Science.gov

Sample records for reporter gene fusion

  1. Multimodality imaging of reporter gene expression using a novel fusion vector in living cells and animals

    DOEpatents

    Gambhir; Sanjiv , Pritha; Ray

    2009-04-28

    Novel double and triple fusion reporter gene constructs harboring distinct imageable reporter genes are provided, as well as applications for the use of such double and triple fusion constructs in living cells and in living animals using distinct imaging technologies.

  2. Multimodality imaging of reporter gene expression using a novel fusion vector in living cells and animals

    DOEpatents

    Gambhir, Sanjiv; Pritha, Ray

    2011-06-07

    Novel double and triple fusion reporter gene constructs harboring distinct imagable reporter genes are provided, as well as applications for the use of such double and triple fusion constructs in living cells and in living animals using distinct imaging technologies.

  3. Multimodality imaging of reporter gene expression using a novel fusion vector in living cells and animals

    SciTech Connect

    Gambhir, Sanjiv; Pritha, Ray

    2015-07-14

    Novel double and triple fusion reporter gene constructs harboring distinct imagable reporter genes are provided, as well as applications for the use of such double and triple fusion constructs in living cells and in living animals using distinct imaging technologies.

  4. The fusion Vibrio campbellii luciferase as a eukaryotic gene reporter.

    PubMed

    Tinikul, Ruchanok; Thotsaporn, Kittisak; Thaveekarn, Wichit; Jitrapakdee, Sarawut; Chaiyen, Pimchai

    2012-12-31

    Bacterial luciferase from Vibrio campbellii is a thermostable enzyme with an in vitro thermal inactivation half-life of ~1020 min at 37°C. The enzyme also binds tightly to reduced FMN. In this study, a V. campbellii fusion luciferase construct in which the α and β subunits are linked with a decapeptide was made and characterized. In general, the overall enzymatic properties of the two enzymes are similar. Expression of the enzymes in Escherichia coli demonstrated that the V. campbellii fusion luciferase emits less light than the native luciferase, but still emits a much greater amount of light than native luciferase from Vibrio harveyi and Photobacterium leiognathi TH1. The intensity of light emitted by the V. campbellii fusion luciferase was more than 80-fold greater than that from the V. harveyi native luciferase when expressed at 37°C. Biochemical characterization has shown that the V. campbellii fusion luciferase also retains a high binding affinity for reduced flavin mononucleotide and high thermostability. The levels of bioluminescence emitted by the V. campbellii fusion luciferase expressed in HEK293T cells reached ~1×10(6) Relative Light Units/mg total protein. These findings suggest that the V. campbellii fusion luciferase is a promising candidate for further development as a luciferase-based reporter for eukaryotic systems.

  5. Xp11 neoplasm with melanocytic differentiation of the prostate harbouring the novel NONO-TFE3 gene fusion: report of a unique case expanding the gene fusion spectrum.

    PubMed

    Wang, Xiao-Tong; Xia, Qiu-Yuan; Ni, Hao; Wang, Zi-Yu; Ye, Sheng-Bing; Li, Rui; Wang, Xuan; Lv, Jing-Huan; Shi, Shan-Shan; Ma, Heng-Hui; Lu, Zhen-Feng; Shen, Qin; Zhou, Xiao-Jun; Rao, Qiu

    2016-09-01

    Recently, an increasing number of TFE3 rearrangement-associated tumours have been reported, such as TFE3 rearrangement-associated perivascular epithelioid cell tumours (PEComas), melanotic Xp11 translocation renal cancers and melanotic Xp11 neoplasms. We have suggested that these tumours belong to a single clinicopathological spectrum. 'Xp11 neoplasm with melanocytic differentiation' or 'melanotic Xp11 neoplasm' have been proposed to designate this unique neoplasm. Herein, we describe the first case of an Xp11 neoplasm with melanocytic differentiation to be described in the prostate, bearing the novel NONO-TFE3 gene fusion. This study both adds to the spectrum regarding melanotic Xp11 neoplasms and expands its gene fusion spectrum. Moreover, we discuss the relationship of these rare tumours to neoplasms such as conventional PEComas, alveolar soft part sarcomas, malignant melanomas, clear cell sarcomas and Xp11 translocation renal cancers.

  6. Dual-therapeutic reporter genes fusion for enhanced cancer gene therapy and imaging.

    PubMed

    Sekar, T V; Foygel, K; Willmann, J K; Paulmurugan, R

    2013-05-01

    Two of the successful gene-directed enzyme prodrug therapies include herpes simplex virus-thymidine kinase (HSV1-TK) enzyme-ganciclovir prodrug and the Escherichia coli nitroreductase (NTR) enzyme-CB1954 prodrug strategies; these enzyme-prodrug combinations produce activated cytotoxic metabolites of the prodrugs capable of tumor cell death by inhibiting DNA synthesis and killing quiescent cells, respectively. Both these strategies also affect significant bystander cell killing of neighboring tumor cells that do not express these enzymes. We have developed a dual-combination gene strategy, where we identified HSV1-TK and NTR fused in a particular orientation can effectively kill tumor cells when the tumor cells are treated with a fusion HSV1-TK-NTR gene- along with a prodrug combination of GCV and CB1954. In order to determine whether the dual-system demonstrate superior therapeutic efficacy than either HSV1-TK or NTR systems alone, we conducted both in vitro and in vivo tumor xenograft studies using triple negative SUM159 breast cancer cells, by evaluating the efficacy of cell death by apoptosis and necrosis upon treatment with the dual HSV1-TK genes-GCV-CB1954 prodrugs system, and compared the efficiency to HSV1-TK-GCV and NTR-CB1954. Our cell-based studies, tumor regression studies in xenograft mice, histological analyses of treated tumors and bystander studies indicate that the dual HSV1-TK-NTR-prodrug system is two times more efficient even with half the doses of both prodrugs than the respective single gene-prodrug system, as evidenced by enhanced apoptosis and necrosis of tumor cells in vitro in culture and xenograft of tumor tissues in animals.

  7. Regulation of Sinorhizobium meliloti 1021 rrnA-reporter gene fusions in response to cold shock.

    PubMed

    Gustafson, Ann M; O'Connell, Kevin P; Thomashow, Michael F

    2002-09-01

    We previously reported that mutants of Sinorhizobium meliloti 1021 carrying luxAB insertions in each of the three 16S rRNA genes exhibited a dramatic (> or = 28-fold) increase in luminescence following a temperature downshift from 30 to 15 degrees C. These results raised the possibility that the rRNA operons (rrn) of S. meliloti were cold shock loci. In testing this possibility, we found that fusion of the S. meliloti 1021 rrnA promoter to two different reporter genes, luxAB and uidA, resulted in hybrid genes that were transiently upregulated (as measured by transcript accumulation) about four- to sixfold in response to a temperature downshift. These results are consistent with the hypothesis that the rrn promoters are transiently upregulated in response to cold shock. However, much of the apparent cold shock regulation of the initial luxAB insertions was due to an unexpected mechanism: an apparent temperature-dependent inhibition of translation. Specifically, the rrnA sequences from +1 to +172 (relative to the start of transcription) were found to greatly decrease the ability of S. meliloti to translate hybrid rrn-luxAB transcripts into active protein at 30 degrees C. This effect, however, was largely eliminated at 15 degrees C. Possible mechanisms for the apparent transient increase in rrnA promoter activity and temperature-dependent inhibition of translation are discussed.

  8. Gene Fusion: A Genome Wide Survey

    NASA Technical Reports Server (NTRS)

    Liang, Ping; Riley, Monica

    2001-01-01

    As a well known fact, organisms form larger and complex multimodular (composite or chimeric) and mostly multi-functional proteins through gene fusion of two or more individual genes which have independent evolution histories and functions. We call each of these components a module. The existence of multimodular proteins may improves the efficiency in gene regulation and in cellular functions, and thus may give the host organism advantages in adaptation to environments. Analysis of all gene fusions in present-day organisms should allow us to examine the patterns of gene fusion in context with cellular functions, to trace back the evolution processes from the ancient smaller and uni-functional proteins to the present-day larger and complex multi-functional proteins, and to estimate the minimal number of ancestor proteins that existed in the last common ancestor for all life on earth. Although many multimodular proteins have been experimentally known, identification of gene fusion events systematically at genome scale had not been possible until recently when large number of completed genome sequences have been becoming available. In addition, technical difficulties for such analysis also exist due to the complexity of this biological and evolutionary process. We report from this study a new strategy to computationally identify multimodular proteins using completed genome sequences and the results surveyed from 22 organisms with the data from over 40 organisms to be presented during the meeting. Additional information is contained in the original extended abstract.

