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Background Dialysis membranes that release nitric oxide (NO) from their surface, mimicking one of the functions of endothelial cells, may suppress platelet reactivity during hemodialysis treatment. The aim of the present study was to examine whether the addition of a NO donor to the dialysis fluid can suppress platelet reactivity during dialysis. Methods Porcine whole blood was circulated for 4 h through a polysulfone (PS) dialyzer or polymethylmethacrylate (PMMA) dialyzer. After the blood was circulated through the blood circuit and dialyzer, sodium nitroprusside was added to the dialysis fluid as a NO donor. The changes in the platelet reactivity, measured by the platelet aggregation activity by the addition of adenosine diphosphate or collagen in the blood sample, were evaluated during ex vivo dialysis experiments in the presence of a dialysis fluid containing or not containing a NO donor. Results The platelet aggregation activity was significantly decreased at 30 min after the start of the experiment in the case where nitroprusside was added to the dialysis fluid (the NO (+) condition) as compared to the case where no nitroprusside was added to the dialysis fluid (the NO (−) condition), for both the PS and PMMA membranes. The suppression of the platelet reactivity in the NO (+) condition was sustained until the end of the experimental period (240 min). The platelet cyclic guanosine monophosphate level was also significantly increased in the NO (+) condition as compared to the NO (−) condition. Conclusions NO in the dialysis fluid appears to be capable of suppressing the increase of the platelet reactivity observed during dialysis.

Background and Aims Activation of platelets and leukocytes when blood comes in contact with a dialysis membrane surface leads to increased oxidative stress and release of proinflammatory cytokines, which may result in complications in dialysis patients. Within the blood vessels, nitric oxide (NO) generated by nitric oxide synthase on the endothelial cells suppresses both aggregation/activation of platelets and migration/adhesion of leukocytes. We previously showed in a rat hemodialysis model that NO (800 ppm) gas added to the dialysate can suppress blood coagulation on the surface of a dialysis membrane. The aim of the present study was to determine, using a rat hemodialysis model, the appropriate concentration of NO that would most effectively prevent blood coagulation without causing any adverse effects. Method Male Sprague Dawley rats weighing 300 to 400 g were used for the experiments. After the rats were anesthetized (isoflurane, 1.5%-3.0%), extracorporeal circulation was established by removal of blood from the carotid artery and returning it to the tail vein. During the experiment, the blood flow rate was kept at 1.0 mL/min, and the dialysate flow rate at 3 mL/min. Heparin was administered by bolus injection (0.7 unit/g-mouse) at the start of the hemodialysis, and no further heparin was added during the 4-hour hemodialysis session. A cellulose triacetate membrane and polymethylpentene membrane were used for making the small dialyzer and gas exchanger, respectively (the prototype module was manufactured by NIPRO, Co., Japan). NO (200 ppm, 400 ppm, or 800 ppm) was added to the dialysate via the gas exchanger in the NO groups, and N2 gas was added to the dialysate in the control group. During the experiment, the arterial pressure, dialyzer inlet pressure, venous pressure, activated whole blood clotting time (ACT) and methemoglobin (met-Hb) concentration were measured. After circulation, the amount of coagulated blood was measured by the hemoglobin (Hb) content and lactate dehydrogenase (LDH) activity in the eluate from the residual blood clotting in the dialyzer. Results The 4-hour hemodialysis session achievement rate was highest in the NO-800 ppm group (control, 54%; NO-200 ppm, 66%; NO-400 ppm, 77%; NO-800 ppm, 85%). The blood met-Hb content was below 10% in all the NO groups. The Hb content and LDH activity in the eluate were significantly lower in the NO groups than in the control group. These results indicate that addition of higher concentrations of NO was associated with a lower likelihood of coagulation on the dialyzer and in the dialysis circuit and a higher likelihood of smooth and successful 4-hour dialysis sessions. Addition of NO was associated with little risk of adverse effects such as met-Hb accumulation and reduction of blood pressure. Therefore, 800 ppm is considered as the most appropriate concentration of NO to be added to the dialysate for effectively inhibiting blood coagulation without causing any adverse effects. Conclusion Our experiment in a rat hemodialysis model showed that 800 ppm was the most appropriate concentration of NO to be added to the dialysate to suppress blood coagulation on the surface of the dialysis membrane without causing any adverse effects.

