Sieving Coefficient Of Albumin Research Articles

Current dialyzer classification in Japan and mortality risk in patients undergoing hemodialysis

Dialyzers are classified into five types based on their β2-microglobulin clearance rate and albumin sieving coefficient: Ia, Ib, IIa, and IIb. In addition, a new classification system introduced a type S dialyzer. However, limited information is available regarding the impact of dialyzer type on patient outcomes. A cohort study was conducted using data from the Japanese Society for Dialysis Therapy Renal Data Registry database. Total 181,804 patients on hemodialysis (HD) were included in the study, categorized into four groups (type Ia, IIa, IIb, and S). The associations between each group and two-year all-cause mortality were assessed using Cox proportional hazard models. Furthermore, propensity score-matching analysis was performed. By the end of 2019, 34,185 patients on dialysis had died. After adjusting for all confounders, the risk for all-cause mortality was significantly lower in the type IIa, and S groups than in the type Ia group. These significant findings were consistent after propensity score matching. In conclusion, our findings suggest that super high-flux dialyzers, with a β2-microglobulin clearance of ≥ 70 mL/min, may be beneficial for patients on HD, regardless of their albumin sieving coefficient. In addition, type S dialyzers may be beneficial for elderly and malnourished patients on dialysis.Trial registration number: UMIN000018641

The Glomerular Sieving Coefficient of Albumin Is Really Very Low

The Glomerular Sieving Coefficient of Albumin Is Really Very Low

Structure design and performance study on anticoagulant-separation dual functional PVC plasma separation membrane

It is necessary to inject anticoagulant continuously during plasma exchange therapy due to the poor anticoagulant performance of the plasma separation membrane. However, it can easily lead to a risk of bleeding or thrombosis. Herein, a novel anticoagulation-separation dual functional polyvinyl chloride (PVC) plasma separation membrane structure is proposed. The small molecular itaconic acid was grafted onto the PVC plasma separation membrane, which can bind Ca2+ in blood to extend coagulation time. When blood flows through the membrane, the blood coagulation time will be prolonged. Meanwhile, plasma containing toxins can be filtrated from the inside of the membrane to the outside. Toxins are removed from human blood effectively. The results proved that itaconic acid was grafted on the PVC plasma separation membrane without reducing membrane pores, membrane sieving coefficient, membrane flux, and membrane tensile strength, successfully. The maximum pores of the adopted anticoagulant-separation dual function PVC plasma separation membrane were 0.30 µm. The bovine serum albumin sieving coefficient reached 95 %. The flux exceeded 300 L·m−2·h−1. The tensile strength was above 4 MPa. Meanwhile, it exhibited excellent hemocompatibility by suppressed bovine serum albumin adsorption, platelets adsorption, erythrocytes adsorption, hemolysis, and strengthened anticoagulant activity. Thus, the anticoagulation-separation dual functional PVC plasma separation membrane designed in this work may have a great clinical application prospect.

Using 2-Photon Microscopy to Quantify the Effects of Chronic Unilateral Ureteral Obstruction on Glomerular Processes.

Applying novel microscopy methods to suitable animal disease models to explore the dynamic physiology of the kidney remains a challenge. Rats with surface glomeruli provide a unique opportunity to investigate physiological and pathophysiological processes using intravital 2-photon microscopy. Quantification of glomerular capillary blood flow and vasoconstriction and dilatation in response to drugs, permeability, and inflammation are just some of the processes that can be studied. In addition, transgenic rats, i.e., podocytes labeled with fluorescent dyes and other molecular biomarker approaches, provide increased resolution to directly monitor and quantify protein-protein interactions and the effects of specific molecular alterations. In mice, which lack surface glomeruli after four weeks of age, unilateral ureteral obstruction (UUO) for several weeks has been used to induce surface glomeruli. As this induction model does not allow for baseline studies, we quantified the effects of UUO on glomerular processes in the UUO model in Munich Wistar Frömter (MWF) rats, which have surface glomeruli under physiologic conditions. The UUO model for five weeks or more induced significant alterations to gross renal morphology, the peritubular and glomerular microvasculature, as well as the structure and function of tubular epithelia. Glomerular and peritubular red blood cell (RBC) flow decreased significantly (p < 0.01), probably due to the significant increase in the adherence of white blood cells (WBCs) within glomerular and peritubular capillaries. The glomerular sieving coefficient of albumin increased from 0.015 ± 0.002 in untreated MWFs to 0.045 ± 0.05 in 5-week-old UUO MWF rats. Twelve weeks of UUO resulted in further increases in surface glomerular density and glomerular sieving coefficient (GSC) for albumin. Fluorescent albumin filtered across the glomeruli was not reabsorbed by the proximal tubules. These data suggest that using UUO to induce surface glomeruli limits the ability to study and interpret normal glomerular processes and disease alterations.

