophils after backfiltration of endotoxin contaminated dialysate.

Lipopolysaccharide (LPS) from gram negative bacteria has been shown to prime human polymorphonuclear neutrophil (PMN) production of platelet activating factor (PAF). PAF is a lipid mediator of inflammation and endotoxic shock and is also involved in leukocyte activation occurring during hemodialysis; PAF induces leukopenia, degranulation of lysosomal granules, and adherence to hemodialysis membranes. Transmembrane passage of LPS has also been shown to occur. To evaluate the relevance of transmembrane passage of LPS on the priming of PMN derived production of PAF, we designed in vitro studies using an experimental circuit equipped with different membranes (Cuprophan, polysulfone, polymethylmethacrylate, polyamide) and recirculation of purified human PMNs. At different time intervals, PMNs were stimulated with FMLP (10 microM) after back-filtration of sterile and LPS contaminated dialysate. The results of these studies suggest that priming of PMN derived production of PAF was related to the percent of backfiltered LPS, and they emphasize the need for careful assessment of microbiologic quality to improve biocompatibility.

Hemodiafiltration without replacement fluid. An experimental study.

Paired filtration dialysis (PFD) is the only hemodiafiltration (HDF) technique in which the ultrafiltrate is continuously available but not mixed with the dialysate. As is the case during all convective or predominantly convective techniques, use of a replacement fluid is necessary in an amount equal to the difference between the ultrafiltrate and the desired patient weight loss. This replacement fluid must have an adequate electrolytic composition (Na+, Ca++, and buffer), and must be sterile and pyrogen free. Using an uncoated adsorbent charcoal cartridge (130 g), the ultrafiltrate obtained in PFD was regenerated, eliminating both the small (except for urea, glucose, and phosphates) and medium-to-large solutes but not the electrolytes and bicarbonate. This verified the ultrafiltrate's possible use as replacement fluid. This technique experimentally studied during 24 standard PFD sessions, with a total mean ultrafiltrate of 9,950 +/- 860 ml, allowed a replacement solution to be obtained with the following mean +/- SD composition: pH 7.467 +/- 0.122, HCO3- 27.0 +/- 2.12 mmol/L, Na+ 137.4 +/- 2.6 mmol/L, K+ 4.1 +/- 0.83 mmol/L, Ca++ 1.12 +/- 0.19 mmol/L, urea 68.3 +/- 16.2 mg/dl, creatinine 0.08 +/- 0.02 mg/dl, uric acid 0.05 mg/dl, phosphates 2.77 +/- 0.71 mg/dl, beta-2 microglobulin 0.5 +/- 0.4 mg/L, and atrial natriuretic peptide 4.41 +/- 5.6 pg/ml.(ABSTRACT TRUNCATED AT 250 WORDS)

Continuous pH and HCO3 monitoring during hemodiafiltration without blood sampling.

The correction of acid-base balance during hemodialysis, especially in high-efficiency techniques, could present some problem related to the lack of an adequate monitoring of pH and blood gases. During hemodiafiltration (HDF), performed with the two-chamber technique (paired filtration dialysis, PFD), the ultrafiltrate (Uf) is continuously available, unmixed with the dialysate. Connecting a pH electrode (as Ag/C1Ag) to the Uf circuit, the authors made 40 determinations on 16 different PFD patients, and they correlated the Uf values obtained with those measured on arterial blood with standard methods. The one sample analysis gave a t = 10.145 (p = 0.0), and the linear regression analysis an r = 0.931 (p = 0.0). At 30 min, in 8 PFD patients, the HCO3 values obtained from Uf, pH and transcutaneous PCO2, gave a t = 6.37 (p = 0.0004), and an r = 0.939 (p = 0.00052). In conclusion, during HDF performed with PFD, continuous pH monitoring of the patient is possible without blood sampling. Moreover, correlation with the transcutaneous PCO2 measurement could provide HCO3 values in real time.

Use of the ultrafiltrate obtained in two-chamber (PFD) hemodiafiltration as replacement fluid. Experimental ex vivo and in vitro study.

