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Biomedical subjects

K Werdan

Publications and source records attributed to K Werdan.

At least 181 records · Page 10Linked to original sources
PubMed1983

Consequences of specific [3H]ouabain binding to guinea pig left atria and cardiac cell membranes.

An analysis of [3H]ouabain binding to electrically stimulated, contracting guinea pig left atria gave the following results. (1) A non-linear Scatchard plot with at least two binding sites: a high-affinity site (KD 1.1 X 10(-6) M) with about 430 receptors/micron2 related to positive inotropy, and a low-affinity site (KD' 2.1 X 10(-4) M) with about 18,000 receptors/micron2, possibly related to (Na+ + K+)ATPase inhibition. A crude left atrial homogenate gave about 530 receptors/micron2. (2) Half-maximal positive inotropic effects occurred at about 4 X 10(-7) M. (3) 86Rb+-uptake was significantly increased at all inotropic ouabain concentrations (10(-7) - 10(-6) M). Toxic concentrations (above 2 X 10(-6) M) inhibited 86Rb+-uptake (half-maximal inhibition at about 5 X 10(-6) M). [3H]Ouabain binding to partly purified guinea pig cardiac cell membranes showed: (a) linear Scatchard plots for (Mg2+, Pi)- and (Na+, ATP, Mg2+)-supported binding (KD 1.18 X 10(-7) M and 1.49 X 10(-7) M, respectively); (b) non-linear Scatchard plots for (Tyrode + ATP)-supported binding (KD 4.7 X 10(-7) M; KD' 6 X 10(-6) M); and (c) half-maximal [3H]ouabain binding occurred at a lower concentration (about 3.2 X 10(-7) M) than half-maximal inhibition of (Na+ + K+)ATPase activity (about 7.2 X 10(-7) M). Thus, we conclude that there may be more than one type of ouabain binding site in guinea pig left atria, and that measurable inhibition of (Na+ + K+)ATPase is not necessarily related to positive inotropy in the guinea pig.

Animals↗
PubMed1982

Effect of vanadium in the +5, +4 and +3 oxidation states on cardiac force of contraction, adenylate cyclase and (Na+ + K+)-ATPase activity.

The influence of vanadium in the nominally +5 (NH4VO3; referred to as V5+), +4 (C10H14O5V and VOSO4; V4+) and +3 oxidation states (VCl3; V3+) on cardiac force of contraction, adenylate cyclase and (Na+ + K+)-ATPase activity was investigated in order to determine which form of vanadium mediates the cardiac effects. V5+, V4+ and V3+ (300 microM each) increased the force of contraction of isolated electrically driven cat papillary muscles by about 100%. In the presence of the reducing agent ascorbic acid (5 mM) none of the three compounds led to any distinct increase in force of contraction. On the particulate adenylate cyclase preparation from feline right ventricles only V5+ stimulated the enzyme activity by about 100%, whereas V4+ and V3+ were ineffective. In the presence of 5 mM ascorbic acid all three compounds were ineffective. In contrast, in the presence of the oxidizing agent diamide (azodicarboxylic acid-bis-dimethylamide; 1 mM) all three compounds became stimulatory. On the isolated (Na+ + K+)-ATPase V5+ (500 microM) alone reduced the basal activity by about 95%. In the presence of ascorbic acid the inhibitory effect of V5+ was greatly diminished. Similar results were obtained with V4+, V3+ (100 microM) alone inhibited (Na+ + K+)-ATPase activity only by about 40%. In the presence of ascorbic acid V3+ was ineffective. From the results it is concluded that positive inotropism, stimulation of adenylate cyclase and inhibition of (Na+ + K+)-ATPase by vanadium compounds likewise result from an action of vanadium in the +5 oxidation state.

Adenylyl Cyclases↗
PubMed1982

Stimulatory (insulin-mimetic) and inhibitory (ouabain-like) action of vanadate on potassium uptake and cellular sodium and potassium in heart cells in culture.

(1) The influence of vanadate (Na3VO4) on sodium and potassium uptake as well as on cellular ion contents of sodium and potassium has been studied in heart muscle and non-muscle cells obtained from various species. An ouabain-like inhibition of potassium uptake (up to 50%), combined with a decrease of cellular potassium (up to 20%) has been observed by vanadate (10(-4)-10(-3) M) in heart non-muscle cells obtained from neonatal guinea pigs and chick embryos. In heart muscle and non-muscle cells prepared from neonatal rats, as well as in Girardi human heart cells, a vanadate-induced stimulation of potassium uptake (up to 100%), combined with a rise in cellular potassium (up to 20%) and without significant alteration of cellular sodium, has been found. A slight increase of 22Na+ influx can be measured in rat heart muscle cells and in Girardi human heart cells in the presence of vanadate (10(-4)--10(-3) M). (2) In beating rat heart muscle cells in culture, detrimental effects of serum deprivation--concerning beating properties, potassium uptake and cellular potassium--can at least in part be overcome by addition of vanadate. Furthermore, this compound prevents ouabain-induced signs of toxicity (contractures) in these cells. (3) The stimulatory effects of vanadate on potassium can be mimicked by insulin (1-10 mU/ml). Furthermore, vanadate produces an insulin-like stimulation of 2-deoxy-D-glucose uptake in rat heart muscle and non-muscle cells as well as in Girardi human heart cells. (4) The experimental data demonstrate an ouabain-like inhibition as well as an insulin-mimetic stimulation of potassium-uptake in various heart cells. The reason for this antagonistic mode of action may be due to the different capabilities of the heart cell types to reduce vanadium in the V-valence state of vanadium in the IV-valence state, thereby favouring either ouabain-like inhibition (vanadium V) or insulin-mimetic stimulation (vanadium IV) of potassium transport.