Using 2-Photon Microscopy to Quantify the Effects of Chronic Unilateral Ureteral Obstruction on Glomerular Processes

Applying novel microscopy methods to suitable animal disease models to explore the dynamic physiology of the kidney remains a challenge. Rats with surface glomeruli provide a unique opportunity to investigate physiological and pathophysiological processes using intravital 2-photon microscopy. Quantification of glomerular capillary blood flow and vasoconstriction and dilatation in response to drugs, permeability, and inflammation are just some of the processes that can be studied. In addition, transgenic rats, i.e., podocytes labeled with fluorescent dyes and other molecular biomarker approaches, provide increased resolution to directly monitor and quantify protein-protein interactions and the effects of specific molecular alterations. In mice, which lack surface glomeruli after four weeks of age, unilateral ureteral obstruction (UUO) for several weeks has been used to induce surface glomeruli. As this induction model does not allow for baseline studies, we quantified the effects of UUO on glomerular processes in the UUO model in Munich Wistar Frömter (MWF) rats, which have surface glomeruli under physiologic conditions. The UUO model for five weeks or more induced significant alterations to gross renal morphology, the peritubular and glomerular microvasculature, as well as the structure and function of tubular epithelia. Glomerular and peritubular red blood cell (RBC) flow decreased significantly (p < 0.01), probably due to the significant increase in the adherence of white blood cells (WBCs) within glomerular and peritubular capillaries. The glomerular sieving coefficient of albumin increased from 0.015 ± 0.002 in untreated MWFs to 0.045 ± 0.05 in 5-week-old UUO MWF rats. Twelve weeks of UUO resulted in further increases in surface glomerular density and glomerular sieving coefficient (GSC) for albumin. Fluorescent albumin filtered across the glomeruli was not reabsorbed by the proximal tubules. These data suggest that using UUO to induce surface glomeruli limits the ability to study and interpret normal glomerular processes and disease alterations.

Rapid Removal of Myeloma Light Chains By Continuous Veno-Venous Hemofiltration in a Patient with Acute Cast Nephropathy As a Result of Lambda Light Chain Myeloma

