Cyclic adenosine-3',5'-monophosphate level in donor plasma]

The method of competitive protein binding was applied to the study of content of cyclic adenosine-3',5'-monophosphate (cAMP) in donor plasma one day before and immediately after the donation of 200 ml of blood. The work was performed by Gilman's radiochemical method. Two types of reaction of donors to donorship were revealed: without any changes and with the changes of the cAMP level. In accordance with these reactions the donors were divided into two subgroups--the stable and the reactive ones. In repeated donors the cAMP level was higher than in the primary ones, and the reactive ones. In repeated donors the cAMP level was higher than in the primary ones, and at the time of blood recovery it increased even more particularly among persons of "reactive" type. In primary donors of reactive type the cAMP content before the blood donation was either below or over the mean value and either increased or decreased to the mean level after the blood loss.     

Cyclic adenosine-3',5'-monophosphate and folate transport in rat jejunum

The intestinal transport of 5-methyltetrahydrofolate and pteroylmonoglutamate was examined in everted sacs of rat jejunum exposed to compounds which increase intracellular cyclic adenosine-3', 5'-monophosphate. Adenyl cyclase stimulators (hydrocortisone and prostaglandin), phosphodiesterase inhibitors (3-isobutyl-l-methylxanthine, aminophylline and papaverine), and dibutyryl adenosine-3',5'-cyclicmonophosphate added to the mucosal medium inhibit the mucosal-to-serosal transport of physiological concentrations of 5-methyltetrahydrofalate and pteroylmonoglutamate. Transport inhibition is correlated with the ability of these agents to increase cellular cyclic adenosine-3', 5'-monophosphate. The active, carrier-mediated transport system of folate compounds is highly sensitive to the increase in cyclic adenosine-3', 5'-monophosphate level, while the diffusion system is insensitive. These data indicate that the active transport system of folates is modulated by cellular cyclic adenosine-3', 5'-monophosphate.     

Differential effects of cyclic adenosine-3',5'-monophosphate on phosphorylation of rat liver nuclear acidic proteins

The effects of cyclic AMP on the phosphorylation of different acidic proteins of rat liver nuclei were examined in vivo and in vitro. N⁶,O²′-dibutyryl cyclic AMP selectively stimulated in vivo phosphorylation of specific nuclear proteins more than twofold within 15 min after injection. Cyclic AMP caused only a small stimulation of phosphorylation of acidic proteins in isolated nuclei but the stimulation was selective for specific proteins. When isolated nuclear acidic proteins were incubated with a soluble cyclic AMP-dependent protein kinase, the cyclic nucleotide stimulated total phosphorylation about 1.7-fold. These results support the view that the regulatory effects of cyclic AMP may involve phosphorylation of acidic proteins associated with DNA in the chromatin.     

Lack of adenosine-3',5'-monophosphate receptor protein and apparent lack of expression of adenosine-3',5'-monophosphate functions in Mycobacterium smegmatis CDC 46

The presence of adenosine-3',5'-monophosphate (cAMP), adenylate cyclase and the effect of glucose on cAMP levels in Mycobacterium spp have been reported earlier. To understand the role(s) of cAMP in these organisms, the induction of various enzyme systems was studied. Beta-galactosidase and L-tryptophanase were present at low levels of activity and could not be induced. Glycerokinase was inducible but the induction was not affected by glucose. The fructose uptake system was inducible, and the induction was lowered in the presence of glucose, but cAMP could not reverse the inhibition. cAMP binding protein was not detectable under a variety of conditions. On the basis of the lack of active cAMP binding protein, a model has been proposed to explain the apparent lack of expression of cAMP function in Mycobacterium smegmatis CDC 46.     

Formation of cyclic adenosine-3',5'-monophosphate in phagocytosis]

The content of cAMP in the phagocytizing macrophages increased, especially in the phagocytosis of live microbes. cAMP formed in phagocytosis was determined in the incubation medium, whereas in the cells its content remained practically unchanged. In the administration of E. coli 055 to the germ-free guinea pigs the concentration of cAMP was found to be increased in the mucosa cells of the small intestine and in the blood serum; this fact indicated that adenylcyclase system participated in the reactions of the interrelations of the microorganisms with the epithelial cells of the smal intestiine.     

The adnosine 3',5'-monophosphate and guanosine 3',5'-monophosphate phosphodiesterases from human lung tissue.

Human lung tissue contains phosphodiesterase enzymes capable of hydrolyzing both adenosine 3′,5′-monophosphate (cyclic AMP) and guanosine 3′,5′-monophosphate (cyclic GMP). The cyclic AMP enzyme exhibits three distinct binding affinities for its substrate (apparent Km = 0.4μM, 3μM, and 40μM) while the cyclic GMP enzyme reveals only two affinities (Km = 5μM and 40μM). The pH optima for the cyclic AMP and cyclic GMP phosphodiesterase are similar (pH 7.6–7.8). Both are inhibited by known inhibitors of phosphodiesterase activity (aminophylline, caffeine, and 3-isobutyl-1-methylxanthine). The divalent cations Mg²⁺ and Mn²⁺ stimulate cyclic AMP phosphodiesterase activity (in the absence of Mg²⁺) while Ca²⁺, Ni²⁺, and Cu²⁺ inhibit the enzyme. Histamine and imidazole slightly stimulate cyclic AMP hydrolytic activity. Thus, human lung tissue does contain multiple forms of both the cyclic AMP and cyclic GMP phosphodiesterase which are influenced by a variety of effectors.     