  9. Combining random gene fission and rational gene fusion to discover near-infrared fluorescent protein fragments that report on protein-protein interactions.

    PubMed

    Pandey, Naresh; Nobles, Christopher L; Zechiedrich, Lynn; Maresso, Anthony W; Silberg, Jonathan J

    2015-05-15

    Gene fission can convert monomeric proteins into two-piece catalysts, reporters, and transcription factors for systems and synthetic biology. However, some proteins can be challenging to fragment without disrupting function, such as near-infrared fluorescent protein (IFP). We describe a directed evolution strategy that can overcome this challenge by randomly fragmenting proteins and concomitantly fusing the protein fragments to pairs of proteins or peptides that associate. We used this method to create libraries that express fragmented IFP as fusions to a pair of associating peptides (IAAL-E3 and IAAL-K3) and proteins (CheA and CheY) and screened for fragmented IFP with detectable near-infrared fluorescence. Thirteen novel fragmented IFPs were identified, all of which arose from backbone fission proximal to the interdomain linker. Either the IAAL-E3 and IAAL-K3 peptides or CheA and CheY proteins could assist with IFP fragment complementation, although the IAAL-E3 and IAAL-K3 peptides consistently yielded higher fluorescence. These results demonstrate how random gene fission can be coupled to rational gene fusion to create libraries enriched in fragmented proteins with AND gate logic that is dependent upon a protein-protein interaction, and they suggest that these near-infrared fluorescent protein fragments will be suitable as reporters for pairs of promoters and protein-protein interactions within whole animals.

  10. Adamantinoma-like Ewing's sarcoma with EWS-FLI1 fusion gene: a case report.

    PubMed

    Fujii, Hiromasa; Honoki, Kanya; Enomoto, Yasunori; Kasai, Takahiko; Kido, Akira; Amano, Itsuto; Kumamoto, Makiko; Morishita, Toru; Mii, Yoshio; Nonomura, Akitaka; Takakura, Yoshinori

    2006-11-01

    Recent studies have advocated the genotypic and phenotypic delineation of a novel Ewing's sarcoma histologic variant showing epithelial features defined as "adamantinoma-like Ewing's sarcoma". We described an 18-year-old girl with a primary small round-cell sarcoma of the right tibia showing polyphenotypic differentiation with epithelioid features. The neoplastic cells had mainly round or oval nuclei with fine chromatin with a portion of epithelial arrangements. The immunohistochemical analysis showed the epithelial markers of cytokeratin 5/6/18, AE1/AE3, and cytokeratin high molecular weight were stained especially in the foci with epithelioid features, as well as MIC2, S100, and NSE. The diagnosis of the lesion was confirmed as Ewing's sarcoma by the presence of the EWS-FLI1 fusion transcript, and could be defined as the so-called "adamantinoma-like Ewing's sarcoma". After wide excision and high-dose chemotherapy with peripheral blood stem cell transfusion, the patient has been well and continuously event-free for 3 years since the initial diagnosis.

  11. Fusion Simulation Project Workshop Report

    NASA Astrophysics Data System (ADS)

    Kritz, Arnold; Keyes, David

    2009-03-01

    The mission of the Fusion Simulation Project is to develop a predictive capability for the integrated modeling of magnetically confined plasmas. This FSP report adds to the previous activities that defined an approach to integrated modeling in magnetic fusion. These previous activities included a Fusion Energy Sciences Advisory Committee panel that was charged to study integrated simulation in 2002. The report of that panel [Journal of Fusion Energy 20, 135 (2001)] recommended the prompt initiation of a Fusion Simulation Project. In 2003, the Office of Fusion Energy Sciences formed a steering committee that developed a project vision, roadmap, and governance concepts [Journal of Fusion Energy 23, 1 (2004)]. The current FSP planning effort involved 46 physicists, applied mathematicians and computer scientists, from 21 institutions, formed into four panels and a coordinating committee. These panels were constituted to consider: Status of Physics Components, Required Computational and Applied Mathematics Tools, Integration and Management of Code Components, and Project Structure and Management. The ideas, reported here, are the products of these panels, working together over several months and culminating in a 3-day workshop in May 2007.

  12. Fusion Breeder Program interim report

    SciTech Connect

    Moir, R.; Lee, J.D.; Neef, W.

    1982-06-11

    This interim report for the FY82 Fusion Breeder Program covers work performed during the scoping phase of the study, December, 1981-February 1982. The goals for the FY82 study are the identification and development of a reference blanket concept using the fission suppression concept and the definition of a development plan to further the fusion breeder application. The context of the study is the tandem mirror reactor, but emphasis is placed upon blanket engineering. A tokamak driver and blanket concept will be selected and studied in more detail during FY83.

  13. Construction of a cytosolic firefly luciferase reporter cassette for use in PCR-mediated gene deletion and fusion in Saccharomyces cerevisiae.

    PubMed

    Ainsworth, W B; Rome, C M; Hjortsø, M A; Benton, M G

    2012-12-01

    Monitoring promoter response to environmental changes using reporter systems has provided invaluable information regarding cellular state. With the development of in vivo luciferase reporter systems, inexpensive, sensitive and accurate promoter assays have been developed without the variability reported between in vitro samplings. Current luciferase reporter systems, however, are largely inflexible to modifications to the promoter of interest. To overcome problems in flexibility and stability of these expression vectors, we report the creation of a novel vector system which introduces a cytosol-localized Photinus pyralis luciferase [LUC*(-SKL)] capable of one-step, in vivo measurements into a promoter-reporter system via PCR-based gene deletion and fusion. After introduction of the reporter under HUG1 promoter control, cytosolic localization was confirmed by fluorescence microscopy. The dose-response of this novel construct was then compared with that of a similar HUG1Δ::yEGFP1 promoter-reporter system and shown to give a similar response pattern.

  14. INTEGRATE: gene fusion discovery using whole genome and transcriptome data

    PubMed Central

    Zhang, Jin; White, Nicole M.; Schmidt, Heather K.; Fulton, Robert S.; Tomlinson, Chad; Warren, Wesley C.; Wilson, Richard K.; Maher, Christopher A.

    2016-01-01

    While next-generation sequencing (NGS) has become the primary technology for discovering gene fusions, we are still faced with the challenge of ensuring that causative mutations are not missed while minimizing false positives. Currently, there are many computational tools that predict structural variations (SV) and gene fusions using whole genome (WGS) and transcriptome sequencing (RNA-seq) data separately. However, as both WGS and RNA-seq have their limitations when used independently, we hypothesize that the orthogonal validation from integrating both data could generate a sensitive and specific approach for detecting high-confidence gene fusion predictions. Fortunately, decreasing NGS costs have resulted in a growing quantity of patients with both data available. Therefore, we developed a gene fusion discovery tool, INTEGRATE, that leverages both RNA-seq and WGS data to reconstruct gene fusion junctions and genomic breakpoints by split-read mapping. To evaluate INTEGRATE, we compared it with eight additional gene fusion discovery tools using the well-characterized breast cell line HCC1395 and peripheral blood lymphocytes derived from the same patient (HCC1395BL). The predictions subsequently underwent a targeted validation leading to the discovery of 131 novel fusions in addition to the seven previously reported fusions. Overall, INTEGRATE only missed six out of the 138 validated fusions and had the highest accuracy of the nine tools evaluated. Additionally, we applied INTEGRATE to 62 breast cancer patients from The Cancer Genome Atlas (TCGA) and found multiple recurrent gene fusions including a subset involving estrogen receptor. Taken together, INTEGRATE is a highly sensitive and accurate tool that is freely available for academic use. PMID:26556708

  15. INTEGRATE: gene fusion discovery using whole genome and transcriptome data.

    PubMed

    Zhang, Jin; White, Nicole M; Schmidt, Heather K; Fulton, Robert S; Tomlinson, Chad; Warren, Wesley C; Wilson, Richard K; Maher, Christopher A

    2016-01-01

    While next-generation sequencing (NGS) has become the primary technology for discovering gene fusions, we are still faced with the challenge of ensuring that causative mutations are not missed while minimizing false positives. Currently, there are many computational tools that predict structural variations (SV) and gene fusions using whole genome (WGS) and transcriptome sequencing (RNA-seq) data separately. However, as both WGS and RNA-seq have their limitations when used independently, we hypothesize that the orthogonal validation from integrating both data could generate a sensitive and specific approach for detecting high-confidence gene fusion predictions. Fortunately, decreasing NGS costs have resulted in a growing quantity of patients with both data available. Therefore, we developed a gene fusion discovery tool, INTEGRATE, that leverages both RNA-seq and WGS data to reconstruct gene fusion junctions and genomic breakpoints by split-read mapping. To evaluate INTEGRATE, we compared it with eight additional gene fusion discovery tools using the well-characterized breast cell line HCC1395 and peripheral blood lymphocytes derived from the same patient (HCC1395BL). The predictions subsequently underwent a targeted validation leading to the discovery of 131 novel fusions in addition to the seven previously reported fusions. Overall, INTEGRATE only missed six out of the 138 validated fusions and had the highest accuracy of the nine tools evaluated. Additionally, we applied INTEGRATE to 62 breast cancer patients from The Cancer Genome Atlas (TCGA) and found multiple recurrent gene fusions including a subset involving estrogen receptor. Taken together, INTEGRATE is a highly sensitive and accurate tool that is freely available for academic use.

