Humoral factors of the spleen in the regulation of the calcium content in blood plasma of rats]

Introduction of protein-free peptides-enriched spleen extract increases the calcium content in blood plasma of rats. After the effect of stress factor (long swimming) it falls. The value of the index under study increases in the splenectomized rats and remains unchanged after stress and introduction of the spleen factors. The most pronounced increase of the calcium concentration is observed in the case of experimental hypersplenism induced by methylcellulose introduction. The calcium-regulating effects of introduction of the enriched protein-free spleen extract and pharmacopoeial splenin preparation are compared. It is supposed that spleen contains two humoral factors of different chemical nature which are able to influence somewhat differently the calcium content in blood plasma.     

Role of the spleen in the regulation of the calcium content of the blood plasma in health and in stress

Splenectomy is found to be followed by an increase of the calcium level in blood. Administration of the spleen extract (splenin) decreases the calcium concentration in blood plasma of splenectomized animals. The normalizing splenin effect is observed in the presence of stress when the concentration of the studied trace element sharply increases as well. It is supposed that the humoral spleen factors participate in maintenance of the constant calcium level in blood preventing its elimination from tissues.     

Stimulating effect of biologically active substances from the spleen on the functional activity of hepatocytes

Studies in the effect of both splenin and spleen "protein-free extract" on the monolayer culture of chick hepatic embryos have revealed that small doses exert a stimulating effect on hepatocytes, while the large ones induce degenerative changes. Hepatotrophic characters of the "protein-free extract" are determined, while utilizing lower doses than those of splenin.     

Role of splenin and its active factor in regulating liver monooxygenase system in experimental immunodeficiency and hepatosohepatitis

An influence of splenin and its non-peptide factor of splenin (NFS) on the state of cytochrome P-450 dependent monooxygenase system (MOS) of liver microsomes in healthy animals under immunodeficiency (splenectomy, administration of gamma-aminobutyric acid (GABA) and toxic hepatosohepatitis was studied. The stimulating action of splenin and NFS on cytochrome P-450 content and MOS activity of liver microsomes in healthy animals has been established. The indices studied markedly decreased after splenectomy. The splenin or NFS administrations promote the recovery of these indices up to starting level in asplenic animals. A decrease in thymic mass dependent in GABA administration is prevented by NFS pretreatment of animals; there is no any effect of mediator acid on cytochrome P-450 content and MOS activity was noted. The preliminary administration NFS potentiates hepatotoxic effect of carbon tetrachloride and increases its inhibitory effect on P-450 dependent MOS of liver microsomes. Under the NFS action the effect in activity of the last is caused by the factor influence on the reparative processes in the liver.     

Effect of splenin and splenic protein-free extracts on the adenosine deaminase activity in various organs of the rat

It has been shown that the adenosine deaminase activity of intact rats increases in such lymphoid organs as the thymus and spleen under the influence of splenic protein-free extracts dissolved in the ratio of 1:100. The enzyme activity in testes does not change but its decrease (by 26.2%) is observed in the adrenal glands under the influence of the splenic protein-free extract. An analogous effect is revealed in splenin and its fractions. The splenic protein-free extract increases (by 83%) the enzyme activity in the thymus of splenectomized rats as compared to intact animals but does not change it in a homogenate of testes.     

Serum cholinesterase activity in hypothermic and posthypothermic goats

When goats are cooled down to 22–23 °C, the activity of nonspecific serum cholinesterase remains unchanged during refrigeration and in deep lethargic hypothermia, and then begins to diminish on the first day of posthypothermia. It remains at the depressed level up to the seventh day of posthypothermia, and then begins to return gradually to the initial level. Hence it is concluded that protein biosynthesis in the cells of liver parenchyma becomes suppressed under the influence of hypothermia, and that this inhibitory effect lasts considerably longer than hypothermia itself.     

Effect of biologically active splenic factors on the functional state of mast cells in rats]

The effects of "splenic protein-free extract" and its fractions as well as splenin on the functional activity of the rat mast cells have been studied. It is established that this extract unlike splenin has no histamine-releasing activity, however it is able to inhibit histamine release from mast cels under the influence of specific liberator--substance 48/80. The found effect is associated with biologically active substances contained in fraction III of splenic protein-free extract.     

