Effect of hollow fiber length on solute removal and quantification of internal filtration rate by Doppler ultrasound.

Renal replacement therapy with dialyzers capable of enhanced internal filtration (IF) can be an alternative to standard hemodiafiltration, as it provides convective solute removal comparable to that of hemodiafiltration by a simple procedure. In this study, we clinically evaluated the effect of the hollow fiber length in the dialyzer, a crucial factor influencing the rate of IF, by comparing two commercial dialyzers (BS-1.6U, BS-1.6UL, Toray, Japan) which differed in the fiber length, but had the same surface area and inner diameter of their hollow fibers. We showed that in the dialyzer with the longer fibers, the pressure profile along the dialyzer was significantly altered, and the solute clearance tended to be increased. In addition, we successfully quantified the IF rate with a Doppler ultrasound in the experimental circuit, by measuring the blood flow velocities along the bundle of fibers. We showed that the changes in the blood flow velocity were more marked in the dialyzer with the longer fibers; the calculated IF rates in the dialyzers with the shorter and longer fibers were 11.1 mL/min and 37.7 mL/min, respectively, which seemed to be compatible with the solute clearances. This simple and readily applicable method is expected to be useful in the development of modified dialyzers to fully exploit the benefits of IF in renal replacement therapy.

Hemodialysis paradox: survey on the incidence rate of hepatocellular carcinoma in antihepatitis virus C-antibody-positive chronic hemodialysis patients.

The prevalence of antihepatitis virus C (HCV) antibodies in chronic hemodialysis (HD) patients is higher than in normal populations, and yet hepatocellular carcinoma (HCC) is infrequent in chronic HD patients who are HCV antibody positive. In this study, we investigated the characteristics of HCV-antibody-positive patients with HCC on chronic HD. A total of 6,366 cases of HCV-antibody-positive patients on chronic HD therapy was analyzed on the basis of answers to questionnaires on the incidence rate of HCC in 314 Japanese dialysis institutions. HCC was a complication in 114 of 6,222 (1.8%) HCV-antibody-positive patients, and cirrhosis was a complication in 536 of 6,242 (8.6%). The incidence rate of both complications was significantly higher in males than in females, and the incidence rate in the chronic HD patients was much lower than in normal populations. Specific immunological status in patients on chronic HD therapy may be an important key for preventing the progression of chronic HCV hepatitis. However, further research is needed because this study was preliminary and excluded the type of HCV virus, pathological findings, and laboratory data.

Effects of operating conditions on selectivity of a plasma fractionator in double filtration plasmapheresis.

In a typical double filtration plasmapheresis treatment, plasma fractionation between albumin and some immunoglobulins associated with toxins is limited because none of the currently available plasma fractionators has a strict cutoff property for these proteins. Selectivity of immunoglobulins over albumin depends not only on the cutoff properties of the membrane but on the operating conditions such as the flow rate of the supplied plasma (Q(P)) and retained plasma to be discarded (Q(D)) in the plasma fractionator. We carried out an in vitro study using human plasma harvested by single plasma exchange treatments to assess the selectivity of a plasma fractionator, Evaflux 2A-F (Kawasumi Laboratories, Inc., Tokyo, Japan), under various operating conditions. The results of rate-constant filtration experiments showed that the concentrations in the feed tank and the sieving coefficient (SC) values of every protein were decreased slightly within 2 h after the start of the experiment because of membrane trapping, adsorption, and/or plugging. The time-averaged SC value of albumin increased with flow rate ratio (Q(P)/Q(D)) due to increasing filtration fraction (FF), but relative removal efficiency (mD/mP*) for albumin decreased with Q(P)/ Q(D) due to decreasing Q(D). For immunoglobulins, on the other hand, the SC values were almost unchanged, and the mD/mP* values increased with Q(P)/Q(D) due to an increase in FF. Both increasing Q(P) and decreasing Q(D) are effective means of improving selectivity between these proteins in the plasma fractionator. Membrane fouling is, however, obvious beyond a Q(P)/Q(D) value that is thought to be a critical point. Operation should be conducted below the critical Q(P)/Q(D) value, which depends on the patient's plasma components and the cutoff property of the membrane.