  16. Characterization of fusion genes and the significantly expressed fusion isoforms in breast cancer by hybrid sequencing.

    PubMed

    Weirather, Jason L; Afshar, Pegah Tootoonchi; Clark, Tyson A; Tseng, Elizabeth; Powers, Linda S; Underwood, Jason G; Zabner, Joseph; Korlach, Jonas; Wong, Wing Hung; Au, Kin Fai

    2015-10-15

    We developed an innovative hybrid sequencing approach, IDP-fusion, to detect fusion genes, determine fusion sites and identify and quantify fusion isoforms. IDP-fusion is the first method to study gene fusion events by integrating Third Generation Sequencing long reads and Second Generation Sequencing short reads. We applied IDP-fusion to PacBio data and Illumina data from the MCF-7 breast cancer cells. Compared with the existing tools, IDP-fusion detects fusion genes at higher precision and a very low false positive rate. The results show that IDP-fusion will be useful for unraveling the complexity of multiple fusion splices and fusion isoforms within tumorigenesis-relevant fusion genes.

  17. ETS fusion genes in prostate cancer.

    PubMed

    Gasi Tandefelt, Delila; Boormans, Joost; Hermans, Karin; Trapman, Jan

    2014-06-01

    Prostate cancer is very common in elderly men in developed countries. Unravelling the molecular and biological processes that contribute to tumor development and progressive growth, including its heterogeneity, is a challenging task. The fusion of the genes ERG and TMPRSS2 is the most frequent genomic alteration in prostate cancer. ERG is an oncogene that encodes a member of the family of ETS transcription factors. At lower frequency, other members of this gene family are also rearranged and overexpressed in prostate cancer. TMPRSS2 is an androgen-regulated gene that is preferentially expressed in the prostate. Most of the less frequent ETS fusion partners are also androgen-regulated and prostate-specific. During the last few years, novel concepts of the process of gene fusion have emerged, and initial experimental results explaining the function of the ETS genes ERG and ETV1 in prostate cancer have been published. In this review, we focus on the most relevant ETS gene fusions and summarize the current knowledge of the role of ETS transcription factors in prostate cancer. Finally, we discuss the clinical relevance of TMRPSS2-ERG and other ETS gene fusions in prostate cancer.

  18. Mammary Analogue Secretory Carcinoma of Salivary Glands: Molecular Analysis of 25 ETV6 Gene Rearranged Tumors With Lack of Detection of Classical ETV6-NTRK3 Fusion Transcript by Standard RT-PCR: Report of 4 Cases Harboring ETV6-X Gene Fusion.

    PubMed

    Skálová, Alena; Vanecek, Tomas; Simpson, Roderick H W; Laco, Jan; Majewska, Hanna; Baneckova, Martina; Steiner, Petr; Michal, Michal

    2016-01-01

    ETV6 gene abnormalities are well described in tumor pathology. Many fusion partners of ETV6 have been reported in a variety of epithelial and hematological malignancies. In salivary gland tumor pathology, however, the ETV6-NTRK3 translocation is specific for mammary analogue secretory carcinoma (MASC), and has not been documented in any other salivary tumor type. The present study comprised a clinical and molecular analysis of 25 cases morphologically and immunohistochemically typical of MASC. They all also displayed the ETV6 rearrangement as visualized by fluorescent in situ hybridization but lacked the classical ETV6-NTRK3 fusion transcript by standard reverse-transcriptase-polymerase chain reaction. In 4 cases, the classical fusion transcript was found by more sensitive, nested reverse-transcription-polymerase chain reaction. Five other cases harbored atypical fusion transcripts as detected by both standard and nested reverse-transcription-polymerase chain reaction. In addition, fluorescent in situ hybridization with an NTRK3 break-apart probe was also performed; rearrangement of NTRK3 gene was detected in 16 of 25 cases. In 3 other cases, the tissue was not analyzable, and in 2 further cases analysis could not be performed because of a lack of appropriate tissue material. Finally, in the 4 remaining cases whose profile was NTRK3 split-negative and ETV6 split-positive, unknown (non-NTRK) genes appeared to fuse with ETV6 (ETV6-X fusion). In looking for possible fusion partners, analysis of rearrangement of other kinase genes known to fuse with ETV6 was also performed, but without positive results. Although numbers were small, correlating the clinico-pathologic features of the 4 ETV6-X fusion tumors and 5 MASC cases with atypical fusion transcripts raises the possibility of that they may behave more aggressively.

  19. FGFR-TACC gene fusions in human glioma.

    PubMed

    Lasorella, Anna; Sanson, Marc; Iavarone, Antonio

    2016-11-16

    Chromosomal translocations joining in-frame members of the fibroblast growth factor receptor-transforming acidic coiled-coil gene families (the FGFR-TACC gene fusions) were first discovered in human glioblastoma multiforme (GBM) and later in many other cancer types. Here, we review this rapidly expanding field of research and discuss the unique biological and clinical features conferred to isocitrate dehydrogenase wild-type glioma cells by FGFR-TACC fusions. FGFR-TACC fusions generate powerful oncogenes that combine growth-promoting effects with aneuploidy through the activation of as yet unclear intracellular signaling mechanisms. FGFR-TACC fusions appear to be clonal tumor-initiating events that confer strong sensitivity to FGFR tyrosine kinase inhibitors. Screening assays have recently been reported for the accurate identification of FGFR-TACC fusion variants in human cancer, and early clinical data have shown promising effects in cancer patients harboring FGFR-TACC fusions and treated with FGFR inhibitors. Thus, FGFR-TACC gene fusions provide a "low-hanging fruit" model for the validation of precision medicine paradigms in human GBM.

  20. Gene Fusion Markup Language: a prototype for exchanging gene fusion data

    PubMed Central

    2012-01-01

    Background An avalanche of next generation sequencing (NGS) studies has generated an unprecedented amount of genomic structural variation data. These studies have also identified many novel gene fusion candidates with more detailed resolution than previously achieved. However, in the excitement and necessity of publishing the observations from this recently developed cutting-edge technology, no community standardization approach has arisen to organize and represent the data with the essential attributes in an interchangeable manner. As transcriptome studies have been widely used for gene fusion discoveries, the current non-standard mode of data representation could potentially impede data accessibility, critical analyses, and further discoveries in the near future. Results Here we propose a prototype, Gene Fusion Markup Language (GFML) as an initiative to provide a standard format for organizing and representing the significant features of gene fusion data. GFML will offer the advantage of representing the data in a machine-readable format to enable data exchange, automated analysis interpretation, and independent verification. As this database-independent exchange initiative evolves it will further facilitate the formation of related databases, repositories, and analysis tools. The GFML prototype is made available at http://code.google.com/p/gfml-prototype/. Conclusion The Gene Fusion Markup Language (GFML) presented here could facilitate the development of a standard format for organizing, integrating and representing the significant features of gene fusion data in an inter-operable and query-able fashion that will enable biologically intuitive access to gene fusion findings and expedite functional characterization. A similar model is envisaged for other NGS data analyses. PMID:23072312

  1. INTRODUCTION: Status report on fusion research

    NASA Astrophysics Data System (ADS)

