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Articles de revues sur le sujet "UCOE"

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Nelson, Everette J. R., Laura M. Tuschong et Dennis D. Hickstein. « Lentiviral Vectors Incorporating Ubiquitous Chromatin Opening Element Driving Canine CD18 Expression ». Blood 128, no 22 (2 décembre 2016) : 5890. http://dx.doi.org/10.1182/blood.v128.22.5890.5890.

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Abstract Leukocyte adhesion deficiency type 1 (LAD1) in humans is caused due to mutations in the ITGB2 gene encoding the leukocyte CD18 subunit (b2 integrin). This results in defective leukocyte adhesion and migration leading to recurrent episodes of life-threatening bacterial infection. Canine leukocyte adhesion deficiency (CLAD) represents a disease-specific large animal model of LAD1 in which new therapeutic approaches could be tested. Our previous studies have demonstrated variable efficiency of CD18 expression under the control of several promoters. These include cellular promoters such as those of human elongation factor 1a (hEF1a): long (1169bp) and short (248bp) fragments, human phosphoglycerate kinase (hPGK), human CD11b and human CD18 genes. In addition, murine stem cell virus (MSCV) promoter has also been demonstrated to lead to very high levels of CD18 expression in CLAD CD34+ cells thereby reversing the CLAD phenotype in dogs previously treated with both foamy and lentiviral vectors. But, due to potential genotoxicity associated with the use of viral promoters, we continued our efforts in search of novel cellular promoters. One such promoter is the ubiquitous chromatin opening element (UCOE) from the human heterogeneous ribonucleoprotein A2/B1 and chromobox homolog 3 (HNRPA2B1-CBX3) loci. UCOE has been previously shown to display reproducible and stable transgene expression within the context of a self-inactivating (SIN) lentiviral vector in the absence of classical enhancer activity (Zhang et al., Blood 2007).It has also been shown to confer resistance to DNA methylation-mediated transgene silencing even upon integration into the heterochromatin regions of the host chromosome (Zhang et al., Mol Ther. 2010). Since the full-length element is about 2.6 kb, we cloned and tested different fragment lengths of the UCOE promoter in a SIN lentiviral vector (pCL20) in CLAD CD34+ cells in vitro. Efficiency of expression of CD18 obtained with the six promoter fragments of UCOE (in bp), namely U3'631, U3'1262, U3'652, U5'1357, U5'723 and U5'655 were compared to those obtained with an MSCV promoter. Functional viral titers were first determined using a human LAD EBV-transformed B-cell line that lacks endogenous human CD18. When comparable titers of each vector were used in an overnight transduction of CLAD CD34+ cells after a 24h cytokine prestimulation in vitro, the percentage of CD18+ cells 5 days after transduction were as follows: U3'631 - 8.49%, U3'1262 - 15.9%, U3'652 - 21.3% (tested at MOI 100), U5'1357 - 2.05% (tested at MOI 30), U5'723 - 2.44% (tested at MOI 20), U5'655 - 3.01% (tested at MOI 50) and MSCV - 35.3% (tested at MOI 100). The CD18 expression levels driven by some of these promoter fragments were comparable to those driven by cellular promoters mentioned previously. The UCOE is promising in that it could overcome possible gene silencing effects when used in vivo, unlike promoters such as EF1a and PGK which were largely subjected to post-transcriptional gene silencing with sub-therapeutic levels of CD18 as previously tested in the dog model. Hence, functional correction of the CD18 defect could be achieved with candidate UCOE-incorporating SIN lentiviral vector(s) when used in the treatment of CLAD dogs. Disclosures No relevant conflicts of interest to declare.
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Ikawa, Yasuhiro, Toru Uchiyama, Guridevi Jayashree Jagadeesh et Fabio Candotti. « Comparison of Immortalization Potential of Gamma-Retroviral, Lentiviral and Foamy Virus Gene Transfer Vectors ». Blood 118, no 21 (18 novembre 2011) : 3116. http://dx.doi.org/10.1182/blood.v118.21.3116.3116.

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Abstract Abstract 3116 Therapeutic gene transfer has been used successfully to treat a variety of human genetic diseases. Although protocols have shown positive clinical outcomes, the success of clinical trials were tempered by adverse events, in which integration of the viral vectors increased transcription of cancer-related genes and thereby contributed to development of leukemia. In all documented cases of insertional mutagenesis, the viral vectors used contained full-length, gamma-retrovirus long terminal repeats (LTRs) that are able to exert strong promoter and enhancer activity driving not only the expression of the transgene carried by the vector, but also that of genes neighboring the insertion site. Assessing safety of integrating viral vectors for future clinical use is therefore of paramount importance. In preparation for gene therapy approaches for the Wiskott-Aldrich syndrome (WAS), we used an in vitro assay of murine bone marrow (BM) cell immortalization to compare the consequences of transduction by four different kinds of viral vectors, including a full-length LTR Moloney leukemia virus (MLV), a self-inactivating MLV carrying a 3' LTR deleted of the viral enhancer region (SIN), a lentivirus (LV), and a foamy virus (FV) construct. All vectors carried EGFP under the control of a ubiquitously acting chromatin opening element (UCOE) or the WAS endogenous promoter (WASp). In this assay, BM cells are harvested from C57BL6 mice, exposed to retroviral supernatants and cultured long-term. Derived lines are considered immortalized based on their ability to continue to grow in vitro for more than 6 weeks in the presence of interleukin-3 and stem cell factor. To date, MLV and SIN transduction of 123 and 132 cultures, respectively, gave rise to 48 and 43 immortalized lines (39.0% and 32.6% immortalization rate). The difference in immortalization rate between MLV and SIN vectors was not statistically significant (Chi square: p=0.30). As expected, immortalized cells were negative/low for IgER, cKit and Sca1 expression, and positive to different degrees for expression of the myeloid markers CD11b/Mac1 and Ly6g/Gr1. Transduction of 114 and 62 cultures with LV and FV vectors, respectively, resulted in no immortalized lines. Real-time PCR was performed to evaluate transduction efficiency of bone marrow cells and immortalized lines. Integrated vector sequences in bone marrow cells transduced by LV and FV were detected in significantly higher quantities than in cells transduced with MLV and SIN vectors. However, expression of the EGFP transgene was markedly reduced in LV- and FV-transduced cells compared to cells exposed to MLV vectors (MFI: 14.0, 1.88, 93.2, respectively). These preliminary results confirm that gamma-retroviral gene transfer vectors are prone to causing immortalization of hematopoietic cells and suggest the vectors based on LV and FV backbones may be safer alternatives for WAS and other genetic disorders, provided that effective gene expression levels can be achieved in the biological model system under study.Table.Summary of immortalization results using MLV, SIN, LV and FV vectorsVirusMOITransduction efficiencyTransduction experiments% ImmortalizationMLV/UCOE/EGFP2067%5643MLV/WASp/EGFP2065%6736SIN/UCOE/EGFP532%7236SIN/WASp/EGFP537%6028LV/UCOE/EGFP1040%570LV/WASp/EGFP1049%570FV/UCOE/EGFP1027%280FV/WASp/EGFP1022%340negativeN.A.N.A.720 Disclosures: No relevant conflicts of interest to declare.
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Boscolo, Sabrina, Francesca Mion, Marta Licciulli, Paolo Macor, Luca De Maso, Martina Brce, Michael N. Antoniou, Roberto Marzari, Claudio Santoro et Daniele Sblattero. « Simple scale-up of recombinant antibody production using an UCOE containing vector ». New Biotechnology 29, no 4 (mai 2012) : 477–84. http://dx.doi.org/10.1016/j.nbt.2011.12.005.

