A bridge from bioimpedance spectroscopy to 50 kHz bioimpedance analysis: application to total body water measurements.

This paper presents a method for extrapolating the total body water (TBW) resistance R(t50) from the resistance measured at 50 kHz (R(50)). A DXA examination and impedance measurements were carried out in a 1st group of 57 healthy volunteers with a Xitron 4200 multifrequency impedancemeter, in order to determine their values of R(t50) by comparison with resistances extrapolated at an infinite frequency by the Xitron (R(infinity)). TBW volumes were calculated using our modified BIS method (Jaffrin et al 2006 Med. Biol. Eng. Comput. 44 873-82) from R(infinity), R(t50) and from the fat-free mass measured by DXA, assuming a hydration rate of 73.2%. The same protocol and calculations were also carried out on a 2nd group of 21 subjects for independent validation. Data of the 1st group showed that values of R(t50), not significantly different from those of R(infinity), could be obtained by dividing R(50) by 1.231 in men and by 1.224 in women. Applying this method to the 2nd group yielded also values of R(t50) not significantly different from R(infinity). TBW volumes V(t50) obtained from R(t50) were not significantly different from those of our modified BIS method V(tn), or from TBW volumes obtained from DXA in both groups. A comparison with three BIA methods of TBW determination showed that our new method gave results in better agreement with TBW from DXA and from our modified BIS method.

Mass transfers in a fluidized bed bioreactor using alginate beads for a future bioartificial liver.

Fluidized bed bioreactor with alginate beads may be an alternative to hollow fiber cartridge to host hepatocytes for bioartificial liver purposes. After the bioreactor design and the characterization of fluid mechanics, the present study was aimed at analyzing bi-directional mass transfers of calibrated species between external fluid and empty beads. Static (batch) and dynamic (fluidized bed bioreactor) experimental conditions were analyzed. A simple modelling approach permitted the definition of mass transfer coefficients. The motion of beads within the bioreactor clearly enhanced mass transfer kinetics, but did not alter the amount exchanged. The shear enhanced diffusion coefficient for VitB12 was 20 times higher in the fluidized bed bioreactor than under batch conditions, proving the efficiency of such a device.

Microfiltration and ultrafiltration of polysaccharides produced by fermentation using a rotating disk dynamic filtration system.

The recovery of exopolysaccharides (EPS) produced by Sinorhizobium meliloti bacteria by dynamic microfiltration was investigated using a rotating disk device designed in our laboratory, equipped with a 0.2 microm nylon membrane. This system differs from commercially available systems by the presence of vanes on the disk which produce a very important increase in permeate flux while yielding excellent EPS transmission. For polymers produced under standard fermentation conditions (70 h at 30 degrees C), the mass flux rose to 650 g h(-1) m(-2) using a disk equipped with 2 mm vanes rotating at 2000 rpm against 380 g h(-1) m(-2) with a smooth disk at the same speed. The maximum flux observed was 1560 g h(-1) m(-2) with a 6-mm vanes disk rotating at 3000 rpm and a 36 degrees C broth. An interesting finding was that the permeate flux J(f) for various disks can be correlated by the same function of the mean shear stress at the membrane tau(wm) according to J(f) = 4.6 tau(wm) (0.717) for a 30 degrees C broth, showing that the effect of vanes is merely to increase the shear stress by raising the fluid core velocity between the membrane and the disk. With 6-mm vanes the core angular velocity was found to be 84% of disk velocity vs. 45% for a smooth disk. When the fermentation temperature was increased to 36 degrees C to produce a lower molecular weight polymer, the permeate flux rose by about 250%, much more than what could be expected from the reduction in permeate viscosity and followed the same power law with membrane shear stress as for 30 degrees C. The same device was equipped with a PES 50 kDa membrane to concentrate EPS by ultrafiltration. Permeate fluxes were of the order of 160 L h(-1) m(-2) at 2000 rpm and 30 degrees C with nearly complete EPS rejection. Finally, the net electrical power consumed by the disk was measured by subtracting the power consumed without fluid from the power during filtration at the same speed. This power increases with speed and with the presence of vanes, but since the gain provided by the vanes is very high, the specific energy per m(3) of permeate is minimal with the highest vanes tested (6 mm) and maximal for smooth disks.

Reversibility of artifacts of fluid volume measurements by bioimpedance caused by position changes during dialysis.