Pharmacology of SCH00013: a novel Ca2+ sensitizer.

Cardiotonic agents that facilitate cardiac pump function by direct improvement of contractile dysfunction are indispensable for the treatment of hemodynamic disorders in acute myocardial failure and the aggravating phase of congestive heart failure. Cardiotonic agents currently available for the treatment of hemodynamic crisis in congestive heart failure are catecholamines, selective phosphodiesterase (PDE) III inhibitors and digitalis, all of which are Ca2+ mobilizers. Considering the number of serious adverse effects of these clinically available cardiotonic agents, development of agents that act via a novel mechanism of action may contribute to the progress of pharmacotherapy of congestive heart failure. Ca2+ sensitizers that act by increasing in myofilament Ca2+ sensitivity may be able to overcome the disadvantage of Ca2+ mobilizers. Ca2+ sensitizers do not increase activation energy, do not produce Ca2+ overload and may be effective even under pathophysiological states such as acidosis, myocardial stunning and heart failure. SCH00013 ((4,5-dihydro-6-[1-[2-hydroxy-2-(4-cyanophenyl)ethyl]-1,2,5,6-tetrahydropyrido-4-yl]pyridazin-3(2H)-one)) is a novel Ca2+ sensitizer that elicits a moderate positive inotropic effect without significant alteration of Ca2+ transients. SCH00013 does not have a positive chronotropic effect and has a weak PDE III inhibitory action and class III antiarrhythmic action. SCH00013 prolonged the survival in a animal heart failure model with genetic cardiomyopathy. The oral bioavailability of SCH00013 is high and equivalent to that via intravenous administration. The unique pharmacological profiles of SCH00013 imply that this agent may be potentially beneficial for pharmacotherapy of contractile dysfunction in congestive heart failure.

Effects of internal filtration on the solute removal efficiency of a dialyzer.

To improve solute removal efficiency, several types of dialyzers with enhanced internal filtration were introduced for clinical application. In these dialyzers, enhanced internal filtration increased convective transport of the solute, in addition to diffusive transport. In this study, the effects of internal filtration on solute removal efficiency were examined by both analytic and experimental studies. Internal filtration is affected by blood (Q(B)) and dialysate (Q(D)) flow rates; the patient's hematocrit and plasma level of total protein; and the effective length (L(eff)), inner diameter (D), and density ratio (DR) of the hollow fibers. An analytic model was introduced for the estimation of the changes in mass and momentum along the dialyzer. It clarified the effects of these parameters on maximum internal filtration flow rate (Q(IF)) and clearance (K) of urea (60 daltons), vitamin B(12) (1,355), and myoglobin (17,000). As a result of the analytic study, Q(IF) was increased, resulting in a smaller D, a longer L(eff), and a larger DR value. Several types of dialyzers with the same cellulose triacetate membrane, produced by Toyobo Co, Ltd., Ohtsu, Japan, and Nissho Corporation, Kusatsu, Japan, were used for the experimental study. An in vitro evaluation using myoglobin solution showed the same trends as found in the analytic study. For example, a dialyzer with 150 microm of D has a 72.0 ml/min myoglobin K value, much higher than that of 53.7 ml/min for a dialyzer with 200 microm of D under constant Q(B) (300 ml/min) and DR (50%) values. Development of a dialyzer with enhanced internal filtration, however, should take the patient's safety into account, and hemolysis and endotoxin invasion from the dialysate to the patient should be avoided.

Can cytokines be removed by hemofiltration or hemoadsorption?

To study the removability of pro-inflammatory cytokines by hemofiltration (HF), we performed experimental HF with various high-flux membranes (HFM) using a closed circuit system filled with monocyte-free human plasma, which contained TNFalpha, IL-1beta, and IL-6. Plasma and filtrate samples were taken before and 1, 2, 3, and 4 hours after the initiation of HF, and each cytokine was determined by enzyme-linked immunosorbent assay. IL-1beta was well removed through filtration during experimental HF using HFM (PAN>CTA>PMMA>PS). TNFalpha and IL-6 were only minimally filtered out by HF using HFM. TNFalpha was removed to some extent by using PS, and IL-6 was partially removed by using PMMA during experimental HF through other mechanisms, such as adsorption, than the filtration. IL-1beta and IL-6 were effectively removed by HA using charcoal adsorbent column, especially during the first 2 hours, while TNFalpha was only partly removed.

