ADENOSINE REGULATES VIA TWO DIFFERENT TYPES OF RECEPTORS, THE ACCUMULATION OF CYCLIC AMP IN CULTURED BRAIN CELLS

Abstract— In cell cultures of glial character derived from perinatal mouse brain adenosine elicits two effects. (a) At submicromolar concentrations It inhibits the increase in the intracellular level of cyclic AMP caused by β-adrenoceptor agonists. (b) At concentrations above micromolar it increases the level of cyclic AMP in the cultures. These two effects are mediated by two different adenosine receptors present on the outer surface of the cells. This is concluded from the following evidence. (a) Both effects are antagonized by methylxanthines but not by blockage of adenosine uptake or inhibition of phosphodiesterase activity. (b) In both cases activity depends on the integrity of the ribose moiety of the nucleotide. Substituents of the purine system are tolerated comparatively well. (c) The order of potency of adenosine analogues is different for the two effects. We suggest the name A1 receptors for those that mediate the inhibition and A2 for those that mediate the stimulation of cyclic AMP accumulation.     

Mutagenicity of plant flavonols in the Salmonella/mammalian microsome test: activation of flavonol glycosides by mixed glycosidases from rat cecal bacteria and other sources.

Over 70 naturally occurring and synthetic flavonoids were screened for mutagenicity with 5 tester strains in the Salmonella/mammalian microsome assay: TA1535, TA100, TA1537, TA1538 and TA98. Frameshift mutagenicity was confined to the flavonols (flavon-3-ols) in strain TA98, TA1537 and TA100. The two most mutagenic falvonols, namely, quercetin (3,3',4',5,7-pentahydroxyflavone) and kaempferol (3,4',5,7-tetrahydroxyflavone), exhibiting 12 and 7 revertants/nmol in TA98 respectively, are also the most common flavonols occurring in plants. Other flavonols exhibited less activity (revertants/nmol): galangin (2.0), rhamnetin (0.45), kaempferide (0.24), fisetin (0.14), myricetin (0.12), robinetin (0.06) and morin (0.05). All of these flavonols apparently exhibited significant activation by Aroclor 1254 induced rat-liver microsome preparations (S9). However, subsequent study revealed that only those flavonols either lacking or possessing one B ring hydroxyl group had an absolute requirement for microsomal activation. Alternatively, quercetin with two B-ring OH groups is not activated by microsomal enzymes, but by soluble (S100) enzymes from liver which are apparently constitutive and not subject to the usual chemical induction. 3 flavonol glycosides, namely, quercetrin (quercetin-3-O-rhamnoside), rutin (quercetin-3-O-rutinoside) and robinin (kaempferol-3-O-galactosido-rhamnoside-7-O-rhamnoside), were found to be nonmutagenic. They could, however, be activated by a variety of mixed glycosidases incorporated in the usual pour plate procedure. The most effective enzyme mixtures were obtained from rat cecal bacteria and from the snail Helix pomatia.     

Connections of the olfactory bulb in the piranha (Serrasalmus nattereri)

Summary The connections of the olfactory bulb were studied in the piranha using the Nauta and horseradish-peroxidase methods. Three olfactory tracts project to seven terminal fields in the telencephalon and one in the diencephalon, all of them bilaterally. The contralateral olfactory bulb also receives a small input. All contralateral projections decussate in the anterior commissure and are relatively weak compared to the ipsilateral projections. HRP-containing cells were found in all of the ipsilateral telencephalic aggregates receiving an olfactory tract projection; the contralateral side was free of labeled cell bodies. Although only about one fourth of the entire telencephalon receives a direct olfactory input, the high degree of differentiation of the olfactory system suggests that the piranha depends substantially on the sense of olfaction and that this species may be a good model for further studies on olfactory mechanisms.     

Utilization of limiting concentrations of ortho-phosphate and production of extracellular organic phosphates in cultures of marine diatoms

The utilization of ortho-phosphate by two coastal marine diatomspecies, Nitzschia closterium and Cyclotella cryptica, was studiedin batch cultures. The hypothesis was tested that thresholdconcentrations in the phosphate uptake determine the lower limitof environmental phosphate, permitting the existence of species.The turn-over time of residual medium phosphate in culturesis {small tilde}10 min, indicating a rapid equilibration ofconcentration dependent on uptake with leakage of ortho-phosphate.Increasing phosphate starvation in cultures diminished the residualortho-phosphate in the range of {small tilde}60–<2nmol l^–1, as measured radiochemically after elution onSephadex® G-10 gel. These concentrations encompass the rangeof limiting phosphate concentration in continuous cultures ofthe few microalgae, for which these concentrations are actuallymeasured. The diatoms excreted {small tilde}20–100 nmolI^–1 of organic phosphate. One dominating compound, probablyan unusual nucleotide, was incompletely or not resorbed underphosphate starvation. In contrast, Nitzschia closterium excretedunder ample phosphate supply a series of three related compounds,probably phospholipids, that were resorbed under depletion.The association of the organic phosphates with macromolecularexudates is interpreted, along with the other observations,as an indication for a hardly explored periplasmatic phosphatemetabolism in these algae. ³Dedicated to Prof. Dr. H.-A. von Stosch in honour of his 75thbirthday. ⁴This study was conducted at the University of Marburg undersupport of the Humboldt Foundation Publication no. 64 of theproject "Biological Research of the Eems-Dollard Estuary".     