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Zhang, Fang, Susannah I. Thornhill, Steven J. Howe, Meera Ulaganathan, Axel Schambach, Joanna Sinclair, Christine Kinnon, H. Bobby Gaspar, Michael Antoniou et Adrian J. Thrasher. « Lentiviral vectors containing an enhancer-less ubiquitously acting chromatin opening element (UCOE) provide highly reproducible and stable transgene expression in hematopoietic cells ». Blood 110, no 5 (1 septembre 2007) : 1448–57. http://dx.doi.org/10.1182/blood-2006-12-060814.

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AbstractUbiquitously acting chromatin opening elements (UCOEs) consist of methylation-free CpG islands encompassing dual divergently transcribed promoters of housekeeping genes that have been shown to confer resistance to transcriptional silencing and to produce consistent and stable transgene expression in tissue culture systems. To develop improved strategies for hematopoietic cell gene therapy, we have assessed the potential of the novel human HNRPA2B1-CBX3 UCOE (A2UCOE) within the context of a self-inactivating (SIN) lentiviral vector. Unlike viral promoters, the enhancer-less A2UCOE gave rise to populations of cells that expressed a reporter transgene at a highly reproducible level. The efficiency of expression per vector genome was also markedly increased in vivo compared with vectors incorporating either spleen focus-forming virus (SFFV) or cytomegalovirus (CMV) promoters, suggesting a relative resistance to silencing. Furthermore, an A2UCOE-IL2RG vector fully restored the IL-2 signaling pathway within IL2RG-deficient human cells in vitro and successfully rescued the X-linked severe combined immunodeficiency (SCID-X1) phenotype in a mouse model of this disease. These data indicate that the A2UCOE displays highly reliable transcriptional activity within a lentiviral vector, largely overcoming insertion-site position effects and giving rise to therapeutically relevant levels of gene expression. These properties are achieved in the absence of classic enhancer activity and therefore may confer a high safety profile.
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Pfaff, Nils, Nico Lachmann, Mania Ackermann, Saskia Kohlscheen, Christian Brendel, Michael Flasshove, Axel Schambach et Thomas Moritz. « The Ubiquitous Chromatin Opening Element (UCOE) Enhances Lentiviral Cytidine Deaminase (CDD) Expression and Drug Resistance During Hematopoietic Differentiation of Murine Induced Pluripotent Stem Cells (iPSCs) »,. Blood 118, no 21 (18 novembre 2011) : 4179. http://dx.doi.org/10.1182/blood.v118.21.4179.4179.

