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Biomedical subjects

M Wolgast

Publications and source records attributed to M Wolgast.

At least 37 records · Page 2Linked to original sources
PubMed1990

Nephron function in the early phase of ischemic renal failure. Significance of erythrocyte trapping.

Trapping of red blood cells (RBCs) in renal medulla vasculature in postischemic acute renal failure (ARF) was found to depend upon the length of the ischemic period. Thus trapping occurred after 45 minutes but not 25 minutes of ischemia. By prior hemodilution to a hematocrit (hct) of 30%, RBC trapping after 45 minutes of ischemia could be completely prevented. Likewise hemo-concentration (hct = 60%) before 25 minutes of ischemia resulted in extensive RBC trapping. By increasing or decreasing the hct, the contribution of RBC trapping to the functional defects and decrease in renal blood flow that follows minor (25 min) and more substantial (45 min) ischemia was investigated. Renal blood flow (RBF) was measured by microspheres, and vascular and tubular pressure by the micropuncture technique. Glomerular filtration rate (GFR) was estimated from inulin clearance, and tubular function from urine osmolality and sodium and potassium excretion. It was found that postischemic RBF was not correlated to RBC trapping but depended on the length of ischemia. After both 25 and 45 minutes of ischemia tubular obstructions occurred in the proximal tubules and/or loops of Henle, causing an increase in proximal tubular pressure. These obstructions were dependent on the length of ischemia but not on RBC trapping. After hemoconcentration and 25 minutes of ischemia there was an increment in distal tubular pressure, indicating that abundant RBC trapping may contribute to an increase in tubular pressure by compression of medullary tubules and thereby reduce GFR. When the damage was more severe other factors came into play and the contribution of RBC trapping to the decrease in GFR was minimal.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Kidney Injury↗
PubMed1990

Mechanism of erythrocyte trapping in ischaemic acute renal failure.

Forty-five minutes of warm ischaemia and 20 min of recirculation in the rat kidney was found to result in (1) a massive transient extravasation of plasma upon recirculation and (2) an increase in plasma-lymph transport of proteins during the first hours after onset of circulation. This was accompanied by trapping of erythrocytes, as determined with 51Cr-labelled erythrocytes, in the capillaries, mainly in the inner stripe of the outer medulla. At scanning electron microscopy of vibratome sections, the trapping appeared as aggregates of polygonally shaped erythrocytes. It is concluded that 45 min of ischaemia and 20 min of recirculation results in an increase in the permeability of the renal capillaries. This increase leads to extravasation of capillary plasma with consequent local haemoconcentration, causing an increase in vascular resistance and in capillary hydrostatic pressure. This elevated pressure will, in turn, lead to perpetuating extravasation of plasma, further haemoconcentration and so on, eventually resulting in dense packing of polygonal erythrocytes, obstructing the blood flow. It is believed that oxygen-derived free radicals generated in the early recirculation phase contribute to the increase in macromolecular permeability, since the scavenger bovine superoxide dismutase and allopurinol, a xanthine oxidase inhibitor, were found to prevent this unfavourable chain of events.

Acute Kidney Injury↗
PubMed1990

The long-term outcome of post-ischaemic acute renal failure in the rat. I. A functional study after treatment with SOD and sucrose.

The long-term outcome in rat kidneys subjected to 45 min of warm ischaemia with no treatment and after administration of 20 mg superoxide dismutase (SOD) and of SOD combined with 2 ml of a 12% sucrose solution was studied by the micropuncture technique. It was found that, although in the acute phase SOD prevented trapping of erythrocytes in the medullary vasculature and that SOD + sucrose also prevented the formation of tubular obstruction, the long-term results as studied 1 week and 1 month after the primary ischaemic insult were virtually identical. This was due to the formation of new obstructions, during the first week, mainly in the thick ascending limb of Henle's loop. After 1 month the proximal tubular free-flow pressure and the single-nephron filtration rate had returned to normal. However, since the total glomerular filtration was only one-third of that under normal conditions, it would seem that two-thirds of the nephrons had undergone complete degeneration. This degeneration was probably the result of persistent tubular obstruction. The tubular degeneration was also accompanied by a reduction in urine osmolality and potassium secretion.

Acute Kidney Injury↗
PubMed1990

The long-term outcome of post-ischaemic acute renal failure in the rat. II. A histopathological study of the untreated kidney.