Anomalous carbonylation of [Pd(dppm)(O2CCF3)]2 to give an asymmetric μ-CO complex

The reactive palladium dimer, [Pd(dppm)(O₂CCF₃)]₂, is carbonylated to [Pd(dppm)(O₂CCF₃)]₂(μ-CO) in a reversible reaction with K = c. 7.2(2)x10⁴ atm⁻¹ (P_1/2 = c. 2.4 Torr). This is significantly larger than is expected based on the λ_max = 280 nm in the electronic spectrum. The product can be isolated in analytically pure form by crystallization under a CO atmosphere. It forms crystals in the monoclinic space group Cc with a = 18.584(5), b = 28.65(1), c = 11.164(3) Å and β = 95.16(2)°. The structure is significantly distorted. The bonding about the two palladium atoms is quite asymmetric. While one is close to a square planar geometry with a Pd---C(O) distance of 1.90(2) Å, the other is significantly pyramidalized and has a longer (2.00(2) Å) bond to the bridging CO. The Pd---Pd distance is only 2.896(2) Å, much shorter than that usually observed for formally non-bonded Pd atoms.     

Pnoepyga immaculata n. sp., eine neue bodenbewohnende Timalie aus dem Nepal-Himalaya

Zusammenfassung Pnoepyga immaculata n. sp. lebt im Areal der beiden anderenPnoepyga-Artenalbiventer (Hodgson 1837) undpusilla Hodgson 1845 und wurde durch ihren markant abweichenden Territorialgesang entdeckt. Sie besitzt eine kurze silberhelle Strophe aus kurzen, in gleichmäßigen Abständen gereihten Pfiff-Elementen, die in der Frequenz leicht abfällt. Neben der Stimme liegen die wesentlichen diagnostischen Merkmale nach jetzigem Kenntnisstand gegenüber sympatrischenalbiventer in wenig größerem Schnabel, der Fleckenlosigkeit von Oberseite, Kopf und Flügeldecken, gegenüberpusilla in den größeren Körperabmessungen, gegenüber sympatrischenalbiventer undpusilla im schwachen grünlichen Schimmer der Oberseite und in der länglichen Schuppung der Unterseite. Es bestehen keine Proportionsunterschiede zwischenimmaculata undalbiventer/pusilla einerseits wie auch nicht zwischen diesen beiden andererseits. Bis jetzt wurden 4 Belegstücke (SammlungenKoelz, Diesselhorst, Martens) und eine durch Bandaufnahme gesicherte Beobachtung zwischen Dhaulagiri in Mittelwest-Nepal bis nahe der Ost-Grenze Nepals bekannt.P. immaculata lebt zur Brutzeit in enger Nachbarschaft mitP. albiventer undP. pusilla, ist jedoch vonalbiventer vertikal getrennt bei nachgewiesenem Kontakt, vonpusilla durch Bevorzugung trockeneren Waldunterwuchses abseits von Bächen. Schon jetzt mußimmaculata als gefährdete Art eingestuft werden, da Waldvegetation in der zur Brutzeit bevorzugten Höhenstufe zwischen 2100 und 3100 m in Nepal weitgehend vernichtet worden ist.

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Pnoepyga immaculata n. sp., a new ground living wren-babbler from the Nepal Himalayas (Timaliidae)