    Burkart, Werner

    2005-10-01

    A major milestone on the path to fusion energy was reached in June 2005 on the occasion of the signing of the joint declaration of all parties to the ITER negotiations, agreeing on future arrangements and on the construction site at Cadarache in France. The International Atomic Energy Agency has been promoting fusion activities since the late 1950s; it took over the auspices of the ITER Conceptual Design Activities in 1988, and of the ITER Engineering and Design Activities in 1992. The Agency continues its support to Member States through the organization of consultancies, workshops and technical meetings, the most prominent being the series of International Fusion Energy Conferences (formerly called the International Conference on Plasma Physics and Controlled Nuclear Fusion Research). The meetings serve as a platform for experts from all Member States to have open discussions on their latest accomplishments as well as on their problems and eventual solutions. The papers presented at the meetings and conferences are routinely published, many being sent to the journal it Nuclear Fusion, co-published monthly by Institute of Physics Publishing, Bristol, UK. The journal's reputation is reflected in the fact that it is a world-renowned publication, and the International Fusion Research Council has used it for the publication of a Status Report on Controlled Thermonuclear Fusion in 1978 and 1990. This present report marks the conclusion of the preparatory phases of ITER activities. It provides background information on the progress of fusion research within the last 15 years. The International Fusion Research Council (IFRC), which initiated the report, was fully aware of the complexities of including all scientific results in just one paper, and so decided to provide an overview and extensive references for the interested reader who need not necessarily be a fusion specialist. Professor Predhiman K. Kaw, Chairman, prepared the report on behalf of the IFRC, reflecting

  2. Molecular pathways: targeting ETS gene fusions in cancer.

    PubMed

    Feng, Felix Y; Brenner, J Chad; Hussain, Maha; Chinnaiyan, Arul M

    2014-09-01

    Rearrangements, or gene fusions, involving the ETS family of transcription factors are common driving events in both prostate cancer and Ewing sarcoma. These rearrangements result in pathogenic expression of the ETS genes and trigger activation of transcriptional programs enriched for invasion and other oncogenic features. Although ETS gene fusions represent intriguing therapeutic targets, transcription factors, such as those comprising the ETS family, have been notoriously difficult to target. Recently, preclinical studies have demonstrated an association between ETS gene fusions and components of the DNA damage response pathway, such as PARP1, the catalytic subunit of DNA protein kinase (DNAPK), and histone deactylase 1 (HDAC1), and have suggested that ETS fusions may confer sensitivity to inhibitors of these DNA repair proteins. In this review, we discuss the role of ETS fusions in cancer, the preclinical rationale for targeting ETS fusions with inhibitors of PARP1, DNAPK, and HDAC1, as well as ongoing clinical trials targeting ETS gene fusions.

  3. Myeloid Neoplasms with t(5;12) and ETV6-ACSL6 Gene Fusion, Potential Mimickers of Myeloid Neoplasm with PDGFRB Rearrangement: Case Report with Imatinib Therapy and Review of the Literature

    PubMed Central

    Ninfea, Jose I. Ruades; Pearson, Lauren; Conant, Joanna; Bryant, Ronald; Zakai, Neil A.; Tang, Mary E.

    2016-01-01

    We report the second case of ETV6-ACSL6 associated myeloproliferative neoplasm that has received a full course of imatinib therapy. The patient was a 51-year-old previously healthy man who presented with three months of worsening dyspnea and was found to have a white count of 216,000/cmm, of which 84% were eosinophil lineage. Cytogenetic analysis revealed a t(5;12)(q31~33;p13). FISH was negative for PDGFRB rearrangement but additional FISH testing demonstrated an ACSL6 rearrangement. ETV6-ACSL6 gene fusion is a rare abnormality that most often presents as a myeloproliferative-type disorder with prominent eosinophilia or basophilia. Review of the literature yielded a total of 11 previous cases. This gene fusion results in a t(5;12)(q31~33;p13) that mimics the t(5;12) found in ETV6-PDGFRB neoplasms. Identification of the fusion genes involved in t(5;12) in eosinophilia-associated myeloproliferative disorders is crucial to direct an effective treatment plan. In particular, while tyrosine kinase inhibitor therapy is effective in patients with PDGFRB rearrangement, there is little information on imatinib efficacy in patients with ETV6-ACSL6 gene fusion. Our patient was found to be nonresponsive to imatinib therapy. PMID:27746819

  4. Repeated evolution of chimeric fusion genes in the β-globin gene family of laurasiatherian mammals.

    PubMed

    Gaudry, Michael J; Storz, Jay F; Butts, Gary Tyler; Campbell, Kevin L; Hoffmann, Federico G

    2014-05-09

    The evolutionary fate of chimeric fusion genes may be strongly influenced by their recombinational mode of origin and the nature of functional divergence between the parental genes. In the β-globin gene family of placental mammals, the two postnatally expressed δ- and β-globin genes (HBD and HBB, respectively) have a propensity for recombinational exchange via gene conversion and unequal crossing-over. In the latter case, there are good reasons to expect differences in retention rates for the reciprocal HBB/HBD and HBD/HBB fusion genes due to thalassemia pathologies associated with the HBD/HBB "Lepore" deletion mutant in humans. Here, we report a comparative genomic analysis of the mammalian β-globin gene cluster, which revealed that chimeric HBB/HBD fusion genes originated independently in four separate lineages of laurasiatherian mammals: Eulipotyphlans (shrews, moles, and hedgehogs), carnivores, microchiropteran bats, and cetaceans. In cases where an independently derived "anti-Lepore" duplication mutant has become fixed, the parental HBD and/or HBB genes have typically been inactivated or deleted, so that the newly created HBB/HBD fusion gene is primarily responsible for synthesizing the β-type subunits of adult and fetal hemoglobin (Hb). Contrary to conventional wisdom that the HBD gene is a vestigial relict that is typically inactivated or expressed at negligible levels, we show that HBD-like genes often encode a substantial fraction (20-100%) of β-chain Hbs in laurasiatherian taxa. Our results indicate that the ascendancy or resuscitation of genes with HBD-like coding sequence requires the secondary acquisition of HBB-like promoter sequence via unequal crossing-over or interparalog gene conversion.

  5. Identification of Targetable FGFR Gene Fusions in Diverse Cancers

    PubMed Central

    Wu, Yi-Mi; Su, Fengyun; Kalyana-Sundaram, Shanker; Khazanov, Nick; Ateeq, Bushra; Cao, Xuhong; Lonigro, Robert J.; Vats, Pankaj; Wang, Rui; Lin, Su-Fang; Cheng, Ann-Joy; Kunju, Lakshmi P.; Siddiqui, Javed; Tomlins, Scott A.; Wyngaard, Peter; Sadis, Seth; Roychowdhury, Sameek; Hussain, Maha H.; Feng, Felix Y.; Zalupski, Mark M.; Talpaz, Moshe; Pienta, Kenneth J.; Rhodes, Daniel R.; Robinson, Dan R.; Chinnaiyan, Arul M.

    2013-01-01

    Through a prospective clinical sequencing program for advanced cancers, four index cases were identified which harbor gene rearrangements of FGFR2 including patients with cholangiocarcinoma, breast cancer, and prostate cancer. After extending our assessment of FGFR rearrangements across multiple tumor cohorts, we identified additional FGFR gene fusions with intact kinase domains in lung squamous cell cancer, bladder cancer, thyroid cancer, oral cancer, glioblastoma, and head and neck squamous cell cancer. All FGFR fusion partners tested exhibit oligomerization capability, suggesting a shared mode of kinase activation. Overexpression of FGFR fusion proteins induced cell proliferation. Two bladder cancer cell lines that harbor FGFR3 fusion proteins exhibited enhanced susceptibility to pharmacologic inhibition in vitro and in vivo. Due to the combinatorial possibilities of FGFR family fusion to a variety of oligomerization partners, clinical sequencing efforts which incorporate transcriptome analysis for gene fusions are poised to identify rare, targetable FGFR fusions across diverse cancer types. PMID:23558953

  6. Identification of targetable FGFR gene fusions in diverse cancers.

    PubMed

    Wu, Yi-Mi; Su, Fengyun; Kalyana-Sundaram, Shanker; Khazanov, Nickolay; Ateeq, Bushra; Cao, Xuhong; Lonigro, Robert J; Vats, Pankaj; Wang, Rui; Lin, Su-Fang; Cheng, Ann-Joy; Kunju, Lakshmi P; Siddiqui, Javed; Tomlins, Scott A; Wyngaard, Peter; Sadis, Seth; Roychowdhury, Sameek; Hussain, Maha H; Feng, Felix Y; Zalupski, Mark M; Talpaz, Moshe; Pienta, Kenneth J; Rhodes, Daniel R; Robinson, Dan R; Chinnaiyan, Arul M