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Abstract Abstract 4179 Introduction: Transfer of drug resistance genes to the hematopoietic system has been advocated for myeloprotection during anti-cancer chemotherapy, however, for malignancies manifestated in blood or bone marrow compartments this strategy carries the risk of inadvertent transduction of tumor cells. Hematopoietic differentiation of induced pluripotent stem cells (iPSCs) has the potential to overcome this problem, and we here have investigated this concept in the context of Cytidine Deaminase (CDD)-mediated myeloprotection. However, epigenetic silencing of transgenic promoter/enhancer elements during differentiation is a major drawback when using genetically modified iPSCs. Therefore, we have transduced iPSCs with lentiviral constructs overexpressing CDD from different constitutive promoter/enhancer elements and have investigated the effects of the ubiquitous chromatin opening element (UCOE) on transgene stability and CDD-mediated drug resistance in hematopoietically differentiated and naïve iPSCs. Materials/Methods: Murine iPSCs were transduced with 3rd-generation self-inactivating (SIN) lentiviral constructs overexpressing CDD and an IRES-coupled dTomato reporter from a truncated elongation factor 1α (EFS) or spleen focus-forming (SFFV) promoter/enhancer. Optionally, the UCOE site was cloned upstream of the respective promoter/enhancer. Transgene expression, Ara-C resistance and selection potential were investigated for naïve iPSCs and after differentiation along the hematopoietic lineage (d 0–8). Ara-C resistance was analyzed for colony-forming cells (d8-16). Expression of transgenic dTomato and CDD was measured by FACS, western blot and qRT-PCR. Bisulfite sequencing was performed to assess promoter methylation for the different lentiviral constructs. Results: Our studies demonstrated efficient transduction and stable EFS-driven CDD expression in undifferentiated iPSCs irrespective of the UCOE site. In contrast, SFFV-driven CDD expression was rapidly silenced. Although transgene expression levels were higher in UCOE.EFS.CDD- versus EFS.CDD-iPSCs (MFI: 89.5 vs 39.0), both, EFS.CDD- and UCOU.EFS.CDD-iPSCs were significantly protected against exposure to Ara-C (2000 nM/ 48 h) and were efficiently selected by continuous exposure to 2000 nM Ara-C (increase of dTomato+ cells from 5–8 % to 75–98 % within 16 days). No influence of CDD expression on iPSC morphology, growth characteristics, and expression of the pluripotency markers Oct4, Sox2, or Nanog was noted. Upon hematopoietic differentiation profound transgene silencing was observed in EFS.CDD-iPSCs. Silencing occurred during the first days of differentiation. Only 5–9% dTomato+ cells were observed on days 4 or 8, and reduced CDD expression levels were detected on days 4, 8, and 16 by Western blot and qRT-PCR analysis. Nevertheless, hematopoietic colony-forming units displayed significant resistance to Ara-C when compared to non-transduced controls. In contrast, UCOE.EFS.CDD-iPSCs only showed minor degrees of differentiation-induced transgene silencing with approx. 50% of the cells still expressing the dTomato transgene on day 8. Moreover, when subjected to clonogenic assays in the presence of 1000nM Ara-C, UCOE.EFS.CDD- in comparison to EFS.CDD-transduced cells exhibited significantly increased drug resistance (colony survival: 74±15 vs. 48±7%, p<0.05, n=3). In addition, bisulfite sequencing demonstrated significantly reduced CpG methylation in UCOE.EFS.CDD transduced cells upon hematopoietic differentiation. Conclusions: Our data suggest that Ara-C resistance in the hematopoietic system can be achieved by hematopoietic differentiation of CDD-overexpressing iPSCs. While EFS and SFFV promoter/enhancer elements upon hematopoietic differentiation are prone to epigenetic silencing, this may be overcome by the use of a UCOE element, which stabilizes transgene expression during hematopoietic differentiation and significantly reduces CpG methylation in regulatory elements of the provirus. Thus, our UCOE.EFS.CDD vector allows for long-term transgene expression in hematopoietic progeny of iPSCs, and hematopoietic differentiation of gene modified, patient-derived iPSCs may be suitable to increase the safety of drug resistance gene therapy in malignant diseases with manifestation in the hematopoietic system. Disclosures: No relevant conflicts of interest to declare.
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Brendel, Christian, Stefan Stein, Michael Antoniou et Manuel Grez. « UCOE (ubiquitous chromatin opening element) mediates copy dependent expression of gp91phox in lentiviral vectors ». Blood Cells, Molecules, and Diseases 40, no 2 (mars 2008) : 257–58. http://dx.doi.org/10.1016/j.bcmd.2007.10.025.

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Hou, Jeff Jia Cheng, Ben S. Hughes, Matthew Smede, Kar Man Leung, Kara Levine, Susan Rigby, Peter P. Gray et Trent P. Munro. « High-throughput ClonePix FL analysis of mAb-expressing clones using the UCOE expression system ». New Biotechnology 31, no 3 (mai 2014) : 214–20. http://dx.doi.org/10.1016/j.nbt.2014.02.002.

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Betts, Zeynep, Alexandra S. Croxford et Alan J. Dickson. « Evaluating the interaction between UCOE and DHFR-linked amplification and stability of recombinant protein expression ». Biotechnology Progress 31, no 4 (8 avril 2015) : 1014–25. http://dx.doi.org/10.1002/btpr.2083.

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Anakok, O. F., K. N. Bayindirli et P. Kose. « Optimising the new ucoe models as higher direct transgene expression profile for effective gene therapy applications ». Cytotherapy 23, no 5 (mai 2021) : S151—S152. http://dx.doi.org/10.1016/s1465324921005284.

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Seymour, Brenda J., Swati Singh, Hannah M. Certo, Karen Sommer, Blythe D. Sather, Socheath Khim, Courtnee Clough et al. « Effective, safe, and sustained correction of murine XLA using a UCOE-BTK promoter-based lentiviral vector ». Molecular Therapy - Methods & ; Clinical Development 20 (mars 2021) : 635–51. http://dx.doi.org/10.1016/j.omtm.2021.01.007.

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Thèses sur le sujet "UCOE"

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ZUCCHETTI, FIONA. « Transposon based technology in DHFR knockout CHO cell line improves generation of AMH high producing clones for industrial applications ». Doctoral thesis, Università degli Studi di Milano-Bicocca, 2018. http://hdl.handle.net/10281/198952.