Animals↗
PubMed1981

Aging in vitro and D-glucose uptake kinetics of diploid human fibroblasts.

By use of a rapid technique, initial rates of D-glucose transport were obtained during the lifespan in vitro of a commercially available strain of human embryo lung fibroblasts (Flow 2000). The apparent Km of the D-glucose carrier did not change during senescence in vitro: x = 1.8 mM (range 1.3-2.3) in phase II, x = 1.8 mM (range 1.5-2.2) in phase III. Transport rates remained constant in stationary phase II cultures, which had completed between 30% and 80% of their replicative lifespan. A wide variation, however, was observed in terminally differentiated cells (phase III), which showed a two- to threefold increase in average cell size and protein content. In some senescent cultures, glucose transport calculated on a per cell basis was also two- to threefold increased, while it was strongly decreased (-75%) in others. When calculated per unit of cell water, protein, and surface area, respectively, transport rates in phase III cultures ranged from values established for stationary phase II cultures down to very low values. Detaching cells flushed off from senescent cultures did not show measurable rates of glucose transport into the inulin impermeable cell space. Present evidence argues against the idea that an impairment of D-glucose transport might precede loss of replicative potential in aging human fibroblasts. Instead our data indicate that the transport capacity of cell membrane finally decreases during postreplicative senescence in terminally differentiated cells.

Biological Transport↗
PubMed1980

Effects of vanadate in cultured rat heart muscle cells. Vanadate transport, intracellular binding and vanadate-induced changes in beating and in active cation flux.

Cultured rat heart muscle cells have been used to study uptake and intracellular binding of Na483VO4 (vanadate), as well as the influence of vanadate on beating and 86Rb+ uptake of these cells. 1. Vanadate is taken up into cultured rat heart muscle cells in an energy-independent manner by a saturable transport system (Km approximately 60 microM, V approximately 200 pmol per mg protein per min at 37 degrees C). Analysis of intracellular binding of vanadate reveals a curved Scatchard plot indicating more than one binding site. Maximal binding amounts to 3 . 10(9) molecules of vanadate per cell. 2. Vanadate exerts a positive chronotropic and inotropic effect and increases automaticity. First effects can be seen at 1 . 10(-7) M Na3VO4. Concentrations higher than 1. 10(-3) M induce toxic effects (arrhythmias, fibrillation and stand-still of the cell). 3. Vanadate-induced alterations of beating is paralleled by a vanadate-induced stimulation of (86Rb+ + K+) uptake into the cells of up to 75%. Maximal stimulation is obtained at concentrations of 1 . 10(-4)--1 . 10(-3) M vanadate. The stimulation is thought to be due to an increased activity of (Na+ + K+)-ATPase, since it can be inhibited by ouabain. This result is in contrast to in vitro experiments with purified membrane preparations of (Na+ + K+)-ATPase of different organs, where an inhibition of (Na+ + K+)-ATPase by vanadate has been found. 4. The results indicate a possible role of vanadate as an endogenous regulator of active cation flux in heart tissue.

Animals↗
PubMed1980

D-Glucose transport into suspended human fibroblasts. Rapid measurement of uptake by silicone oil filtration centrifugation, and comparison of different cell detachment procedures.

The uptake of 14C-labeled D-glucose into the cellular space of human diploid fibroblasts (Flow 2000)--grown to confluency and detached with trypsin-EDTA--was studied using silicone-oil-layerfiltering centrifugation. This method is rapid enough to enable the determination of initial transport rates, which are not complicated by subsequent metabolism of the hexose taken up into the cells. D-Glucose uptake shows saturation kinetics with a Km of 1.8mM and maximal transport capacity of 4--8 nmol/(10(6) cells x min) at 20 degrees C. This saturable transport system is responsible for at least 80% of the total glucose taken up into the cells in the concentration range tested (0.1--10mM D-glucose in incubation medium). The glucose carrier is stereo-specific, is independent of sodium and potassium ions, and is inhibited by cytochalasin B. Its temperature dependence reveals an activation energy of 3 1 kJ/mol (7.5 kcal/mol; Q10 approximately equal to 1.5). As detachment of the cells from the culture flasks is necessary for applying silicone-layer-filtering centrifugation, various detachment procedures were tested. In the enzymatic procedure cells were treated with either trypsin or pronase. In the chelating method, Ca2+ and Mg2+ ions were chelated by EDTA and K+ ions with sodium tetraphenylborate. For mechanical detachment, cells were grown initially on plastic foil. After each of these detachment procedures the transport of D-glucose was the same. It is therefore concluded that this method of rapid measurement of D-glucose uptake in suspended human fibroblasts may serve as an alternative to the uptake measurement with glucose analogues in attached cells when studying the hexose transport system in human diploid fibroblasts.

