Effect of heparin on the lipid peroxidation reaction of erythrocytes and their stability

Physiological concentration (10 units/ml) of heparin activates ascorbate-dependent lipid peroxidation of erythrocytes and reduces their stability in the citrate-phosphate buffer (pH 3.0). In a concentration of 100 units/ml heparin does not influence the thermal (62 degrees) oxidation of methyloleate. It follows that heparin is not direct prooxidant.     

Effect of sub-inhibitory concentrations of antibiotics on the hemolytic activity of Legionella

An in vitro test for the antiproliferative effect of human leukocyte interferon (IFN-alpha) was performed in primary cultures of tumor cells obtained from 32 patients with either malignant melanoma (13), renal carcinoma (4) or bladder carcinoma (15). Our results demonstrated activity of IFN in all three groups of solid tumors. However, appreciable differences in sensitivity to antiproliferative effect of IFN between individual tumors of the same type were found. The potential of this antiproliferative test for prediction of treatment response in IFN-therapy is discussed.     

Effect of some pesticides/weedicides on cathepsin B activity and lysosomal membrane

An asymptomatic 9-year-old boy was found to have a cardiac murmur in a heart disease survey. Two-dimensional echocardiogram identified a large left ventricular pseudoaneurysm. Color flow imaging showed a transtunnel "to-and-fro" mosaic pattern between the pseudoaneurysm and left ventricle. ECG-gated magnetic resonance imaging demonstrated the exact size and related location of the left ventricular pseudoaneurysm and its ostium. Without cardiac catheterization or angiography, he was successfully operated on by closure of the communicating tunnel.     

In vitro influence of phenylalanine on acetylcholinesterase activity and DNA

Acetylcholinesterase (AChE) is a significant component of the membrane contributing to the permeability changes during synaptic transmission and conduction. Phenylketonuria is a group of metabolic disorders in which phenylalanine (Phe) is highly elevated in blood (up to 0.1 M) resulting in mental retardation etc. AChE activity was measured spectrophotometrically after incubation with various Phe concentrations. Phe interaction with DNA was evaluated with an established HPLC method. Phe was found to inhibit AChE almost 40%, at a concentration of 5 mM, whereas a 62.5% DNA peak exclusion (molecular interaction) was observed when Phe was incubated with DNA at a concentration of 3 mM. In addition the ratio of DNA: Phe determined the potency of the observed molecular effect.     

A mutant toxin of Vibrio parahaemolyticus thermostable direct hemolysin which has lost hemolytic activity but retains ability to bind to erythrocytes

A mutant toxin, R7, of thermostable direct hemolysin (TDH) with a single amino acid substitution at glycine 62 was analyzed. The hemolytic activity of R7 decreased to less than 1/1,000 of that of wild-type TDH, and its mouse lethality was undetectable. This mutant toxin, however, showed a marked inhibitory effect on hemolysis by wild-type TDH. Enzyme immunoassay and flow cytometric analysis demonstrated that R7 retained approximately 50% of the ability to bind to erythrocytes compared with that of wild-type TDH, suggesting that its inhibition of hemolysis by wild-type TDH might be due to blocking the binding sites on the erythrocyte membrane. Wild-type TDH affected the erythrocyte membrane by causing an influx of calcium and propidium iodide, while R7 showed no detectable effects of these kinds. These results suggest that hemolysis by TDH consists of at least two steps, binding and postbinding, and that R7 is likely to be a postbinding activity-deficient mutant toxin of TDH.     

Effect of hydration on the water content of human erythrocytes

An ideal, hydrated, nondilute pseudobinary salt-protein-water solution model of the RBC intracellular solution has been developed to describe the osmotic behavior of human erythrocytes during freezing and thawing. Because of the hydration of intracellular solutes (mostly cell proteins), our analytical results predict that at least 16.65% of the isotonic cell water content will be retained within RBCs placed in hypertonic solutions. These findings are consistent not only with the experimental measurements of the amount of isotonic cell water retained within RBCs subjected to nonisotonic extracellular solutions (20-32%) but also with the experimental evidence that all of the water within RBCs is solvent water. By modeling the RBC intracellular solution as a hydrated salt-protein-water solution, no anomalous osmotic behavior is apparent.     

Effect of Glycine max lectin on the interaction of prothrombin and erythrocytes

Native porcine erythrocytes do not initiate blood coagulation, though even the weak association (Kd = 4,25 +/- 9,35 microM) of prothrombin with their surface which is limited to the projection of two phospholipid polar head groups onto the external cell membrane, exerts a slight but authentic influence on the acceleration of blood clotting. There are 11.9 x 10(7) sites for 125I-prothrombin on one erythrocytes. At physiological concentration of prothrombin the surface of erythrocytes is far from saturation. The binding of 125I-labelled prothrombin to erythrocytes after their surface was covered by lectin Glycine max by means of the accessible residues of N-acetyl-D-galactosamine and D-galactose, glycoproteins and dlycolipids can never become saturated with any used concentration of the ligand. In the presence of the suspension of erythrocytes treated lectin platelet free plasma coagulates at the same speed as the control plasma.     