Histopathological changes in kidneys subjected to 45 min of isothermic unilateral ischaemia in the acute phase and 1 week and 1 month after primary damage were studied at the electron microscopic level. During the first week after recirculation long homogeneous cylinders, probably consisting of Tamm-Horsfall protein, developed in the medullary parts of the nephron, and after 1 month of recirculation there were two types of nephrons: (1) nephrons with a normal histological appearance and (2) degenerated nephrons. The latter group gave rise to crypts in the outer cortical area. It is hypothesized that the generation of the long Tamm-Horsfall cylinders in the thick ascending limb of the loop of Henle plays an important role in the long-term outcome of the kidney after the primary damage. The persistent blockade caused by these cylinders will lead to precipitation of the ultrafiltrate, resulting in long cell-protein cylinders in the proximal parts of the nephron. This precipitation will proceed in the retrograde direction, reaching the mother glomeruli, eventually leading to total degeneration of the nephron.

Acute Kidney Injury↗
PubMed1990

The effect of loop diuretics on the long-term outcome of post-ischaemic acute renal failure in the rat.

The effects of continuous treatment with loop-acting diuretics on the long-term functional and histopathological outcome in kidneys subjected to 45 min of warm ischaemia were studied. One month after the primary damage the inulin clearance in the untreated kidneys was 0.44 +/- 0.05 ml min-1, improving significantly to 0.69 +/- 0.11 ml min-1 in furosemide-treated animals and to 0.75 +/- 0.09 ml min-1 in those treated with piretanide. Urine osmolality increased from 986 +/- 89 mosmol kg-1 in the untreated animals to 1479 +/- 195 mosmol kg-1 in the furosemide-treated ones. At the same time the total area of the outer medulla occupied by Tamm-Horsfall protein cylinders decreased from 7.0 +/- 1.2% in the untreated animals to 3.6 +/- 0.52% in the treated ones. It is concluded that by decreasing the number of nephrons blocked by Tamm-Horsfall cylinders an improvement in the function of ischaemically damaged kidneys can be achieved. This blockade, also called secondary damage, is of critical prognostic importance for the long-term outcome of the ischaemic renal failure. Treatment of the animals with loop diuretics decreased the occurrence of these cylinders, leading to an improvement of kidney function I month after the primary damage, this despite the fact that the primary damage seen in the early recirculation period was not treated specifically.

Acute Kidney Injury↗
PubMed1990

Peritubular capillary permeability and intravascular RBC aggregation after ischemia: effects of neutrophils.

The influence of neutrophils on peritubular capillary permeability and intravascular red blood cell (RBC) aggregation after renal ischemia was studied in anesthetized Sprague-Dawley rats. Intraperitoneal administration of antineutrophil serum (ANS) reduced the number of neutrophils in the blood to 3% of normal. The control group received an equal volume of inactive serum. Renal macromolecular capillary permeability was studied from 1) extravasation of albumin and 2) plasma to lymph transport of plasma proteins and of neutral and negatively charged lactate dehydrogenase (LDH). The net driving force (NDF) for fluid transfer over the peritubular capillary membrane was determined by the micropuncture technique. The intrarenal distributions of neutrophils and RBC were measured by a histochemical method and 51Cr-labeled RBC, respectively. Under preischemic control conditions neither macromolecular permeability nor renal clearance of inulin was affected by ANS. However, the steep increase in the macromolecular transport from plasma to lymph resulting from 45 min of ischemia and reperfusion was blunted by ANS, and preischemic control values were restored after 1 h of recirculation. In the control group the mass transport of plasma proteins increased twofold and that of both neutral and negatively charged LDH fourfold. NDF was equal in the two groups. In the ANS-treated animals the intrarenal neutrophil content was only 2% of the control. Neutrophils were found mainly in the cortex, whereas RBC aggregation was observed only in the renal medulla. It is concluded that neutrophils mediate postischemic capillary leakage. It is suggested that this leakage underlies RBC aggregation and incomplete return of blood flow in the renal medulla after ischemia.

Animals↗
PubMed1989

The net electric charge of proteins. A comparison of determinations by Donnan potential measurements and by gel electrophoresis.

We compare a new method for the determination of the net charge of proteins based on Donnan potential measurements, as described briefly by Ojteg, G., Nygren, K. and Wolgast, M. (1987) Acta Physiol. Scand. 129, 277-286, with a conventional method using polyacrylamide gel electrophoresis. The new technique utilizes the Donnan potential, which develops over a semipermeable membrane that separates the non-permeating protein from the surrounding bath of the same ionic composition as the protein solution, to determine the net valency. The advantages of this method, besides its simplicity, are that it can determine the charge of, e.g., a protein in a free-fluid phase and that the pH and ionic composition of the bathing fluid can be varied over a broad range. The Donnan potential decreased to half its original value when the ionic strength was doubled. Usually a protein concentration of 1-10 mg.ml-1 must be used. The Donnan potential method was applied to determine the net charges of a series of proteins with different isoelectric points. The values showed close agreement with the data obtained by gel electrophoresis.