The effect of temporary position changes, sitting up from supine, on extracellular (ECW) and intracellular (ICW) resistances and fluid volumes calculated from whole body bioimpedance using a Xitron 4200 impedancemeter was investigated on 8 patients during dialysis for a total of 11 tests. It was found that ECW resistance decreased instantaneously by an average of 2.3% when the patient sits up, due to plasma and interstitial fluid shift into the legs which decreases leg resistance, the major contributor to total resistance. This drop in resistance is incorrectly interpreted by the device as an increase in ECW volume which averages 235 ml. But this effect is completely reversible and both ECW resistance and fluid volume rapidly resume their normal course when the patient returns to his initial position. No significant variation in ICW resistance was observed in any of the patients at the position change. We conclude that segmental impedance, which has been proposed to minimize this artifact, is not advisable in dialysis monitoring and that it is simpler to ignore or switch off measurements during the position change so that later data are not affected by it.

Fluid volumes determination by impedance spectroscopy and hematocrit monitoring: application to pediatric hemodialysis.

A method for extracting fluid volumes from multifrequency bioimpedance, which takes into account the body geometry and the presence of nonconducting elements, was tested on 12 young dialyzed patients against correlations for total body water volumes (TBW) from Watson et al. and Humes et al. Our calculations of TBW from impedance were found to overestimate Humes' values by 0.25 L (0.8%) postdialysis and by 2.08 L (6.5%) predialysis. Extracellular water (ECW) was found to contribute an average of 93% of ultrafiltered volume. Intracellular water volume (ICW) determination from impedance was found to be too imprecise to predict its variation during dialysis; therefore, ICW variations were calculated as the difference between ultrafiltration and ECW changes. The continuous recording of hematocrit by an optical device monitored changes in plasma and interstitial volumes. In most cases, ultrafiltration was compensated mainly by a contribution from interstitial fluid, and the drop in plasma volume was generally moderate.

Influence of fermentation conditions and microfiltration processes on membrane fouling during recovery of glucuronane polysaccharides from fermentation broths.

We have investigated the recovery of exopolysaccharides produced by Sinorhizobium meliloti M5N1 CS bacteria from fermentation broths using different membrane filtration processes: cross-flow filtration with a 7 mm i.d. tubular ceramic membrane of 0.5-microm pores under fixed transmembrane pressure or fixed permeate flux and dynamic filtration with a 0.2 microm nylon membrane using a 16-cm rotating disc filter. With the tubular membrane, the polysaccharide mass flux was mainly limited by polymer transmission that decayed to 10% after 90 min. The mass flux of polymer produced under standard fermentation conditions (70 h at 30 degrees C) stabilized after 70 min to 15 g/h/m(2). This mass flux rises to 36 g/h/m(2) when the mean stirring speed during fermentation is increased and to 123 g/h/m(2) when fermentation is extended to 120 h. In both cases, the mean molecular weight of polysaccharides drops from 4.0 10(5) g/mol under standard conditions to 2.7 10(5) g/mol. A similar reduction in molecular weight was observed when the fermentation temperature was raised to 36 degrees C without benefit to the mass flux. These changes in fermentation conditions have little effect on stabilized permeate flux, but raise significantly the sieving coefficient, due probably to molecular weight reduction and the filamentous aspect of the polymer as observed from SEM photographs. The polymer-mass flux was also increased by reducing transmembrane pressure (TMP) and raising the shear rate by inserting a rod in the membrane lumen. Operation under fixed permeate flux instead of constant TMP inhibited fouling during the first 4 h, resulting in higher sieving coefficients and polymer mass fluxes. The most interesting results were obtained with dynamic filtration because it allows operation at high-shear rates and low TMP. Sieving coefficients remained between 90 and 100%. With a smooth disc, the polysaccharide mass flux remained close to 180 g/h/m(2) at 1500 rpm and cell concentrations from 1 to 3 g/L. When radial rods were glued to the disc to increase wall shear stress and turbulence, the mass flux rose to 275 g/h/m(2) at the same speed and cell concentration.

Comparison of optical, electrical, and centrifugation techniques for haematocrit monitoring of dialysed patients.

