A gel-membrane model of glomerular charge and size selectivity in series

2001 ◽  
Vol 280 (3) ◽  
pp. F396-F405 ◽  
Author(s):  
Maria Ohlson ◽  
Jenny Sörensson ◽  
Börje Haraldsson
Keyword(s):  
Negative Charge ◽  
Careful Analysis ◽  
Size Selectivity ◽  
Gel Structure ◽  
Experimental Conditions ◽  
Ion Interactions ◽  
In Series ◽  
A Charge ◽  
Glomerular Barrier

We have analyzed glomerular sieving data from humans, rats in vivo, and from isolated perfused rat kidneys (IPK) and present a unifying hypothesis that seems to resolve most of the conflicting results that exist in the literature. Particularly important are the data obtained in the cooled IPK, because they allow a variety of experimental conditions for careful analysis of the glomerular barrier; conditions that never can be obtained in vivo. The data strongly support the classic concept of a negative charge barrier, but separate components seem to be responsible for charge and size selectivity. The new model is composed of a dynamic gel and a more static membrane layer. First, the charged gel structure close to the blood compartment has a charge density of 35–45 meq/l, reducing the concentration of albumin to 5–10% of that in plasma, due to ion-ion interactions. Second, the size-selective structure has numerous functional small pores (radius 45–50 Å) and far less frequent large pores (radius 75–115 Å), the latter accounting for 1% of the total hydraulic conductance. Both structures are required for the maintenance of an intact glomerular barrier.

Puromycin aminonucleoside damages the glomerular size barrier with minimal effects on charge density

2001 ◽  
Vol 281 (3) ◽  
pp. F503-F512 ◽  
Author(s):  
Clara Hjalmarsson ◽  
Maria Ohlson ◽  
Börje Haraldsson
Keyword(s):  
Charge Density ◽  
Size Selectivity ◽  
Puromycin Aminonucleoside ◽  
Pore Analysis ◽  
Small Pore ◽  
Glomerular Size ◽  
Glomerular Barrier ◽  
Wall Charge

Puromycin aminonucleoside (PAN) has been suggested to reduce glomerular charge density, to create large glomerular “leaks,” or not to affect the glomerular barrier. Therefore, we analyzed glomerular charge and size selectivity in vivo and in isolated kidneys perfused at 8°C (cIPK) in control and PAN-treated rats. The fractional clearances (θ) for albumin and Ficoll of similar hydrodynamic size were 0.0017 ± 0.0004 and 0.15 ± 0.02, respectively, in control cIPKs. Two-pore analysis gave similar results in vivo and in vitro, with small- and large-pore radii of 47–52 and 85–105 Å, respectively, in controls. Puromycin increased the number of large pores 40–50 times, the total pore area over diffusion distance decreased by a factor of 25–30, and the small-pore radius increased by 33% ( P < 0.001 for all comparisons of size selectivity and θ). The effect of PAN was less dramatic on the estimated wall charge density, which was 73% of that of controls. We conclude that puromycin effectively destroys the glomerular size barrier with minimal effects on charge density.

scholarly journals Ultrastructural model for size selectivity in glomerular filtration

1999 ◽  
Vol 276 (6) ◽  
pp. F892-F902 ◽  
Author(s):  
Aurélie Edwards ◽  
Barbara S. Daniels ◽  
William M. Deen
Keyword(s):  
Structural Characteristics ◽  
Capillary Wall ◽  
Slit Diaphragm ◽  
Homogeneous Material ◽  
Size Selectivity ◽  
Electron Microscopic ◽  
Sieving Coefficient ◽  
Ultrastructural Model ◽  
Glomerular Barrier