Cholinesterases in Single Nerve Cells Isolated from the Locus Ceruleus and from Nucleus of the Facial Nerve of the Rat: A Microgasometric Study

Cholinesterase activity in single nerve cell bodies isolated from the locus ceruleus and nucleus of the facial nerve of the rat was analyzed by the microgasometric method. Acetylcholinesterase activity is about the same in both types of cells. Nonspecific cholinesterase is present in noradrenergic cells of the locus ceruleus but not in the cholinergic cells of the nucleus of the facial nerve. The total activity of cholinesterases and the activity of acetylcholinesterase in nerve cell bodies isolated from the locus ceruleus remains practically unchanged from the tenth postnatal day until the age of 24 months. Depletion of noradrenaline by a high dose of reserpine does not influence the total activity of cholinesterases in nerve cell bodies of locus ceruleus.     

Acetylcholine induced reciprocal changes in the Na, K-ATPase and acetylcholinesterase activity of nerve and membrane preparations of Na, K-ATF-azy]

ATPase and cholinesterase activities in the homogenate of the frog nerve and membrane Na,K-ATPase preparation of the bovine brain were investigated. Preliminary treatment of the nerve and the preparation by acetylcholine solution (10(-6)--10(-7) M) enhanced their Na,K-ATPase activity and reduced their cholinesterase activity. Possible mechanisms of this phenomenon are discussed.     

Participation of the adenylate cyclase system in inductive synthesis of acetylcholinesterase in the brain]

The enhancement of the brain acetyl cholinesterase (ACHE) activity in rats by the intraventricular injection of adrenaline or dibutiryl analogue cyclic adenosine-3',5'-monophosphate (cAMP) was shown to be due to the enzyme inductive synthesis. ACHE induction manifests itself more in the subcortical white matter than in the cortex. The stimulating effect of adrenaline on the ACHE activity is suppressed under the beta-adrenoreceptor block, while the cAMP effect remains unchanged. On the contrary, the block of the alpha-adrenoreceptors stimulates the enzyme synthesis induction. The effects of adrenaline and cAMP are of the same direction and are realized through the beta-adrenoreceptors. The enhancement of ACHE activity during the block of the alpha-adrenoreceptors is accounted for by the elimination of their inhibitory influence on the beta-adrenoreceptors.     

The effect of nitroxyl radicals and sodium nitrite on the methemoglobin content of the blood and on the enzymatic activity of erythrocyte antioxidant protection]

Preliminary introduction of nitroxyl radicals to mice decreases methaemoglobin-forming effect of sodium nitrite and diminishes the content of total SH-groups and restored glutathione as well as the activity of glutathione reductase and total activity of dehydrogenases of the pentosophosphate path of erythrocytes. High level of lipids peroxidation in case of sodium nitrite intoxication remains unchanged under preliminary administration of nitroxyl radicals as well. Activity of the key enzymes of antioxidant protection of erythrocytes, superoxide dismutase and catalase, is not recovered with sodium nitrite intoxication in presence of nitroxyl radicals.     

CHOLINESTERASE ACTIVITY OF THE MOTOR ENDPLATE IN ISOLATED MUSCLE MEMBRANE

Abstract— The cholinesterase activity of motor endplates in tibialis anterior muscle of rats accounted for about 20 per cent of the total cholinesterase activity of the muscle. In the isolated muscle membrane preparation of rat intercostal muscle, the cholinesterase activity was localized solely in the motor endplate, as shown by cholinesterase staining. The cholinesterase activity of the membrane per unit of nitrogen was 26·9 times that of the muscle homogenate. The membrane (endplate) cholinesterase had an optimal pH of 8, K_m value of 3·1 m m , and was stable at 4° for at least 13 days. Cholinesterase of a motor endplate hydrolysed 2·69 × 10⁸ acetylcholine molecules in 1 msec. Since it is estimated that 10⁸ cholinesterase active sites are present in a motor endplate, the turnover time (time necessary for one enzyme site to hydrolyse one acetylcholine molecule) is calculated to be 372 μ sec, and the turnover number (molecules of acetylcholine hydrolysed by one enzyme site/min) to be 1·61 × 10⁵. From studies with cholinesterase inhibitors, cholinesterase activity was estimated to be due mostly to acetylcholinesterase, and only a minor part to pseudocholinesterase. The muscle membrane preparation seems to be useful for the study of other properties of the motor endplate.     

