Обеспечение безопасности ядерных материалов на гипотетическом объекте

Предметом исследования являются инсайдерские угрозы, угрозы со стороны, инсайдер в сговоре с угрозами со стороны, уязвимость атомных электростанций, анализ угроз, беспилотные летательные аппараты (беспилотные летательные аппараты), АЭС ВВЭР, категоризация и анализ наиболее уязвимых районов этого объекта , учет и контроль ядерных материалов, проектирование и функции ППС.The subject of the study are insider threats, outsider threats, insider in collusion with outsider threats, nuclear power plant vulnerabilities, analysis of threats that unmanned aerial vehicles (drones) impose on nuclear power plant, design of hypothetical nuclear facility for VVER NPP, categorization

Impact of central nervous system involvement in adult patients with Philadelphia-negative acute lymphoblastic leukemia: a GRAALL-2005 study

Whereas the prognosis of adult patients with Philadelphia-negative acute lymphoblastic leukemia (ALL) has greatly improved since the advent of pediatric-inspired regimens, the impact of initial central nervous system (CNS) involvement has not been formerly re-evaluated. We report here the outcome of patients with initial CNS involvement included in the pediatric-inspired prospective randomized GRAALL-2005 study. Between 2006 and 2014, 784 adult patients (aged 18-59 years) with newly diagnosed Philadelphia-negative ALL were included, of whom 55 (7%) had CNS involvement. In CNSpositive patients, overall survival was shorter (median 1.9 years vs. not reached, HR=1.8 [1.3-2.6], P<0.001). While there was no statistical difference in cumulative incidence of relapse between CNS+ and CNS- patients (HR=1.5 [0.9-2.5], P=0.11), non-relapse mortality was significantly higher in those with initial CNS disease (HR=2.1 [1.2-3.5], P=0.01). This increase in toxicity was mostly observed in patients randomized to the high-dose cyclophosphamide arm and in those who received allogeneic stem cell transplantation. Exploratory landmark analyses did not show any association between either cranial irradiation or allogeneic stem cell transplantation and outcome. Despite improved outcome in young adult ALL patients with pediatric-inspired protocols, CNS involvement is associated with a worse outcome mainly due to excess toxicity, without improved outcome with allogeneic SCT

B7‐H3 protein expression in acute myeloid leukemia

Costimulatory molecules are essential regulators of the immunological synapse and enable the fine‐tuning of the immune response. These mechanisms are subverted by cancer cells to evade immunosurveillance. The B7 family of costimulatory molecules comprises several ligands that may contribute to immunoescape. B7‐H3 [B7‐homolog 3 or CD276] remains poorly investigated in hematological malignancies. To determine the role B7‐H3, we analyzed the expression of this molecule in blast cells from a cohort of 111 acute myeloid leukemia (AML) patients. B7‐H3 was expressed in blast cells with a mean fluorescence intensity ratio >3 in 30 (27%) of the 111 patients. B7‐H3 expression was higher in the M3 and M5 FAB subtypes and in cases with mutated NPM1 and wild type CEBPA. There were no significant differences found for the FLT3‐ITD or cytogenetic risk groups. The complete remission (CR) rate between the 17 B7‐H3‐positive and 58 negative patients who were treated intensively was not different. The event free survival was longer in B7‐H3‐positive patients (P = 0.014), and there was a trend toward better overall survival. However, this difference was not statistically significant (P = 0.053). In conclusion, B7‐H3 is one of the most strongly expressed B7‐family molecules in AML and merits further investigation

Transgenic rabbit as a model to study prion diseases

National audienc

Abstract 1342: RIP3 is downregulated in human myeloid leukemia cells and modulates apoptosis and caspase-mediated p65/RelA cleavage