Solute removal characteristics of continuous recirculating peritoneal dialysis in experimental and clinical studies.

Continuous recirculating peritoneal dialysis (CRPD) was introduced to enhance solute removal efficiency in conventional peritoneal dialysis (PD) therapies such as continuous ambulatory peritoneal dialysis (CAPD). In CRPD, a portion of the dwell dialysate in the patient's peritoneal cavity is drained through a double-lumen catheter and purified by an extracorporeal dialyzer. In this study, solute removal characteristics and safety of CRPD are examined in ex vivo and clinical studies. Recirculation dialysis experiments using nine dogs (13.6 +/- 2.5 kg of body weight) were carried out for 240 min in the ex vivo study, whereas another seven dogs (12.1 +/- 2.8 kg) received conventional peritoneal dialysis (CPD) (120 min dwelling x 2) and six additional dogs (11.9 +/- 2.7 kg) received a Tidal PD (20 min dwelling x 12; 50% of tidal volume ratio) as controls. The ex vivo study revealed that CRPD has a higher efficiency for solute removal than CPD and is equivalent to Tidal PD. In the BUN reduction rate, the 19.4 +/- 5.5% in 240 min CRPD (n = 9) was significantly higher (p < 0.05) than the 3.5 +/- 3.6% in 240 min CPD (n = 7) and equivalent to the 17.3 +/- 4.7% in 240 min Tidal PD (n = 6). Continuous recirculating peritoneal dialysis maintained a low UN level in the peritoneal cavity due to dialysis with an extracorporeal dialyzer. This tendency was also seen in creatinine removal. In the clinical study, CRPD (n = 10) and CPD (n = 5) treatments were used in three renal failure patients. Higher solute removal efficiency was shown in CRPD than in CPD treatments, and the urea peritoneal clearance was 14.1 +/- 4.4 ml/min in CRPD (n = 10), significantly higher (p < 0.05) than the 7.3 +/- 2.1 ml/min in CPD (n = 5). No fibrin formation occurred during CRPD treatments.

Continuous monitoring of blood volume in double filtration plasmapheresis.

A continuous hematocrit (HCT) monitor, Crit-Line, was introduced to examine the change in patients' blood volume (BV) due to albumin loss during double filtration plasmapheresis (DFPP) treatments. Nine patients with autoimmune diseases or ABO incompatible renal transplantation received 15 DFPP treatments under Crit-Line monitoring. In these patients, plasma albumin concentration (C(P)) changed from 3.7 +/- 0.6 g/dl to 3.5 +/- 0.5 g/dl and HCT from 28.7% +/- 3.3% to 31.3% +/- 4.3% (change ratio [CR] of BV = -8.1%) during treatment with albumin concentrations (C(S)) of 9.5 +/- 1.0 g/dl and 500 ml volumes (V(S)) of supplementation fluid. Although the apparent CR value of C(P) was -5.3%, on average, the CR of albumin in the patients' plasma (M(P)) was -16.1%, which means a corrected CR value of C(P) by the HCT value to eliminate the influence of the patient's blood volume contraction during treatment. Albumin loss usually occurred in DFPP treatments. The decrease in BV was induced by an oncotic pressure drop due to albumin loss, and often resulted in a blood pressure drop. The amount of albumin loss during DFPP treatments strongly depends on sieving coefficients of the plasma separator (SC(PS)) and the plasma fractionator (SC(PF)), the filtration fraction of the plasma fractionator (FF(PF)), pretreatment C(P) value, and C(S) and V(S) values of the supplementation fluid. To determine the optimum C(S) and V(S) values for each patient, the authors introduced a variable blood volume model for albumin transport in DFPP. In this model, changes in C(P), HCT, and BV values could be estimated during treatment. For example, a patient with an HCT of 31.2%, body weight of 61.1 kg, and pretreatment C(P) of 4.4 g/dl received a DFPP treatment using a plasma separator, OP-05 (SC(PS) of 0.99), and a plasma fractionator, Evaflux 2A (SC(PF) of 0.40), under FF(PF) of 0.8 with a V(S) of 500 ml. A value for C(S) of about 10 g/dl is required for the patient to maintain a normal C(P) level during treatment by an estimation from the model. As a result of the treatment with a C(S) of 10 g/dl, the patient had no adverse reactions, such as a blood pressure decrease, during treatment under these conditions.