Haematocrits were measured as a function of ultrafiltration in a simulated haemodialysis circuit using bovine blood (plasma conductivity 12 mS cm-1) and hypotonic (8.6 mS cm-1) or hypertonic (16 mS cm-1) dialysates as well as in the absence of dialysate. A comparison was made between measurements by light absorption due to haemoglobin, by impedance in the blood line at 5 kHz using Hanai's model of blood conductivity, by conductivity measurements of blood samples at 1.2 kHz using a conductimeter, by centrifugation of blood samples and by calculations using fluid conservation. The validity of Hanai's model was verified to be satisfactory by direct blood and plasma conductivity measurements. In the absence of ionic transfer the impedance device underestimated the haematocrit by 5 to 7%. This underestimation reached 18% in the case of hypertonic dialysate, but this effect can be minimised if the haematocrit necessary for calibration is measured by centrifugation after 15 min of dialysate circulation when ionic balance is achieved. It was found that the optical method monitors haemoglobin concentration rather than red cell volume changes and is not affected by osmotic red cell swelling in the case of hypotonic dialysate. It can be concluded that the light absorption technique is both more accurate and more convenient to use than impedance.

Continuous measurements by impedance of haematocrit and plasma volume variations during dialysis.

A technique for continuous measurements of haematocrit and plasma volume in the arterial line of dialysed patients has been tested in vitro and in vivo. This method uses impedance measurements at 5 kHz and requires a single haematocrit measurement. It relies on two assumptions: that plasma resistivity does not change during dialysis and that blood resistivity obeys Hanai's model. Both assumptions are verified during in vitro tests. Haematocrits measured in vivo by this method are found to be in good agreement with direct measurements from blood samples. The haematocrit variation is then used to monitor changes in plasma volume, assuming conservation of erythrocyte volume. In addition, it is possible to obtain the variation in interstitial volume by combining these data with body impedance measurements.

Extra- and intracellular volume monitoring by impedance during haemodialysis using Cole-Cole extrapolation.

A method is presented for monitoring the relative variation of extracellular and intracellular fluid volumes using a multifrequency impedance meter and the Cole-Cole extrapolation technique. It is found that this extrapolation is necessary to obtain reliable data for the resistance of the intracellular fluid. The extracellular and intracellular resistances can be approached using frequencies of, respectively, 5 kHz and 1000 kHz, but the use of 100 kHz leads to unacceptable errors. In the conventional treatment the overall relative variation of intracellular resistance is found to be relatively small.

Pulsed flow cascade filtration. Long-term experience in low density lipoprotein and lipoprotein a removal.

The authors present the results of pulsed flow cascade filtration (PFCF) for low density lipoprotein (LDL) and lipoprolein a (La) removal in four patients with familial hypercholesterolemia. Forty-six treatments were performed. Pressure and flow pulsations were superimposed on the retentate of the secondary filter using a modified single roller peristaltic pump. The pulsation frequency was adjusted to achieve, on average, zero retentate flow (dead end filtration). The transmembrane pressure and sieving coefficients of immunoglobulin G, apolipoprotein A1, and apolipoprotein B of the secondary filter remained constant during filtration periods of 150 min. The PFCF technique was found to remove more LDL and La than did adsorption on dextran sulfate columns, while maintaining adequate albumin recovery (90%).

Extracellular and intracellular fluid volume monitoring during dialysis by multifrequency impedancemetry.

Impedance techniques can, in principle, permit non-invasive monitoring of extracellular (Ve) and intracellular (Vi) fluid volumes during dialysis. The authors present a method that determines the resistances Re and Ri of extracellular and intracellular compartments, respectively, by extrapolating impedance measurement toward zero, as well as infinite frequencies, according to the Cole-Cole model of biologic issues. These measurements were made using a XITRON 4000 B impedance meter (Xitron Technologies Inc., San Diego, CA) at frequencies ranging from 5 kHz to 1 MHz. The fact that the body is not a cylindric, homogeneous conductor is taken into account by introducing shape factors ke and ki and different resistivities pe and pi for the extracellular and intracellular fluid compartments. Assuming that these four unknown parameters can be regarded as constant during dialysis, the authors obtain: [formula: see text] where subscript o denotes initial values at start of dialysis. Impedances were measured at 30 min intervals on 11 pediatric patients and two adults, whereas total body water volume was determined by measuring urea in collected dialysate. Without ultrafiltration, Ve and Vi do not change significantly in percentage whereas, in the presence of ultrafiltration, Ve decreased by 15% to 25%. In cases when Vi does not change, it is possible to determine Ve and its variation during dialysis.

Convective mass transfer in hemodialysis.