Biological Transport, Active↗
PubMed1979

[Investigations on combined injuries, 26th communication: investigations on the changes in the state of resistance following sublethal whole-body irradiation (author's transl)].

The risk of a bacterial invasion by pseudomonas, e. coli, proteus, citrobacter and gaffkya via an open skin wound was investigated in combined injured mice (infliction of a skin wound and irradiations). Suspensions of different bacterial strains were applied on the wound and then the lethality was determined. Besides of apathogenic (citrobacter, proteus, gaffkya) and high pathogenic bacteria (e. coli and pseudomonas) an e. coli strain of the 0-Group 81 was found that seems to induce a decrease of lethality even after sublethal whole body irradiation.

Animals↗
PubMed1979

[Investigations on combined injuries, 24th communication: response of blood circulation and respiratory function in mice upon whom was inflicted whole-body irradiation in combination with open skin wounds (author's transl)].

Combined injuries were inflicted upon NMRI-mice, hurting each with an open skin wound subsequent to sublethal exposition to X-rays. Lethality among animals with combined injuries was between 40 and 60%, while animals only irradiated and those with only a skin wound had lethalities between 10 and 20% and 0%, respectively. Blood circulation and respiration of animals injured with combined lesions were studied in an attempt to understand the cause of the high lethality in this group. The blood volume and the oxygen transport capacity were drastically reduced in animals with combined injuries as compared to those in animals only irradiated, although plasma volume, vascular permeability and distribution of the blood volume remained similar in both groups. Analyses of gases and acid/base composition of blood revealed neither respiratory nor metabolic acidosis. These findings indicate that combined injuries hardly impair blood circulation and respiratory function.

Acid-Base Equilibrium↗
PubMed1979

[Investigations on combined injuries, 25th communication: renal function and energy metabolism in mice with injuries combined of whole-body irradiation and an open skin wound (author's transl)].

In NMRI-mice injured with combined lesions (whole-body irradiation with 500 R and, two or seven days later, an open skin wound), lethality increased from 10 or 20% following sole irradiation up to 60%. Investigations of urinary excretion and serum levels in animals with combined injuries did not show undue elevations of the levels of electrolytes, creatinine, urea, uric acid and acid or alkaline equivalents. Analysis of lactate, pyruvate, ATP and ADP in liver tissue and blood serum indicated no significant changes in the energy metabolism in animals with combined injuries in comparison with mice irradiated only. These findings and those of our last communication (24. Mitteilung) seem to be incompatible with the presence of a shock state as cause of the high lethality of mice with combined injuries.

Animals↗
PubMed1978

Specific transport of inorganic phosphate, 3-phosphoglycerate and triosephosphates across the inner membrane of the envelope in spinach chloroplasts.

The uptake of phosphate and phosphorylated compounds into the chloroplast stroma has been studied by silicone layer filtering centrifugation. 1. Inorganic phosphate, 3-phosphoglycerate, dihydroxyacetone phosphate and glyceraldehyde phosphate are transported across the envelope leading to an accumulation in the chloroplast stroma. This uptake proceeds by a counter exchange with phosphate and phosphorylated compounds present there. 2. The transport shows saturation characteristics allowing the determination of Km and V. 3. The phosphorylated compounds transported act as competitive inhibitors of the transport. From measurements of the Km and Ki the specificity of the transport is described. 4. The transport of inorganic phosphate and 3-phosphoglycerate is inhibited by p-chloromercuriphenyl sulfonate, pyridoxal 5'-phosphate and trinitrobenzene sulfonate. 5. The activation energy of phosphate transport as determined from the temperature dependence is evaluated to be 16 kcal (0--12 degrees C). 6. It is concluded that inorganic phosphate, 3-phosphoglycerate, dihydroxy-acetone phosphate and glyceraldehyde phosphate are transported by the same carrier, which has been nominated phosphate translocator. 7. Simultaneous measurements of the proton concentration in the medium and the transport into the chloroplasts show that the transfer of 3-phosphoglycerate involves a transfer of a proton into the same direction. 8. Measurements of the pH dependence of the transport indicate that all substances including 3-phosphoglycerate are transported by the phosphate translocator as divalent anions. 9. The physiological function of the phosphate translocator is discussed.

Biological Transport↗