Optimum albumin concentration of supplementation fluid for double filtration plasmapheresis.

Until recently, the albumin concentration of supplementation fluid for double filtration plasmapheresis (DFPP) has been empirically determined. Inadequate albumin infusion often leads to hypoproteinemic symptoms such as edema. In the current study, an aimed condensation coefficient (CCaimed) was introduced in an attempt to estimate the appropriate plasma albumin level for each patient. This coefficient is theoretically derived from a one-compartment model for the patient's plasma albumin: CCaimed = CS/CD = 1 - (1 - CR)/[1 - exp(- CC.VR)] where CD and CS are albumin concentrations in discarded plasma and supplementation fluid. CR is the change ratio of albumin concentration in the patient's plasma during a DFPP treatment, and VR(= VS/VP) is the ratio of supplementation fluid volume (VS) to the patient's total plasma volume (VP). And CC denotes the albumin condensation coefficient in a DFPP line, which depends on the filtration fraction of the plasma fractionator (FFPF) and the sieving coefficients of both the plasma separator (SCPS) and the plasma fractionator (SCPF): CC = CD/CP = SCPS.(1 - FFPF.SCPF)/(1 - FFPF) where CP is the albumin concentration of the patient's plasma. From the above relations, CS can be determined as follows: CS = CC.CCaimed.CP Because many kinds of proteins are removed during a single DFPP treatment, a slightly higher albumin concentration in the supplementation fluid is needed to maintain an appropriate plasma level. Therefore, the CR value should be more than unity. For a patient with hematocrit (HCT) of 30%, body weight (BW) of 50 kg, and CP of 3.0 g/dl, who is receiving a DFPP treatment using AP-05H (SCPS of 0.970) and Evaflux 2A (SCPF of 0.526) under FFPF of 0.8 with VS of 500 ml, VP = BW(1- HCT/100)/13 = 50 x (1 - 30/100)/13 = 2.69 L, VR = 500/(2.69 x 1,000) = 0.186, CC = 2.81, and CCaimed = 1.25 assuming 1.1 for CR. Therefore, CS = 2.81 x 1.25 x 3.0 = 10.5 g/dl using the above equations.

Development of continuous recirculating peritoneal dialysis using a double lumen catheter.

Continuous recirculating peritoneal dialysis (CRPD) was newly introduced to improve solute removal efficiency in conventional dialysis therapies such as hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD). In CRPD, a part of the dialysate in the peritoneal cavity was drained through a double-lumen catheter and purified by an extracorporeal dialyzer. Urea removal characteristics in CRPD were examined in a canine study. In this study, a recirculation-dialysis experiment using a dog weighing 9.0 kg was carried out under 100 and 200 ml/min of flow for recirculating and delivered dialysates, respectively. An FB-50H (Nipro Medical Industries, Ltd., Osaka, Japan) composed of cellulose diacetate membrane with 0.5 m2 of surface area and Dianeal-1.5 (Baxter Limited Laboratories, Tokyo, Japan) containing urea were used as the extracorporeal dialyzer and dialysate. Urea peritoneal and dialyzer dialysances (DBP and DBD) were 3.05 and 33.3 ml/min by computer simulation using a compartment model for CRPD. This DBP value can be estimated as 20.3 ml/min for a 60 kg human. From this result, time-averaged value for BUN over an 8 hr/day CRPD, combined with three exchanges/day as CAPD is estimated to be 34.3 mg/dl, which is much lower than 45.2 mg/dl for a 12 hr/week HD, or 53.0 mg/dl for conventional CAPD.