Myeloma cast nephropathy or myeloma kidney (MK) is a complication observed in up to one third of patients with multiple myeloma (MM). MM patients who recover kidney function have longer remissions and increased overall survival. Previous attempts at light chain reduction with the use of plasma exchange in addition to chemotherapy have not consistently improved patient outcomes and remains an area of research. The inefficiency of plasma exchange is due to the high volume of distribution of light chains in the extravascular space which will repopulate the vascular space once the exchange procedure is complete.Continuous Veno-Venous Hemofiltration (CVVH) is a form of continuous hemodialysis (HD). The use of CVVH in addition to chemotherapy in MK is rational, as continuous filtration allows for ongoing equilibrium between the extravascular and intravascular compartments, thus reducing the total body MM protein load. The Purema 500 (NxStage Medical, Lawrence, MA) filter used for CVVH has an average pore size of 65 kDa, and albumin Sieving co-efficient of 0.05. The myeloma kappa light chain is approximately 25 kDa whereas the lambda light chain (LLC), circulating as a dimer, is approximately 45 kDa and the average IgG complex is approximately 150 kDa. These data suggest that CVVH filtration could reduce the light chain MM protein in patients with MK, but would likely be ineffective for IgG, IgA or IgM MM.This hypothesis was tested in a 60 year-old-man with known normal baseline kidney function and LLC MM, who developed MK in the perioperative period, of a hip stabilization procedure, at the time of MM recurrence. Biopsy showed the kidney failure was due to MK. The LLC concentration at the time of kidney failure was 3,750 mg/L. We employed CVVH, run 24 hours a day for 72 hours, with concurrent high dose intravenous dexamethasone 40mg days 1-4 and IV bortezomib 1.3mg/m2 days 1,4,8, 11.Serum protein electrophoresis (SPEP) obtained prior to CVVH showed a monoclonal spike of 6.7 g/L or 11.5% of the total protein (figure 1A). Protein electrophoresis (PEP) of the first bag of spent dialysate showed a restricted peak 1.1g/L or 2.1% of the total protein with a similar appearance to the monoclonal spike on the SPEP, demonstrating removal of the LLC by CVVH (figure 1B). 72 hours after the initiation of CVVH and chemotherapy, the LLC concentration decreased 77% from 3,750 to 850 mg/L (figure 1D), SPEP showed the monoclonal spike had decreased to 0.5 g/L or 1% of the total protein (figure 1C). The patient became dialysis independent three months following CVVH.Previous series have reported on 11 patients with acute MK, who have been treated with as specialized high cut-off hemodialysis (HCO-HD) filter, which has a pore size of 45 kDa, and concurrent chemotherapy. Seven of these patients (63%) became dialysis independent. These studies showed an overall reduction of serum light chain protein in their patients, but did not report if the light chains were present in the dialysate. The EUropean trial of free LIght chain removal by exTEnded haemodialysis in cast nephropathy (EuLITE) reported an improved rate of renal recovery 33% vs 43% with the use of HCO-HD versus standard HD.By way of comparison, CVVH, which is widely available in the United States, rapidly decreased the LLC myeloma protein. Our data supports the depletion of the LLC stores in the extravascular compartment as we did not see a rebound of LLC after stopping CVVH (figure 1D). As kappa light chains have a lower molecular weight than LLC, we would expect the same result with kappa light chain MM. Patients with IgG MK may also benefit from CVVH, as we saw proteins with a molecular weight higher than 65 kDa in the spent dialysate PEP, including proteins in the IgG range (figure 1C). CVVH filters lead to less protein wasting as compared to the HCO-HD filters which would negate the need for albumin replacement. The observed rapid rate of decline of the light chain concentration in this case supports the addition of CVVH to chemotherapy, as the clearance of the myeloma protein was more rapid than has been described in previous trials using bortezomib and dexamethasone alone. Randomized clinical trials would need to be conducted to know if this approach is superior to chemotherapy alone. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

MO676COMPLEMENT ACTIVATION BY DIALYSIS MEMBRANES AND ITS ASSOCIATION WITH SECONDARY MEMBRANE FORMATION AND SURFACE CHARGE

Abstract Background and Aims Activation of the complement system may occur during blood-membrane interactions in hemodialysis and contribute to chronic inflammation of patients with end-stage renal disease (ESRD). Hydrophilic modification with polyvinylpyrrolidone (PVP) has been suggested to increase the biocompatibility profile of dialysis membranes. In the present study we compared complement activation of synthetic and cellulose-based membranes, including the polysulfone membrane with α-tocopherol-stabilized, PVP-enriched inner surface of the novel FX CorAL dialyzer, and linked the results to their physical characteristics. Method Eight synthetic and cellulose-based dialyzers (FX CorAL, FX CorDiax [Fresenius Medical Care]; Polyflux, THERANOVA [Baxter]; ELISIO, SUREFLUX [Nipro]; xevonta [B. Braun]; FDX [Nikkisio Medical]) were investigated in the present study. Complement activation (C3a, C5a, sC5b-9) was evaluated in a 3h ex vivo recirculation model with human blood. Albumin sieving coefficients were determined over a 4h ex vivo recirculation model with human plasma as a surrogate of secondary membrane formation. Zeta potential was measured as an indicator for the surface charge of the membranes. Results The FX CorAL dialyzer induced the lowest activation of the three complement factors (C3a: -39.4%; C5a: -57.5%; sC5b-9: -58.9% compared to the reference). Highest complement activation was found for the cellulose-based SUREFLUX (C3a: +154.0%) and the FDX (C5a: +335.0%; sC5b-9: +287.9%) dialyzers. Moreover, the FX CorAL dialyzer had the nearest-to-neutral zeta potential (-2.38 mV) and the lowest albumin sieving coefficient decrease over time. Albumin sieving coefficient decrease was associated with complement activation by the investigated dialyzers. Conclusion Our present results indicate that the surface modification implemented in the FX CorAL dialyzer reduces secondary membrane formation and improves the biocompatibility profile. Further clinical studies are needed to investigate whether these observations will result in a lower inflammatory burden of hemodialysis patients.