Acetylcholinesterase in normal and malignant human cells

Acetylcholinesterase (AChE) activity was determined in normal and malignant human cell lines by histochemical methods. In normal human fibroblasts, no AChE activity could be demonstrated by any histochemical technique or substrate. Enzymic activity was observed in HT-1080 human fibrosarcoma cells, RD 2 human rhabdomyosarcoma cells, and SW 311 human colon carcinoma cells. Activity was localized around the nuclear envelope, in the cytoplasm and associated with the cortical region of most cells. The specificity of the reaction was shown through the use of specific cholinesterase inhibitors.     

Status of cholinesterase activities in blood in neuropsychiatric disorders

The availability of easily accessible biochemical trait markers of central cholinergic activity would assist in the identification of homogeneous subgroups of neuropsychiatric patients within specific diagnostic categories. In addition to a refinement of nosology, these measures could also help to design specific treatment interventions. The activities of cholinesterase isoenzymes in blood have been reported to be abnormal in neuropsychiatric disorders with proven or hypothesized abnormalities of central cholinergic transmission. However, the relevance of these peripheral cholinesterase abnormalities to disordered central cholinergic transmission remains uncertain. Future studies examining specific cholinesterase isoenzymes in homogeneous diagnostic groups are needed to determine the potential utility of these measures as peripheral trait markers of central cholinergic activity.     

Determination of cytoplasmic 5'-nucleotidase which preferentially hydrolyses 6-hydroxypurine nucleotides in pig, rat and human tissues by immunotitration.

1. Activity of a 5'-nucleotidase which preferentially hydrolyses IMP and GMP was determined by immunotitration in various mammalian tissues. 2. Activity per g of rat tissue was high in testis and spleen and low in skeletal muscle. 3. In human cells, the activity was high in fibroblasts and low in erythrocytes.     

Brain enzyme activity in irradiated sympathectomized rats

The results are reviewed from studies of activity of hexokinase (2.7.1.1.EC), dehydrogenase glucose-6-phosphate (1.1.1.49 EC), and cholinesterase (3.1.1.7 EC) in subcellular fractions of rat brain at the background of chemical sympathectomy induced by long-term administration of guanethidine and subsequent irradiation with a dose of 7 Gy. In conditions of sympathectomy, the enzyme activity is inhibited; in irradiated sympathectomized rats, activity of hexokinase and cholinesterase increases to reach the level of that of intact animals while dehydrogenase remains inhibited.     

Effect of dextran gelatin on cholinesterase of rats

This study has been carried out by measuring the cholinesterase (ChE) activity in blood serum and in some organs (brain, liver, spleen, kidney, small intestine, lung, and cardiac muscle) of rats before and at different time intervals after infusion of 65 dextran 70, and 5% gelatin 40 solutions (1 ml/100 g body weight). The controls received infusions of the diluent of the gelatin preparation. The data obtained showed that the infusion of the diluent in rats has no effect on ChE activity neither in blood serum nor in other organs at any time intervals after infusion. In case of dextran and gelatin, a significant increase in ChE activity in blood serum and the tested organs was observed at different time intervals after infusion. The increase in case of dextran was more marked than in case of gelatin. These measurements returned to base-line values during 72 h after infusion. Furthermore, the study failed to disclose any untoward reactions, either immediate or delayed, which could be attributed to the infusion solutions.     