Correlation between the proliferative response to granulocyte colony-stimulating factor and the positivity of transferrin receptor in acute myeloblastic leukemia cells.

The use of granulocyte colony-stimulating factor (G-CSF) after chemotherapy for acute myeloblastic leukemia (AML) has been reported. However, there is a drawback in that G-CSF may stimulate the proliferation of AML progenitors. To determine the parameter(s) indicative of responsiveness of AML blasts to G-CSF, various surface phenotypes of blasts were examined in relation to the blast colony formation stimulated by G-CSF in 39 AML patients. A correlation was found only with transferrin receptor positivity among the various phenotypes studied. The population mean of percentages of transferrin receptor-positive blasts in the group responding to G-CSF in vitro was significantly higher than that of blasts in the group not responding to G-CSF. A further correlation was found between transferrin receptor positivity and the number of G-CSF receptors on the blasts; that is, blasts expressing more G-CSF receptors have greater transferrin receptor positivity. In our previous study, we observed that blasts with a large number of G-CSF receptors produce more colonies in response to G-CSF. These results indicated that blasts expressing more transferrin receptors have a larger number of G-CSF receptors and may show more active proliferation in response to G-CSF. Therefore, the proliferative response of blasts to G-CSF can be predicted by examining transferrin receptor positivity. The clinical use of G-CSF in AML patients may be recommended when the patient's blasts have a low level of transferrin receptor expression. The measurement of transferrin receptors on blasts, instead of the rather complicated G-CSF receptor determination, would be a useful indicator for the safer application of G-CSF in AML patients.

Non-heparin hemodialysis with oral administration of newly developed antiplatelet agent.

Non-heparin hemodialysis (HD) was successfully done in anuric dogs with the oral administration of a newly developed antiplatelet agent, 4-cyano-5,5-bis(4-methoxyphenyl)-4-pentenoic acid, (E5510, Eisai Pharmaceutical Co., Japan). In the current study, the antithrombotic effect of E5510 during HD was investigated. Eleven mongrel dogs with bilateral ureteral ligation were given 0.1 mg/kg of E5510 orally 1 hr before undergoing 4 hr HD using hollow fiber dialyzers, (PMMA 5, regenerated cellulose 6) without heparin and under general anesthesia. Blood samples were taken before the administration of E5510 and before and 1, 2, 3, and 4 hr after starting HD; blood counts, hematocrits, blood chemistries, and plasma thromboxane levels (TxB2) were examined. Platelet aggregation, activated clotting times (ACT), and activated partial thrombin times (APTT) were also measured, and sequential plasma E5510 concentrations were determined. In 10 of 11 anuric dogs, non-heparin HD was successfully done with minimal clotting in the dialyzer and drip chambers. The maximum aggregation rate was depressed to less than 20% of the initial value throughout HD. Plasma TxB2 concentration was depressed, and ACT and APTT were mildly, but not significantly prolonged. Neither hemorrhagic complications nor other side effects of E5510 were observed.

Difference in beta 2-microglobulin removal between cellulosic and synthetic polymer membrane dialyzers.

Several kinds of dialyzers, with highly permeable membranes (HPM), have been designed to specifically remove beta 2-microglobulin (BMG). To clarify their solute transport characteristics, nine types of HPM dialyzers were evaluated during in vivo and in vitro studies using human plasma and aqueous solutions. No BMG membrane adsorption and/or plugging was seen with cellulosic membrane dialyzers during in vitro experiments using human plasma. On the other hand, all synthetic polymer membrane dialyzers had adsorptive properties, and in a dialyzer with a polymethylmethacrylate membrane, a large amount of BMG was removed by adsorption alone. Dialyzers with cellulose triacetate and polyacrylonitril membranes showed higher values of BMG diffusive dialysance (greater than 20 ml/min) and sieving coefficient (greater than 0.9). From in vitro experiments using an aqueous solution containing several solutes with relatively small or middle molecular weights, all HPM dialyzers had a higher overall mass transfer coefficient than any conventional membrane dialyzer.