Allosteric control of acetylcholinesterase activity by monoclonal antibodies

The aim of this study was to assess the effect of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ)-induced alpha2-adrenoceptor inactivation on regulatory G proteins and the recovery of agonist and antagonist binding sites. EEDQ induced a rapid increase in the abundance of rat brain cortical Galphai1/2 proteins (30% at 6 h) which reached a maximum at 4 days (45%) and which then slowly returned (7-30 days) to control values. EEDQ did not alter significantly the levels of Galphai3 and Galphao proteins. By using the standard monoexponential model, the analysis of the recovery of alpha2-adrenoceptor density (6 h-30 days) with [3H]UK 14304 (bromoxidine) and [3H]RX 821002 (2-metoxy idazoxan) in the cerebral cortex did not reveal differences in receptor turnover parameters. However, the recovery of [3H]UK 14304 binding fitted best to a new biphasic recovery model, suggesting the existence of two distinct phases of recovery of agonist sites (r1 and r2 = 15.7 and 7.4 fmol mg protein(-1) day(-1); k1 and k2 = 0.51 and 0.25 day(-1); (t1/2)1 and (t1/2)2 = 1.4 and 2.7 days). In contrast, the recovery of [3H]RX 821002 antagonist sites did not fit to the biphasic model (r = 8.1, k = 0.14, t1/2 = 4.9). Because agonist binding requires coupling to G proteins, the present results suggest that the rapid over-expression of Galphai1/2 proteins induced by EEDQ is related to the biphasic recovery of [3H]UK 14304 binding. The possible implication of the faster recovery of alpha2-adrenoceptor function after EEDQ inactivation is discussed.     

Morphometry and acetylcholinesterase activity of the myenteric plexus of the wild mouse Calomys callosus

The myenteric plexus of the digestive tract of the wild mouse Calomys callosus was examined using a histochemical method that selectively stains nerve cells, and the acetylcholinesterase (AChE) histochemical technique in whole-mount preparations. Neuronal density was 1,500 +/- 116 neurons/cm2 (mean +/- SEM) in the esophagus, 8,900 +/- 1,518 in the stomach, 9,000 +/- 711 in the jejunum and 13,100 +/- 2,089 in the colon. The difference in neuronal density between the esophagus and other regions was statistically significant. The neuron profile area ranged from 45 to 1,100 microns2. The difference in nerve cell size between the jejunum and other regions was statistically significant. AChE-positive nerve fibers were distributed within the myenteric plexus which is formed by a primary meshwork of large nerve bundles and a secondary meshwork of finer nerve bundles. Most of the nerve cells displayed AChE activity in the cytoplasm of different reaction intensities. These results are important in order to understand the changes occurring in the myenteric plexus in experimental Chagas' disease.     

Light and electron microscopic localization of acetylcholinesterase activity in the rat renal nerves

Acetylcholinesterase activity is shown in the renal nerves of the rat with the technique of Karnovsky and Roots. By light microscopy, the acetylcholinesterase-positive nerves are seen in association with blood vessels, including the glomerular arterioles, and occasionally with renal tubules. By electron microscopy the precipitate appears extracellularly around axons and varicosities. DFP inhibits the deposition of precipitate. Previous demonstration by serial section electron microscopy in the rat revealed that all nerves around the glomerular arterioles contain small dense-cored vesicles characteristic of adrenergic nerves, indicating that the acetylcholinesterase-positive nerves demonstrated here are likely to be adrenergic nerves containing acetylcholinesterase.     

Adenosine deaminase activity in human erythrocytes and lymphocytes

A radiochromatographic method is described for measuring enzymatic activity of adenosine deaminase in human erythrocytes and lymphocytes. [8-14C]-adenosine is converted into inosine and hypoxanthine; after chromatographic separation of the products, the radioactivity is determined. The kinetic properties of the enzyme have been studied. The Km values for the erythrocyte and lymphocyte enzymes are higher as compared with purified deaminase. Optimum conditions for substrate concentration for assay were established. The mean normal activity (+/- S.E. of mean) is: for erythrocytes, 494 +/- 61; nmol min-1 ml-1; for lymphocytes- 147 +/- 0.18 nmol min-1 10(6) cellules. The mean values are higher than that given by other methods working at a lower (non-staurating) substrate concentration.     

The role of acetylcholine receptors and acetylcholinesterase activity in the development of denervation supersensitivity

Strips of muscle from innervated and denervated rat hemidiaphragm were tested for sensitivity to acetylcholine and to carbachol. For both agonists, denervation (6-8 days) produced notable supersensitivity. However, the increase in sensitivity to acetylcholine (ca. 600-fold) was much greater than that to carbachol (ca. 51-fold). Denervation also produced an increase in [3H]alpha-bungarotoxin binding (ca. 20-fold), presumably indicative of an increase in the number of acetylcholine receptors. In addition to causing increases in tissue sensitivity and receptor number, denervation caused a marked loss of acetylcholinesterase activity (ca. 70%) and a modest loss of butyrylcholinesterase activity (ca. 20%). When innervated muscle was pretreated with eserine (5 X 10(-5) M), there was a loss of acetylcholinesterase activity (ca. 86%) and butyrylcholinesterase activity (ca. 36%). Simultaneously, there was an increase in tissue sensitivity to acetylcholine (ca. 26-fold). When denervated muscle was pretreated with eserine, there was no loss of enzyme activity beyond that caused by denervation. Furthermore, eserine pretreatment did not increase denervated muscle sensitivity to acetylcholine. The data suggest that both an increase in acetylcholine receptors and a decrease in acetylcholinesterase activity contribute to the phenomenon of denervation supersensitivity.