    2013-06-01

    Through a prospective clinical sequencing program for advanced cancers, four index cases were identified which harbor gene rearrangements of FGFR2, including patients with cholangiocarcinoma, breast cancer, and prostate cancer. After extending our assessment of FGFR rearrangements across multiple tumor cohorts, we identified additional FGFR fusions with intact kinase domains in lung squamous cell cancer, bladder cancer, thyroid cancer, oral cancer, glioblastoma, and head and neck squamous cell cancer. All FGFR fusion partners tested exhibit oligomerization capability, suggesting a shared mode of kinase activation. Overexpression of FGFR fusion proteins induced cell proliferation. Two bladder cancer cell lines that harbor FGFR3 fusion proteins exhibited enhanced susceptibility to pharmacologic inhibition in vitro and in vivo. Because of the combinatorial possibilities of FGFR family fusion to a variety of oligomerization partners, clinical sequencing efforts, which incorporate transcriptome analysis for gene fusions, are poised to identify rare, targetable FGFR fusions across diverse cancer types.

  7. Gene fusions with lacZ by duplication insertion in the radioresistant bacterium Deinococcus radiodurans

    SciTech Connect

    Lennon, E.; Minton, K.W. )

    1990-06-01

    Deinococcus radiodurans is the most-studied species of a eubacterial family characterized by extreme resistance to DNA damage. We have focused on developing molecular biological techniques to investigate the genetics of this organism. We report construction of lacZ gene fusions by a method involving both in vitro splicing and the natural transformation of D. radiodurans. Numerous fusion strains were identified by expression of beta-galactosidase. Among these fusion strains, several were inducible by exposure to the DNA-damaging agent mitomycin C, and four of the inducible fusion constructs were cloned in Escherichia coli. Hybridization studies indicate that one of the damage-inducible genes contains a sequence reiterated throughout the D. radiodurans chromosome. Survival measurements show that two of the fusion strains have increased sensitivity to mitomycin C, suggesting that the fusions within these strains inactivate repair functions.

  8. Secretory carcinoma of the breast containing the ETV6-NTRK3 fusion gene in a male: case report and review of the literature

    PubMed Central

    Arce, C; Cortes-Padilla, D; Huntsman, DG; Miller, MA; Dueñnas-Gonzalez, A; Alvarado, A; Pérez, V; Gallardo-Rincón, D; Lara-Medina, F

    2005-01-01

    Background Secretory carcinoma (SC) of the breast is a rare and indolent tumor. Although originally described in children, it is now known to occur in adults of both sexes. Recently, the tumor was associated with the ETV6-NTRK3 gene translocation. Case presentation A 52-year-old male was diagnosed with secretory breast carcinoma and underwent a modified radical mastectomy. At 18 months the tumor recurred at the chest wall and the patient developed lung metastases. He was treated concurrently with radiation and chemotherapy without response. His tumor showed the ETV6-NTRK3 translocation as demonstrated by fluorescent in situ hybridization (FISH). Conclusion SC is a rare slow-growing tumor best treated surgically. There are insufficient data to support the use of adjuvant radiation or chemotherapy. Its association with the ETV6-NTRK3 fusion gene gives some clues for the better understanding of this neoplasm and eventually, the development of specific therapies. PMID:15963235

  9. A Plan for the Development of Fusion Energy. Final Report to Fusion Energy Sciences Advisory Committee, Fusion Development Path Panel

    SciTech Connect

    None, None

    2003-03-05

    This report presents a plan for the deployment of a fusion demonstration power plant within 35 years, leading to commercial application of fusion energy by mid-century. The plan is derived from the necessary features of a demonstration fusion power plant and from the time scale defined by President Bush. It identifies critical milestones, key decision points, needed major facilities and required budgets.

  10. Origin and Ascendancy of a Chimeric Fusion Gene: The β/δ-Globin Gene of Paenungulate Mammals

    PubMed Central

    Opazo, Juan C.; Sloan, Angela M.; Campbell, Kevin L.

    2009-01-01

    The δ-globin gene (HBD) of eutherian mammals exhibits a propensity for recombinational exchange with the closely linked β-globin gene (HBB) and has been independently converted by the HBB gene in multiple lineages. Here we report the presence of a chimeric β/δ fusion gene in the African elephant (Loxodonta africana) that was created by unequal crossing-over between misaligned HBD and HBB paralogs. The recombinant chromosome that harbors the β/δ fusion gene in elephants is structurally similar to the “anti-Lepore” duplication mutant of humans (the reciprocal exchange product of the hemoglobin Lepore deletion mutant). However, the situation in the African elephant is unique in that the chimeric β/δ fusion gene supplanted the parental HBB gene and is therefore solely responsible for synthesizing the β-chain subunits of adult hemoglobin. A phylogenetic survey of β-like globin genes in afrotherian and xenarthran mammals revealed that the origin of the chimeric β/δ fusion gene and the concomitant inactivation of the HBB gene predated the radiation of “Paenungulata,” a clade of afrotherian mammals that includes three orders: Proboscidea (elephants), Sirenia (dugongs and manatees), and Hyracoidea (hyraxes). The reduced fitness of the human Hb Lepore deletion mutant helps to explain why independently derived β/δ fusion genes (which occur on an anti-Lepore chromosome) have been fixed in a number of mammalian lineages, whereas the reciprocal δ/β fusion gene (which occurs on a Lepore chromosome) has yet to be documented in any nonhuman mammal. This illustrates how the evolutionary fates of chimeric fusion genes can be strongly influenced by their recombinational mode of origin. PMID:19332641

  11. Transforming fusions of FGFR and TACC genes in human glioblastoma.

    PubMed

    Singh, Devendra; Chan, Joseph Minhow; Zoppoli, Pietro; Niola, Francesco; Sullivan, Ryan; Castano, Angelica; Liu, Eric Minwei; Reichel, Jonathan; Porrati, Paola; Pellegatta, Serena; Qiu, Kunlong; Gao, Zhibo; Ceccarelli, Michele; Riccardi, Riccardo; Brat, Daniel J; Guha, Abhijit; Aldape, Ken; Golfinos, John G; Zagzag, David; Mikkelsen, Tom; Finocchiaro, Gaetano; Lasorella, Anna; Rabadan, Raul; Iavarone, Antonio

    2012-09-07

    The brain tumor glioblastoma multiforme (GBM) is among the most lethal forms of human cancer. Here, we report that a small subset of GBMs (3.1%; 3 of 97 tumors examined) harbors oncogenic chromosomal translocations that fuse in-frame the tyrosine kinase coding domains of fibroblast growth factor receptor (FGFR) genes (FGFR1 or FGFR3) to the transforming acidic coiled-coil (TACC) coding domains of TACC1 or TACC3, respectively. The FGFR-TACC fusion protein displays oncogenic activity when introduced into astrocytes or stereotactically transduced in the mouse brain. The fusion protein, which localizes to mitotic spindle poles, has constitutive kinase activity and induces mitotic and chromosomal segregation defects and triggers aneuploidy. Inhibition of FGFR kinase corrects the aneuploidy, and oral administration of an FGFR inhibitor prolongs survival of mice harboring intracranial FGFR3-TACC3-initiated glioma. FGFR-TACC fusions could potentially identify a subset of GBM patients who would benefit from targeted FGFR kinase inhibition.