Convective mass transfer in hemodialysis is associated with ultrafiltration (UF). In the absence of diffusion as in hemofiltration, the convective clearance is equal to S.QF where S is the apparent solute sieving coefficient and QF the UF flow rate, but the convective contribution significantly decreases when diffusion is present. A rigorous calculation of the combined diffusion-convection mass transfer for partially rejected solutes is very complex. In this paper we review various models of mass fluxes found in the literature. Since all these models express the mass flux through the membrane as a linear function of blood and dialysate concentrations with different coefficients, we present a general expression for the hemodiafiltration clearance combining diffusion and convection which can be adapted to each model of mass flux. A surprising result is that the convective contribution to the clearance is, in the limit of dominant ultrafiltration, independent of the solute sieving coefficient, in contrast to the model of Villaroel et al. This is due to the effect of increased solute concentration at the membrane which compensates exactly for the effect of the sieving coefficient. This effect is overlooked in the Villaroel et al. model which assumes well mixed blood and dialysate compartments. Comparison with in vitro clearance measurements for urea, creatinin, vitamin B12, and myoglobin (16,000 daltons) supports this observation even when diffusion dominates as in the case of clinical conditions for hemodiafiltration. An empirical correlation for the overall clearance valid for all solutes and blood flows between 200 and 500 ml/min is found to be K = KD + 0.43 QF + 8.3 x 10(-3) Q2F when clearances and QF are in ml/min.

A high selectivity cascade filtration technique for LDL-cholesterol and Lp(a) removal.

This work proposes an improvement of the cascade filtration technique in the treatment of familial hypercholesterolemia. A model of the whole process permitted the definition of the parameters that could influence the selectivity of the fractionation: the pore size, the sieving coefficients of both fractionation and plasmapheresis membrane, and the final retentate flow rate. In vivo studies have shown that the dead-end mode for the secondary filter was not always practical because of severe membrane plugging except when a pulsatile pump was included in the extracorporeal circuit. This pump generated hydrodynamic instabilities which decreased membrane fouling and retarded the build up of the polarization concentration layer. Optimization of these specific operating conditions permitted increase in the selectivity index from 1.15 to 2.24. The performances of cascade filtration were then comparable to those of adsorption on dextran sulfate columns.

Urea, creatinine and phosphate kinetic modeling during dialysis: application to pediatric hemodialysis.

The kinetics of urea, creatinine and phosphate removal during dialysis were investigated in pediatric patients using a two-pool model taking into account fluid shifts and mass transfer between the two compartments. It is found that even urea must be described by a two-pool model since it presents a post dialysis rebound due to equilibration between the two compartments. Phosphate plasma concentration drops very sharply during the first hour of dialysis and rises rapidly during the rebound period. This pattern cannot be accounted for by the classical two-pool model with constant generation rate and mass transfer coefficients, but corresponds to a large time-dependent phosphate influx from the intracellular compartment in which phosphate is generated by biochemical reactions or liberated from the bones. This influx was calculated for four patients representing 8 dialysis sessions and was found to reach a plateau after 90 minutes of dialysis, dropping rapidly during the rebound period.

Hemolysis during membrane plasma separation with pulsed flow filtration enhancement.

The use of pulsed blood flow in membrane plasmapheresis permits enhancement of plasma filtration yet may result in high levels of hemolysis due to large increases in instantaneous transmembrane pressure (TMP). This work investigates the occurrence of hemolysis as a function of TMP and wall shear rates (gamma w) for both steady and pulsed blood flow conditions. Two types of hollow fiber filters with identical polypropylene membranes but different lengths and membrane areas (0.1 and 0.25 m2) were tested. Fresh citrated bovine blood was circulated through the fibers at various blood flowrates and TMP in a single pass circuit using a pulsation generator, made of a single roller peristaltic pump. The free hemoglobin concentration of the plasma, Hbm, was measured from permeate samples collected at each set of TMP and gamma w conditions. It was found that the net hemolysis generated by the filtration was proportional to the membrane area. This justified the introduction of an hemolysis index, IH, equal to the plasma hemoglobin per unit membrane area. The boundary for the occurrence of hemolysis was thus defined by setting IH = 30 mg/dl.m2. For both steady and pulsed flow conditions the hemolysis boundaries were found to be straight lines in the TMP-gamma w plane. They were identical for the two filters under steady flow but different for pulsed flow. At the same time mean wall shear rates hemolysis occurred at a lower time mean TMP under pulsed flow conditions than under steady flow conditions.

