In vitro hypoxia-conditioned colon cancer cell lines derived from HCT116 and HT29 exhibit altered apoptosis susceptibility and a more angiogenic profile in vivo

Hypoxia is an important selective force in the clonal evolution of tumours. Through HIF-1 and other transcription factors combined with tumour-specific genetic alterations, hypoxia is a dominant factor in the angiogenic phenotype. Cellular adaptation to hypoxia is an important requirement of tumour progression independent of angiogenesis. The adaptive changes, insofar as they alter hypoxia-induced apoptosis, are likely to determine responsiveness to antiangiogenic strategies. To investigate this adaptation of tumour cells to hypoxia, we recreated in vitro the in vivo situation of chronic intermittent exposure to low-oxygen levels. The colon carcinoma cell lines HT29 and HCT116 were subjected to 40 episodes of sublethal hypoxia (4 h) three times a week. The resulting two hypoxia-conditioned cell lines have been maintained in culture for more than 2 years. In both cell lines changes in doubling times occurred: in HT29 an increase, and in HCT116 a decrease. Cell survival in response to hypoxia and to DNA damage differed strikingly in the two cell lines. The HT29 hypoxia-conditioned cells were more resistant than the parental line to a 24 h hypoxic challenge, while those from HCT116 surprisingly were more sensitive. Sensitivity to cisplatin in vitro was also significantly different for the hypoxia-conditioned compared with the parental lines, suggesting a change in pathways leading to apoptosis following DNA damage signaling. The growth of both conditioned cell lines in vivo as xenografts in immunodeficient (SCID) mice was more rapid than their parental lines, and was accompanied in each by evidence of enhanced vascular proliferation as a consequence of the hypoxia-conditioning. Thus the changes in apoptotic susceptibility were independent of altered angiogenesis. The derivation of these lines provides a model for events within hypoxic regions of colon cancers, and for the acquisition of resistance and sensitivity characteristics that may have therapeutic implications for the use of antiangiogenesis drugs

Search for the Phi(1860) Pentaquark at COMPASS

Narrow Xi-pi+- and Xi-bar+pi+- resonances produced by quasi-real photons have been searched for by the COMPASS experiment at CERN. The study was stimulated by the recent observation of an exotic baryonic state decaying into Xi-pi-, at a mass of 1862 MeV, interpreted as a pentaquark. While the ordinary hyperon states Xi(1530)^0 and Xi-bar(1530)^0 are clearly seen, no exotic baryon is observed in the data taken in 2002 and 2003.Comment: 10 pages, 5 figure

Development of portable Liquid Scintillation counters for on-site primary measurement of radionuclides using the Triple-To-Double Coincidence Ratio method

Conference of 2013 3rd International Conference on Advancements in Nuclear Instrumentation, Measurement Methods and Their Applications, ANIMMA 2013 ; Conference Code:102802International audienceThe Triple-to-Double Coincidence Ratio (TDCR) method in Liquid Scintillation counting (LSC) is a primary radionuclide standardization method widely used in National Metrology laboratories and was primarily developed for the activity measurement of beta emitters. It is based on liquid scintillation: the light is detected by three photomultipliers (PM) and the detection efficiency is evaluated by using a model which uses the ratio of triple-to-double coincidences between the PM tubes.Up to now, most of current TDCR systems were locally-made metrology instruments neither aimed at nor suitable for in-situ measurements. In the framework of the European Metrofission project, a work package was dedicated to the realisation of miniature self-calibrated primary TDCR systems, which are state-of-the-art, for use on-site. The challenge was to develop a versatile portable, table-top designed instrument, from this metrology device. This implied improvements for the miniaturisation of the detection chamber, for the miniaturisation of electronic modules by exploring the possibilities of digital treatment, and for the validation of models and extension of them to nuclides with special beta spectrum shapes, to nuclides with complex decay schemes including many gamma-rays and to nuclides with higher atomic number decaying by electron capture. Four prototypes of counters were built by the Metrofission partners ENEA (Italy), LNHB (France), NPL (UK) and PTB (Germany) using various technical approaches.The paper describes these prototypes and provides some details on the choice of the technical options concerning the design of the optical chamber, of the photodetectors and of the acquisition system