Microscopic observation of leukocyte kinesis in the vascular bed during hemodialysis using the rabbit ear chamber technique.

Leukocyte kinesis in the capillary vascular bed during hemodialysis (HD) was investigated to elucidate the mechanism of transient leukopenia. Leukocyte movement was observed microscopically during HD using the rabbit ear chamber (REC) technique, which permits visualization of the movement of blood corpuscles in capillaries. Blood was drawn from the femoral artery and returned into the auricular and/or carotid artery so that the blood passing through the hollow fiber artificial kidney (HFAK) flowed into capillaries in the REC. Leukocyte counts of blood samples taken from the afferent and efferent limbs of the HD circuit, the right jugular vein and the right atrium were determined consecutively during HD. The difference in the leukocyte count was observed between the afferent and efferent limbs for the first 15 minutes and thereafter between the efferent limb and the jugular vein. The "transpulmonary" difference in the leukocyte count was not noticed throughout HD. Between 15 and 90 minutes after the start of HD, scarcely any circulating leukocytes were found in capillaries in the REC and some leukocytes were attached to the endothelial surface. Thereafter circulating leukocytes were seen again and detachment of leukocytes from the endothelial surface was observed. No leukocyte aggregation or embolization of aggregating leukocytes was noticed. This evidence suggests that leukopenia may be attributed to the transient shift of leukocytes to the marginal pool of the vessel lumen and this process may not be specific for the pulmonary vasculature, but may occur in the first capillary bed into which the blood passing through the HFAK flows.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in electrical charge of leukocyte surface membranes during hemodialysis. Possible role in transient leukopenia.

Change in the electrical charge of leukocyte surface membranes during hemodialysis (HD) using a cellulose membrane was studied by examining adsorbability of leukocytes taken from patients undergoing HD to cation and anion exchange resins. Leukocytes of HD patients were well adsorbed to both anion and cation exchange resins, whereas normal leukocytes were only minimally adsorbed to a cation exchange resin, suggesting that the surface membranes of normal leukocytes were charged electronegatively. Adsorbability to anion exchange resin by patients' leukocytes taken from the venous limb of the HD circuit line 15 minutes after starting HD was significantly decreased when using a cellulosic membrane. This suggests that the decrease in electronegative leukocyte surface membrane charge, which may facilitate the attachment of leukocytes to electronegatively charged endothelial cell membranes, is the factor leading to leukopenia. Adsorbability to anion exchange resin by normal leukocytes incubated with plasma taken from HD patients 10 to 60 minutes after starting HD was increased, suggesting an increase in electronegatively charged substances in patients' plasma; these recombine with leukocyte surface membranes thereby increasing the electronegative charges of the leukocyte surface membranes and enabling subsequent release of leukocytes from endothelial cells, consistent with the transient nature of the leukopenia.

Effect of membrane trapping in plasma fractionator on separative characteristics.

Several kinds of plasma fractionators have been introduced to actively separate protein fractions between albumin and globulins in double filtration plasmapheresis. However, relatively large molecular weight proteins are known to be partially trapped by the membrane in a plasma fractionator. In this paper, effects of membrane trapping on separation characteristics in plasma fractionators were examined during in vitro and in vivo studies. All in vitro experiments were done with a closed circuit under constant-flow rate filtration. Protein concentration in feed tank kept constant at no filtration in AS-14H, Evaflux 4A and 2A, while 20-40% of IgG and 40-60% of beta-lipoprotein in 2 liter plasma were removed by membrane trapping for 300 min when filtration fraction equaled 0.87. Protein plugging to the membrane seems to be a major factor in these proteins. And, Dead-end and Partially Discarded modalities with relatively high filtration fraction are effective for the separation between albumin and globulin. Twenty-seven DFPP treatments in 13 patients with autoimmune diseases were done to allow us to estimate the effects of membrane trapping during an in vivo study. All treatments using 6 types of plasma fractionator were performed under constant operating conditions with Partially Discarded modality. In any plasma fractionator, beta-lipoprotein was fairly trapped at 30 min after the start of treatment.