The brachymorphic mutation of mice and altered developmental patterns of limb bud 3′∶5′ cyclic adenosine monosphosphate

Summary Concentrations of cyclic AMP (cAMP) were determined in paired fore and hind limbs from day 12–16 of development in murine fetuses homozygous for the brachymorphic (bm) mutation and normal controls. A developmental rise in cAMP occurred 1 day earlier inbm/bm than in +/+ hind limbs and cAMP was higher in day-13bm/bm than in +/+ fore limbs. Since cAMP is well documented to stimulate chondrogenic differentiation, premature cartilage determination secondary to altered levels of cAMP could play a role inbm/bm short-limbed dwarfism.This work was supported by a grant from the National Foundation-March of Dimes.     

Regional and strain variation in brain 3':5' cyclic adenosine monophosphate of inbred mice.

Regional variations were found in cAMP levels in flash frozen mouse brains with the pons and cerebellum having higher levels than the cerebrum. There were also strain variations with CBA/J and BALB/cJ having higher levels than C57B1/6J in the pons and cerebellum.     

Freisetzung von Kalium aus denerviertem und innerviertem Skelettmuskel durch Acetylcholin

Summary Both hind limbs of cats (one being normally innervated, the other 8–12 days after denervation) were perfused and the quantities of potassium liberated from the two limbs in response to intraarterial injections of varying amounts of acetylcholine were compared by collecting the venous outflow from both sides separately; considerably larger quantities of potassium were found to be liberated from the denervated than from the innervated muscle after equal doses of ACH; the threshold dose of ACH for eliciting a measurable liberation of potassium being much lower on the denervated side. The dose response relation between injected ACH and potassium-liberation is of a different nature for innervated and denervated muscle. The results obtained are compatible with expectations derived from the short circuit theory of the transmitter action on the neuromuscular endplate (Fatt andKatz). The reasons of the difference in the dose relations for the innervated and denervated muscle are discussed.     

Erzeugung von Rigor der Rattenxtremitäten durch Hemmung der Glyzin-Synthese

Summary Inhibition of glycine synthesis by aminopterine, an antagonist of pteroilglutaminic acid, causes rigidity of hind limbs in rats. This rigidity can be abolished by the injection of glycine.     

3'5'-Cyclic AMP phosphodiesterase activity in the internal and external layers of human myometrium at the end of pregnancy]

The hydrolysis of cAMP by phosphodiesterase was studied in whole homogenate from human myometrium at the end of the pregnancy before onset of labour. Tissue samples were taken from outer and inner layers of placental and anti-placental sites. Kinetic analysis shows in every case two apparent Km values for low and high cAMP concentrations in the order of 1 x 10(-5) M and 1 x 10(-4) M. On the other hand Vmax values are lower for the enzyme isolated from the placental site than for the one isolated from the anti-placental area. In the 4 zones studied, an appreciable proportion of the low Km enzyme is present.     

Drug susceptibility of PCA in WBB6F1-W/Wv mice

Our previous study revealed that passive cutaneous anaphylaxis (PCA) can be produced in congenitally mast cell-deficient WBB6F1-W/W^v (abbreviated as W/W^v) mice on sensitization with undiluted or slightly diluted allogeneic and xenogeneic antisera but not on sensitization with allogeneic monoclonal immunoglobulin (Ig)E and IgG1 antibodies regardless of the antibody concentration [1]. In view of these findings, the present study was conducted to characterize PCA in this strain from its drug susceptibilities using mast cell-bearing WBB6F1-+/+ (abbreviated as +/+) and B6D2F1 mice as references. PCA in W/W^v mice mediated by a low dilution (1  4) of hyperimmune serum to bovine serum albumin of the B6D2F1 mouse origin was markedly suppressed by CV-6209, an antagonist of platelet-activating factor (PAF), but not by antihistamines such as cyproheptadine and oxatomide. In contrast, PCA in +/+ and B6D2F1 mice mediated by a high dilution (1  128) of the anti-serum (virtually by IgG1 antibody) was nearly completely suppressed by antihistamines but not by CV-6209. A remarkable difference between PCA in W/W^v and reference mice was also observed in the susceptibility to monoclonal anti mouse granulocyte (Gr-1) antibody PCA in W/W^v mice was potently suppressed by the 1- to 3-day pretreatment with this antibody but that in references was not at all. Putting these present results together with the previous finding that anti-granulocyte antibody greatly reduces circulatory Gr-1⁺ leukocytes, 1 to 3 days after the treatment [2], it is highly probable that PCA in W/W^v mice mediated by some antibody isotypes other than IgE and IgG1 is produced by PAF mainly released from Gr-1⁺ cells, while IgG1 antibody-mediated PCA in mast cell-bearing reference mice is evoked by histamine derived from mast cells. PCA homologous to that in W/W^v mice could also be produced in the reference mice on sensitization with undiluted or slightly diluted antiserum, when generalized blueing due to excess IgG1 antibody was removed by the oxatomide treatment be fore the antigen challenge. Received 10 December 1997; received after revision 2 February 1998; accepted 23 February 1998     

Physiological studies on the effects of nutritional imbalance on the central nervous system. II. Effects of thiamine deficiency on oxidative enzymes in the brain of chicken, Gallus domesticus.

