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Aurora-A, -B, and -C are members of a small family of mitotic serine/threonine kinases that regulate centrosome maturation, chromosome segregation, and cytokinesis. They are often overexpressed in different human tumor types and have been identified as attractive targets for anticancer drug development. As specific inhibitors of the Aurora kinases are entering phase I clinical trials, there is a high need for appropriate Aurora-A biomarkers to follow mechanism of action or response. To identify novel Aurora-A substrates potentially useful as specific biomarkers we applied several modifications to the original KESTREL (Kinase Substrate Tracking and Elucidation) method in conjunction with gel electrophoresis and MALDI−MS and LC−MS/MS. The major modifications to the method included the introduction of a heating step to inactivate endogenous kinases after cell lysis and the execution of the in vitro kinase reaction in the presence of 5 mM Mg²⁺ and at high (1 mM) ATP concentration. Total and fractionated extracts from nocodazole-treated HeLa cells were used as a source of Aurora-A substrates. Using this approach, we were able to detect a number of Aurora-A specific phospholabeled signals and to identify vimentin as a putative Aurora-A substrate. Vimentin was then confirmed as an in vitro substrate of Aurora-A by the phosphorylation of the recombinant protein followed by MS and antibody detection. Keywords: aurora • substrate • phosphorylation • KESTREL • vimentin

We present an identification list for the visible spectra of three B stars: p Cet (B7V), 36 Lyn B8 IIIp (?)),and 134 Tau (B9 IV). Equivalent widths have been measured on medium dispersion plates (7.0 and 12.4 Å mm-1) taken with the 152 cm coudé reflector at the Observatoire de Haute Provence (France). These results are also presented.

This note demonstrates the application of a rigorous procedure for estimating the limiting standard deviation in the absolute timing, of periodic sources of known period, to the problem of the evaluation of the effects of the main source and instrumental parameters. The procedure is used to examine the case of the optical pulsar NP 0532. The results are given in diagrammatic form.

Chemical lesion of serotonergic afferents to the hippocampus resulted in a potentiation of phenylephrine (PHE) stimulation of cyclic AMP (cAMP) formation in rat hippocampal slices. The concentration-response curve of the alpha-1-agonist in lesioned rats was shifted markedly to the left with the peak increase occurring at a concentration 200 times lower (500 nM) than that required to raise cAMP to the same extent in slices of the sham-operated controls. Adenylate cyclase activity measured on crude homogenate preparations was increased by the lesion, too. The alpha-1 antagonist WB-4101 (dimethyloxyphenoxy-ethylamino-methyl benzo-1,4-dioxan) was the most potent inhibitor of PHE in lesioned rats with an IC50 of 125 nM, 110 times lower than that of prazosin (14 microM); yohimbine and propranolol, respectively, alpha-2 and beta-antagonists caused inhibition only at very high concentrations. Serotonin (10 microM) induced a marked rightward shift in the concentration response of PHE in lesioned rats, without any effect of its own on the cAMP level. PHE stimulated inositol phosphates accumulation in hippocampal slices in a concentration-dependent manner with an EC50 of 20 microM. Chemical lesion of the serotonergic median raphe nuclei caused a significant reduction of PHEs maximal effect on inositol phosphate accumulation by 50% with no change detectable in the EC50. The results imply that both second messenger responses to PHE stimulation are modulated by serotonin. Whether cAMP and phosphatidylinositol-4,5-bisphosphate generated second messengers act in a cooperative manner, or independently, is discussed.

Abstract: This paper contains the doscription of a method of computation of the power spectrum of a stochastic signal that minimizes the error intrinsic with the direct way of computation. The method is a generalization of the mean value rejection and prewhitening techniques. It grounds itself on the adaptive construction of a filter which transforms the true spectrum in a white one by means of the linear filtering of the signal given, whichever the true spectrum may be. The method is suitable of automatic computation in digital form. Various examples of the results obtained by means of a Fortran IV program, also given in the paper, are furnished and discussed.