Summary P. immaculata n. sp. was discovered by its voice which strongly differs from the two other sympatric species,albiventer (Hodgson 1837) andpusilla Hodgson 1845. As a territorial song, it displays a silvery strophe arranged of short, more or less regularly spaced whistling notes (fig. 7a–d), slightly descending in pitch. The strophe is about 2 s long. Besides the distinct voice, the essential diagnostic characters in comparison toP. albiventer are the slightly larger bill, plain head, upper side and wing coverts (fig. 1a–d, 2, 4a–d), in comparison topusilla the larger body size (fig. 1i–m), in comparison to sympatricalbiventer andpusilla the slightly olive tinge of the upper side (against warm dark brown in both others, fig. 1e–m), and the more longish scaly feathers of the lower side (against stout scaly appearance in both others, fig. 1 a–d). These characters hold true at least in Central and Eastern Himalayan populations of the respective species. There do not exist proportional differences in the relations of any part of the body betweenimmaculata andpusilla and not between the latter ones as well. Heretofore, 4 museum specimens are known (Koelz Coll.,Diesselhorst Coll.,Martens Coll.) from West-central (Thakkhola, type locality) and East Nepal (Ting Sang La), 3 of them from the presumed breeding grounds (2100–3100 m), one from the winter quarters in the Terai lowlands. A field observation close to the Darjeeling border in far East Nepal is verified by a tape-recording.P. immaculata embarasses the biologist and systematist because it lives in close neighbourhood of the similar speciesalbiventer andpusilla. It must have been overlooked by its apparent scarcity and overall similarity to both other species of the genus. Slight ecological differences rather correspond to the vertical distribution ofalbiventer, which populates a higher forest belt. But both have already been found on territory at close range.P. pusilla inhabits different microbiotopes, especially the close proximity of running water in the same altitudinal belt asimmaculata. When discovered,P. immaculata is to be classified already as an endangered species. Forest vegetation of its preferred altitude is already greatly reduced in the Central and Eastern Nepal Himalayas. To get a more detailed information on its general distribution and biology in order to quickly arrange protected areas and thus to guarantee the survival ofP. immaculata, should be the urgent next step.

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Results of the Himalaya Expeditions ofJ. Martens, No. 163. — For No. 162 see: Stuttgarter Beitr. Naturkde. (A) 453: 1–46, 1990.     

Differing Neurochemical and Morphological Sequelae of Global Ischemia: Comparison of Single- and Multiple-Insult Paradigms

The purpose of this investigation was to investigate pathomechanisms responsible for the deleterious effects of repeated episodes of brief forebrain ischemia. Halothane-anesthetized male Wistar rats were subjected to either (a) a single 15-min period or (b) three 5-min periods (separated by 1 h) of global forebrain ischemia by bilateral carotid artery occlusions plus hypotension (50 mm Hg), followed by various periods of recirculation. Brain temperature was normothermic throughout. In one series of rats, extracellular levels of glutamate, glycine, and gamma-aminobutyric acid (GABA) were measured in the dorsolateral striatum (n = 6-8 per group) and lateral thalamus (n = 4-6 per group) by microdialysis and HPLC before and during ischemia and during 3-5 h of recirculation. In a parallel series of rats (n = 6 per group), ischemic cell change was quantified at 2 (dark neurons), 24, or 72 h following either single or multiple ischemic insults. A single 15-min ischemic period led to massive glutamate release (13-fold increase; p = 0.001), which returned to normal by 20-30 min of recirculation and remained normal thereafter. By contrast, in rats with three 5-min periods of ischemia, the glutamate level rise with each repeated insult (four- to 4.5-fold; p < or = 0.02) was smaller than that observed during the single 15-min insult, but a late sustained rise (five- to six-fold; p < 0.05) occurred at 2-3 h of recirculation. Brief ischemia-induced elevations of glycine and GABA levels were detected in both the single- and multiple-insult groups, with normalization during recirculation. In contrast, the excitotoxic index, a composite measure of neurotransmitter release ([glutamate] x [glycine]/[GABA]), differed markedly following single versus multiple insults (p = 0.002 by repeated-measures analysis of variance) and increased by seven- to 12-fold (p < 0.05) at 1-3 h following the third insult. The total amount of glutamate released was 3.3-fold higher in the multiple-insult than in the single-insult group (p < 0.02). At 2 h of recirculation, histopathological analysis of dorsolateral striatum showed a significantly greater frequency of dark neurons in the multiple- than in the single-insult group (p < 0.05 by analysis of variance). In the thalamus, a higher frequency of ischemic neurons was seen in the multiple-than in the single-insult group at all intervals studied. Thus, in rats with multiple ischemic insults, accelerated ischemic damage was found in the striatum, and severe ischemic injury was documented in the thalamus.(ABSTRACT TRUNCATED AT 400 WORDS)     

Survival of phage M13 with uracils on one or both DNA strands

Summary The survival of M13 DNA was studied after partial replacement of thymine by uracil in the bacteriophage. Uracils carry the same genetic information as the thymines. Nevertheless in Escherichia coli wild-type cells, uracils in DNA are replaced by thymines by excision repair initiated by uracil-DNA glycosylase (UDG). Thus inactivation of uracil-containing phage DNA is solely due to repair initiated by UDG. Incorporation of uracils was achieved in one or in both strands, either randomly or site-specifically using differently uracylated oligonucleotides. The results show that up to 580 uracils can be repaired without a significant decrease in survival if the uracils are localized in the (–) strand only. Incorporation of 246 uracils into the (+) strand leads to 30% or 10% survival when expressed in Escherichia coli strains CMK and JM103, respectively. However, when uracils are distributed over both strands a sharp decrease in survival occurs. This shows that the repair of two uracils localized in close proximity on opposite strands of the DNA by the excision repair mechanism is difficult, whereas uracils occurring in one strand are repaired more efficiently, irrespective of their number.