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In ricerca ed in applicazione industriale la richiesta di proteine ricombinanti è in continua crescita; vengono investite sempre più risorse per sviluppare un sistema efficace in grado di ottenere un’elevata espressione proteica.Le cellule di mammifero sono l’ospite di elezione per lo sviluppo di proteine ricombinanti in quanto ne garantiscono la qualità.Linee cellulari CHO deficitarie dell’enzima diidrofolato reduttasi(DHFR) sono molto diffuse come sistema di espressione.In tali linee, il DHFR, un enzima essenziale coinvolto nella sintesi di purine, viene sfruttato come marker di selezione. È possibile aumentare la pressione selettiva con il metotrexate, molecola che presenta un’attività inibitoria sul DHFR.L’inibizione spinge le cellule ad amplificare il gene DHFR come meccanismo di resistenza, ne consegue una contestuale amplificazione del gene di interesse presente nella cassetta di espressione del plasmide. Grazie al progresso della tecnologia del DNA ricombinante, sono state recentemente sviluppate, linee cellulari knockout per il gene DHFR. Dati preliminari, ottenuti nel nostro laboratorio, hanno dimostrato che utilizzando il sistema del trasposone piggyBac(PB) è possibile migliorare la frequenza di cloni alto-producenti rispetto al metodo di trasfezione convenzionale. Il progetto di ricerca ha avuto come primo obiettivo quello di valutare se la potenzialità del trasposone si esprimesse anche nella linea cellulare DHFR knockout e di analizzare gli effetti del processo di amplificazione sul transgene integrato, comparandolo con i vettori convenzionali. Lo scopo finale era l’ottenimento di cloni alto-producenti dell'ormone anti-Mulleriano(AMH). AMH è una glicoproteina dimerica che causa la regressione dei dotti Mulleriani negli embrioni maschili ed è utilizzata come marker diagnostico nella riproduzione assistita. I dati indicano che il sistema del trasposone risulta essere più efficiente rispetto al vettore standard nell’ottenere un’elevata espressione proteica e consentendo l’integrazione di un numero maggiore di copie del transgene e la stabilità genetica nel tempo. Nonostante il miglioramento ottenuto, la resa proteica nello scale-up non è stata soddisfacente. Pertanto, ci siamo concentrati sui costrutti per aumentare la produttività e potenziare l’amplificazione genica. Abbiamo sperimentato strategie volte a ridurre l'attività del DHFR agendo sulla sua via di espressione. Anche se l'inserimento di un IRES meno efficiente ha permesso di ottenere dati incoraggianti, la resa proteica non è risultata paragonabile a quella riportata in letteratura. Inoltre, l’indebolimento della via di espressione del DHFR ha reso difficile la crescita delle colture cellulari scoraggiando un'ulteriore esplorazione. Abbiamo quindi valutato gli elementi regolatori epigenetici per migliorare l’efficienza trascrizionale.Ottimi risultati sono stati ottenuti in pool di cellule trasfettate con i vettori contenenti gli UCOE.Sono stati infatti raggiunti elevati livelli di espressione proteica dimostrando il sostanziale effetto di queste sequenze. Per caratterizzare ulteriormente gli UCOE, abbiamo isolato alcuni cloni. Lo scopo era evidenziare differenze tra questi nuovi vettori ed il trasposone precedentemente utilizzato. L'obiettivo finale è stato raggiunto.Abbiamo, infatti, isolato un clone ad alta produttività caratterizzato sia dalla stabilità genetica sia da quella dell’espressione nel tempo.In conclusione, l’utilizzo di una linea cellulare DHFR knockout in associazione con il sistema del trasposone PB ha permesso di ottenere una buona espressione della proteina ricombinante AMH.Il trasposone si è confermato un sistema efficiente per incrementare l'espressione proteica; l'utilizzo dell’elemento regolatore epigenetico UCOE non solo ha permesso un ulteriore e significativo aumento della resa proteica, ma anche un rigoroso mantenimento della stabilità genetica e dell’espressione nel tempo
The claim of recombinant protein continues to increase for both research and industrial application. More and more resources are invested to develop the most efficient system for high protein expression. Mammalian cells are the best host for quality and reliability of expressed recombinant proteins. A diffuse protein production technology is based on dihydrofolate reductase (DHFR) deficient CHO cell line expression system. DHFR is an essential enzyme involved in purines synthesis and it is used as a selection marker. This technology has the potentiality to strength the selection process by inhibiting DHFR activity with methotrexate (MTX), forcing cells to amplify the DHFR gene and the gene of interest carried by the plasmid used for transfection. Thanks to the continuous progression in recombinant DNA technology, DHFR knockout cell lines are recently developed using different molecular tools. Preliminary data showed that by using the piggyBac (PB) transposon system it is possible to enhance the frequency of high producing clones compared to conventional transfections. In order to evaluate if PB transposon potentiality also occur in DHFR knockout cell line and to analyze the effects of amplification process on integrated transgene, research plan involved the comparison of the conventional gene transfer method to the PB transposon system. Our aim was to explore PB transposon in the DHFR knockout CHO expression system to obtain clones producing high yield of anti-Mullerian hormone (AMH) protein for industrial purposes. AMH is a dimeric glycoprotein, member of the TGFβ superfamily that causes regression of Mullerian ducts in male embryos. It has a fundamental role as diagnostic marker in assisted reproduction to predict ovarian reserve. The PB transposon system resulted a more efficient method to obtain high protein expression compared to standard vector. Moreover, it allowed higher performance in gene copy number integration and genetic stability than standard transfection method. In spite of the substantial improvement obtained by this new approach, AMH protein yield in scale-up was unsatisfactory. Therefore, we focused on construct engineering to increase productivity, since a higher MTX gene amplification was expected. Strategies to reduce the DHFR activity acting on its expression pathway were performed to enhance the AMH protein production. Even though encouraging data were obtained with the insertion of a less efficient IRES, achieved AMH protein yield was not comparable to what reported in literature exploiting the same approaches. Moreover, difficulties in culturing cells due to DHFR expression pathway impairments discouraged a further exploration. Thus, we decided to shift our focus on epigenetic regulatory elements in order to force transcriptional efficiency. Impressive results were obtained in cell pools transfected with vectors carrying the UCOEs, in which high protein expression levels were reached, thus demonstrating the valid impact of these sequences. To further characterize the UCOE we decided to isolate single cell clones from the cell pool derived from the transposon with one UCOE sequence. We aimed to highlight differences between this new vector and our previous PB transposon. We reached the final goal to isolate a high-producing clone, which guaranteed expression and genetic stability over time. In conclusion, in this work a new generated DHFR knockout cell line was exploited in association with the piggyBac transposon system to achieve high expression of recombinant protein. The PB transposon system was confirmed to be a powerful method to enhance the expression of AMH protein allowing higher performances in gene copy number integration and genetic stability compared to the standard transfection method. Employing epigenetic regulatory elements, such as the UCOE, not only a substantial increase in AMH protein yield was achieved, but also expression and genetic stability was strictly conserved over time
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Wong, Chi-kin Felix, et 黃子鍵. « Ubiquitous chromatin opening element (UCOE)-mediated human coagulationFactor IX secretion by lentiviral transduction of human mesenchymalstem cells ». Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2013. http://hub.hku.hk/bib/B50713255.