A theoretical model was developed to relate the size selectivity of the glomerular barrier to the structural characteristics of the individual layers of the capillary wall. Thicknesses and other linear dimensions were evaluated, where possible, from previous electron microscopic studies. The glomerular basement membrane (GBM) was represented as a homogeneous material characterized by a Darcy permeability and by size-dependent hindrance coefficients for diffusion and convection, respectively; those coefficients were estimated from recent data obtained with isolated rat GBM. The filtration slit diaphragm was modeled as a single row of cylindrical fibers of equal radius but nonuniform spacing. The resistances of the remainder of the slit channel, and of the endothelial fenestrae, to macromolecule movement were calculated to be negligible. The slit diaphragm was found to be the most restrictive part of the barrier. Because of that, macromolecule concentrations in the GBM increased, rather than decreased, in the direction of flow. Thus the overall sieving coefficient (ratio of Bowman’s space concentration to that in plasma) was predicted to be larger for the intact capillary wall than for a hypothetical structure with no GBM. In other words, because the slit diaphragm and GBM do not act independently, the overall sieving coefficient is not simply the product of those for GBM alone and the slit diaphragm alone. Whereas the calculated sieving coefficients were sensitive to the structural features of the slit diaphragm and to the GBM hindrance coefficients, variations in GBM thickness or filtration slit frequency were predicted to have little effect. The ability of the ultrastructural model to represent fractional clearance data in vivo was at least equal to that of conventional pore models with the same number of adjustable parameters. The main strength of the present approach, however, is that it provides a framework for relating structural findings to the size selectivity of the glomerular barrier.

Influence of Sensor Size on the Accuracy of In-Vivo Ligament and Tendon Force Measurements

1998 ◽  
Vol 120 (6) ◽  
pp. 764-769 ◽  
Author(s):  
M. Rupert ◽  
E. Grood ◽  
T. Byczkowski ◽  
M. Levy
Keyword(s):  
Stress Distribution ◽  
Measurement Errors ◽  
Force Transducer ◽  
Load Cell ◽  
Experimental Conditions ◽  
Calibration Equation ◽  
In Series ◽  
Tendon Force ◽  
Selection Of

In-vivo tendon forces are commonly measured using transducers, which detect tension in the tendon fibers. A poorly understood source of measurement errors is the difference in stress distribution within the tendon between experimental and transducer calibration conditions. The objective of this study was to investigate this source of error, and to determine whether these errors could be minimized by proper selection of transducer size. The study was conducted using the infrapatellar ligament (patellar tendon) of New Zealand White rabbits. Tendon force was measured with two different size implantable force transducers (IFTs), one Wide and one Narrow, and by a strain gaged load cell in series with the tendon. Tests were conducted at five different loading conditions selected to produce five different stress distributions within the tendon. One loading condition corresponded to a typical post-experiment calibration, and the data from that condition were used to develop a calibration equation for the transducer. The errors that resulted from using this calibration were determined by comparing the tendon force measured by the in-series load cell with the force predicted from the IFT output using the calibration equation. Changes in stress distribution produced measurement errors up to 64 N with the Narrow IFT but only 24 N with the Wide IFT. We found the measurement error was dependent on sensor width. Our results support the hypothesis that measurement errors can be caused by differences in tendon stress distribution between calibration and experimental conditions. We further showed that these errors can be minimized by using an IFT, which samples the tension in a large percentage of the tendon fibers. Information from this study can be used for selection of an appropriately sized implantable force transducer for measuring tendon and ligament force.

Mild renal ischemia-reperfusion reduces charge and size selectivity of the glomerular barrier

2007 ◽  
Vol 292 (6) ◽  
pp. F1802-F1809 ◽  
Author(s):  
Maria Andersson ◽  
Ulf Nilsson ◽  
Clara Hjalmarsson ◽  
Börje Haraldsson ◽  
Jenny Sörensson Nyström
Keyword(s):  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Cell Activity ◽  
Renal Ischemia ◽  
Endothelial Glycocalyx ◽  
Size Selectivity ◽  
Fractional Clearance ◽  
Tentative Explanation ◽  
Glomerular Barrier