Complement activation by dialysis membranes and its association with secondary membrane formation and surface charge

Activation of the complement system may occur during blood-membrane interactions in hemodialysis and contribute to chronic inflammation of patients with end-stage renal disease. Hydrophilic modification with polyvinylpyrrolidone (PVP) has been suggested to increase the biocompatibility profile of dialysis membranes. In the present study we compared the complement activation of synthetic and cellulose-based membranes, including the polysulfone membrane with α-tocopherol-stabilized PVP-enriched inner surface of the novel FX CorAL dialyzer, and linked the results to their physical characteristics. Eight synthetic and cellulose-based dialyzers (FX CorAL, FX CorDiax [Fresenius Medical Care]; Polyflux, THERANOVA [Baxter]; ELISIO, SUREFLUX [Nipro]; xevonta [B. Braun]; FDX [Nikkisio Medical]) were investigated in the present study. Complement activation (C3a, C5a, and sC5b-9) was evaluated in a 3hours ex vivo recirculation model with human blood. Albumin sieving coefficients were determined over a 4hours ex vivo recirculation model with human plasma as a surrogate of secondary membrane formation. Zeta potential was measured as an indicator for the surface charge of the membranes. The FX CorAL dialyzer induced the lowest activation of the three complement factors (C3a: -39.4%; C5a: -57.5%; and sC5b-9: -58.9% compared to the reference). Highest complement activation was found for the cellulose-based SUREFLUX (C3a: +154.0%) and the FDX (C5a: +335.0% and sC5b-9: +287.9%) dialyzers. Moreover, the FX CorAL dialyzer had the nearest-to-neutral zeta potential (-2.38mV) and the lowest albumin sieving coefficient decrease over time. Albumin sieving coefficient decrease was associated with complement activation by the investigated dialyzers. Our present results indicate that the surface modification implemented in the FX CorAL dialyzer reduces the secondary membrane formation and improves the biocompatibility profile. Further clinical studies are needed to investigate whether these observations will result in a lower inflammatory burden of hemodialysis patients.

Validity of Three-Dimensional Tortuous Pore Structure and Fouling of Hemoconcentration Capillary Membrane Using the Tortuous Pore Diffusion Model and Scanning Probe Microscopy.

Hemoconcentration membranes used in cardiopulmonary bypass require a pore structure design with high pure water permeability, which does not allow excessive protein adsorption and useful protein loss. However, studies on hemoconcentration membranes have not been conducted yet. The purpose of this study was to analyze three-dimensional pore structures and protein fouling before and after blood contact with capillary membranes using the tortuous pore diffusion model and a scanning probe microscope system. We examined two commercially available capillary membranes of similar polymer composition that are successfully used in hemoconcentration clinically. Assuming the conditions of actual use in cardiopulmonary bypass, bovine blood was perfused inside the lumens of these membranes. Pure water permeability before and after bovine blood perfusion was measured using dead-end filtration. The scanning probe microscopy system was used for analysis. High-resolution three-dimensional pore structures on the inner surface of the membranes were observed before blood contact. On the other hand, many pore structures after blood contact could not be observed due to protein fouling. The pore diameters calculated by the tortuous pore diffusion model and scanning probe microscopy were mostly similar and could be validated reciprocally. Achievable pure water permeabilities showed no difference, despite protein fouling on the pore inlets (membrane surface). In addition, low values of albumin sieving coefficient are attributable to protein fouling that occurs on the membrane surface. Therefore, it is essential to design the membrane structure that provides the appropriate control of fouling. The characteristics of the hemoconcentration membranes examined in this study are suitable for clinical use.