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Haemophilia B is a bleeding disorder caused by various mutations of the coagulation Factor IX gene (F9) resulting in qualitative or quantitative Factor IX protein (FIX) deficiency. Factor replacement therapy is the current standard of care. Cure may be possible in the near future by gene therapy — the transfer of normal copies of F9 to patients with haemophilia, causing establishment of FIX production and correction of the bleeding phenotype. Mesenchymal stem cells (MSC) are potential vehicles for gene delivery through ex vivo gene transfer and subsequent transplantation to the patient. Lentiviral vectors can transduce MSC effectively and mediate long term gene expression. However, gene expression may decline with time due to transgene silencing. Ubiquitous Chromatin Opening Element (UCOE) is a set of genetic sequences cloned from housekeeping genes that can maintain a transcriptionally competent, open chromatin structure and was shown to prevent gene silencing by resisting DNA methylation. We tested human F9 expression and FIX protein secretion by transducing MSC with lentiviral vectors that carry the FIX gene under the control of A2UCOE (A2UCOE-hF9). A2UCOE is a 2.2 kb sequence cloned from the HNRPA2B1–CBX3 gene loci that harbour UCOE function. A2UCOE-eGFP, an enhanced Green Fluorescent Protein (eGFP) gene expression construct, was used to assist in vector titration of A2UCOE-hF9 by flow cytometry and quantitative reverse transcription-polymerase chain reaction (qRT-PCR). MSC were transduced at various Multiplicities of Infection (MOIs) by A2UCOE-hF9 lentiviral vector. Upon transduction, F9 mRNA expression and FIX secretion were measured by qRT-PCR and ELISA respectively. Osteogenic and adipogenic differentiation assay were performed to compare differentiation potential before and after transduction at an MOI of 1. F9 mRNA expression and FIX secretion were both undetectable in untransduced MSC. Upon transduction, vector dose-dependent increase in F9 mRNA expression and FIX secretion were detected at MOIs of 1, 2, 4 and 8. The level of secreted FIX ranges from 20 to 150 μIU in 72 hours. Osteogenic and adipogenic differentiation were not affected post-transduction at an MOI of 1. In conclusion, FIX secretion by MSC was detected upon A2UCOE-hF9 lentiviral transduction. However, the level of FIX appeared to be low compared to published studies. Further studies are required to determine the cause of low FIX expression, develop methods to maximize FIX expression and confirm whether A2UCOE can prevent gene silencing and maintain sustainable gene expression.
published_or_final_version
Paediatrics and Adolescent Medicine
Master
Master of Research in Medicine
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Hassinger, Elena. « Compétition d'états fondamentaux dans URu2Si2 et UCoGe ». Phd thesis, Université de Grenoble, 2010. http://tel.archives-ouvertes.fr/tel-00533732.

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Dans cette thèse, deux composés à fermions lourds ont été étudiés sous pression. L'état fondamental en dessous de T0 = 17.5 K dans URu2Si2 est appelé "ordre caché (HO), parce que le paramètre d'ordre n'a pas encore été trouvé. A 1.5 K, le système devient en plus supraconducteur. Sous pression, le système devient antiferromagnetique (AF) au dessus d'une pression critique. Des mesures Shubnikov-de Haas sous pression montre, que la surface de Fermi ne change pas entre les deux phases. Dans la phase AF, le dédoublement de la maille implique une reconstruction de la surface de Fermi. Vu que celle-ci ne change pas sous pression, ce dédoublement doit avoir lieu déjà dans l'HO. Nos mesures de la dépendance angulaire des fréquences d'oscillation supportent des nouveau calcules de bande dans le cas d'électrons plutôt itinérant. Comme deuxième partie de ma thèse, j'ai étudié le diagramme de phase sous pression du supraconducteur (SC) ferromagnétique (FM) UCoGe (TC = 2.8 K,Tsc = 0.6 K). Les mesures de résistivité, ac calorimétrie et ac susceptibilité montrent que la phase FM est supprimé a environs 1 GPa mais la phase SC existe jusque dans la phase paramagnétique induite par la pression. Ce diagramme de phase est unique dans la classe des supraconducteurs ferromagnétiques.
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Wu, Beilun. « Unconventional superconductivity in the ferromagnetic superconductor UCoGe ». Thesis, Université Grenoble Alpes (ComUE), 2017. http://www.theses.fr/2017GREAY010/document.

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Cette thèse discute essentiellement sur le champ critique supérieur du supraconducteur ferromagnétique UCoGe. La conductivité thermique et d'autres méthodes expérimentales ont été utilisées pour confirmer les nombreux comportements particuliers de Hc2 dans UCoGe, précédemment observés dans des études de résistivité. Ces caractéristiques, y compris une anisotropie forte et des courbures anormales, ne peuvent pas être interprétées en termes de théories classiques pour Hc2. Au lieu de cela, un phénomène spécifique aux supraconducteurs ferromagnétiques - la dépendance en champ de l'interaction d'appariement doit être considéré. Nous montrons que cet effet peut être analysé de façon cohérente avec des propriétés de la phase normales et peut être aussi comparé quantitativement avec une théorie existante. Ceci conduit à une clarification nette pour le cas de H//c dans UCoGe, et explique en même temps le comportement différent de Hc2 dans UCoGe et URhGe. Ces résultats soutiennent fortement l'origine magnétique de la supraconductivité dans ces systèmes. Pour H//b, nous montrons que certaines observations expérimentales convergentes suggèrent un possible changement d'état supraconducteur induit par le champ magnétique transversal dans UCoGe. Indépendamment du reste de l'étude, le dernier chapitre présente quelques résultats expérimentaux sur la phase normale de UCoGe et sur l'autre système de fermions lourds UBe13
This thesis mainly discuss the upper critical field of the ferromagnetic superconductor UCoGe.Thermal conductivity and other experimental methods have been used to confirm the numerous particularbehaviors of Hc2 in UCoGe, previously observed in resistivity studies. These features, including the stronganisotropy and the anomalous curvatures, cannot be interpreted in terms of classical theories for Hc2.Instead, a phenomenon specific to the ferromagnetic superconductors - the field dependence of the pairinginteraction, needs to be considered. We show that this effect can be consistently analyzed with normalphase properties, and is quantitatively compared with existing theory. This leads to a net clarificationfor the case of H//c in UCoGe, and at the same timeexplains the different behavior of Hc2 in UCoGe and URhGe. These resultsstrongly support the magnetic origin of superconductivity in these systems. For H//b, we showconvergent experimental observations that suggest a possible change of the superconducting state inducedby the transverse magnetic field in UCoGe. Independent from the rest of the study, the last chapter presents someexperimental results on the normal phase of UCoGe and on the other heavy-fermion system UBe13
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Li, Feng. « Bewertung von Ausbaumaßnahmen zur Engpassbeseitigung im UCTE-Verbundnetz / ». Aachen : Klinkenberg Verl, 2005. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=014188256&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.

