Extracellular guanosine increases astrocyte cAMP: inhibition by adenosine A2 antagonists.

Both extracellular guanosine and adenosine stimulated astrocyte proliferation in vitro and increased intracellular cAMP 6-fold within 2 min. The effects of both guanosine and adenosine on proliferation and cAMP levels were inhibited by antagonists of adenosine A2 receptors but augmented by A1 receptor antagonists. The correlation between cAMP accumulation and stimulation of cell proliferation by adenosine and guanosine indicates that increased intracellular cAMP may be one of the second messengers involved in these effects. Guanosine is not an adenosine A2 receptor agonist and does not activate adenylate cyclase. It may exert its effects indirectly by increasing the endogenous extracellular adenosine concentration.

Purinergic stimulation of cell division and differentiation: mechanisms and pharmacological implications.

Extracellular purine nucleosides and nucleotides in micromolar concentrations stimulate proliferation of a variety of cell types in vitro and in vivo. As well they act synergistically with NGF to stimulate neurite outgrowth from PC12 cells. A variety of purine nucleosides and deoxyribonucleosides promote cell proliferation and increase intracellular cAMP. Their activities are inhibited by adenosine A2 receptor antagonists. Only adenosine interacts with the A2 receptor. We propose that the other nucleosides and deoxyribonucleosides inhibit extracellular adenosine deaminase, thereby increasing the extracellular concentration of adenosine. The nucleotides apparently act by stimulating P2y receptors coupled to inositol phosphate metabolism. We propose that the nucleosides and nucleotides act synergistically with other growth factors because each has distinct but complementary second messenger systems. If our hypotheses are correct, it should prove possible to modulate the growth and morphogenesis in several cell types using drugs that inhibit or stimulate adenosine A2 or purine P2y receptor agonists or the second messenger systems coupled to these receptors.

Extracellular purine nucleosides stimulate cell division and morphogenesis: pathological and physiological implications.

Extracellular purine nucleosides and nucleotides are ubiquitous, phylogenetically ancient, intercellular signals. Purines are released from hypoxic, damaged or dying cells. Purine nucleosides and nucleotides are potent mitogens for several types of cells such as fibroblasts, endothelial cells and neuroglia. They also induce other cell types to differentiate. For example, they act synergistically with nerve growth factor to stimulate neurite outgrowth from a pheochromocytoma cell line (PC12). We propose that after injury to tissues, including the central nervous system, purine nucleosides and nucleotides interact synergistically with other growth factors. They stimulate proliferation and morphological changes in the various cell types involved in the wound healing response. In the central nervous system this response includes glial proliferation, capillary endothelial cell proliferation, and sprouting of nerve axons. Since many actions of extracellular purines are mediated through specific cell surface receptors, this hypothesis has broad pharmacological implications.

Neostigmine in the hippocampus increases thermogenesis and T4-to-T3 conversion.

The firing rate of the nerves innervating interscapular brown adipose tissue (IBAT), IBAT and colonic temperatures (TIBAT and TC), and O2 consumption were monitored in urethan-anesthetized male Sprague-Dawley rats. These variables were measured for 40 min before (baseline values) and 40 min after a neostigmine (5 x 10(-7) mol) or saline injection in the hippocampus. The blood level of 3,5,3'-triiodothyronine and L-thyroxine (T3 and T4) and the 5'-deiodinating activity of IBAT, liver, and kidneys were determined in other rats with neostigmine or saline injection. The results showed that neostigmine injection increased firing rate, TIBAT, TC, O2 consumption, blood level of T3, and 5'-deiodinating activity of IBAT. No change was found in the T4 level and in 5'-deiodinating activity of the liver and kidneys. These findings suggest that neostigmine injection in the hippocampus increases heat production by stimulating sympathetic nerves to IBAT and by elevating the blood level of T3.

Effect of zinc concentration in Mueller-Hinton agar on susceptibility of Pseudomonas aeruginosa to imipenem.

The susceptibility of Pseudomonas aeruginosa to imipenem has been shown to vary according to zinc concentration in the media. MICs of imipenem for 68 unique clinical isolates of P. aeruginosa were determined in media supplemented with zinc at concentrations between 0.5 and 6.0 micrograms/ml. In agar containing up to 3 micrograms of zinc/ml, 75 to 82% of the strains were susceptible to imipenem at an MIC of < or = 4 micrograms/ml. In agar supplemented to contain 6 micrograms of zinc/ml, however, only 40% of the strains were susceptible to imipenem. Manufacturers should ensure that the concentration of zinc in commercial media is below 3 micrograms/ml to avoid false classification of isolates as resistant to imipenem.

Imipenem stability in a predried susceptibility panel.

We performed a 15-month study using 11 clinical strains and 1 control strain (ATCC 27853) of Pseudomonas aeruginosa to determine whether changes in the manufacturing process of Sensititre predried panels result in a reliable test of susceptibility to imipenem. MIC and breakpoint susceptibility results remained stable during the manufacturer's recommended shelf life of 18 months and compared well with standard agar disk diffusion and broth macrodilution results. Imipenem concentrations measured by high-pressure liquid chromatography were acceptable through 15 months but declined in the breakpoint panels by approximately 50% at 18 months. Between 9 months and panel expiration, 13 of 141 (9%) of the MIC panel packages had moisture entry, as indicated by pink desiccants, with a resultant loss of imipenem activity of 32 to 100%. It appears that the new manufacturing process produces MIC panels that are reliable for imipenem susceptibility testing until the labeled expiration date, provided that packages containing pink desiccants are not used.