Animals↗
PubMed1988

Extensive tubular secretion and reabsorption of creatinine in humans.

The validity of creatinine as a marker for the glomerular filtration rate was studied in 8 healthy volunteers in different stages of hydration and during large variations in urinary flow rates. The urine flow was 8.4 ml/min in a dehydrated state (due to furosemide infusion; 8 mg/h) and raised to 23.2 ml/min after rapid rehydration. The creatinine to inulin clearance ratio changed considerably from 1.47 in rehydrated state, indicating a substantial tubular secretion of creatinine, to 1.05 in dehydrated state, indicating a reabsorption of creatinine almost equal to secretion. Thus, substantial tubular secretion and reabsorption of creatinine, changing in relative importance in relation to the degree of hydration, make creatinine clearance an unreliable marker for the glomerular filtration rate.

Blood Urea Nitrogen↗
PubMed1988

Tissue distribution of 125I-labelled bovine superoxide dismutase (SOD) in the rat.

Bovine copper/zinc superoxide dismutase (SOD) was labelled with 125I using the chloramine-T method. The tissue distribution of 125I-SOD (dose of SOD 5 mg/kg) was studied with whole-body and microautoradiography at various times after an intravenous injection. The distribution of 125I-SOD showed a remarkable organ specificity in that the localization of the enzyme to the kidneys and the urinary tract completely dominated the autoradiograms. The time pattern of localization of 125I-SOD also gives a clear picture of the renal handling of the enzyme in that, as a consequence of the renal elimination, the enzyme rapidly disappears from the circulation with an elimination half time of about 6 min. Up to 20 min. after the injection, there were high concentrations of 125I-SOD in the renal pelvis, ureter and urinary bladder showing that in addition to renal uptake there was an initial substantial urinary excretion of the enzyme. From the microautoradiography it is clear that the grains were exclusively localized over proximal tubular cells and tended to be concentrated at the luminal rather than the peritubular side of tubule. This would be compatible with renal uptake secondary to glomerular filtration of 125I-SOD, which is what one would expect from the renal handling of a protein with a molecular weight around 31,000 and an isoelectric point around pH 5.4. Pretreatment with a large dose of SOD (88 mg/kg) tended to competitively decrease the renal uptake of labelled SOD after 5 min. and apparently further increase its renal excretion. However, a noticeable renal uptake of 125I-SOD was still apparent.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Permeability of renal capillaries. III. Theoretical analysis of hydraulic conductivity, pore structure and electric properties.

The functional characteristics of the capillary membranes of rat kidneys were analysed in a computer-based model utilizing previous data on the hydraulic conductivity of the membranes and transport by diffusion, hydraulic flow and ion migration of: inulin, myoglobin (negative, neutral and positive), horseradish peroxidase (negative and neutral), lactate dehydrogenase (negative, neutral and positive) and albumin. The results showed that in the peritubular capillaries the main fluid reabsorption (26.8 x 10(-8) ml nephron-1 100 g body wt-1) occurs through a set of 20 A pores with a total pore area over pore length of 30 cm x (nephron 100 g body wt)-1, whereas the plasma proteins enter the renal interstitium through a few 140 A large pores with a total pore area over pore length of 7.2 x 10(-4) cm and a fluid reabsorption of 1.2 x 10(-10) ml nephron-1 100 g body wt-1. The intramembranous concentration of negative fixed charges in the large pore system was estimated at 20 mM (range 15-22 mM). Since this was accompanied by a potential difference of 0.3 mV, a net driving electro-osmotic force, favouring reabsorption, of 4 mmHg developed. The charges and the electro-osmotic force was found to be essential for protein transport, since if the membrane was uncharged, the transport and interstitial concentration of e.g. albumin was twice as high. The glomerular capillaries seem to have a more homogeneous structure, that is in essence a one-pore system; the pore radius was 44 A, the total pore area over pore length 2.8 cm and the concentration of negative fixed charges 40 mM (range 37-43 mM).

Animals↗
PubMed1988

Plasma elimination kinetics and renal handling of copper/zinc superoxide dismutase in the rat.

