Using IMAP technology to identify kinase inhibitors: comparison with a substrate depletion approach and analysis of the nature of false positives

IMAP is a fluorescence polarisation-based assay method which can be applied to the measurement of protein kinase activity. Using a model serine/threonine kinase we found that IMAP generated a good assay window (Z' > 0.8), was very tolerant of DMSO, and was flexible with respect to sample processing (stopped reactions were stable over a period of several days). Using a set of six low molecular weight inhibitors of the kinase, we found a good correlation between IMAP and scintillation proximity assay (SPA) potency data. IMAP, which measures product accumulation, was compared in an HTS setting with a substrate depletion method (luminescence-based measurement of ATP concentration). There was a reasonable (approximately 50%) overlap in primary hits from a 17,000 compound set, but more apparent false positives were generated from the IMAP method. We followed up the compounds that showed activity in the IMAP method but not in the luminescence assay. Approximately 10% of these compounds displayed intrinsic fluorescence, suggesting that they were false actives by virtue of intrinsic spectroscopic properties. Compound activity by competition of phosphopeptide binding to IMAP beads can occur with high concentrations of chelating compounds, but did not occur with any of the false actives, suggesting that this form of interference is rare.     

Removal of actinides and fission products activity from intermediate alkaline waste using inorganic exchangers

Hydrated iron oxide or amorphous-Fe₂O₃·3.5 H₂O (HFeO), hydrated titanium oxide (HTiO) and hydrated thorium oxide (HThO) were synthesized and their applicability for the decontamination of intermediate level liquid wastes (ILLW) was tested. The sorption of a few actinides like plutonium and americium on HFeO, ¹³⁷Cs and ¹⁰⁶Ru on HTiO and ⁹⁰Sr on HThO was investigated as a function of pH, time and loading capacity of the hydrous oxide with metal ions. The influence of the total dissolved salt content was also monitored. Some of these parameters influenced the sorption behavior significantly. The radiation stability of these inorganic sorbents were studied by irradiating them up to 48 Mrad. Adsorbed actinides and fission products were successfully eluted from HFeO and from the mix-bed of HTiO and HThO by 0.5M nitric acid.The authors wish to thank Shri R. D. Changarani, Chief Superintendent NRG Facilities and Shri P. K. Dey, Head FRD for their valuable advice and constant support.     

Micropolar flow past a stretching sheet

Summary This paper studies the flow of an incompressible, constant density micropolar fluid past a stretching sheet. The governing boundary layer equations of the flow are solved numerically using a globally convergent homotopy method in conjunction with a least change secant update quasi-Newton algorithm. The flow pattern depends on three non-dimensional parameters. Some interesting results are illustrated graphically and discussed.

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Résumé Nous étudions l'écoulement d'un fluid micropolaire et incompressible, de densité constante, le long d'une surface qui l'étend. Les équations de la couche limite qui régissent l'écoulement sont résolues numériquement. On utilise un algorithme quasi-Newtonien de type least change secant update avec une méthode homotopique /`a convergence globale. Certains résultats intéressants sont discutés et illustrés graphiquement.

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This work performed at Sandia National Laboratories supported by the U. S. Department of Energy under contract number DE-AC04-76DP00789.     

Electrochromatography in the separation of ions: Separation of alkaline earth metals

Complete Separation of ternary mixtures of Ca, Mg and Sr or Ba ions with ammonium chloride and less complete but detectable separation of the same mixtures with ammonium nitrate have been achieved, as well as a few binary separations of the type Ca-Ba, Ca-Sr, Ca-Mg, Mg-Sr and Mg-Ba.     

Compatibility studies on polyblends of PVC with chlororubber-20 and its graft polyblends by ultrasonics

Compatibilities of various PVC blends with chlororubber-20 and its graft polyblends have been studied on the basis of our previous findings. It has been found that chlororubber-20 makes compatible blends with PVC, while grafting of styrene, ethyl acrylate or acrylonitrile onto chlororubber-20 generates heterogeneity in their blends with PVC. The behaviour of these blends at the molecular level is reflected in the changes of ultrasonic velocity and absolute viscosity with composition.     

Mössbauer effect of ferrous zirconium double sulphate

The Mössbauer spectrum of ferrous zirconium double sulphate FeZrO(SO₄)₂·8H₂O shows a quadrupole split doublet at room temperature. The results are discussed in terms of high-spin Fe(II) in distorted octahedral symmetry.     

Complexometric estimation of copper

Summary Copper(II) or Copper(II) and iron(III) in a mixture are determined by titration with EDTA (disodium salt) solution as complexometric reagent at p_H 4.0 using potassium thiocyanate-p-anisidine as an indicator.Fe^III is estimated in the mixture by titration with EDTA (disodium salt) solution using potassium thiocyanate-ether as an indicator.Cu^II present in the mixture is, therefore, equivalent to the difference in amounts of the EDTA (disodium salt) used for the two sets of titrations of the mixture using potassium thiocyanate-p-anisidine and potassium thiocyanate-ether as indicators respectively.Strontium, magnesium, manganese, calcium, barium and mercuric ions do not interfere in these estimations.

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Zusammenfassung Kupfer(II) oder Kupfer(II) und Eisen(III) zusammen werden komplexometrisch mit ÄDTA-Lösung bei p_H 4,0 unter Verwendung von Kaliumthiocyanat-p-Anisidin als Indicator titriert. Eisen(III) wird in dem Gemisch durch komplexometrische Titration gegen Kaliumthiocyanat-Äther als Indicator bestimmt. Der Kupfer(II)-gehalt ergibt sich aus der Differenz der beiden Titrationen. Strontium, Magnesium, Mangan, Calcium, Barium und Quecksilber(II) stören nicht.

     

Chromatographic study of anionic complexes

Summary In this paper we have reported our results on the attempted separation of copper(II), nickel(II), cobalt(II) and iron(III) in mixtures by filter paper strip chromatography, using aqueous ethanol as solvent. The effect of the presence of varying concentrations of citrate ion as a complexing agent in the mixtures has been studied. The complexing agent has been added in the metal solution, and not in the solvent as usually done by previous workers.