Abstract Introduction. Impairment in cell death pathways represents a general characteristic of most cancer cells. The receptor-interacting protein kinase 3 (RIP3) associates with RIP1 in a necrosome complex that can induce necroptosis, apoptosis, or cell proliferation. The role of RIP3 in necroptosis and inflammation has been extensively studied, but its role in cancer remains poorly understood Methods. We analyzed the expression of RIP1 and RIP3 in CD34+ leukemia cells from a cohort of patients with acute myeloid leukemia (AML) and CD34+ cells from healthy donors. To analyze the potential advantages for myeloid malignant cells due to reduced RIP3 expression, we induced the expression of RIP3 in the DA1-3b mouse leukemia cell line. Results. RIP3 expression was significantly reduced in most AML samples, whereas the expression of RIP1 did not differ significantly. When re-expressed in the mouse DA1-3b leukemia cell line, RIP3 induced apoptosis, and necroptosis in the presence of caspase inhibitors. Surprisingly, the re-expression of a RIP3 mutant with an inactive kinase domain (RIP3-KD) induced significantly more and earlier apoptosis than wild-type RIP3 (RIP3 WT), indicating that the RIP3 kinase domain is an essential regulator of apoptosis/necroptosis in leukemia cells. The induced in vivo expression of RIP3-KD, but not RIP3 WT prolonged the survival of mice injected with leukemia cells. RIP3-KD-induced cell death but not RIP3 WT was significantly antagonized by an IKKβSSEE constitutively active mutant, showing that RIP3-KD-induced apoptosis, but not RIP3 WT-induced apoptosis, was dependent on NF-κB activity. The expression of RIP3-KD induced p65/RelA NF-κB subunit caspase-dependent cleavage, and a non-cleavable p65/RelA D361E mutant rescued cells from apoptosis. The protective effect of the p65/RelA D361E mutant against apoptosis was specific to RIP3-KD-induced cell death because no change in cell death was observed when apoptosis was instead induced by treatment with imatinib or DMSO. The p65/RelA D361E mutant was generated by mutating the INFD putative consensus recognition site for caspase-6. The caspase-6 inhibitor Z-VEID-fmk partially reduced the cell death induced by RIP3-KD and slightly reduced p65/RelA cleavage. p65/RelA cleavage appears to be at least partially mediated by caspase-6. Conclusions. These data indicate that RIP3 silencing in leukemia cells results in suppression of the complex regulation of the apoptosis/necroptosis switch and the modulation of the NF-κB pathway through the caspase-mediated cleavage of p65/RelA. Citation Format: Anne-Lucie Nugues, Hassiba Bouafia, Dominique Hetuin, Celine Berthon, Anne Loyens, Elisabeth Bertrand, Nathalie Jouy, Thierry Idziorek, Bruno Quesnel. RIP3 is downregulated in human myeloid leukemia cells and modulates apoptosis and caspase-mediated p65/RelA cleavage. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1342. doi:10.1158/1538-7445.AM2014-1342</jats:p

Mitochondrial Spare Reserve Capacity : A New Predictive Metabolic Biomarker for Aggressiveness of Acute Myeloid Leukemia