The renal handling of bovine and human superoxide dismutase (SOD) was investigated in Sprague-Dawley and Munich-Wistar rats. Under normal physiological conditions the half-time of the major rapid component of the plasma elimination curve was estimated at 6.0 +/- 0.5 min, the volume of distribution at 35.7 +/- 3.3 ml kg-1, i.e. the plasma volume and the corresponding plasma clearance at 4 ml min-1 kg-1. After a single intravenous dose, most of the enzyme was distributed to and eliminated by the two kidneys, whereas the non-renal clearance was low, 0.5 ml min-1 kg-1. The single nephron filtration of SOD, as assessed from micropuncture of Bowman's space, was 10.4 +/- 1.0 nl min-1, which was 26 +/- 2% of that for inulin. The total elimination of SOD by glomerular filtration would thereby be 2.5 ml min-1 kg-1, i.e. the glomerular ultrafiltration process would account for the largest part of the elimination of SOD from circulating plasma. After the dosage of 20 mg kg-1, about two-thirds of the injected SOD was excreted as the intact molecule into the urine, whereas one-third was found to be reabsorbed and metabolized by the proximal tubular epithelial cells. In animals suffering from unilateral post-ischaemic acute renal failure, the elimination half-time was 13 +/- 1.9 min, a value which increased by 55 +/- 5 min after bilateral functional nephrectomy. By contrast, the distribution volume remained essentially unchanged.

Acute Kidney Injury↗
PubMed1988

Electrophysiology of renal capillary membranes: gel concept applied and Starling model challenged.

In the classical Starling model the hydrostatic pressure in the pores is generally lower than that in capillary plasma, a phenomenon that necessitates the assumption of a rigid porous membrane. In flexible gel membranes, the capillary pressure is suggested to be balanced by a gel swelling pressure generated by negative fixed charges. Regarding the fluid transfer, the transmembranous electrical potential gradient will generate a net driving electroosmotic force. This force will be numerically similar to the net driving Starling force in small pores, but distinctly different in large pores. From previous data on the hydrostatic and colloid osmotic forces, the fixed charge density at the two interfaces of 1) the glomerular and 2) the peritubular capillary membrane were calculated and used to predict the flux of a series of charged protein probes. The close fit to the experimental data in both the capillary beds is in line with the gel concept presented. The gel concept (but hardly a rigid membrane) explains the ability of capillary membranes to alter their permeability in response to external forces. Gel membranes can furthermore be predicted to have a self-rinsing ability, as entrapped proteins will increase the local fixed charge density, leading to fluid entry into the region between the particle and the pore rim, which by consequent widening of the channel will facilitate extrusion of trapped proteins.

Animals↗
PubMed1987

Permeability of renal capillaries. I. Preparation of neutral and charged protein probes.

This paper describes the preparation of charged and uncharged protein molecular probes for study of the permselectivity of renal capillaries. Horse heart myoglobin was used as a neutral myoglobin. Since it contained several fractions with different isoelectric points, it was purified by fast protein liquid chromatography (FPLC). To obtain a negatively charged myoglobin, the original horse heart myoglobin was treated with cyanate, resulting in net charge of -5.7 +/- 0.3 at physiological pH (mean +/- SEM). The charge was determined from the Donnan potential which develops over a semipermeable membrane separating the inside solution in which the protein was dissolved from a surrounding bath of equal ionic strength. Sperm whale myoglobin was similarly purified by FPLC and used as a positively (+1.7 +/- 0.2) charged isomer. Horseradish peroxidase (HRP) was purified by means of gel and ion-exchange chromatography and found to be neutral at physiological pH. Negatively charged (-14.0 +/- 0.5) HRP was obtained by succinylation. Two isomers of lactate dehyrogenase (LDH) were used, namely the slightly positive (+2) LDH-M4 and the strongly negative (-19) LDH-H4. These isomers, which occur naturally, did not require further purification. The Stokes-Einstein radii, as measured by gel chromatography, of inulin, myoglobin, HRP and LDH were 11, 17.5, 32 and 46 A, respectively. The chemical modifications did not alter the Stokes-Einstein radii. In biological studies on rat kidneys samples of both plasma and renal hilar lymph were found to contain radioactive low molecular weight degradation products in addition to the intact proteins. This necessitated separation of all individual samples on small Sephadex columns prior to analysis.

Animals↗
PubMed1987

Permeability of renal capillaries. II. Transport of neutral and charged protein molecular probes.