Mesangial Cells Regulate the Glomerular Filtration Rate: A Multiphoton Microscopy Study

The intra‐glomerular mesangial cells fill the space between the glomerular capillaries and may support the structural integrity of the glomerulus. Data from cultured mesangial cells suggest a contractile cell phenotype. However, the in vivo relevance of mesangial cells for the function of the glomerular filtration barrier and the regulation of glomerular filtration remains unclear.Here we hypothesized that contractile mesangial cells actively regulate the single nephron glomerular filtration rate (snGFR) and that mesangial cells support the integrity of the glomerular filtration barrier. To test this hypothesis, we assessed the function of mesangial cells in the life animal by intravital multiphoton microscopy. Mesangial cells were depleted in Munich Wistar Froemter rats using the Thy1.1 antibody model. Administration of the Thy1.1 antibody (1 mg/kg BW) caused the loss of approximately 80% of mesangial cells. Immune‐staining of kidneys for Thy1.1 showed an exclusive binding of the antibody to mesangial cells. To assess if mesangial cells contribute to the regulation of snGFR by vasoconstrictor agents, angiotensin II (12.8 ng/kg/min) was infused during intravital imaging. In response to angiotensin II, the systolic arterial blood pressure rose from 100.5 to 125.1 mm Hg mm Hg in Thy1.1‐treated rats (n=17; p&lt;.0001), and, similarly, from 98.8 mm Hg to 125.2 mm Hg in controls (n=19; p&lt;.0001). SnGFR, as determined by flow measurements in the proximal tubule after bolus injections of Lucifer Yellow, decreased after angiotensin II infusion by 35±12% in controls (n=5; p=.004), whereas it remained unchanged in Thy1.1‐treated rats (−12±10%, n=5). In control rats, changes in snGFR after angiotensin II infusion were accompanied by a marked rotating movement of the capillary loops within Bowman’s space. This phenomenon was absent in Thy1.1‐treated rats. In addition, glomerular capillaries at baseline were dilated, and blood flow velocity was reduced in Thy1.1‐treated rats compared with controls, whereas the calculated capillary flow was similar in both groups (5.8±.6 nl/min in controls vs. 4.9±.5 nl/min in Thy1.1‐treated rats; n=6; n.s.). Glomerular capillary flow declined in response to angiotensin II in controls but it remained unchanged in Thy1.1‐treated rats (2.7±.2 nl/min vs. 3.7±.4 nl/min, respectively).Next, to determine the relevance of mesangial cells for the integrity of the glomerular barrier function, we measured the sieving coefficient for albumin (GSC). Albumin GSC was low in control rats averaging 0.00059±.00006. After angiotensin II infusion, the GSC for albumin increased to 0.00112±.00015 in control rats (n=8; p&lt;.0001). In contrast, in Thy1.1‐treated rats, baseline albumin GSC was elevated (0.0033±.00080; n=7; p&lt;.0001) and did not change in response to angiotensin II (0.0047±.001; n.s.).In summary, the depletion of mesangial cells by Thy1.1 antibody application abolished the regulation of the snGFR by angiotensin II and markedly increased the permeability of the filtration barrier for plasma proteins. Or data suggest that mesangial cells contribute to the regulation of the snGFR in vivo. Furthermore, mesangial cells are crucially involved in the maintenance of the integrity of the glomerular filtration barrier.Support or Funding InformationSFB 1350/B2

Aldosterone induces albuminuria via matrix metalloproteinase–dependent damage of the endothelial glycocalyx

Aldosterone contributes to end-organ damage in heart failure and chronic kidney disease. Mineralocorticoid-receptor inhibitors limit activation of the receptor by aldosterone and slow disease progression, but side effects, including hyperkalemia, limit their clinical use. Damage to the endothelial glycocalyx (a luminal biopolymer layer) has been implicated in the pathogenesis of endothelial dysfunction and albuminuria, but to date no one has investigated whether the glomerular endothelial glycocalyx is affected by aldosterone. Invitro, human glomerular endothelial cells exposed to 0.1 nM aldosterone and 145 mMol NaCl exhibited reduced cell surface glycocalyx components (heparan sulfate and syndecan-4) and disrupted shear sensing consistent with damage of the glycocalyx. Invivo, administration of 0.6 μg/g/d of aldosterone (subcutaneous minipump) and 1% NaCl drinking water increased glomerular matrix metalloproteinase 2 activity, reduced syndecan 4 expression, and caused albuminuria. Intravital multiphoton imaging confirmed that aldosterone caused damage of the glomerular endothelial glycocalyx and increased the glomerular sieving coefficient for albumin. Targeting matrix metalloproteinases 2 and 9 with a specific gelatinase inhibitor preserved the glycocalyx, blocked the rise in glomerular sieving coefficient, and prevented albuminuria. Together these data suggest that preservation of the glomerular endothelial glycocalyx may represent a novel strategy for limiting the pathological effects of aldosterone.