  12. Magnetized Target Fusion Collaboration. Final report

    SciTech Connect

    Slough, John

    2012-04-18

    Nuclear fusion has the potential to satisfy the prodigious power that the world will demand in the future, but it has yet to be harnessed as a practical energy source. The entry of fusion as a viable, competitive source of power has been stymied by the challenge of finding an economical way to provide for the confinement and heating of the plasma fuel. It is the contention here that a simpler path to fusion can be achieved by creating fusion conditions in a different regime at small scale (~ a few cm). One such program now under study, referred to as Magnetized Target Fusion (MTF), is directed at obtaining fusion in this high energy density regime by rapidly compressing a compact toroidal plasmoid commonly referred to as a Field Reversed Configuration (FRC). To make fusion practical at this smaller scale, an efficient method for compressing the FRC to fusion gain conditions is required. In one variant of MTF a conducting metal shell is imploded electrically. This radially compresses and heats the FRC plasmoid to fusion conditions. The closed magnetic field in the target plasmoid suppresses the thermal transport to the confining shell, thus lowering the imploding power needed to compress the target. The undertaking described in this report was to provide a suitable target FRC, as well as a simple and robust method for inserting and stopping the FRC within the imploding liner. The FRC must also survive during the time it takes for the metal liner to compress the FRC target. The initial work at the UW was focused on developing adequate preionization and flux trapping that were found to be essential in past experiments for obtaining the density, flux and most critically, FRC lifetime required for MTF. The timescale for testing and development of such a source can be rapidly accelerated by taking advantage of a new facility funded by the Department of Energy. At this facility, two inductive plasma accelerators (IPA) were constructed and tested. Recent experiments with

  13. A Search for Gene Fusions/Translocations in Breast Cancer

    DTIC Science & Technology

    2013-11-01

    specific expression patterns, including pseudogenes derived from AURKA (kidney samples), RHOB (colon samples), and HMGB1 ( myeloproliferative neoplasms ...the scale indicated at the bottom). The key clusters are labeled with their corresponding parental gene symbols. MPN, myeloproliferative neoplasms . See...gene fusions of FGFR1 in myeloproliferative disorder ( 35 ) and 3′ FGFR3 fusions in peripheral T-cell lymphoma ( 36 ) and multiple myeloma ( 35

  14. Inertial Confinement Fusion Annual Report 1997

    SciTech Connect

    Correll, D

    1998-06-01

    The ICF Annual Report provides documentation of the achievements of the LLNL ICF Program during the fiscal year by the use of two formats: (1) an Overview that is a narrative summary of important results for the fiscal year and (2) a compilation of the articles that previously appeared in the ICF Quarterly Report that year. Both the Overview and Quarterly Report are also on the Web at http://lasers.llnl.gov/lasers/pubs/icfq.html. Beginning in Fiscal Year 1997, the fourth quarter issue of the ICF Quarterly was no longer printed as a separate document but rather included in the ICF Annual. This change provided a more efficient process of documenting our accomplishments with-out unnecessary duplication of printing. In addition we introduced a new document, the ICF Program Monthly Highlights. Starting with the September 1997 issue and each month following, the Monthly Highlights will provide a brief description of noteworthy activities of interest to our DOE sponsors and our stakeholders. The underlying theme for LLNL's ICF Program research continues to be defined within DOE's Defense Programs missions and goals. In support of these missions and goals, the ICF Program advances research and technology development in major interrelated areas that include fusion target theory and design, target fabrication, target experiments, and laser and optical science and technology. While in pursuit of its goal of demonstrating thermonuclear fusion ignition and energy gain in the laboratory, the ICF Program provides research and development opportunities in fundamental high-energy-density physics and supports the necessary research base for the possible long-term application of inertial fusion energy for civilian power production. ICF technologies continue to have spin-off applications for additional government and industrial use. In addition to these topics, the ICF Annual Report covers non-ICF funded, but related, laser research and development and associated applications. We also

  15. Fusion Power Program biannual progress report, April-September 1979

    SciTech Connect

    Not Available

    1980-02-01

    This biannual report summarizes the Argonne National Laboratory work performed for the Office of Fusion Energy during the April-September 1979 quarter in the following research and development areas: materials; energy storage and transfer; tritium containment, recovery and control; advanced reactor design; atomic data; reactor safety; fusion-fission hybrid systems; alternate applications of fusion energy; and other work related to fusion power. Separate abstracts were prepared for three sections. (MOW)

  16. Primary intracerebral angiomatoid fibrous histiocytoma: report of a case with a t(12;22)(q13;q12) causing type 1 fusion of the EWS and ATF-1 genes.

    PubMed

    Dunham, Christopher; Hussong, Jerry; Seiff, Michael; Pfeifer, John; Perry, Arie

    2008-03-01

    Angiomatoid fibrous histiocytoma (AFH) is generally considered a soft tissue sarcoma of low malignant potential that occurs in children/young adults and most frequently affects the extremities. AFH infrequently recurs and rarely metastasizes. AFH has a characteristic histomorphology, and immunohistochemical reactivities for desmin and CD68 have led to myofibroblastic and fibrohistiocytic histogenetic hypotheses, respectively. Although only a limited number of AFH cases have been molecularly characterized, many have demonstrated evidence of an underlying translocation event. Reverse transcription-polymerase chain reaction and fluorescence in situ hybridization studies suggest that chromosomal rearrangement in AFH most frequently involve the EWS, CREB, ATF-1, and FUS genes. We report the first pathologically confirmed case of an AFH presenting as an intracerebral primary in a previously healthy 25-year-old man. Genetic analyses revealed a t(12;22)(q13;q12) and a unique underlying clear cell sarcomalike type 1 EWS/ATF-1 gene fusion.

  17. Pinpointing disease genes through phenomic and genomic data fusion

    PubMed Central

    2015-01-01

    Background Pinpointing genes involved in inherited human diseases remains a great challenge in the post-genomics era. Although approaches have been proposed either based on the guilt-by-association principle or making use of disease phenotype similarities, the low coverage of both diseases and genes in existing methods has been preventing the scan of causative genes for a significant proportion of diseases at the whole-genome level. Results To overcome this limitation, we proposed a rigorous statistical method called pgFusion to prioritize candidate genes by integrating one type of disease phenotype similarity derived from the Unified Medical Language System (UMLS) and seven types of gene functional similarities calculated from gene expression, gene ontology, pathway membership, protein sequence, protein domain, protein-protein interaction and regulation pattern, respectively. Our method covered a total of 7,719 diseases and 20,327 genes, achieving the highest coverage thus far for both diseases and genes. We performed leave-one-out cross-validation experiments to demonstrate the superior performance of our method and applied it to a real exome sequencing dataset of epileptic encephalopathies, showing the capability of this approach in finding causative genes for complex diseases. We further provided the standalone software and online services of pgFusion at http://bioinfo.au.tsinghua.edu.cn/jianglab/pgfusion. Conclusions pgFusion not only provided an effective way for prioritizing candidate genes, but also demonstrated feasible solutions to two fundamental questions in the analysis of big genomic data: the comparability of heterogeneous data and the integration of multiple types of data. Applications of this method in exome or whole genome sequencing studies would accelerate the finding of causative genes for human diseases. Other research fields in genomics could also benefit from the incorporation of our data fusion methodology. PMID:25708473

  18. Particle beam fusion progress report for 1989

    SciTech Connect

    Sweeney, M.A.

    1994-08-01

    This report summarizes the progress on the pulsed power approach to inertial confinement fusion. In 1989, the authors achieved a proton focal intensity of 5 TW/cm{sup 2} on PBFA-II in a 15-cm-radius applied magnetic-field (applied-B) ion diode. This is an improvement by a factor of 4 compared to previous PBFA-II experiments. They completed development of the three-dimensional (3-D), electromagnetic, particle-in-cell code QUICKSILVER and obtained the first 3-D simulations of an applied-B ion diode. The simulations, together with analytic theory, suggest that control of electromagnetic instabilities could reduce ion divergence. In experiments using a lithium fluoride source, they delivered 26 kJ of lithium energy to the diode axis. Rutherford-scattered ion diagnostics have been developed and tested using a conical foil located inside the diode. They can now obtain energy density profiles by using range filters and recording ion images on nuclear track recording film. Timing uncertainties in power flow experiments on PBFA-II have been reduced by a factor of 5. They are investigating three plasma opening switches that use magnetic fields to control and confine the injected plasma. These new switches provide better power flow than the standard plasma erosion switch. Advanced pulsed-power fusion drivers will require extraction-geometry applied-B ion diodes. During this reporting period, progress was made in evaluating the generation, transport, and focus of multiple ion beams in an extraction geometry and in assessing the probable damage to a target chamber first wall.

  19. Patient Reported Outcomes from Sacroiliac Joint Fusion

    PubMed Central

    McGowan, Shane M.; Audley, Brittany N.; Sokunbi, Gbolabo; Puccio, Steven T.

