Geological and Geomorphological study of the original hill at the base of Fourth Dynasty Egyptian monuments. Etude géologique et géomorphologique de la colline originelle à la base des monuments de la quatrième dynastie égyptienne

22 pages; 20 figuresRock foundations of the Kephren and Kheops pyramids are examined in comparison with other Fourth Dynasty monuments: the Sphinx, Queen Kentkawes' mastaba and the Abu Rawash pyramid. This study is based on geological and geomorphological observations, visual observation, and photomontages. Results, correlated with those of former studies, demonstrate the existence of natural hills used as substrata in the construction of the two great pyramids. The minimum volume of these hills can be estimated at 12% and 23% respectively of the volumes of the Kephren and Kheops pyramids. The use of worked rock hills appears to be a characteristic of the construction methods under the Fourth Dynasty.Le substratum rocheux des pyramides de Kheops et Khephren est étudié en comparaison avec celui d'autres monuments de la quatrième dynastie de l'ancienne Egypte : le Sphinx, le mastaba de la reine Kentkawes et la pyramide d'Abu Rawash. Cette étude est basée sur des observations géologiques et géomorphologiques, l'observation visuelle et des photomontages, ainsi que des mesures réalisées sur le terrain. Les résultats, corrélés avec ceux d'études antérieures, démontrent l'existence de collines naturelles utilisées comme assises pour la construction des deux grandes pyramides. Le volume minimum de ces collines peut être estimé par rapport au volume total à 12% pour Khephren et 23% pour Kheops. L'utilisation de collines rocheuses pour asseoir la construction d'un monument apparaît être caractéristique des méthodes de construction utilisées sous la quatrième dynastie

Étude pétrographique et géochimique comparative du support en mortier de deux tables à marqueterie de pierres

Les tables à marqueterie de pierre ont été créées comme catalogues pour présenter les pierres marbrières disponibles sur le marché au xviie et xviiiesiècle. Notre étude porte sur deux tables dont le plateau est une marqueterie de pierres marbrières provenant très majoritairement de la région Languedoc-Roussillon. Dans les deux cas, la marqueterie de pierre est montée sur un support en mortier, technique de fabrication connue dans les ateliers français. Le plus célèbre de ces ateliers est l’atelier des Gobelins, ouvert entre 1645 et 1715, et dont on pense que la table de Versailles dite «table du Roi» est issue. Les résultats des observations et des analyses des mortiers des deux tables à notre disposition (observation macroscopique, microscope polarisant, microsonde électronique, diffraction X, éléments en traces par ICP-MS) montrent qu’il s’agit de mortiers dont l’agrégat est formé de tufs volcaniques importés de la province magmatique romaine et toscane en Italie, lié par de la phillipsite. La similitude extrême des résultats obtenus permet de conclure à la fabrication contemporaine et dans un même atelier des tables étudiées.Stones inlaid tables have been created to present the available marbles on the market at 17th or 18th centuries. Our study deals with two tables from which tops are inlaid of marbles coming in the majority of cases from Languedoc-Roussillon. The two tables show a mortar made support; a technique applied in French workshop the Atelier des Gobelins between 1645 and 1715. The table call “king table” localised to the Versailles castle was probably made in the Atelier des Gobelins. Results of mortars analysis show that the aggregate is made of volcanic tuffs imported from Roman and Toscan magmatic province, bound with Phillipsite. The extreme similarity of the results allows concluding that the two tables making have been contemporaneous and done in the same workshop

Metabolomic changes of the multi (-AGC-) kinase inhibitor AT13148 in cells, mice and patients are associated with NOS regulation

introduction: To generate biomarkers of target engagement or predictive response for multi-target drugs is challenging. One such compound is the multi-AGC kinase inhibitor AT13148. Metabolic signatures of selective signal transduction inhibitors identified in preclinical models have previously been confirmed in early clinical studies. This study explores whether metabolic signatures could be used as biomarkers for the multi-AGC kinase inhibitor AT13148.Objectives: To identify metabolomic changes of biomarkers of multi-AGC kinase inhibitor AT13148 in cells, xenograft / mouse models and in patients in a Phase I clinical study.Methods: HILIC LC–MS/MS methods and Biocrates AbsoluteIDQ™ p180 kit were used for targeted metabolomics; followed by multivariate data analysis in SIMCA and statistical analysis in Graphpad. Metaboanalyst and String were used for network analysis.Results: BT474 and PC3 cells treated with AT13148 affected metabolites which are in a gene protein metabolite network associated with Nitric oxide synthases (NOS). In mice bearing the human tumour xenografts BT474 and PC3, AT13148 treatment did not produce a common robust tumour specific metabolite change. However, AT13148 treatment of non-tumour bearing mice revealed 45 metabolites that were different from non-treated mice. These changes were also observed in patients at doses where biomarker modulation was observed. Further network analysis of these metabolites indicated enrichment for genes associated with the NOS pathway. The impact of AT13148 on the metabolite changes and the involvement of NOS-AT13148- Asymmetric dimethylarginine (ADMA) interaction were consistent with hypotension observed in patients in higher dose cohorts (160-300 mg).Conclusion: AT13148 affects metabolites associated with NOS in cells, mice and patients which is consistent with the clinical dose-limiting hypotension