PFD (Paired Filtration Dialysis) is the only hemodiafiltration (HDF) technique in which the ultrafiltrate (UF) is continuously available not mixed with the dialysate. As with all convective or prevailingly convective techniques, a replacement fluid is necessary in an amount equal to the difference between the UF and the desired weight loss. This replacement fluid (R) must have an adequate electrolytic balance (Na+, Ca++, and buffer), and must be sterile and pyrogen-free. Using an uncoated adsorbent charcoal cartridge, we "regenerated" the UF obtained in PFD, eliminating the small (except for urea, which was later eliminated by diffusion in the dialyzing section of the PFD system) and the medium-to-large molecules (vit B12 and myoglobin in vitro and beta-2-microglobulin (B2m) and (hANP) in vivo), but not the electrolytes and the endogenous bicarbonate, so as to verify its possible use as R. This technique, experimentally performed in 12 patients under HDF treatment with standard PFD, with a total mean UF of 9650 +/- 875 ml and the use of 130 g of uncoated charcoal, produced a solution with the following composition: Na+ 135.4 +/- 2.4 mmol/l, K+ 3.4 +/- 1.23 mmol/l, Ca++ 1.18 +/- 0.14 mmol/l, HCO3- 26.7 +/- 2.3 mmol/l, phosphates 2.88 +/- 0.81 mg/dl, urea 63 +/- 14 mg/dl, creatinine 0.08 +/- 0.02 mg/dl, uric acid 0.05 +/- 0.0 mg/dl, beta-2 microglobulin 0.5 +/- 0.5 mg/l, and hANP 4.15 +/- 5 pg/l.(ABSTRACT TRUNCATED AT 250 WORDS)

In vivo solute elimination of paired filtration dialysis.

Paired filtration dialysis (PFD) is a new dialysis strategy whereby a hemofilter and a hemodialyzer are coupled in series. It has been assumed that this approach allows a better solute elimination than conventional hemodialysis (HD), allowing a shortening of dialysis time. To evaluate this hypothesis, solute elimination with PFD either with 0.4 m2 polysulphone (PS) and 1.36 m2 cuprophan (CU) 3x3 h/wk, or with 0.4 m2 PS and 1.06 m2 CU 3x4 h/wk, was compared to HD with 1.36 m2 CU, 3x4 hours weekly in the same patients. During PFD, 10L were ultrafiltered and substituted by saline. Overall extraction of UV absorbing solutes (MW less than 10,000 Dalton), and extraction of individual solutes identified by high performance liquid chromatography (HPLC) were compared as well as urea kinetics. For PFD 3x3 h overall extraction of UV-absorbing compounds and of hippuric acid was significantly higher than for HD 3x4 h (p less than 0.05). Overall extraction of UV-absorbing compounds and of all but one individual compound under study was markedly higher for PFD 3x4 h vs conventional HD 3x4 h (p less than 0.01), in spite of a higher diffusive area with the latter technique. No differences in urea kinetics were observed for the 3 strategies. It is concluded that solute extraction during PFD is higher than during HD, if the treatment time is the same. Even if treatment is shortened to 3x3 h weekly, solute extraction with PFD is at least as efficient as with HD.

[Hemodiafiltration: choice or necessity in high-efficiency treatment?]. / Emodiafiltrazione: scelta o obbligo nel trattamento ad alta efficienza?

The risks of back-filtration that occur with the use of high hydraulic permeability membranes with haemodialytic techniques in the course of which the difference between forced and necessary ultrafiltration is compensated for by correcting transmembrane pressure in favour of the dialyser compartment. In this way a form of concealed haemodiafiltration is attained in which the replacement fluid is the dialysing solution, annulling, owing to the possible consequences of the transit of bacterial endotoxins into the circulation, all the advantages linked to the use of these membranes. It is concluded by suggesting the implementation of well controlled haemodiafiltration through the careful quali-quantitative evaluation of the replacement fluid.