Despite recent discoveries of molecules in podocytes, the mechanisms behind most conditions of proteinuria are still poorly understood. To understand more about this delicate barrier, we studied the functional and morphological effects of mild (15 min) renal ischemia-reperfusion injury (IRI). Renal function was studied in rats in vivo, followed by a more detailed analysis of the glomerular barrier in cooled (8°C) isolated perfused kidneys (cIPK). Renal blood flow was quickly restored, whereas the glomerular filtration rate remained halved 30 min after IRI. Tubular cell activity was intact as judged from the unaffected Cr-EDTA U/P concentration ratio. In vivo, the fractional clearance (θ) for albumin increased 16 times. In rats subjected to cIPK starting 30 min after in vivo IRI, θalbumin was 15 times and θFicoll_36Å 1.8 times higher than in control cIPKs. According to the heterogeneous charged fiber model, IRI reduced the fiber charge density to 38% of control ( P < 0.01, n = 7). Morphometric analysis with electron microscopy did not reveal any changes in the podocytes or the glomerular basement membrane (GBM) after IRI, suggesting more subtle changes of the GBM and/or the endothelial glycocalyx. We conclude that mild renal IRI induces formation of reactive oxygen species, massive proteinuria, and loss of charged fibers with no apparent change in morphology. These novel findings stress the importance of other components of the barrier, such as proteoglycans produced by the glomerular cells, and provide a tentative explanation for the mechanisms behind proteinuria in glomerulonephritis, for example.

Superoxide, Xanthine Oxidase and Platelet Reactions: Further Studies on Mechanisms by which Oxidants Influence Platelets

1981 ◽  
Vol 45 (03) ◽  
pp. 290-293 ◽  
Author(s):  
Peter H Levine ◽  
Danielle G Sladdin ◽  
Norman I Krinsky
Keyword(s):  
Superoxide Dismutase ◽  
Cytochrome C ◽  
Xanthine Oxidase ◽  
Direct Effect ◽  
Platelet Function ◽  
Experimental Conditions ◽  
Release Reaction

SummaryIn the course of studying the effects on platelets of the oxidant species superoxide (O- 2), Of was generated by the interaction of xanthine oxidase plus xanthine. Surprisingly, gel-filtered platelets, when exposed to xanthine oxidase in the absence of xanthine substrate, were found to generate superoxide (O- 2), as determined by the reduction of added cytochrome c and by the inhibition of this reduction in the presence of superoxide dismutase.In addition to generating Of, the xanthine oxidase-treated platelets display both aggregation and evidence of the release reaction. This xanthine oxidase induced aggreagtion is not inhibited by the addition of either superoxide dismutase or cytochrome c, suggesting that it is due to either a further metabolite of O- 2, or that O- 2 itself exerts no important direct effect on platelet function under these experimental conditions. The ability of Of to modulate platelet reactions in vivo or in vitro remains in doubt, and xanthine oxidase is an unsuitable source of O- 2 in platelet studies because of its own effects on platelets.

Desmopressin (DDAVP) Enhances Platelet Adhesion to the Extracellular Matrix of Cultured Human Endothelial Cells through Increased Expression of Tissue Factor

1997 ◽  
Vol 77 (05) ◽  
pp. 0975-0980 ◽  
Author(s):  
Angel Gálvez ◽  
Goretti Gómez-Ortiz ◽  
Maribel Díaz-Ricart ◽  
Ginés Escolar ◽  
Rogelio González-Sarmiento ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Tissue Factor ◽  
Platelet Adhesion ◽  
Umbilical Vein ◽  
Procoagulant Activity ◽  
Experimental Conditions ◽  
Chromogenic Assay