    2017-01-01

    Study Design Retrospective, case series. Purpose The purpose of this study is to determine morbidity, complications, and patient reported outcomes from minimally invasive sacroiliac joint (SIJ) fusion. Overview of Literature Lumbar back pain emanating from the SIJ can be surgically treated via a percutaneous approach in the appropriately selected patient with minimal morbidity and acceptable functional outcomes. Methods Patients diagnosed by >2 physical examination maneuvers and subjective relief from a computed tomography–guided lidocaine-bupivacaine-steroid injection underwent SIJ fusion after failing conservative management with a combination of oral anti-inflammatory medications, physical therapy, and pelvic belt stabilization. Perioperative data collected include estimated blood loss (EBL) and operative time. Oswestry disability index, 12-item short form health survey (SF-12), visual analogue score, and functional status were analyzed. All complications were noted. Results The study cohort of 45 cases (69% female) achieved postoperative survey follow-up at 9.9 and 32.3 months. SF-12 physical component summary statistically improved while all other scores were equivalent. Mean EBL and operative time were 22 mL and 36 minutes, respectively. Initial survey showed that 64% of patients discontinued narcotics (29/45), 71% do not use an assistive device (32/45), and 15.6% do not work due to pain (7/45). 73% of patients stated they would have the surgery again (33/45). For the second survey, 65% of patients discontinued narcotics (26/40), 70% did not use an assistive device (28/40), and 17.5% did not work due to pain (7/40). A history of thoracolumbar instrumentation (16/45) did not significantly affect outcomes. Three complications described by screw malposition with neurologic deficit (6.7%) were treated with screw repositioning (1 case) and removal of a single superior implant (2 cases) with time to revision of 2.2 months. All three ultimately had resolution of

  20. Analysis of mammary specific gene locus regulation in differentiated cells derived by somatic cell fusion

    SciTech Connect

    Robinson, Claire; Kolb, Andreas F.

    2009-02-01

    The transcriptional regulation of a gene is best analysed in the context of its normal chromatin surroundings. However, most somatic cells, in contrast to embryonic stem cells, are refractory to accurate modification by homologous recombination. We show here that it is possible to introduce precise genomic modifications in ES cells and to analyse the phenotypic consequences in differentiated cells by using a combination of gene targeting, site-specific recombination and somatic cell fusion. To provide a proof of principle, we have analysed the regulation of the casein gene locus in mammary gland cells derived from modified murine ES cells by somatic cell fusion. A {beta}-galactosidase reporter gene was inserted in place of the {beta}-casein gene and the modified ES cells, which do not express the reporter gene, were fused with the mouse mammary gland cell line HC11. The resulting cell clones expressed the {beta}-galactosidase gene to a similar extent and with similar hormone responsiveness as the endogenous gene. However, a reporter gene under the control of a minimal {beta}-casein promoter (encompassing the two consensus STAT5 binding sites which mediate the hormone response of the casein genes) was unable to replicate expression levels or hormone responsiveness of the endogenous gene when inserted into the same site of the casein locus. As expected, these results implicate sequences other than the STAT5 sites in the regulation of the {beta}-casein gene.

  1. Mammary analogue secretory carcinoma of salivary glands with high-grade transformation: report of 3 cases with the ETV6-NTRK3 gene fusion and analysis of TP53, β-catenin, EGFR, and CCND1 genes.

    PubMed

    Skálová, Alena; Vanecek, Tomas; Majewska, Hanna; Laco, Jan; Grossmann, Petr; Simpson, Roderick H W; Hauer, Lukas; Andrle, Pavel; Hosticka, Lubor; Branžovský, Jindrich; Michal, Michal

    2014-01-01

    Mammary analogue secretory carcinoma of salivary gland origin (MASC) is a recently described tumor resembling secretory carcinoma of the breast characterized by strong S-100 protein, mammaglobin, and vimentin immunoexpression and which harbors a t(12;15) (p13;q25) translocation resulting in ETV6-NTRK3 fusion product. Histologically, conventional MASC displays bland histomorphology and a lobulated growth pattern and is often composed of microcystic, tubular, and solid structures with abundant eosinophilic homogenous or bubbly secretions. Colloid-like secretory material stains positively for periodic acid-Schiff with and without diastase as well as for Alcian Blue. We present for the first time, 3 patients with MASC of the parotid gland in which high-grade (HG) transformation developed in each case characterized by an accelerated clinical course and poor outcome. The HG component revealed strong membrane staining for EGFR and β-catenin, cytoplasmic/nuclear staining for S-100 protein, and nuclear staining for cyclin-D1, whereas HER-2/neu was absent. Analysis for the presence of the ETV6-NTRK3 fusion transcript revealed positivity in both HG and low-grade component of MASC in 2 of the 3 studied cases. The tumor in case 2 was negative in both its elements for the t(12;15) translocation, but ETV6 gene rearrangement was detected in both components in all 3 cases. Analysis of TP53 and CTNNB1 gene mutations in the HG component of MASCs as well as detection of copy number aberration of EGFR and CCND1 gene did not harbor any abnormalities. All 3 patients with HG-transformed MASC died of disseminated disease within 2 to 6 years after diagnosis. Recognizing HG-transformed MASC and testing for ETV6 rearrangement may be of potential value in patient treatment, because the presence of the ETV6-NTRK3 translocation may represent a therapeutic target in MASC.

  2. ETS Gene Fusions as Predictive Biomarkers of Resistance to Radiation Therapy for Prostate Cancer

    DTIC Science & Technology

    2013-08-01

    University of Michigan Ann Arbor, MI 48109 REPORT DATE: August 2013 TYPE OF REPORT: Annual ummary PREPARED FOR: U.S. Army Medical...2. REPORT TYPE Annual 3. DATES COVERED 15 July 2012 to 14 July 2013 4. TITLE AND SUBTITLE ETS Gene Fusions as Predictive Biomarkers of...wild- type prostate cancer cells and human prostate cancer samples (data not shown). Unfortunately, this level of homogenous diffuse expression, as well

  3. Inference of gene function based on gene fusion events: the rosetta-stone method.

    PubMed

    Suhre, Karsten

    2007-01-01

    The method described in this chapter can be used to infer putative functional links between two proteins. The basic idea is based on the principle of "guilt by association." It is assumed that two proteins, which are found to be transcribed by a single transcript in one (or several) genomes are likely to be functionally linked, for example by acting in a same metabolic pathway or by forming a multiprotein complex. This method is of particular interest for studying genes that exhibit no, or only remote, homologies with already well-characterized proteins. Combined with other non-homology based methods, gene fusion events may yield valuable information for hypothesis building on protein function, and may guide experimental characterization of the target protein, for example by suggesting potential ligands or binding partners. This chapter uses the FusionDB database (http://www.igs.cnrs-mrs.fr/FusionDB/) as source of information. FusionDB provides a characterization of a large number of gene fusion events at hand of multiple sequence alignments. Orthologous genes are included to yield a comprehensive view of the structure of a gene fusion event. Phylogenetic tree reconstruction is provided to evaluate the history of a gene fusion event, and three-dimensional protein structure information is used, where available, to further characterize the nature of the gene fusion. For genes that are not comprised in FusionDB, some instructions are given as how to generate a similar type of information, based solely on publicly available web tools that are listed here.

  4. InFusion: Advancing Discovery of Fusion Genes and Chimeric Transcripts from Deep RNA-Sequencing Data

    PubMed Central

    Okonechnikov, Konstantin; Imai-Matsushima, Aki; Seitz, Alexander; Meyer, Thomas F.; Garcia-Alcalde, Fernando

    2016-01-01

    Analysis of fusion transcripts has become increasingly important due to their link with cancer development. Since high-throughput sequencing approaches survey fusion events exhaustively, several computational methods for the detection of gene fusions from RNA-seq data have been developed. This kind of analysis, however, is complicated by native trans-splicing events, the splicing-induced complexity of the transcriptome and biases and artefacts introduced in experiments and data analysis. There are a number of tools available for the detection of fusions from RNA-seq data; however, certain differences in specificity and sensitivity between commonly used approaches have been found. The ability to detect gene fusions of different types, including isoform fusions and fusions involving non-coding regions, has not been thoroughly studied yet. Here, we propose a novel computational toolkit called InFusion for fusion gene detection from RNA-seq data. InFusion introduces several unique features, such as discovery of fusions involving intergenic regions, and detection of anti-sense transcription in chimeric RNAs based on strand-specificity. Our approach demonstrates superior detection accuracy on simulated data and several public RNA-seq datasets. This improved performance was also evident when evaluating data from RNA deep-sequencing of two well-established prostate cancer cell lines. InFusion identified 26 novel fusion events that were validated in vitro, including alternatively spliced gene fusion isoforms and chimeric transcripts that include intergenic regions. The toolkit is freely available to download from http:/bitbucket.org/kokonech/infusion. PMID:27907167