Structure-based optimization of potent, selective, and orally bioavailable CDK8 inhibitors discovered by high-throughput screening

The mediator complex-associated cyclin dependent kinase CDK8 regulates beta-catenin-dependent transcription following activation of WNT signaling. Multiple lines of evidence suggest CDK8 may act as an oncogene in the development of colorectal cancer. Here we describe the successful optimization of an imidazo-thiadiazole series of CDK8 inhibitors that was identified in a high-throughput screening campaign and further progressed by structure-based design. In several optimization cycles, we improved the microsomal stability, potency, and kinase selectivity. The initial imidazo-thiadiazole scaffold was replaced by a 3-methyl-1H-pyrazolo[3,4-b]-pyridine which resulted in compound 25 (MSC2530818) that displayed excellent kinase selectivity, biochemical and cellular potency, microsomal stability, and is orally bioavailable. Furthermore, we demonstrated modulation phospho-STAT1, a pharmacodynamic biomarker of CDK8 activity, and tumor growth inhibition in an APC mutant SW620 human colorectal carcinoma xenograft model after oral administration. Compound 25 demonstrated suitable potency and selectivity to progress into preclinical in vivo efficacy and safety studies

Plasma Metabolomic Changes following PI3K Inhibition as Pharmacodynamic Biomarkers: Preclinical Discovery to Phase I Trial Evaluation.

PI3K plays a key role in cellular metabolism and cancer. Using a mass spectrometry-based metabolomics platform, we discovered that plasma concentrations of 26 metabolites, including amino acids, acylcarnitines, and phosphatidylcholines, were decreased in mice bearing PTEN-deficient tumors compared with non-tumor-bearing controls and in addition were increased following dosing with class I PI3K inhibitor pictilisib (GDC-0941). These candidate metabolomics biomarkers were evaluated in a phase I dose-escalation clinical trial of pictilisib. Time- and dose-dependent effects were observed in patients for 22 plasma metabolites. The changes exceeded baseline variability, resolved after drug washout, and were recapitulated on continuous dosing. Our study provides a link between modulation of the PI3K pathway and changes in the plasma metabolome and demonstrates that plasma metabolomics is a feasible and promising strategy for biomarker evaluation. Also, our findings provide additional support for an association between insulin resistance, branched-chain amino acids, and related metabolites following PI3K inhibition. Mol Cancer Ther; 15(6); 1412-24. ©2016 AACR.The Institute of Cancer ResearchThis is the author accepted manuscript. The final version is available from the American Association for Cancer Research via http://dx.doi.org/10.1158/1535-7163.MCT-15-081

2,8-Disubstituted-1,6-Naphthyridines and 4,6-Disubstituted-Isoquinolines with Potent, Selective Affinity for CDK8/19

We demonstrate a designed scaffold-hop approach to the discovery of 2,8-disubstituted-1,6-naphthyridine- and 4,6-disubstituted-isoquinoline-based dual CDK8/19 ligands. Optimized compounds in both series exhibited rapid aldehyde oxidase-mediated metabolism, which could be abrogated by introduction of an amino substituent at C5 of the 1,6-naphthyridine scaffold or at C1 of the isoquinoline scaffold. Compounds 51 and 59 were progressed to in vivo pharmacokinetic studies, and 51 also demonstrated sustained inhibition of STAT1SER727 phosphorylation, a biomarker of CDK8 inhibition, in an SW620 colorectal carcinoma human tumor xenograft model following oral dosing

Signalling involving MET and FAK supports cell division independent of the activity of the cell cycle-regulating CDK4/6 kinases.