OK-432 stimulates primary production and activity of murine natural killer cells

Although many immunostimulants have been shown to increase the lytic activity of natural killer (NK) cells in the periphery, little is known about their effects on NK cells in the bone marrow, the primary site of NK production. In the experiments reported here, we tested OK-432, a pharmaceutical preparation of Streptococcus pyogenes, for its effects on both the primary production and lytic activity of NK cells in C57BL/6J mice. NK activity in bone marrow cells (BMC) and spleen cells (SC) was significantly increased following intravenous administration of OK-432, peaking on day 2 in BMC and on day 3 in SC. Concomitantly, there were marked changes in the cellularity in the two compartments. Bone marrow cellularity fell significantly on day 1 post-OK-432 and then gradually returned to normal, whereas spleen cellularity rose rapidly and remained elevated. As a consequence, the total NK activity (per femur or per spleen) was significantly increased at 48-96 h after administration of OK-432. The target specificity was unchanged. The phenotype of NK cells in BMC as determined by cytotoxic depletion was unchanged by OK-432, but splenic NK activity shifted to a 'less mature' phenotype, intermediate between that of normal BMC and SC. Cytokinetic studies using 3H-TdR revealed an increase in the production of NK cells in the bone marrow following administration of OK-432. Proliferating NK cells also appeared in the spleen. Whether these were recently produced NK cells from the bone marrow that still retained the ability to proliferate or mature NK cells that were stimulated into cell cycle cannot be determined from these experiments. These data are the first to directly demonstrate the modulation of the primary production of NK cells by an immunologically active drug.     

Cholinesterase activity during embryonic development in the blood-feeding bug Triatoma patagonica

Triatoma patagonica Del Ponte (Hemiptera: Reduviidae), a vector of Chagas' disease, is widely distributed in Argentina and is found in sylvatic and peridomiciliary ecotopes, as well as occasionally in human dwellings after the chemical control of Triatoma infestans. Anti-cholinesteratic products can be applied in peridomiciliary areas and thus knowledge of cholinesterase activity during embryonic development in this species might contribute further information relevant to effective chemical control. Cholinesterase activity was characterized by reactions to eserine 10(-5) m, to increasing concentrations of substrate and to varying centrifugal speeds. Acetylcholinesterase activity was detected on day 4 and was significant from day 5. A reduction in cholinesterase activity towards acetylthiocholine (ATC) was observed on days 9 and 10 of development. Cholinesterase activity towards ATC and butyrylthiocholine (BTC) in homogenates of eggs was inhibited by eserine 10(-5) m. The shape of the curve indicating levels of inhibition at different concentrations of ATC was typical of acetylcholinesterase. Activity towards BTC did not appear to be inhibited by excess substrate, which parallels the behaviour of butyrylcholinesterases. Cholinesterase activity towards ATC was reduced in supernatant centrifuged at 15 000 g compared with supernatant centrifuged at 1100 g. The cholinesterase system that hydrolyzes mainly ATC seems to belong to the nervous system, as indicated by its behaviour towards the substrates assayed, its greater insolubility and the fact that it evolves parallel to the development of the nervous system. Knowledge of biochemical changes associated with the development and maturation of the nervous system during embryonic development would contribute to the better understanding of anti-cholinesteratic compounds with ovicidal action that might be used in control campaigns against vectors of Chagas' disease.     

Human spleen dihydroorotate dehydrogenase: properties and partial purification

Human spleen dihydroorotate dehydrogenase is associated with the mitochondrial membrane and is linked to the respiratory chain via ubiquinone. The enzyme activity was unaffected by pyridine nucleotides. The product of the reaction, orotate, was a potent inhibitor. However, a range of other naturally occurring pyrimidines or purines had no significant effect on the activity. No evidence for the involvement of a complexed metal ion or for an active sulfhydryl group was obtained. Purification of the enzyme was achieved by preparation of an acetone powder and extraction with Triton X-100, followed by preparative polyacrylamide gel electrophoresis. Activity was observed by the addition of the artificial electron acceptors, ubiquinone 50 or PMS. Purification resulted in alteration of the pH optimum and of other kinetic characteristics. Two molecular-weight species, of molecular weight 88,000 and 98,000, were consistently observed. The properties of the human spleen enzyme were similar in principle to those for the rat liver enzyme. Differences in the mode of linkage to the respiratory chain for the mitochondrially bound enzyme, and in the characteristics of the purified enzyme, were observed.