Increase of Total Nephron Albumin Filtration and Reabsorption in Diabetic Nephropathy

The amount of albumin filtered through the glomeruli and reabsorbed at the proximal tubules in normal and in diabetic kidneys is debated. The megalin/cubilin complex mediates protein reabsorption, but genetic knockout of megalin is perinatally lethal. To overcome current technical problems, we generated a drug-inducible megalin-knockout mouse line, megalin(lox/lox);Ndrg1-CreERT2 (iMegKO), in which megalin expression can be shut off at any time by administration of tamoxifen (Tam). Tam administration in adult iMegKO mice decreased the expression of renal megalin protein by 92% compared with that in wild-type C57BL/6J mice and almost completely abrogated renal reabsorption of intravenously injected retinol-binding protein. Furthermore, urinary albumin excretion increased to 175 μg/d (0.46 mg albumin/mg creatinine) in Tam-treated iMegKO mice, suggesting that this was the amount of total nephron albumin filtration. By comparing Tam-treated, streptozotocin-induced diabetic iMegKO mice with Tam-treated nondiabetic iMegKO mice, we estimated that the development of diabetes led to a 1.9-fold increase in total nephron albumin filtration, a 1.8-fold increase in reabsorption, and a significant reduction in reabsorption efficiency (86% efficiency versus 96% efficiency in nondiabetic mice). Insulin treatment normalized these abnormalities. Akita;iMegKO mice, another model of type 1 diabetes, showed equivalent results. Finally, nondiabetic iMegKO mice had a glomerular sieving coefficient of albumin of 1.7×10-5, which approximately doubled in diabetic iMegKO mice. This study reveals actual values and changes of albumin filtration and reabsorption in early diabetic nephropathy in mice, bringing new insights to our understanding of renal albumin dynamics associated with the hyperfiltration status of diabetic nephropathy.

First Implantation of Silicon Nanopore Membrane Hemofilters.

An implantable hemofilter for the treatment of kidney failure depends critically on the transport characteristics of the membrane and the biocompatibility of the membrane, cartridge, and blood conduits. A novel membrane with slit-shaped pores optimizes the trade-off between permeability and selectivity, enabling implanted therapy. Sustained (3-8) day function of an implanted parallel-plate hemofilter with minimal anticoagulation was achieved by considering biocompatibility at the subnanometer scale of chemical interactions and the millimeter scale of blood fluid dynamics. A total of 400 nm-thick polysilicon flat sheet membranes with 5-8 nm × 2 micron slit-shaped pores were surface-modified with polyethylene glycol. Hemofilter cartridge geometries were refined based on computational fluid dynamics models of blood flow. In an uncontrolled pilot study, silicon filters were implanted in six class A dogs. Cartridges were connected to the cardiovascular system by anastamoses to the aorta and inferior vena cava and filtrate was drained to collection pouches positioned in the peritoneum. Pain medicine and acetylsalicylic acid were administered twice daily until the hemofilters were harvested on postoperative days 3 (n = 2), 4 (n = 2), 5 (n = 1), and 8 (n = 1). No hemofilters were thrombosed. Animals treated for 5 and 8 days had microscopic fractures in the silicon nanopore membranes and 20-50 ml of transudative (albumin sieving coefficient θalb ~ 0.5 - 0.7) fluid in the collection pouches at the time of explant. Shorter experimental durations (3-4 days) resulted in filtration volumes similar to predictions based on mean arterial pressures and membrane hydraulic permeability and (θalb ~ 0.2 - 0.3), similar to preimplantation measurements. In conclusion, a detailed mechanistic and materials science attention to blood-material interactions allows implanted hemofilters to resist thrombosis. Additional testing is needed to determine optimal membrane characteristics and identify limiting factors in long-term implantation.