Introduction The persistence of leukemic cells after treatment limits the effectiveness of anticancer drugs and is the cause of relapse in patients with acute myeloid leukemia (AML). After exposure to chemotherapeutic drugs, the survival of leukemic cells is mainly supported by mitochondrial energy metabolism. Several preclinical studies have shown that the combination of mitochondrial oxidative phosphorylation inhibitors with various anticancer treatments constitutes an effective therapeutic combination in vitro to eradicate the surviving leukemic cells. Evaluating the mitochondrial bioenergetic activity of blasts from AML patients could therefore provide predictive information on treatment response. The basal oxygen consumption of cells varies according to hematopoietic differentiation and depends on the energy needs in the in vitro condition of measurement. But it is necessary to treat the cells with uncoupling agents (eg FCCP) to assess the maximum activity that the respiratory chain could reach to respond to energy stress. Then, the switch from a basal level of oxygen consumption to a maximum level defines the mitochondrial spare reserve capacity (SRC). In this study, we propose to determine whether spare reserve capacity of blasts is a potential biomarker of AML aggressiveness in patients and to characterize the biochemical processes involved in the control of SRC in leukemic cells. Results Using the XFe24 Seahorse fluorometric oximeter, we first determined the mitochondrial oxygen consumption and glycolytic activity in hematopoietic cells (monocytes, lymphocytes, dendritic cells) of healthy donors, in AML patient blasts at diagnosis or at relapse and in AML cell lines (HL-60, MOLM-13, THP-1, KG1, OCI-AML3, MV-4-11, U-937). All measures have been assessed from freshly collected samples of peripheral blood and of bone marrow. As expected, AMLs are characterized by low oxidative phosphorylation activity compared to normal hematopoietic cells. From all the OXPHOS values obtained we defined a SRC threshold above which the SRC is considered high. This threshold has been set at a capacity to increase basal respiration by 250%. From patients blasts, we have therefore defined two groups characterized by high (n=14) or low (n=21) mitochondrial spare reserve capacity. Blasts with high SRC exhibit high glycolytic activity suggesting a link between spare reserve capacity and glucose metabolism. Using U-13C6 glucose and pharmacological inhibitors, we have demonstrated that the utilization of the mitochondrial spare reserve capacity of leukemic cells is supported through glycolysis and that mitochondrial oxidation of pyruvate is a key element for SRC recruitment. Mitochondrial pyruvate carrier inhibitors (as UK-5099) or gene silencing of BRP44 abolish the SRC of leukemic cells highlighting the importance of pyruvate oxidation to increase oxygen consumption. Since high mutation rate is recognized as an unfavorable prognostic factor in AML, we have also sequenced 45 commonly genes mutated in AMLs characterized by high or low SRC blasts. Interestingly, DNA sequencing analysis showed that AML with low SRC blasts have a higher mutation rate than high SRC blasts and also exhibited exclusive mutations such as ASXL1 (25%), IDH2 (25%), NPM1 (25%), IDH1 (13%), JAK2 (13%) and SF3B1 (13%). Conclusion Currently, most of the clinical biomarkers used to predict AML aggressiveness are based on DNA analysis, but the emergence of mutations is not always associated with phenotypic changes. This study shows that the mitochondrial spare reserve capacity of blasts represents a new functional biomarker based on the assessment of the energetic phenotype and could help the clinicians to determine the prognosis of AML. Moreover we have showed that altering pyruvate metabolism highly decrease spare reserve capacity of blasts and then could be evaluated as metabolic strategies to improve the therapeutic response in patients with AML. Disclosures Kluza: Daiichi-Sankyo: Research Funding. </jats:sec

Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients

Resistant acute myeloid leukemia (AML) exhibits mitochondrial energy metabolism changes compared to newly diagnosed AML. This phenotype is often observed by evaluating the mitochondrial oxygen consumption of blasts, but most of the oximetry protocols were established from leukemia cell lines without validation on primary leukemia cells. Moreover, the cultures and storage conditions of blasts freshly extracted from patient blood or bone marrow cause stress, which must be evaluated before determining oxidative phosphorylation (OXPHOS). Herein, we evaluated different conditions to measure the oxygen consumption of blasts using extracellular flow analyzers. We first determined the minimum number of blasts required to measure OXPHOS. Next, we compared the OXPHOS of blasts cultured for 3 h and 18 h after collection and found that to maintain metabolic organization for 18 h, cytokine supplementation is necessary. Cytokines are also needed when measuring OXPHOS in cryopreserved, thawed and recultured blasts. Next, the concentrations of respiratory chain inhibitors and uncoupler FCCP were established. We found that the FCCP concentration required to reach the maximal respiration of blasts varied depending on the patient sample analyzed. These protocols provided can be used in future clinical studies to evaluate OXPHOS as a biomarker and assess the efficacy of treatments targeting mitochondria