Cytotoxicity and global transcriptional responses induced by zinc oxide nanoparticles NM 110 in PMA-differentiated THP-1 cells

Despite a wide production and use of zinc oxide nanoparticles (ZnONP), their toxicological study is only of limited number and their impact at a molecular level is seldom addressed. Thus, we have used, as a model, zinc oxide nanoparticle NM110 (ZnO110NP) exposure to PMA-differentiated THP-1 macrophages. The cell viability was studied at the cellular level using WST-1, LDH and Alamar Blue® assays, as well as at the molecular level by transcriptomic analysis. Exposure of cells to ZnO110NP for 24 h decreased their viability in a dose-dependent manner with mean inhibitory concentrations (IC50) of 8.1 μg/mL. Transcriptomic study of cells exposed to two concentrations of ZnO110NP: IC50 and a quarter of it (IC50/4) for 4 h showed that the expressions of genes involved in metal metabolism are perturbed. In addition, expression of genes acting in transcription regulation and DNA binding, as well as clusters of genes related to protein synthesis and structure were altered. It has to be noted that the expressions of metallothioneins genes (MT1, MT2) and genes of heat-shock proteins genes (HSP) were strongly upregulated for both conditions. These genes might be used as an early marker of exposure to ZnONP. On the contrary, at IC50 exposure, modifications of gene expression involved in inflammation, apoptosis and mitochondrial suffering were noted indicating a less specific cellular response. Overall, this study brings a resource of transcriptional data for ZnONP toxicity for further mechanistic studies

Extension of the TDCR model to compute counting efficiencies for radionuclides with complex decay schemes

Proceedings of the 19th International Conference on Radionuclide Metrology and its Applications 17–21 June 2013, Antwerp, BelgiumInternational audienceThe triple-to-double coincidence ratio (TDCR) method is frequently used to measure the activity of radionuclides decaying by pure β emission or electron capture (EC). Some radionuclides with more complex decays have also been studied, but accurate calculations of decay branches which are accompanied by many coincident γ transitions have not yet been investigated. This paper describes recent extensions of the model to make efficiency computations for more complex decay schemes possible. In particular, the MICELLE2 program that applies a stochastic approach of the free parameter model was extended. With an improved code, efficiencies for β(-), β(+) and EC branches with up to seven coincident γ transitions can be calculated. Moreover, a new parametrization for the computation of electron stopping powers has been implemented to compute the ionization quenching function of 10 commercial scintillation cocktails. In order to demonstrate the capabilities of the TDCR method, the following radionuclides are discussed: (166m)Ho (complex β(-)/γ), (59)Fe (complex β(-)/γ), (64)Cu (β(-), β(+), EC and EC/γ) and (229)Th in equilibrium with its progenies (decay chain with many α, β and complex β(-)/γ transitions)

Supplementary comparison of the measurement of the alpha and beta particle surface emission rates from large area sources (CCRI(II)-S10 LASCE)

International audienceIn 2009, the Consultative Committee for Ionizing Radiation (CCRI) approved its first supplementary comparison, to be organized by the ENEA (as the pilot laboratory), for the measurement of the alpha and beta particle surface (i.e. 2 solid angle) emission rate from large area sources of the type used for calibrating surface contamination monitors. Five sources were disseminated to the twenty-three participating laboratories consisting of one each of 241Am, 14C, 147Pm and 90Sr for emission rate measurements, with one additional 90Sr source for the evaluation of source uniformity. Measurements of the radionuclide activity and radionuclidic purity were also made although not strictly required. This report describes the organization of this comparison and the material and measurement methods used. The proposed supplementary comparison reference values (SCRV) for each of the comparison measurands are given, together with the Degrees of Equivalence and their associated uncertainties for each participating laboratory. The results of this supplementary comparison may be used as evidence by participating National Metrology Institutes (NMIs) and Designated Institutes (DIs) when submitting calibration and measurement capabilities (CMCs) for the given radionuclides for similar types of large area sources; this is an important aspect of this comparison, given that only one other international supplementary comparison for surface emission rates had been organized before

Polycomb protein EZH2 regulates E2F1-dependent apoptosis through epigenetically modulating Bim expression

10.1038/cdd.2009.162Cell Death and Differentiation175801-81