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Hattori, Taisuke. « Spin-Triplet Superconductivity Induced by Ferromagnetic Fluctuations in UCoGe ». 京都大学 (Kyoto University), 2014. http://hdl.handle.net/2433/188471.

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Araujo, Santos Ana Luisa de. « uCom : spatial displays for visual awareness of remote locations ». Thesis, Massachusetts Institute of Technology, 2009. http://hdl.handle.net/1721.1/55199.

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Thesis (S.M.)--Massachusetts Institute of Technology, School of Architecture and Planning, Program in Media Arts and Sciences, 2009.
Cataloged from PDF version of thesis.
Includes bibliographical references (p. [113]-116).
uCom enables remote users to be visually aware of each other using "spatial displays" - live views of a remote space assembled according to an estimate of the remote space's layout. The main elements of the system design are a 3D representation of each space and a multi-display physical setup. The 3D image-based representation of a space is composed of an aggregate of live video feeds acquired from multiple viewpoints and rendered in a graphical visualization resembling a 3D collage. Its navigation controls allow users to transition among the remote views, while maintaining a sense of how the images relate in 3D space. Additionally, the system uses a configurable set of displays to portray always-on visual connections with a remote site integrated into the local physical environment. The evaluation investigates to what extent the system improves users' understanding of the layout of a remote space.
by Ana Luisa de Araujo Santos.
S.M.
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Camy, Alexandre Rosa. « Aplicação do modelo UCON abc em sistemas de comércio eletrônico B2B ». Florianópolis, SC, 2005. http://repositorio.ufsc.br/handle/123456789/102447.

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Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. Programa de Pós-Graduaçõa em Ciência da Computação.
Made available in DSpace on 2013-07-16T00:58:32Z (GMT). No. of bitstreams: 1 220823.pdf: 1700842 bytes, checksum: 339544c3cbf00d6d06566d73e3518766 (MD5)
Recentemente foi proposto um modelo de controle de acesso, denominado UCONABC, que além de unir alguns dos principais conceitos de controle de acesso ainda propõe novos conceitos como: obrigações, condições, continuidade e mutabilidade. Apesar de abrangente, o UCONABC possui limitações e existem ainda muitas melhorias a serem pesquisadas, como por exemplo, a definição de uma forma adequada da aplicação deste modelo em Sistemas de Comércio Eletrônico (CE) Business-to-Business (B2B). Publicações científicas nesta área afirmam que são necessárias pesquisas na especificação, validação e execução de políticas de controle de acesso para sistemas B2B. Esta dissertação possui como principal contribuição científica a proposta de uma forma de aplicação do UCONABC em sistemas de CE B2B que interajam entre si. Além disso, é proposto o Agrupamento Implícito Parcial, uma técnica que facilita o gerenciamento de permissões neste tipo de sistema. A aplicabilidade da proposta desta dissertação é apresentada através de uma descrição detalhada da implementação de um sistema de CE B2B onde o controle de acesso segue as especificações desta proposta. Por fim, é apresentado um estudo de caso em que é possível visualizar, através de um exemplo do mundo real, a aplicação da proposta desta dissertação neste tipo de sistema.
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Taupin, Mathieu. « Etude des fermions lourds magnétiques UCoGe et YbRh2Si2 par mesures de transport ». Phd thesis, Grenoble, 2013. http://tel.archives-ouvertes.fr/tel-01026388.

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Les mesures de conduction thermique ont été effectuées à basses températures dans le supraconducteur ferromagnétique UCoGe et dans le composé faiblement antiferromagnétique YbRh2Si2. Les fluctuations magnétiques sont un élément important dans les propriétés de ces deux composés, et sont responsables d'un canal de chaleur à basses températures. Dans UCoGe, la contribution supplémentaire causée par les fluctuations magnétiques ont la même dépendance en champ magnétique que celles vues par RMN. Étonnamment, un nouveau canal de chaleur apparaît à très basses températures. Les mesures dans l'état supraconducteur ont confirmé le caractère multigap de UCoGe. Des mesures de XMCD ont également faites dans UCoGe. Dans YbRh2Si2, les fluctuations magnétiques sont suspectées d'être responsables d'un canal de chaleur visible à très basses températures, empêchant de pouvoir conclure sur la violation ou la validité de la loi de Wiedemann-Franz au niveau du point critique quantique. Cependant, les résultats peuvent être interprétés sans avoir recours à sa violation.
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君島, 健之. « ucodeを用いたコンクリートの品質トレーサビリティの研究 ». 京都大学 (Kyoto University), 2011. http://hdl.handle.net/2433/151988.

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Livres sur le sujet "UCOE"

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Nduka, Uche. Ijele : Uche Nduka. Booklyn, N.Y : Overpass Books, 2012.

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Okeke, Uche. Uche Okeke : Biodata. Nimo, Anambra State : Documentation Centre, 1993.

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ill, Marszał-Demianiuk Ewa, dir. Ucze sie pisać. Poznań : Olesiejuk, 2015.

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United States. Unemployment Insurance Service. UCFE instructions for federal agencies. [Washington, D.C.?] : U.S. Dept. of Labor, Employment and Training Administration, Unemployment Insurance Service, 1995.