Background and Aims Since hemofilters used for continuous renal replacement therapy (CRRT) come in prolonged contact with blood during treatment, clotting and cell adhesion induced by contact of blood with the membrane often occur, increasing the risk to the patient and burden on the staff. Suppressed platelet adhesion and better biocompatibility have been reported with the use of a dialysis membrane whose surface has been modified with an NV polymer in patients on maintenance hemodialysis. Therefore, it may be advantageous to use the NV membrane as a hemofilter for CRRT. In the present study, we evaluated the solute removal performance change and blood cell adhesion on the NV membrane after it comes in contact with blood, to clarify the characteristics of the NV membrane for long-time use. Methods A new polysulfone membrane containing the NV polymer (SNV) and a conventional polysulfone membrane (SHG) were used for the ex vivo blood filtration experiment. To compare the solute removal performance change and blood cell adhesion after blood comes in contact with the membranes, porcine blood collected from a single animal was divided into two portions, and a 24-hour of continuous hemofiltration (CHF) experiment was performed. The circulation conditions were as follows; blood flow rate, 100 mL/min; filtrate flow rate, 20 mL/min; heparin dose adjusted as appropriate to maintain the activated clotting time from 300 to 400 sec during the experiment. We evaluated the time-courses of the transmembrane pressure (TMP) and pressure drop at the hemofilter and the change in the sieving coefficients of dextran of different molecular weights (molecular weight cutoff curve) before and after the experiment, and blood cell adhesion. The amount of blood cell adhesion was evaluated based on the hemoglobin content and lactate dehydrogenase (LDH) activity in the eluate from the residual blood clots on the hemofilter 24 hours after the CHF experiment. Results The changes in the TMP and the pressure drop were significantly lower with the use of SNV as compared to SHG (n = 9, p < 0.01). From the molecular weight cutoff curves before and after the experiment, the molecular weights when the sieving coefficient was 0.1 to 0.4, which reflect the pore size of the pores through which relatively small molecules can pass, decreased to a lower extent after blood contact with SNV as compared to SHG (n = 3, SC = 0.1-0.3, p < 0.05; SC = 0.4, p < 0.01).The hemoglobin content and LDH activity in the blood clots adhering to the membrane after the experiment were significantly lower in SNV as compared to SHG (n = 9; Hb, p < 0.01; LDH, p < 0.05), indicating that blood cell adhesion on SNV was less pronounced than that on SHG. Conclusion SNV suppressed the increase in the TMP and also the pressure drop, allowed a high rate of solute removal performance to be maintained, and suppresses blood cell adhesion to a greater extent as compared to the conventional polysulfone membrane.

Aims: The aims of the present study were to evaluate the changes in the enterobacterial flora and evaluate the hydrogen gas-producing ability of the enterobacterial flora in a mouse model of severe acute lung injury (ALI mouse model). Methods: The ALI mouse model was prepared by administering lipopolysaccharide (LPS) after activation of natural killer cells with α-galactosylceramide. The exhaled hydrogen concentrations of the mice were measured. The oxidative stress of the lung was evaluated by measuring the myeloperoxidase (MPO) activity. At the same time, feces were collected from the cecum of the mice and the hydrogen gas-producing ability was measured. Metagenomic analysis was also carried out to evaluate the changes in the enterobacterial flora. Results: The MPO activity in the lung of the ALI mouse model was significantly increased as compared to that of the control (n=25; P Conclusion: The ALI mouse model showed an increased hydrogen gas-producing ability of their enterobacterial flora, along with a change in the composition of the enterobacterial flora, which may protect organs against severe inflammation.

The purpose of the present study was to evaluate the adhesiveness of blood cells and the solute removal performance change of modified polysulfone membranes which have increased polyvinylpyrrolidone (PVP) coverage over their surface. Continuous hemofiltration (CHF) experiments for 24 h were carried out using an ex vivo hemofilter evaluation system to compare a modified polysulfone hemofilter (SHG) with the conventional polysulfone hemofilter (SH). The 25 and 50 % cutoff values of the sieving coefficient of dextran after CHF and the protein concentration in the filtrate was higher in SHG, indicating that less fouling occurred in the SHG membrane. Adhesion of blood cells after 24 h of CHF was significantly higher in the case of SH than in the case of SHG. Blood cell adhesion and membrane fouling were reduced with the use of a polysulfone membrane modified with increased PVP coverage over the surface.

The purpose of the present study was to establish a continuous hemofiltration model using porcine blood to compare filter life. Continuous hemofiltration (CHF) experiments were performed using an in vitro hemofilter evaluation system utilizing porcine blood containing trisodium citrate in addition to nafamostat mesilate as anticoagulants. The lifetime of the hemofilter was evaluated using the transmembrane pressure and the pressure drop across the hemofilter at varying trisodium citrate concentrations. The porcine blood used in this experiment was considered to be in a slightly hypercoagulable state because of the continuous contact with non-biological materials and calcium inflow from substitution fluid. Blood containing 7 or 8 mM of trisodium citrate and nafamostat mesilate could be effectively used to compare the lifetimes of hemofilters utilized under the same conditions. In this CHF model using porcine blood, the plugging of the hollow fibers occurred shortly after the plugging of the membrane pores. In conclusion, a CHF model using porcine blood can be established by adjusting the concentration of trisodium citrate added to the blood.