SummaryThe effect of desmopressin (DDAVP) on thrombogenicity, expression of tissue factor and procoagulant activity (PCA) of extracellular matrix (ECM) generated by human umbilical vein endothelial cells cultures (HUVEC), was studied under different experimental conditions. HUVEC were incubated with DDAVP (1, 5 and 30 ng/ml) and then detached from their ECM. The reactivity towards platelets of this ECM was tested in a perfusion system. Coverslips covered with DD A VP-treated ECMs were inserted in a parallel-plate chamber and exposed to normal blood anticoagulated with low molecular weight heparin (Fragmin®, 20 U/ml). Perfusions were run for 5 min at a shear rate of 800 s1. Deposition of platelets on ECMs was significantly increased with respect to control ECMs when DDAVP was used at 5 and 30 ng/ml (p <0.05 and p <0.01 respectively). The increase in platelet deposition was prevented by incubation of ECMs with an antibody against human tissue factor prior to perfusion. Immunofluorescence studies positively detected tissue factor antigen on DDAVP derived ECMs. A chromogenic assay performed under standardized conditions revealed a statistically significant increase in the procoagulant activity of the ECMs produced by ECs incubated with 30 ng/ml DDAVP (p <0.01 vs. control samples). Northern blot analysis revealed increased levels of tissue factor mRNA in extracts from ECs exposed to DDAVP. Our data indicate that DDAVP in vitro enhances platelet adhesion to the ECMs through increased expression of tissue factor. A similar increase in the expression of tissue factor might contribute to the in vivo hemostatic effect of DDAVP.

In vivo Production of Soluble Inorganic Pyrophosphatases in Two Strains of Vibrio cholerae

1970 ◽  
Vol 24 (1) ◽  
pp. 38-41
Author(s):  
Taslima Taher Lina ◽  
Mohammad Ilias
Keyword(s):  
Vibrio Cholerae ◽  
Specific Activity ◽  
Experimental Conditions ◽  
Environmental Strain ◽  
Cell Extracts ◽  
Atcc Strain ◽  
The Family ◽  
Effects Of Ph ◽  
Vibrio Cholerae O1

The in vivo production of soluble inorganic pyrophosphatases (PPases) was investigated in two strains, namely, Vibrio cholerae EM 004 (environmental strain) and Vibrio cholerae O1 757 (ATCC strain). V. cholerae is known to contain both family I and family II PPase coding sequences. The production of family I and family II PPases were determined by measuring the enzyme activity in cell extracts. The effects of pH, temperature, salinity of the growth medium on the production of soluble PPases were studied. In case of family I PPase, V. cholerae EM 004 gave the highest specific activity at pH 9.0, with 2% NaCl + 0.011% NaF and at 37°C. The strain V. cholerae O1 757 gave the highest specific activity at pH 9.0, with media containing 0% NaCl and at 37°C. On the other hand, under all the conditions family II PPase did not give any significant specific activity, suggesting that the family II PPase was not produced in vivo in either strains of V. cholerae under different experimental conditions. Keywords: Vibrio cholerae, Pyrophosphatases (PPases), Specific activityDOI: http://dx.doi.org/10.3329/bjm.v24i1.1235 Bangladesh J Microbiol, Volume 24, Number 1, June 2007, pp 38-41

Validation of a monetary Taylor Aggression Paradigm: Associations with trait aggression and role of provocation sequence

2020 ◽  
Author(s):  
J Konzok ◽  
L Kreuzpointner ◽  
GI Henze ◽  
L Wagels ◽  
C Kärgel ◽  
...  
Keyword(s):  
Random Sequence ◽  
Reactive Aggression ◽  
Reaction Time Task ◽  
Competitive Reaction ◽  
Experimental Conditions ◽  
Fixed Sequence ◽  
Taylor Aggression Paradigm ◽  
Trait Aggression ◽  
Significant Difference ◽  
In Series