The activitiy levels of succinate dehydrogenase, glutamate dehydrogenase and pyruvate dehydrogenase in the fore, mid and hind brain regions of the thiamine deficient chicken, Gallus domesticus were determined. The activity levels of succinate dehydrogenase and glutamate dehydrogenase in all the 3 regions of brain showed augmentation on inducing thiamine deficiency. In contrast the activity levels of pyruvate dehydrogenase decreased in the brain of thiamine deficient animals. It is suggested that these changes in the oxidative enzymes indicate disturbance caused in the operation of the tricarboxylic acid cycle in thiamine deficiency.     

Increase of interferon antiviral activity by exogenous cyclic adenosine-3':5'-monophosphate (cAMP).

Some effects of cAMP on replication of Semliki Forest Virus in chick embryo fibroblast cell cultures are described. Depending on concentration, the incorporation of [3H]-uridine into viral RNA or the formation of plaque-forming units is inhibited; the highest concentration tested was 8 mM. Cyclic AMP has an effect of its own and increases the Interferon action in the lower concentration ranges of Interferon (up to 1 unit/ml). The effect of cyclic AMP is fast, needs no induction and is also visible in late phases of viral replication. However, these experiments do not establish a causal relation between cAMP and Interferon.     

Physiological studies on the effects of nutritional imbalance on the central nervous system. II. Effects of thiamine deficiency on oxidative enzymes in the brain of chicken,Gallus domesticus

Summary The activity levels of succinate dehydrogenase, glutamate dehydrogenase and pyruvate dehydrogenase in the fore, mid and hind brain regions of the thiamine deficient chicken,Gallus domesticus were determined. The activity levels of scccinate dehydrogenase and glutamate dehydrogenase in all the 3 regions of brain showed augmentation on inducing thiamine deficiency. In contrast the activity levels of pyruvate dehydrogenase decreased in the brain of thiamine deficient animals. It is suggested that these changes in the oxidative enzymes indicate disturbance caused in the operation of the tricarboxylic acid cycle in thiamine deficiency.     

Correlation of immunogenicity with suppression of lymphocyte adenosine 3',5'-monophosphate-dependent protein kinase.

Cyclic-AMP-dependent protein kinase activity was depressed in whole spleen as well as in isolated splenic lymphocytes from 3-methylcholanthrene (MCA), R3230 AdCa mammary adenocarcinoma, N-hydroxy-2-acetylaminofluorene, and 4-dimethylaminoazobenzene (DMAAB) tumor-bearing Fischer rats as compared to control animals. The magnitude of depression increased with the immunogenicity of the tumor. The depressed enzyme activity was the result of a reduced Vmax for adenosine 3',5'-monophosphate (cAMP)-stimulated histone phosphorylation.     

Effect of dibutyryl cyclic AMP and analogs on the rate of contractions of myocytes in culture.

2O, 6N-butyryl, 3', 5'-cyclic monophosphate (dibu cAMP) when added to fetal rat heart cells in culture inhibits myocyte contraction. This inhibition is 100, 84 and 51% complete when the dibu cAMP concentration used is 2, 0.2 and 0.02 mM, respectively. The potency of dibu cAMP derivatives in myocyte contraction inhibition follows the order, dibu cAMP greater than 6N-bu cAMP greater than 2O-bu cAMP = AMP greater than butyrate. The inhibition caused by the first three chemicals is greater than 70%.     

Effects of starvation and Ca++ on glucose-induced accumulation of cyclic 3',5'-AMP in pancreatic islets.

Exposure to glucose in the presence of 3-isobutyl-1-methylxanthine leads to accumulation of cAMP in islets microdissected from ob/ob mice. This process is dependent on extracellular Ca++ but differs markedly from the glucose action on insulin release in the same in vitro system in disappearing after 18 h of starvation.     

Fetal calf serum alters cyclic adenosine 3',5'-monophosphate's effect on heme synthesis in vitro.

An investigation of the effect of cAMP on heme synthesis of rat bone marrow cells revealed that at 10(-2) M this cyclic nucleotide inhibits heme synthesis and that optimum stimulation occurs at 10(-4) M. Some unidentified constituent of fetal calf serum in the culture medium modifies the direction and degree of cAMP's effect.     