Effect of L-DOPA on cyclic adenosine-3',5'-monophosphate in neurogenically damaged cardiac muscle

During electric stimulation of the aortal reflexogenic zone in rabbits, administration of L-DOPA prevented the reduction of the level of cyclic adenosine-3',5'-monophosphate (cAMP) in the cardiac muscle and blood plasma. This is likely to be related to L-DOPA ability to participate in the biosynthesis of endogenous catecholamines, and thus to stimulate the synthesis of cAMP.     

Relationship of adenosine-3', 5'-monophosphate to other adenine nucleotides in human platelets

Labeled cyclic AMP and other adenine nucleotides in human platelets were distinctly separated by means of thin-layer chromatography. In lysates of human platelets, ATP was decomposed following the major route: ATP→ADP→AMP→IMP→inosine→hypoxanthine. In contrast, cyclic AMP synthesis occurred rapidly following the breakdown of ATP and leveled off after 30–60 min of incubation. Cyclic AMP synthesis in platelet lysates was 1.02 ± 0.39 nanomoles/hr/mg protein. The level of cyclic AMP formed was related to the 5′-AMP level, although the former did not exceed 5 % of the latter.     

Concentration of prostaglandins and cyclic adenosine-3',5'-monophosphate in the tissues of rats]

The content of prostaglandines (PG) and cyclic 3',5'-adenosine monphosphate (cAMP) was investigated in rat tissues by the radioisotopic method of competitive binding. Maximum quantities of both PG and cAMP were revealed in the same most actively functioning organs: the brain, incretory glands, small intestine. Fatty tissue showed minimum quantities of these substances. Results indicate a close functional relationship between the PG synthesis and adenylatecyclase activity in the body tissues.     

Partial purification and characterization of adenosine- and guanosine-3',5'-monophosphate phosphodiesterases from human lung tissue.

Crude extracts of human lung tissue were examined for cyclic adenosine- and guanosine-3',5'-monophosphate (cAMP and cGMP) phosphodiesterase activities. Nonlinear reciprocal plots were observed for each substrate. DEAE-Sephadex chromatography of the extracts revealed four main fractions of activity, which were further purified by Sephadex gel filtration. The phosphodiesterase activity of the resulting individual fractions was partially characterized with respect to substrate specificity, kinetic parameters, apparent molecular weight (gel filtration), thermal stability at 30 and 37 degrees C, effect of the cyclic nucleotide not utilized as substrate, and the possible influence of Ca2+-dependent protein activator. The results indicate that the tissue contains phosphodiesterases with strict specificity and a high apparent affinity for each of the two cyclic nucleotides (the Km values determined were approximately 0.3-0.4 muM). The high affinity cAMP phosphodiesterase activity was enriched in two of the purified fractions; both activities probably represent fragments of the native high affinity cAMP specific enzyme. A third purified phosphodiesterase showed mixed substrate specificity. The Km value recorded for hydrolysis of either substrate with this enzyme was approximately 25 muM. A fourth, irregularly occurring, phosphodiesterase activity also showed mixed substrate specificity. The Km value registered for hydrolysis of either substrate with this fraction was approximately 0.4 muM. There was no evidence for a Ca2+-dependent specific activation by a boiled lung tissue supernatant of any of the purified enzymes.     

Effect of cyclic adenosine-3', 5'-monophosphate on cells of experimental lympholeukemia L-5178]

A study was made of the effect of cyclic adenosine-3',5'-monophosphate (cAMP) dibutyril-cAMP and theophylline (phosphoesterase inhibitor - an enzyme transforming adenosine-3'-5'-monophosphate into adenosine-5'-monophosphate) on the intensity of proliferation (by the increase in the content of nucleic acids in the culture), DNA synthesis (by the H3-thymidine incorporation) and on the transplantation properties (the capacity to repopulation in the animal organism) of leukemic cells of the L-5178 strain. It was found that cAMP in a concentration of 0.8 mM considerably inhibited the H3-thymidine incorporation, retarded the proliferation and decreased the transplantation capacity of leukemic cells. Theophylline and dibutyril-cAMP had a comparatively low inhibitory capacity on the DNA synthesis, proliferative activity and the transplantation properties of the cells.     

Cyclic adenosine-3',5'-monophosphate content in the limbic system structures of the rat brain on the administration of corticosteroids

The effect of hydrocortisone and DOCA on the cAMP content in the hypothalamus, hippocampus and striate body of the rat brain was investigated. Single (determined after 1 and 24 hours) and repeated (7 days) hydrocortison administration in a dose of 5 mg/100 g body weight was accompanied by an increase in the cAMP concentration in the brain structures under study. Single administration of DOCA in a dose of 0.5 mg/100 g body weight did not produce any changes in the cAMP level in the structures of the rat brain limbic system; however, the dose of 2.5 mg raised the cAMP level. Prolonged administration of the hormone in the above doses dod not change the cAMP level in the brain structures. Only the hippocampus showed a 210% increase in the cAMP level during DOCA administration in a dose of 0.5 mg.