In Vivo Visualization of the Antialbuminuric Effects of the Angiotensin-Converting Enzyme Inhibitor Enalapril

Angiotensin-converting enzyme (ACE) inhibitors are commonly used antiproteinuric drugs. Here we assessed the effect of the ACE inhibitor enalapril on the glomerular sieving coefficient of albumin (GSCA) using intravital multiphoton microscopy. Munich Wistar Frömter (MWF) rats were used as a model of hypertension-related glomerular lesions. Young (9-week-old) MWF rats were nonproteinuric, similar to what was observed in control Wistar rats. However, urinary albumin excretion in the MWF rats gradually increased during aging, averaging 0.00062 ± 0.0001 at age 9 weeks and 0.0054 ± 0.0003 (mg/mOsmol per liter) at age 52 weeks (P < 0.0001). Albuminuria in aged MWF rats was accompanied by structural changes, which were indicative of glomerular lesions. The GSCA was low in young MWF rats but increased markedly during aging, averaging 0.00057 ± 4.7 × 10(-5) (n = 25) in young MWF rats and 0.0027 ± 0.00036 in 52-week-old MWF rats (n = 36; P < 0.0001). Treatment of proteinuric 12-month-old MWF rats with enalapril over a 4-week period reduced the GSCA from 0.0027 ± 0.00036 to 0.00139 ± 0.00013 (P = 0.0005). Similarly, urinary albumin excretion was reduced, averaging 0.0051 ± 0.0003 and 0.0036 ± 0.0005 mg/mOsmol per liter before and after enalapril administration, respectively (P = 0.0089). In parallel, enalapril treatment reduced the mean arterial blood pressure (144.6 ± 6.5 mm Hg in untreated versus 110.9 ± 0.6 mm Hg in enalapril-treated MWF rats) and increased the glomerular filtration rate from 1.64 ± 0.3 ml/min to 3.58 ± 0.3 ml/min (P = 0.0025 versus baseline). In summary, enalapril reduced the GSCA in proteinuric MWF rats, which was paralleled by a similar reduction in urinary albumin excretion. These data suggest that glomerular rather than tubular mechanisms account for the beneficial antiproteinuric effects of the ACE inhibitor.

Multiphoton Imaging of the Glomerular Permeability of Angiotensinogen

Patients and animals with renal injury exhibit increased urinary excretion of angiotensinogen. Although increased tubular synthesis of angiotensinogen contributes to the increased excretion, we do not know to what degree glomerular filtration of systemic angiotensinogen, especially through an abnormal glomerular filtration barrier, contributes to the increase in urinary levels. Here, we used multiphoton microscopy to visualize and quantify the glomerular permeability of angiotensinogen in the intact mouse and rat kidney. In healthy mice and Munich-Wistar-Frömter rats at the early stage of glomerulosclerosis, the glomerular sieving coefficient of systemically infused Atto565-labeled human angiotensinogen (Atto565-hAGT), which rodent renin cannot cleave, was only 25% of the glomerular sieving coefficient of albumin, and its urinary excretion was undetectable. In a more advanced phase of kidney disease, the glomerular permeability of Atto565-hAGT was slightly higher but still very low. Furthermore, unlike urinary albumin, the significantly higher urinary excretion of endogenous rat angiotensinogen did not correlate with either the Atto565-hAGT or Atto565-albumin glomerular sieving coefficients. These results strongly suggest that the vast majority of urinary angiotensinogen originates from the tubules rather than glomerular filtration.

Multiple Factors Influence Glomerular Albumin Permeability in Rats

Different laboratories recently reported incongruous results describing the quantification of albumin filtration using two-photon microscopy. We investigated the factors that influence the glomerular sieving coefficient for albumin (GSC(A)) in an effort to explain these discordant reports and to develop standard operating procedures for determining GSC(A). Multiple factors influenced GSC(A), including the kidney depth of image acquisition (10-20 μm was appropriate), the selection of fluorophore (probes emitting longer wavelengths were superior), the selection of plasma regions for fluorescence measurements, the size and molecular dispersion characteristics of dextran polymers if used, dietary status, and the genetic strain of rat. Fasting reduced the GSC(A) in Simonsen Munich Wistar rats from 0.035±0.005 to 0.016±0.004 (P<0.01). Frömter Munich Wistar rats had a much lower GSC(A) in both the fed and the fasted states. Finally, we documented extensive albumin transcytosis with vesicular and tubular delivery to and fusion with the basolateral membrane in S1 proximal tubule cells. In summary, these results help explain the previously conflicting microscopy and micropuncture data describing albumin filtration and highlight the dynamic nature of glomerular albumin permeability.