© 2020 Elsevier Inc. The Taylor Aggression Paradigm (TAP) is widely used to measure reactive aggression in laboratory settings. While modified versions (mTAPs) with various stimulus characteristics (shocks, noise, pressure, heat) have already been established, a modified version with monetary stimuli has only been introduced very recently. In this experiment, 209 young healthy participants (104 males, 105 females) completed a mock Competitive Reaction Time Task (CRTT) with a fictional opponent with preprogrammed 40 win and 60 lose trials. In lose trials, participants were provoked by subtracting a low (0–20 euro cents), medium (30–60 cents) or high (70–90 cents) amount of money from their fictitious account. Provocation stimuli were either presented randomly or in a fixed sequence (experimental conditions). In contrast to a random sequence, the fixed sequence was generated by repeating trials from the same provocation category in series of three. Linear mixed models (LMMs) considering aggression trajectories revealed significant effects of provocation (low, medium, high) and trait aggression (K-FAF) on reactive aggression. Men showed significantly higher reactive aggression levels than women. In regard to provocation sequence, we found no significant difference in reactive aggression between the random vs. fixed stimulus sequences. The findings provide new evidence supporting the view that the monetary mTAP is able to induce as well as capture reactive aggression in the laboratory. Additionally, we found no advantage of a fixed sequence as the level of reactive aggression in a given trial appeared to be mainly predicted by the preceding provocation trial.

Applicability of Instability Index for In vitro Protein Stability Prediction

2019 ◽  
Vol 26 (5) ◽  
pp. 339-347 ◽  
Author(s):  
Dilani G. Gamage ◽  
Ajith Gunaratne ◽  
Gopal R. Periyannan ◽  
Timothy G. Russell
Keyword(s):  
Protein Stability ◽  
Caulobacter Crescentus ◽  
Effect Of Temperature ◽  
Instability Index ◽  
Dipeptide Composition ◽  
Experimental Conditions ◽  
The Stability ◽  
Vitro Protein

Background: The dipeptide composition-based Instability Index (II) is one of the protein primary structure-dependent methods available for in vivo protein stability predictions. As per this method, proteins with II value below 40 are stable proteins. Intracellular protein stability principles guided the original development of the II method. However, the use of the II method for in vitro protein stability predictions raises questions about the validity of applying the II method under experimental conditions that are different from the in vivo setting. Objective: The aim of this study is to experimentally test the validity of the use of II as an in vitro protein stability predictor. Methods: A representative protein CCM (CCM - Caulobacter crescentus metalloprotein) that rapidly degrades under in vitro conditions was used to probe the dipeptide sequence-dependent degradation properties of CCM by generating CCM mutants to represent stable and unstable II values. A comparative degradation analysis was carried out under in vitro conditions using wildtype CCM, CCM mutants and two other candidate proteins: metallo-β-lactamase L1 and α -S1- casein representing stable, borderline stable/unstable, and unstable proteins as per the II predictions. The effect of temperature and a protein stabilizing agent on CCM degradation was also tested. Results: Data support the dipeptide composition-dependent protein stability/instability in wt-CCM and mutants as predicted by the II method under in vitro conditions. However, the II failed to accurately represent the stability of other tested proteins. Data indicate the influence of protein environmental factors on the autoproteolysis of proteins. Conclusion: Broader application of the II method for the prediction of protein stability under in vitro conditions is questionable as the stability of the protein may be dependent not only on the intrinsic nature of the protein but also on the conditions of the protein milieu.

scholarly journals Interaction of 4-hydroxynonenal-modified low-density lipoproteins with the fibroblast apolipoprotein B/E receptor

1986 ◽  
Vol 234 (1) ◽  
pp. 245-248 ◽  
Author(s):  
W Jessup ◽  
G Jurgens ◽  
J Lang ◽  
H Esterbauer ◽  
R T Dean
Keyword(s):  
Apolipoprotein B ◽  
Negative Charge ◽  
Low Density Lipoprotein ◽  
Ldl Receptor ◽  
Density Lipoprotein ◽  
Low Density ◽  
Lipid Peroxidation Product ◽  
Modified Low Density Lipoproteins ◽  
Peroxidation Product

The incorporation of the lipid peroxidation product 4-hydroxynonenal into low-density lipoprotein (LDL) increases the negative charge of the particle, and decreases its affinity for the fibroblast LDL receptor. It is suggested that this modification may occur in vivo, and might promote atherogenesis.

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