A feasibility study of a filtration type autotransfusion device.

This paper describes a feasibility study of a disposable autotransfusion device for blood salvage during surgery. The goal was to concentrate hemolyzed blood at 20% hematocrit to 50% while reducing the plasma free hemoglobin concentration from 10 to 1.5 g/l. The device should have a total membrane area of less than 0.6 m2 and should be able to process ten 500 ml blood bags. The processing time for each blood bag should not exceed 5 min. The basic idea was to use several polypropylene hollow fibre plasma filters of 0.1 m2 in series with saline addition between them. Since the mean pore size is 0.5 microns, anticoagulant and plasma hemoglobin can pass freely across the membrane and their concentration is reduced by dilution. The process was first modelled using mass balance equations for red blood cells and plasma hemoglobin in order to find the best device configuration (number of filters and dilutions). It was found that a three filter system could theoretically meet the requirements, if the last filter had a larger surface area (0.3 m2). Some experiments permitted us to prove the validity of this model and to define fully the third filtration stage. Finally, it was shown that the treatment of a 500 ml blood bag required three filtration stages (whose surface areas were respectively 0.1, 0.1 and 0.3 m2) and the use of 750 ml of saline solution added between the filters. This configuration also offers the possibility of using a vacuum driving force instead of pumps, so that the device becomes completely disposable.

Hemolysis reduction in plasmapheresis by module design: operating with pulsed flow filtration enhancement.

This study is an investigation into the effects of module design on hemolysis levels during the filtration enhancement of plasmapheresis using pulsed blood flow. Two polypropylene hollow fiber modules (F1 and F2) were compared under steady and pulsed flow conditions. The fiber lengths and membrane areas of the respective filters were 136 mm, 0.1 m2 and 226 mm, 0.25 m2 and were used at wall shear rates of 400 and 600 s-1. Fresh citrated bovine blood was circulated at 90 ml/min through their fibers with permate and retentate being recirculated. Plasma samples were analysed by the Cripps method to obtain the mean free hemoglobin concentration. A comparison of the average hemoglobin released with time per fiber, and per unit length allowed an assessment of the effects of fiber length and shear rate on hemolysis levels. It was concluded that a reduction in area and fiber length would reduce hemolysis levels and design equations are suggested to find the optimum length.

A process for enhancing selectivity and limiting plugging in plasmafractionation for APOB removal.

A new process using pressure and flow pulsations was investigated in the second stage of cascade filtration for ApoB removal. Due to enhanced filtration by the pulsations, the retentate flow vanished naturally without clamping the line, which reduced albumin and ApoA1 losses. The performance of Eval 4A and cellulose PF100 filters in this case were compared with that of classical dead end mode at a plasma flux of 18 ml/(min m2). The apparent sieving coefficient for small proteins (albumin, ApoA1) under pulsed conditions increased with increasing pressure instead of dropping as in dead end mode. In both cases the ApoB sieving coefficients remained below 0.02. These encouraging results were accompanied by a slower rise of transmembrane pressure showing a decrease of membrane plugging. Thus, the recovery of albumin and ApoA1 is increased while ApoB removal remains constant, which improves the selectivity of the fractionation stage.

Dynamic filtration of blood: a new concept for enhancing plasma filtration.

We have shown previously that blood flow pulsations created by intermittent squeezing of the inlet blood line significantly increased the plasma filtration rate in membrane plasmapheresis. However, in order to avoid hemolysis, the filtration increase had to be limited to about 50%. We have now devised a more efficient pulsation generator. By properly matching the tubing compliance and the pulsation amplitude, it is possible to extract 50 ml/min of plasma from 90 ml/min of blood at 36% hematocrit with a 1000 cm2 polypropylene hollow fiber filter without hemolysis. Simultaneous recording of the time course of plasma filtration rate measured by an electromagnetic flow meter and transmembrane showed that the increase in mean plasma flow rate was due to a dynamic filtration process which prevents the establishment of concentration polarization. The transmembrane pressure (Ptm) increases over a 0.5-second interval when the tube is squeezed. The membrane responds with an increase in filtration since the concentration polarization layer takes a few seconds to build up. The Ptm then drops when the tube is released before the polarization layer has time to build up appreciably and a sudden acceleration of the blood flow (velocity spike) helps clean the membrane, reducing the polarization. Tests with bovine show that the system is very efficient in reducing membrane plugging with small area filters.