Testing a model of excitatory interactions between oscillators

The experimentally observed influence of regularly arriving tugs upon the AP discharge of the slowly-adapting stretch receptor organ (SAO) of crayfish was compared to a model of pacemaker excitatory synaptic interactions (Segundo and Kohn 1981). Criteria for compliance referred to facets as A) the excitation, B) the postulates, and C) the behavior. A) Excitation was implied primarily by the tug initially increasing the AP rate (it subsequently decreased it). B) The pacemaker AP discharges, and with more reason the electronically driven tugs, were considered acceptably regular sequence (postulate i). Tugs advanced the next AP (postulate ii); the "delay function" plots of delays vs. phases, i.e. interval shortenings vs. the time from the last AP to the tug, were close to the V of postulate iii, even though the shortest phases tended to postpone the next AP and the longest ones did not trigger immediately but with an around 5 ms latency. These effects were displayed also as "old phase vs. new phase" plots. The interval following that with the tug tended to be lengthened, but the pre-tug timing was not recovered. C) Behavior during a train of excitatory events, both in model and experiments, went through very similar initial settlings and eventual steady-states. The latter were characterized in the model by 1. an average excitatory vs. excited rate display formed by an endless number of segments with all positive rational slopes separated by negative-going discontinuities, 2. locking in the sense of preferential phases, and 3. periodic repetition of the same phases and inter-AP intervals. Experimental results were compatible with this. Such behavior was absent when the tug sequence was highly irregular. The initial settling, in the SAO as in the model, depended jointly on the first phase phi 1 and the intertug interval E. If the former was under lambda, it went through one or two monotonic phase-decreasing stages (one smaller, the other larger, than lambda), or through a single increasing one, depending on E being smaller or greater than, respectively, an estimated but never actually observed E leading to unstable lockings. If the initial phase was greater than lambda, settling with E's under rN + lambda involved jumps between larger than and smaller than lambda phases; with E's over rn + lambda, it involved an intermediate stable locking with phi = E-rN.(ABSTRACT TRUNCATED AT 400 WORDS)     

Humic acids inhibit the formation but not the mutagenicity of N-methyl-N-nitrosourea

Humic acids in the form of potassium humate (KH), at concentrations exerting a strong inhibitory effect on the formation of N-methyl-N-nitrosourea (MNU) when present during the nitrosation of N-methylurea (MU) at pH 3, did not reduce the mutagenicity of preformed MNU in Tradescantia, clone 4430. The inhibitory effect of 20 mg/ml KH corresponds approximately to that of 3.75 mM (0.66 mg/ml) ascorbic acid towards the formation of MNU from the mixture of 7.5 mM MU + 7.5 mM NaNO2.     

Cytochemical localization of calcium in renal sac nephrocytes of Helix aspersa

In renal sac nephrocytes of Helix aspersa, intracellular calcium has been localized using the oxalate-pyroantimonate (OPA) and phosphate-pyroantimonate (PPA) methods. Pyroantimonate precipitates are preferentially localized in the excretory spherule and in vesicles located in the basal and lateral regions of the nephrocyte. Such vesicles appear to release their content into the excretory vacuole. Calcium may interact with diverse types of molecules present in the excretory vacuole, thus favouring stabilization and packaging of the excretory spherule.     

Germination of concentrated suspensions of spores from Aspergillus niger

Summary Germination of condiospores fromA.niger in very concentrated suspension was required to inoculate solid fermentation media, but a germination self-inhibitor was detected in spores. It was found that the inhibitory effect depended on the composition of the medium and was reduced when glucose was used as a carbon source. The effect of the self-inhibitor was eliminated by washing the spores and using glucose and a protein nitrogen source in the germination medium. By this method it was possible to increase about 100 times (10⁶ to 10⁸) spore concentration, keeping more than 90% germination.     

Functional alpha 2-adrenoceptors in rat adipocytes: inhibition of cyclic AMP accumulation

Alpha 2-adrenoceptor activation inhibits cyclic AMP accumulation in fat cells from many species. However, the presence of alpha 2-adrenoceptors in rat adipocytes has been difficult to demonstrate. We observed that alpha 2-adrenergic activation inhibits forskolin-stimulated cyclic AMP accumulation both in rat and hamster adipocytes; UK 14304, p-amino clonidine and clonidine were the agents with higher efficacy. The effect of UK 14304 was blocked by yohimbine but not by prazosin demonstrating the involvement of alpha 2-adrenoceptors. Pertussis toxin blocked the alpha 2-adrenergic effect. Our results demonstrate the presence in rat fat cells of alpha 2-adrenoceptors coupled to adenylate cyclase via "Gi".     

Spin label and microcalorimetric studies of the interaction of DNA with unilamellar phosphatidylcholine liposomes

High-molecular DNA from chicken erythrocytes interacts with 1,2-dipalmitoylphosphatidylcholine in unilamellar liposomes, both in the presence and absence of Mg2+ ions. This interaction results in a phase separation in liposome membranes. The new phase induced by DNA and Mg2+ has a higher gel-liquid crystal phase transition temperature as measured by microcalorimetry. In the liquid crystalline state, the 16- and 5-doxyl stearic acid spin labels indicate changed local bilayer properties at the label position in the new phase.     

Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as solid support.

Approaches to direct solid phase sequencing of genomic and plasmid DNA have been developed using magnetic beads, coated with streptavidin, as solid support. The DNA is immobilized through selective incorporation of biotin into one of the strands. A single stranded template, suitable for sequencing, is obtained through strand-specific elution. Using this concept, in vitro amplified plasmid DNA and chromosomal DNA were sequenced directly from single colonies. The solid phase approach ensures that the amplification and the sequencing reactions can be performed under optimal conditions. The system was found to be suitable for sequencing using both isotope- and fluorescent-labelled primers.     

Identification of a novel 7-cis-11-trans-lipoxin A4 generated by human neutrophils: total synthesis, spasmogenic activities and comparison with other geometric isomers of lipoxins A4 and B4

Addition of (15S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid (15-HETE) and the ionophore A23187 (2.5 microM) to human neutrophils led to the formation of both lipoxin A4 and lipoxin B4 as well as a novel 5,6,15-trihydroxyeicosatetraenoic acid. The new compound was identified using an improved isolation and detection system and its basic structure was determined by physical methods. On the basis of biosynthetic considerations, geometric isomers of lipoxin A4 and lipoxin B4 were prepared by total synthesis. Comparison of these synthetic materials with the neutrophil-derived product showed that the new compound is (5S,6R,15S)-trihydroxy-9,11,13-trans-7-cis-eicosatetraenoic acid or the 7-cis-11-trans-isomer of LXA4 (7-cis-11-trans-LXA4). LXA4, 11-trans-LXA4, 7-cis-LXA4 and 7-cis-11-trans-LXA4 all evoked dose-dependent (0.1-10 microM) contractions of the guinea pig lung strip, whereas 6-cis-LXB4 and 6-cis-8-trans-LXB4 relaxed this preparation. LXA4 and 7-cis-LXA4 were approx. 10-times more potent than the compounds with 11-trans geometry. However, all four double-bond isomers of LXA4 caused contractions which, based upon pharmacological evidence, appeared to involve specific activation of the same site as cysteinyl-containing leukotrienes. In conclusion, 7-cis-11-trans-LXA4 was isolated and identified as a novel biologically active eicosanoid formed by human neutrophils.