Rp-diastereomer of adenosine cyclic 3',5'-phosphorothioate as an antagonist of the stimulatory action of cyclic AMP on the ouabain-insensitive Na efflux in single barnacle muscle fibers

Single muscle fibers of the barnacle Balanus nubilus have been used as a preparation to test the possibility that the Rp-diastereomer of adenosine cyclic 3',5'-phosphorothioate, which is the first available analog of cAMP that acts as an antagonist of cAMP, may reduce the magnitude of cAMP-mediated stimulation of the resting ouabain-insensitive Na efflux. The results obtained show that this antagonist is, in fact, able to reduce stimulation of the Na efflux by injected cAMP in a dose-dependent manner.     

Effect of dibutyryl cyclic AMP and analogs on the rate of contractions of myocytes in culture

Summary ²O,⁶N-butyryl, 3, 5-cyclic monophosphate (dibu cAMP) when added to fetal rat heart cells in culture inhibits myocyte contraction. This inhibition is 100, 84 and 51% complete when the dibu cAMP concentration used is 2, 0.2 and 0.02 mM, respectively. The potency of dibu cAMP derivatives in myocyte contraction inhibition follows the order, dibu cAMP> ⁶N-bu cAMP> ²O-bu cAMP=AMP>butyrate. The inhibition caused by the first three chemicals is greater than 70%.The author was a Moss Heart Fellow and acknowledges the technical assistance of Ms Martha Peet. This work was supported in part by the American Heart Association, National Science Foundation and American Cancer Society.     

Über den Einfluss von Desoxycorticosteron auf Kontraktur und Kaliumfreisetzung durch Acetylcholin an der innervierten und denervierten Skelettmuskulatur

Summary The release of potassium ions from striated muscle, and the changes in mechanical tension developed by the gastrocnemius muscle on intraarterial injection of ACh, were investigated by isolated perfusion of the hind limbs in cats. The reaction of the normal innervated and chronically denervated muscle in the same animal were compared before and during perfusion with 2·10^–4 M/l desoxycorticosterone glucoside (DCG). The following results were obtained: after perfusion with DCG no change in the spontaneous release of potassium ions occurred neither on the innervated nor on the denervated muscle. The potassium release following intra-arterial injections of various doses of ACh was significantly reduced on innervated and denervated muscle. On the denervated muscle there was also a considerable reduction of the height of contractures caused by ACh administration. The results make it probable that DCG acts by an inhibition of depolarisation in the same way asd-tubocurarine.     

Caffeine intake induces an alteration in human neutrophil A2A adenosine receptors

Caffeine is the most widely used drug in the world and acts mainly through antagonism of the effects mediated by the adenosine receptor subtypes A1, A2A, A2B and A3. We determined whether repeated caffeine administration at different doses and for different periods of time (400 or 600 mg/day for 1 week and 400 mg/day for 2 weeks) alters human neutrophil A2A adenosine receptor density and function. Saturation binding assays showed an increase in affinity (K(D)) and density (B(max)) of A2A adenosine receptors after caffeine intake. These changes were accompanied by increases in cAMP accumulation and decreases in superoxide anion production after stimulation of the A2A receptor subtype using the agonist 5'-N-ethylcarboxamidoadenosine (NECA). Binding and functional changes of A2A receptors returned to baseline after 48 h of caffeine withdrawal. The findings are consistent with a potential anti-inflammatory effect of caffeine mediated by neutrophil A2A receptors.     

Modification of theophylline-induced lipolysis in human fat cells after trypsination.

Trypsin-treatment of human fat cells results in the potentiation of the lipolytic response and the cAMP accumulation induced by theophylline (5 . 10(-4) M) but not of those induced by theophylline (5 . 10(-3) M). The amount of cAMP formed after exposure to theophylline (5 . 10(-3) M) plus norepinephrine (5 . 10(-6) M) remains, however, 2.6 fold higher in trypsin-treated human fat cells than in the control ones.     

Desensitization of adrenal tumor cells Y1 by ACTH and dibutyryl cyclic AMP]

The steroidogenic responsiveness of mouse adrenal tumor cell line (Y-1) to ACTH and db cAMP declines after stimulations by these compounds. The desensitization induced by ACTH appears early and is more pronounced than that induced by db cAMP. After removal of the hormone or the nucleotide, the recovery of cell responsiveness to both stimuli isprogressively restored. Full responsiveness is recovered within 24 hrs.     

Regional and strain variation in brain 3′∶5′ cyclic adenosine monophosphate of inbred mice

Summary Regional variations were found in cAMP levels in flash frozen mouse brains with the pons and cerebellum having higher levels than the cerebrum. There were also strain variations with CBA/J and BALB/cJ having higher levels than C57B1/6J in the pons and cerebellum.