Electrical forces determine glomerular permeability.

There is ongoing controversy about the mechanisms that determine the characteristics of the glomerular filter. Here, we tested whether flow across the glomerular filter generates extracellular electrical potential differences, which could be an important determinant of glomerular filtration. In micropuncture experiments in Necturus maculosus, we measured a potential difference across the glomerular filtration barrier that was proportional to filtration pressure (-0.045 mV/10 cm H₂O). The filtration-dependent potential was generated without temporal delay and was negative within Bowman's space. Perfusion with the cationic polymer protamine abolished the potential difference. We propose a mathematical model that considers the relative contributions of diffusion, convection, and electrophoretic effects on the total flux of albumin across the filter. According to this model, potential differences of -0.02 to -0.05 mV can induce electrophoretic effects that significantly influence the glomerular sieving coefficient of albumin. This model of glomerular filtration has the potential to provide a mechanistic theory, based on experimental data, about the filtration characteristics of the glomerular filtration barrier. It provides a unique approach to the microanatomy of the glomerulus, renal autoregulation, and the pathogenesis of proteinuria.

Selective Albumin Exchange: A Novel and Simple Method to Remove Bilirubin

Owing to its non-selectivity, plasma exchange has limited use in the treatment of patients with hepatic failure, although it is effective in removal of protein-bound toxins. This study reports a novel way to perform selective albumin exchange (SAE) by using a secondary plasma separator and aims to study its depurative capacity in the removal of bilirubin. In ex vivo experiments, the sieving coefficients (SCs) of plasma proteins for two secondary plasma separators, EC20W and EC30W (Asahi Medical, Tokyo, Japan), were measured. The EC20W membrane was chosen for use in clinical treatment because of its higher selectivity in separating albumin from plasma than the EC30W. The SCs of albumin, immunoglobumin G, A and M (IgG, IgA, IgM) for the EC20W were 0.3+/-0.021, 0.017+/-0.012, and 0, respectively. Five inpatients with plasma total bilirubin (TB) more than 200 micromol/L were enrolled in the present study and received a total of 10 SAE therapy sessions. Each session lasted 10 hours. SAE using the EC20W was conducted, making it similar to post-dilution continuous veno-venous hemofiltration (CVVH), with a filtrate rate of 2000 ml/h. Replacement fluid was composed by adding human albumin into conventional CVVH replacement fluid, with a final albumin concentration of 0.6%. During each treatment, the parameters of plasma and filtrate TB, direct bilirubin (DB), indirect bilirubin (IDB), and proteins were dynamically monitored. Hemostasis parameters were measured before and after sessions. The reduction ratio of plasma TB, DB, and IDB after a single session was 29.1+/-3.0%, 31.3+/-4.5%, and 18.7+/-10.2%, respectively. The clearance of TB at initiation was 11.1+/-1.3 mL/min and declined to 4.4+/-0.5 ml/min at the end (p<0.01). This decline was accompanied by a reduction in albumin SC from 0.3+/-0.021 to 0.13+/-0.05. The molar ratio of bilirubin to albumin in filtrate was comparable to that in plasma. After a single treatment, plasma protein concentration including total protein, albumin, IgA and IgM remained unaffected, except for globulin and IgG, which were reduced by 11.5+/-7.8% and 11.1+/-2.3%, respectively. An improvement in hemostasis parameters, including plasma fibrinogen, prothrombase time and INR, was found after treatment. No obvious side effects were reported during any of the sessions. Selective albumin exchange is a simple and effective method to remove bilirubin. However, further studies are required to verify its effects on clinical outcome.

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Controversies in nephrology: Response to ‘renal albumin handling, facts, and artifacts’

For 40 years indirect measurements of the glomerular sieving coefficient of albumin yielded very low values. The first direct measurement by 2-photon microscopy by Russo et al (Kidney Int (2007) 71, 504-513) gives values 50-times higher. This demonstrated that relatively large quantities of albumin are normally filtered based on size selectivity alone. Most of this albumin is retrieved and returned to the blood supply. These new discoveries represent a paradigm shift in our understanding of albumin processing by the kidney. They also serve to explain several anomalous aspects of previous studies on glomerular filtration and mechanism of albuminuria and support the fact that glomerular charge selectivity is not a major factor controlling glomerular permselectivity.