A CHOLINERGIC COMPONENT IN THE INNERVATION OF THE LONGITUDINAL SMOOTH MUSCLE OF THE GUINEA PIG VAS DEFERENS : The Fine Structural Localization of Acetylcholinesterase

Acetylcholinesterase (AChE) has been detected on the plasma membrane of about 25% of the axons in the longitudinal smooth muscle tissue of guinea pig vas deferens. These axons are presumably cholinergic. No enzyme was detected in the remaining 75% of axons. These axons are presumably adrenergic. The plasma membrane of the Schwann cells associated with the cholinergic axons also stained for AChE. Some axon bundles contained only cholinergic or adrenergic axons while others contained both types of axon. When a cholinergic axon approached within 1100 A of a smooth muscle cell, there was a patch of AChE activity on the muscle membrane adjacent to the axon. It is suggested that these approaches are the points of effective transmission from cholinergic axons to smooth muscle cells. Butyrylcholinesterase activity was detected on the plasma membranes of all axons and smooth muscle cells in this tissue.     

Some properties of hepatic glycerol kinase and their relation to the control of glycerol utilization

1. Glycerol kinase (EC 2.7.1.30) is shown to catalyse a non-equilibrium reaction in rat liver; and, as it is the first enzyme in the pathway metabolizing glycerol, its properties may be pertinent to the metabolic regulation of glycerol uptake and utilization by this tissue. 2. The properties of hepatic glycerol kinase were studied by using a radiochemical technique to measure the enzyme activity. When the concentration of ATP is low the activity of glycerol kinase is inhibited by high concentrations of glycerol; but when the concentration of ATP is high there is no inhibition and the double-reciprocal plot is linear, providing a K(m) for glycerol of 3.16x10(-6)m. Glycerol kinase is activated by high ATP concentrations provided that the concentration of the second substrate (glycerol) is high; at low concentrations of glycerol ATP does not activate the enzyme so that the double-reciprocal plot is linear, providing a K(m) for ATP of 5.8x10(-5)m. It is suggested that these kinetics may be explained by a model similar to that described by Ferdinand (1966) for phosphofructokinase. 3. Hepatic glycerol kinase is inhibited by ADP and AMP, and raising the Mg(2+) concentration increases the inhibition by these two compounds; this suggests that ADP-Mg(2+) and AMP-Mg(2+) complexes are the inhibitory species. The physiological significance of these inhibitions may be to prevent phosphorylation of glycerol when the hepatic ATP concentration is low. It is suggested that this inhibition may provide an approach to the problem of measurement of rates of lipolysis by glycerol release in tissues that contain glycerol kinase (e.g. liver, kidney, muscle, adipose tissue). 4. Hepatic glycerol kinase is inhibited by l-3-glycerophosphate competitively with respect to glycerol. The physiological significance of this inhibition may be that factors that change the intracellular concentration of l-3-glycerophosphate could change glycerol uptake by the tissue. Thus it is suggested that thyroxine treatment or feeding rats on a diet high in glycerol, which increase the activity of glycerophosphate oxidase in liver and kidney cortex respectively, lead to an increased glycerol uptake through a decrease in the concentration of glycerophosphate in these tissues. It is known that ethanol administration decreases glycerol uptake by liver, and this can be explained by the increased concentration of l-3-glycerophosphate causing inhibition of glycerol kinase.     

The catabolism of plasma glycoproteins in normal and injured rats

The catabolism of (14)C-labelled plasma glycoprotein in rats was studied after injecting homologous plasma protein labelled in the N-acetylglucosamine and sialic acid moieties. In normal animals the catabolism was approximately described by a four-compartment model. The fractional rate of catabolism of the plasma-protein amino sugar was found to be 0.0305hr.(-1), corresponding to the degradation of 2.75mumoles/hr. The (14)C label was eliminated from the animals largely as carbon dioxide with a small proportion appearing in the urine. Freely circulating amino sugars or glycopeptides did not appear in the plasma as a result of the catabolic processes, and there was no evidence that the protein-bound amino sugars were reutilized in biosynthetic processes. A study of the distribution of (14)C label in the carcasses of animals 24hr. after injection provided evidence that the gastrointestinal tract accounted for 25-38% of the total catabolic pool; the lungs, kidneys, spleen and liver also appeared to contribute to catabolism. Studies were conducted with rats that had been treated with turpentine to induce an inflammatory reaction; the results could not be analysed kinetically, since the metabolism of plasma proteins in these animals did not appear to be in a steady state. The injected plasma protein disappeared from the intravascular pool more quickly than in normal animals, but there were no significant differences in the rates of excretion of the (14)C label.     

Lead Poisoning in Gurkha Soldiers in Hong Kong

Investigation of an outbreak of lead-poisoning in 121 Gurkha soldiers showed that this was due to the contamination of chilli powder (cayenne pepper), a constituent of curry powder, with lead chromate. Comprehensive systems of food sampling are needed in developing communities.     

Nitrogenase activity in extracts of heterocystous and non-heterocystous blue-green algae

Summary Cell-free extracts capable of acetylene reduction and cyanide reduction have been prepared from heterocystous (Anabaena cylindrica) and non-heterocystous (Plectonema boryanum 594) blue-green algae. Extracts from Anabaena were obtained from cultures grown in blulk under aerobic conditions, while the Plectonema cultures were grown in bulk on nitrate-nitrogen, then washed free from nitrate and sparged with A/CO₂ for 40 h after which time maximum nitrogenase activity was detected. The nitrogenases of both algae are similar and resemble in many respects nitrogenases from bacteria and legumes. Activity is located primarily in a 40,000xgx15 min supernatant fraction and the rate of C₂H₂ reduction observed is about 10 per cent of whole cell activity. ATP and a source of reducing power (Na₂S₂O₄) are required for efficient functioning of the enzyme. ATP-dependent hydrogen evolution occurs, the extracts are cold labile and highly sensitive to oxygen and the oxygen inhibition is irreversible.     

Distribution of isotopic label after the oral administration of free and bound 14C-labelled glucosamine in rats

The metabolic fate of [1-(14)C]glucosamine, of N-acetyl[1-(14)C]glucosamine and of glycoproteins labelled with [1-(14)C]glucosamine was studied in rats for a period of 24hr. after these materials were given orally or injected. When [1-(14)C]glucosamine was injected 26.3% of the label was excreted in the urine, 19.7% was expired as carbon dioxide and 12.7% was incorporated into plasma proteins. When the same compound was given orally, 49.2% of the label was expired as carbon dioxide, with little appearing in the urine or in the plasma. When N-acetyl[1-(14)C]glucosamine was injected, 51.3% of the label was excreted in the urine with 12.3% appearing in carbon dioxide, but there was little incorporation into plasma protein. When this compound was given orally, 46.5% of the label was expired as carbon dioxide, 7.4% was recovered in the urine and 1.7% was incorporated into plasma protein. After the injection of (14)C-labelled glycoprotein 21.0% of the label was expired as carbon dioxide, whereas when it was given orally 49.8% of the label was recovered in carbon dioxide. The differences observed between the metabolic fate of the amino sugars when they were given orally and their fate when injected could not be accounted for by the action of the intestinal microflora or by the rate of administration of the material. It is concluded that amino sugars undergo metabolic alteration or degradation during absorption.