Deregulation of cyclin-dependent kinases 4 and 6 (CDK4/6) is highly prevalent in cancer; yet, inhibitors against these kinases are currently used only in restricted tumour contexts. The extent to which cancers depend on CDK4/6 and the mechanisms that may undermine such dependency are poorly understood. Here, we report that signalling engaging the MET proto-oncogene receptor tyrosine kinase/focal adhesion kinase (FAK) axis leads to CDK4/6-independent CDK2 activation, involving as critical mechanistic events loss of the CDKI p21CIP1 and gain of its regulator, the ubiquitin ligase subunit SKP2. Combined inhibition of MET/FAK and CDK4/6 eliminates the proliferation capacity of cancer cells in culture, and enhances tumour growth inhibition in vivo. Activation of the MET/FAK axis is known to arise through cancer extrinsic and intrinsic cues. Our work predicts that such cues support cell division independent of the activity of the cell cycle-regulating CDK4/6 kinases and identifies MET/FAK as a tractable route to broaden the utility of CDK4/6 inhibitor-based therapies in the clinic.The work was supported by grants from Cancer Research UK (ref. C309/A11566, ref. C309/A8274 and ref. C309/A8992 (PW), ref. C423/A1421 and ref. C423/A15043 (SM)) and the World Cancer Research Fund (WCRF) (ref. 12-1280). CZ was supported by a Wellcome Trust studentship (ref. 094885/Z/10/Z)

Signalling involving MET and FAK supports cell division independent of the activity of the cell cycle-regulating CDK4/6 kinases.

Deregulation of cyclin-dependent kinases 4 and 6 (CDK4/6) is highly prevalent in cancer; yet, inhibitors against these kinases are currently used only in restricted tumour contexts. The extent to which cancers depend on CDK4/6 and the mechanisms that may undermine such dependency are poorly understood. Here, we report that signalling engaging the MET proto-oncogene receptor tyrosine kinase/focal adhesion kinase (FAK) axis leads to CDK4/6-independent CDK2 activation, involving as critical mechanistic events loss of the CDKI p21CIP1 and gain of its regulator, the ubiquitin ligase subunit SKP2. Combined inhibition of MET/FAK and CDK4/6 eliminates the proliferation capacity of cancer cells in culture, and enhances tumour growth inhibition in vivo. Activation of the MET/FAK axis is known to arise through cancer extrinsic and intrinsic cues. Our work predicts that such cues support cell division independent of the activity of the cell cycle-regulating CDK4/6 kinases and identifies MET/FAK as a tractable route to broaden the utility of CDK4/6 inhibitor-based therapies in the clinic.The work was supported by grants from Cancer Research UK (ref. C309/A11566, ref. C309/A8274 and ref. C309/A8992 (PW), ref. C423/A1421 and ref. C423/A15043 (SM)) and the World Cancer Research Fund (WCRF) (ref. 12-1280). CZ was supported by a Wellcome Trust studentship (ref. 094885/Z/10/Z)

Convective-scale perturbation growth across the spectrum of convective regimes

Convection-permitting ensembles have led to improved forecasts of many atmospheric phenomena. However, to fully utilize these forecasts the dependence of predictability on synoptic conditions needs to be understood. In this study, convective regimes are diagnosed based on a convective timescale which identifies the degree to which convection is in equilibrium with the large-scale forcing. Six convective cases are examined in a convection-permitting ensemble constructed using the Met Office Unified Model. The ensemble members were generated using small-amplitude buoyancy perturbations added into the boundary layer, which can be considered to represent turbulent fluctuations close to the gridscale. Perturbation growth is shown to occur on different scales with an order of magnitude difference between the regimes (O(1 km) for cases closer to non-equilibrium convection and O(10 km) for cases closer to equilibrium convection). This difference reflects the fact that cell locations are essentially random in the equilibrium events after the first 12 h of the forecast, indicating a more rapid upscale perturbation growth compared to the non-equilibrium events. Furthermore, large temporal variability is exhibited in all perturbation growth diagnostics for the nonequilibrium regime. Two boundary condition driven cases are also considered and show similar characteristics to the non-equilibrium cases, implying that caution is needed to interpret the timescale when initiation is not within the domain. Further understanding of perturbation growth within the different regimes could lead to a better understanding of where ensemble design improvements can be made beyond increasing the model resolution and could improve interpretation of forecasts

Structure-Guided Evolution of Potent and Selective CHK1 Inhibitors through Scaffold Morphing

Pyrazolopyridine inhibitors with low micromolar potency for CHK1 and good selectivity against CHK2 were previously identified by fragment-based screening. The optimization of the pyrazolopyridines to a series of potent and CHK1-selective isoquinolines demonstrates how fragment-growing and scaffold morphing strategies arising from a structure-based understanding of CHK1 inhibitor binding can be combined to successfully progress fragment-derived hit matter to compounds with activity in vivo. The challenges of improving CHK1 potency and selectivity, addressing synthetic tractability, and achieving novelty in the crowded kinase inhibitor chemical space were tackled by multiple scaffold morphing steps, which progressed through tricyclic pyrimido[2,3-b]azaindoles to N-(pyrazin-2-yl)pyrimidin-4-amines and ultimately to imidazo[4,5-c]pyridines and isoquinolines. A potent and highly selective isoquinoline CHK1 inhibitor (SAR-020106) was identified, which potentiated the efficacies of irinotecan and gemcitabine in SW620 human colon carcinoma xenografts in nude mice