The permselectivity of the renal capillaries was investigated from the transport of a series of molecular probes: inulin, positive (+2, net charge at pH 7.4), neutral (0), and negative (-6) myoglobin, neutral (0) and negative (-14) horseradish peroxidase (HRP) and two isomers of lactate dehydrogenase (LDH), namely the positively charged (+2) LDH-M4 and the negatively charged (-19) LDH-H4. The determination of the concentration of tracer proteins necessitated gel separation of both plasma and renal hilar lymph. The reason for this is that the proteins, after filtration, will be reabsorbed and degraded by the proximal tubular cells into small molecular compounds (amino acids), which will return to both the renal interstitium and systemic plasma. Even if this degradation is of low degree, as for high-molecular-weight proteins, separation is still required, since the relative lymph concentration (plasma concentration put at 1) is also low, that is, even small amounts of low molecular compounds will distort the relative lymph concentration obtained. The transport from plasma to renal hilar lymph of the tracer molecules fell with increasing Stokes-Einstein radius. The relative lymph concentration of the 11 A inulin was 1.06 +/- 0.03 (mean +/- SEM), of the neutral 17.5 A myoglobin 0.76 +/- 0.05, of the neutral 32 A HRP 0.32 +/- 0.02 and of the neutral 46 A LDH 0.12 +/- 0.01. The data are compatible with a two-pore system. The negative tracer molecules were in general proportionally more restricted than the neutral (or positive) moieties (P less than 0.001) thus suggesting a negatively charged peritubular capillary membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Renal capillary permeability and intravascular red cell aggregation after ischaemia. I. Effects of xanthine oxidase activity.

The macromolecular permeability of renal capillaries and the intravascular red cell aggregation resulting from 45 min of warm ischaemia were investigated. The effects of the xanthine oxidase inhibitor Allopurinol on these factors and also on the post-ischaemic nephron function were also studied. Following ischaemia there was a more than 10-fold increase in the transport from plasma to renal hilar lymph both of plasma proteins and of two isomers of lactate dehydrogenase (LDH)-the nearly neutral LDH-M4 and the negatively charged LDH-H4. The ischaemia also resulted in massive intravascular red cell aggregation, especially in the renal medulla. Through reduction of plasma xanthine oxidase activity from 13.1 +/- 1.1 microU microliter-1 (mean +/- SEM) to essentially zero by Allopurinol, the capillary leakiness was substantially diminished with almost complete normalization after 120 min. At the same time the relative volume of trapped red cells was reduced; in the inner stripe of the outer medulla, for example, it decreased from 11.3 +/- 1.7% in untreated animals to 4.0 +/- 1.1% after treatment with 20 mg of Allopurinol given intravenously 3 h before the ischaemia. Oral feeding with 4 mg of Allopurinol day-1 for one week gave essentially the same result. The net driving force for filtration after treatment with this drug was thus 19 mmHg, as against 26 mmHg in the normal kidney and the resulting SNGFR was half the normal. The total filtration rate was proportionally more reduced to less than 1/3 of the normal. Tubular obstruction was still present but was not as severe as in untreated kidneys (Karlberg et al., 1982b) where the tubular fluid flow and thereby the filtration are essentially zero. It is suggested that oxygen free radicals increased the macromolecular permeability and the adhesiveness of white blood cells and that these two factors combined underlie the aggregation of red blood cells in the medullary vasa recta with consequent persistence of medullary ischaemia.

Allopurinol↗
PubMed1987

Prevention of ischaemic acute renal failure with superoxide dismutase and sucrose.

The preventive effects of intravenously administered superoxide dismutase (SOD) and of SOD combined with sucrose on acute renal failure were investigated in rat kidneys exposed to 45 min of warm ischaemia. Superoxide dismutase (20 mg) given just before primary ischaemia and in the early recirculation phase was found to ameliorate the red cell aggregation in the renal medulla, in particular, in the inner stripe of the outer zone the volume of trapped red cells decreased from 11.2 +/- 1.6% in untreated animals to 0.02 +/- 0.001%, thus allowing improved restoration of medullary blood flow. This was also accompanied by an expected restoration of the urine osmolality reaching almost 400 mOsm kg-1 after administration of SOD + sucrose. Superoxide dismutase also restored the capillary macromolecular permeability as evidenced by normalization of plasma to lymph transport of proteins. Micropuncture studies showed that in ischaemically damaged but untreated kidneys the tubules were obstructed and that the proximal tubular pressure rose to such a level that the net driving force for filtration approached zero. This explains the marked decrease in glomerular filtration rate (GFR) from a normal value of about 1 ml min-1 to 0.01 +/- 0.02 ml min-1. After treatment with SOD the tubules were still largely obstructed, resulting in a depression of the net driving force and a decrease in single nephron glomerular filtration rate (SNGFR) to about 11 nl min-1, that is, to only 25% of the normal SNGFR. The total filtration was 0.09 +/- 0.04 ml min-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Kidney Injury↗