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UCFE instructions for federal agencies. [Washington, D.C.?] : U.S. Dept. of Labor, Employment and Training Administration, Unemployment Insurance Service, 1995.

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United States. Unemployment Insurance Service. UCFE instructions for federal agencies. [Washington, D.C.?] : U.S. Dept. of Labor, Employment and Training Administration, Unemployment Insurance Service, 1995.

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Ford, Gary. Cisco Unified Contact Center Enterprise (UCCE). Indianapolis, IN : Cisco Press, 2012.

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Aḥmad, Ashfāq. Uce burj Lāhaur de. Lāhaur : ʻAzīz Pablisharz, 1993.

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Aḥmad, Ashfāq. Uce burj Lāhaur de. Lāhaur : Sang-i Mīl Pablīkeshanz, 2005.

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Mitawā, Darashana. Uce caṛi kai dekhiā. Ammritasara : Rawī Sāhita Prakāshana, 1991.

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Chapitres de livres sur le sujet "UCOE"

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Villars, P., K. Cenzual, J. Daams, R. Gladyshevskii, O. Shcherban, V. Dubenskyy, N. Melnichenko-Koblyuk et al. « UCo5 ». Dans Structure Types. Part 5 : Space Groups (173) P63 - (166) R-3m, 713. Berlin, Heidelberg : Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/978-3-540-46933-9_585.

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Fioretti, A. « UCOL Baseline ». Dans Ultra-wideband Coherent Optical LANs, 3–7. Berlin, Heidelberg : Springer Berlin Heidelberg, 1993. http://dx.doi.org/10.1007/978-3-642-45733-3_2.

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Forcesi, S. « UCOL Bibliography ». Dans Ultra-wideband Coherent Optical LANs, 87–93. Berlin, Heidelberg : Springer Berlin Heidelberg, 1993. http://dx.doi.org/10.1007/978-3-642-45733-3_8.

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Benton, Trish, T. Chen, M. Mcentee, B. Fox, D. King, R. Crombie, T. Thomas et C. Bebbington. « The Use of UCOE Vectors in Combination with a Preadapted Serum Free, Suspension Cell Line Allows for Rapid Production of Large Quantities of Protein ». Dans Animal Cell Technology : From Target to Market, 52–57. Dordrecht : Springer Netherlands, 2001. http://dx.doi.org/10.1007/978-94-010-0369-8_12.

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Bianchi, A., D. Capolupo, F. del Castello et A. Fantini. « UCOL Network Architecture ». Dans Ultra-wideband Coherent Optical LANs, 9–30. Berlin, Heidelberg : Springer Berlin Heidelberg, 1993. http://dx.doi.org/10.1007/978-3-642-45733-3_3.

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Forcesi, S., E. Neri, J. Bekooij et O. Koning. « UCOL Optical Architecture ». Dans Ultra-wideband Coherent Optical LANs, 31–38. Berlin, Heidelberg : Springer Berlin Heidelberg, 1993. http://dx.doi.org/10.1007/978-3-642-45733-3_4.

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Villars, P., K. Cenzual, R. Gladyshevskii, O. Shcherban, V. Dubenskyy, V. Kuprysyuk, I. Savysyuk et R. Zaremba. « UCo2-xSn2-y ». Dans Landolt-Börnstein - Group III Condensed Matter, 264. Berlin, Heidelberg : Springer Berlin Heidelberg, 2012. http://dx.doi.org/10.1007/978-3-642-22847-6_195.

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Crastan, Valentin. « Einführung, UCTE, ENTSO-E ». Dans Elektrische Energieversorgung 1, 3–16. Berlin, Heidelberg : Springer Berlin Heidelberg, 2015. http://dx.doi.org/10.1007/978-3-662-45985-0_1.

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Crastan, Valentin. « Einführung, UCTE, ENTSO-E ». Dans Elektrische Energieversorgung 1, 3–15. Berlin, Heidelberg : Springer Berlin Heidelberg, 2012. http://dx.doi.org/10.1007/978-3-642-22346-4_1.

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Matsuda, Takahiro, et Goichiro Hanaoka. « Chosen Ciphertext Security via UCE ». Dans Public-Key Cryptography – PKC 2014, 56–76. Berlin, Heidelberg : Springer Berlin Heidelberg, 2014. http://dx.doi.org/10.1007/978-3-642-54631-0_4.

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Actes de conférences sur le sujet "UCOE"

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Santos, Ana Luisa, et V. Michael Bove. « uCom ». Dans the 28th of the international conference extended abstracts. New York, New York, USA : ACM Press, 2010. http://dx.doi.org/10.1145/1753846.1754119.

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Almutairi, Abulgader, et François Siewe. « CA-UCON ». Dans the 5th ACM International Workshop. New York, New York, USA : ACM Press, 2011. http://dx.doi.org/10.1145/2036146.2036153.

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Hu, Hao, Hao Li et Dengguo Feng. « L-UCON : Towards Layered Access Control with UCON ». Dans 2009 International Conference on Computational Science and Engineering. IEEE, 2009. http://dx.doi.org/10.1109/cse.2009.163.

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« Enforcement of CA-UCON Model ». Dans International Academy of Engineers. International Academy of Engineers, 2015. http://dx.doi.org/10.15242/iae.iae0615001.

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Amalfi, Raffaele L., Todd Salamon, Filippo Cataldo, Jackson B. Marcinichen et John R. Thome. « Ultra-Compact Micro-Scale Heat Exchanger for Advanced Thermal Management in Datacenters ». Dans ASME 2020 International Technical Conference and Exhibition on Packaging and Integration of Electronic and Photonic Microsystems. American Society of Mechanical Engineers, 2020. http://dx.doi.org/10.1115/ipack2020-2542.

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Abstract The present study is focused on the experimental characterization of two-phase heat transfer performance and pressure drops within an ultra-compact heat exchanger (UCHE) suitable for electronics cooling applications. In this specific work, the UCHE prototype is anticipated to be a critical component for realizing a new passive two-phase cooling technology for high-power server racks, as it is more compact and lighter weight than conventional heat exchangers. This technology makes use of a novel combination of thermosyphon loops, at the server-level and rack-level, to passively cool an entire rack. In the proposed two-phase cooling technology, a smaller form factor UCHE is used to transfer heat from the server-level thermosyphon cooling loop to the rack-level thermosyphon cooling loop, while a larger form factor UCHE is used to reject the total heat from the server rack into the facility-level cooling loop. The UCHE is composed of a double-side-copper finned plate enclosed in a stainless steel enclosure. The geometry of the fins and channels on both sides are optimized to enhance the heat transfer performance and flow stability, while minimizing the pressure drops. These features make the UCHE the ideal component for thermosyphon cooling systems, where low pressure drops are required to achieve high passive flow circulation rates and thus achieve high critical heat flux values. The UCHE’s thermal-hydraulic performance is first evaluated in a pump-driven system at the Laboratory of Heat and Mass Transfer (LTCM-EPFL), where experiments include many configurations and operating conditions. Then, the UCHE is installed and tested as the condenser of a thermosyphon loop that rejects heat to a pumped refrigerant system at Nokia Bell Labs, in which both sides operate with refrigerants in phase change (condensation-to-boiling). Experimental results demonstrate high thermal performance with a maximum heat dissipation density of 5455 (kW/m3/K), which is significantly larger than conventional air-cooled heat exchangers and liquid-cooled small pressing depth brazed plate heat exchangers. Finally, a thermal performance analysis is presented that provides guidelines in terms of heat density dissipations at the server- and rack-level when using passive two-phase cooling.
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Vališka, Michal, Jiří Pospíšil, Gwilherm Nénert, Anne Stunnault, Karel Prokeš et Vladimír Sechovský. « Evolution of Magnetism in UCo1−xRuxGe ». Dans Proceedings of the International Conference on Strongly Correlated Electron Systems (SCES2013). Journal of the Physical Society of Japan, 2014. http://dx.doi.org/10.7566/jpscp.3.012011.

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Zhou, Larry, Angela Chiu, Michael Satterlee, Dave Mahar, Qiong Zhang, Paparao Palacharla et Ikeuchi Tadashi. « IoT Gateway Edge VNFs on uCPE ». Dans 2018 IEEE Conference on Network Function Virtualization and Software Defined Networks (NFV-SDN). IEEE, 2018. http://dx.doi.org/10.1109/nfv-sdn.2018.8725618.

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LI, Zude, et Xiaojun YE. « ATTRIBUTE ANALYSIS OF USAGE CONTROL (UCON) ». Dans 11th Joint International Computer Conference - JICC 2005. WORLD SCIENTIFIC, 2005. http://dx.doi.org/10.1142/9789812701534_0014.

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Quiles, Francisco J., Manuel Ortiz, Maria Brox, Carlos D. Moreno, Javier Hormigo et Julio Villalba. « UCORE : Reconfigurable Platform for Educational Purposes ». Dans 2010 International Conference on Reconfigurable Computing and FPGAs (ReConFig 2010). IEEE, 2010. http://dx.doi.org/10.1109/reconfig.2010.60.

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Gansterer, Wilfried N., et Michael Ilger. « Analyzing UCE/UBE traffic ». Dans the ninth international conference. New York, New York, USA : ACM Press, 2007. http://dx.doi.org/10.1145/1282100.1282139.

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Rapports d'organisations sur le sujet "UCOE"

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Ishikawa, C. A URN Namespace for ucode. RFC Editor, avril 2012. http://dx.doi.org/10.17487/rfc6588.

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Nekoogar, F. UCom : Ultra-wideband Communications in Harsh Propagation Environments. Office of Scientific and Technical Information (OSTI), mars 2007. http://dx.doi.org/10.2172/902376.

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DeVelasco, R. I. Critical process parameters for UCO kernel production. Office of Scientific and Technical Information (OSTI), septembre 1988. http://dx.doi.org/10.2172/453983.

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Allen, M. D., Catherine Macheret et Mary A. Malloy. C2 Core and UCore Message Design Capstone : Interoperable Message Structure. Fort Belvoir, VA : Defense Technical Information Center, septembre 2009. http://dx.doi.org/10.21236/ada596680.

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Gerhard Strydom. Reactor Physics Characterization of the HTR Module with UCO Fuel. Office of Scientific and Technical Information (OSTI), janvier 2011. http://dx.doi.org/10.2172/1009138.

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Hunn, John D., Tyler J. Gerczak, Fred C. Montgomery, Darren J. Skitt, Charles A. Baldwin, Grant W. Helmreich, Brian D. Eckhart et John A. Dyer. AGR-2 Safety-Tested UCO Compact 6-4-2 PIE Report. Office of Scientific and Technical Information (OSTI), juin 2018. http://dx.doi.org/10.2172/1440824.

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Hunn, John D., Tyler J. Gerczak, Fred C. Montgomery, Darren J. Skitt, Charles A. Baldwin, Grant W. Helmreich, Brian D. Eckhart et John A. Dyer. AGR-2 As-Irradiated UCO Compact 5-4-2 PIE Report. Office of Scientific and Technical Information (OSTI), juin 2018. http://dx.doi.org/10.2172/1454396.

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Demkowicz, Paul. Technical Bases for the Performance Demonstration of TRISO-coated UCO Fuel Particles. Office of Scientific and Technical Information (OSTI), septembre 2018. http://dx.doi.org/10.2172/1471737.

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Hunn, John, Fred Montgomery et Peter Pappano. Data Compilation for AGR-2 UCO Variant Compact Lot LEU09-OP2-Z. Office of Scientific and Technical Information (OSTI), février 2010. http://dx.doi.org/10.2172/1630497.

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Hunn, John, Fred Montgomery et Peter Pappano. Data Compilation for AGR-2 UCO Baseline Compact Lot LEU07-OP1-Z. Office of Scientific and Technical Information (OSTI), décembre 2009. http://dx.doi.org/10.2172/1632080.

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