  5. Gene Prioritization by Compressive Data Fusion and Chaining

    PubMed Central

    Žitnik, Marinka; Nam, Edward A.; Dinh, Christopher; Kuspa, Adam; Shaulsky, Gad; Zupan, Blaž

    2015-01-01

    Data integration procedures combine heterogeneous data sets into predictive models, but they are limited to data explicitly related to the target object type, such as genes. Collage is a new data fusion approach to gene prioritization. It considers data sets of various association levels with the prediction task, utilizes collective matrix factorization to compress the data, and chaining to relate different object types contained in a data compendium. Collage prioritizes genes based on their similarity to several seed genes. We tested Collage by prioritizing bacterial response genes in Dictyostelium as a novel model system for prokaryote-eukaryote interactions. Using 4 seed genes and 14 data sets, only one of which was directly related to the bacterial response, Collage proposed 8 candidate genes that were readily validated as necessary for the response of Dictyostelium to Gram-negative bacteria. These findings establish Collage as a method for inferring biological knowledge from the integration of heterogeneous and coarsely related data sets. PMID:26465776

  6. Fusion safety program Annual report, Fiscal year 1995

    SciTech Connect

    Longhurst, G.R.; Cadwallader, L.C.; Carmack, W.J.

    1995-12-01

    This report summarizes the major activities of the Fusion Safety Program in FY-95. The Idaho National Engineering Laboratory (INEL) is the designated lead laboratory, and Lockheed Idaho Technologies Company is the prime contractor for this program. The Fusion Safety Program was initiated in 1979. Activities are conducted at the INEL, at other DOE laboratories, and at other institutions. Among the technical areas covered in this report are tritium safety, beryllium safety, chemical reactions and activation product release, safety aspects of fusion magnet systems, plasma disruptions, risk assessment failure rate database development, and safety code development and application to fusion safety issues. Most of this work has been done in support of the International Thermonuclear Experimental Reactor (ITER). Also included in the report are summaries of the safety and environmental studies performed by the Fusion Safety Program for the Tokamak Physics Experiment and the Tokamak Fusion Test Reactor and the technical support for commercial fusion facility conceptual design studies. A final activity described is work to develop DOE Technical Standards for Safety of Fusion Test Facilities.

  7. Fundamental studies of fusion plasmas. Final report

    SciTech Connect

    Aamodt, R.E.

    1998-01-30

    Lodestar has carried out a vigorous research program in the areas of rf, edge plasma and divertor physics, with emphasis largely geared towards improving the understanding and performance of ion-cyclotron heating and current drive (ICRF) systems. Additionally, a research program in the field of edge plasma and divertor modeling was initiated. Theoretical work on high power rf sheath formation for multi-strap rf arrays was developed and benchmarked against recent experimental data from the new JET A2 antennas. Sophisticated modeling tools were employed to understand the sheath formation taking into account realistic three-dimensional antenna geometry. A novel physics explanation of an observed anomaly in the low power loading of antennas was applied to qualitatively interpret data on DIII-D in terms of rf sheaths, and potential applications of the idea to develop a near-field sheath diagnostic were explored. Other rf-wave related topics were also investigated. Full wave ICRF modeling studies were carried out in support of ongoing and planned tokamaks experiments, including the investigation of low frequency plasma heating and current drive regimes for IGNITOR. In a cross-disciplinary study involving both MHD and ICRF physics, ponderomotive feedback stabilization by rf was investigated as a potential means of controlling external kink mode disruptions. In another study, the instability of the ion hybrid wave (IHW) in the presence of fusion alpha particles was studied. In the field of edge plasma and divertor modeling studies, Lodestar began the development of a theory of generalized ballooning and sheath instabilities in the scrape off layer (SOL) of divertor tokamaks. A detailed summary of the technical progress in these areas during the contract period is included, as well as where references to published work can be found. A separate listing of publications, meeting abstracts, and other presentations is also given at the end of this final report.

  8. Fusion Simulation Program Definition. Final report

    SciTech Connect

    Cary, John R.

    2012-09-05

    We have completed our contributions to the Fusion Simulation Program Definition Project. Our contributions were in the overall planning with concentration in the definition of the area of Software Integration and Support. We contributed to the planning of multiple meetings, and we contributed to multiple planning documents.

  9. RWCFusion: identifying phenotype-specific cancer driver gene fusions based on fusion pair random walk scoring method

    PubMed Central

    Zhao, Jianmei; Li, Xuecang; Yao, Qianlan; Li, Meng; Zhang, Jian; Ai, Bo; Liu, Wei; Wang, Qiuyu; Feng, Chenchen; Liu, Yuejuan; Bai, Xuefeng; Song, Chao; Li, Shang; Li, Enmin; Xu, Liyan; Li, Chunquan

    2016-01-01

    While gene fusions have been increasingly detected by next-generation sequencing (NGS) technologies based methods in human cancers, these methods have limitations in identifying driver fusions. In addition, the existing methods to identify driver gene fusions ignored the specificity among different cancers or only considered their local rather than global topology features in networks. Here, we proposed a novel network-based method, called RWCFusion, to identify phenotype-specific cancer driver gene fusions. To evaluate its performance, we used leave-one-out cross-validation in 35 cancers and achieved a high AUC value 0.925 for overall cancers and an average 0.929 for signal cancer. Furthermore, we classified 35 cancers into two classes: haematological and solid, of which the haematological got a highly AUC which is up to 0.968. Finally, we applied RWCFusion to breast cancer and found that top 13 gene fusions, such as BCAS3-BCAS4, NOTCH-NUP214, MED13-BCAS3 and CARM-SMARCA4, have been previously proved to be drivers for breast cancer. Additionally, 8 among the top 10 of the remaining candidate gene fusions, such as SULF2-ZNF217, MED1-ACSF2, and ACACA-STAC2, were inferred to be potential driver gene fusions of breast cancer by us. PMID:27506935

  10. RWCFusion: identifying phenotype-specific cancer driver gene fusions based on fusion pair random walk scoring method.

    PubMed

    Zhao, Jianmei; Li, Xuecang; Yao, Qianlan; Li, Meng; Zhang, Jian; Ai, Bo; Liu, Wei; Wang, Qiuyu; Feng, Chenchen; Liu, Yuejuan; Bai, Xuefeng; Song, Chao; Li, Shang; Li, Enmin; Xu, Liyan; Li, Chunquan

    2016-09-20

    While gene fusions have been increasingly detected by next-generation sequencing (NGS) technologies based methods in human cancers, these methods have limitations in identifying driver fusions. In addition, the existing methods to identify driver gene fusions ignored the specificity among different cancers or only considered their local rather than global topology features in networks. Here, we proposed a novel network-based method, called RWCFusion, to identify phenotype-specific cancer driver gene fusions. To evaluate its performance, we used leave-one-out cross-validation in 35 cancers and achieved a high AUC value 0.925 for overall cancers and an average 0.929 for signal cancer. Furthermore, we classified 35 cancers into two classes: haematological and solid, of which the haematological got a highly AUC which is up to 0.968. Finally, we applied RWCFusion to breast cancer and found that top 13 gene fusions, such as BCAS3-BCAS4, NOTCH-NUP214, MED13-BCAS3 and CARM-SMARCA4, have been previously proved to be drivers for breast cancer. Additionally, 8 among the top 10 of the remaining candidate gene fusions, such as SULF2-ZNF217, MED1-ACSF2, and ACACA-STAC2, were inferred to be potential driver gene fusions of breast cancer by us.

  11. Fusion safety program annual report fiscal year 1997

    SciTech Connect

    Longhurst, G.R.; Anderl, R.A.; Cadwallader, L.C.

    1998-01-01

    This report summarizes the major activities of the Fusion Safety Program in FY 1997. The Idaho National Engineering and Environmental Laboratory (INEEL) is the designated lead laboratory, and Lockheed Martin Idaho Technologies Company is the prime contractor for this program. The Fusion Safety Program was initiated in FY 1979 to perform research and develop data needed to ensure safety in fusion facilities. Activities include experiments, analysis, code development and application, and other forms of research. These activities are conducted at the INEEL, different DOE laboratories, and other institutions. The technical areas covered in this report include chemical reactions and activation product release, tritium safety, risk assessment failure rate database development, and safety code development and application to fusion safety issues. Most of this work has been done in support of the International Thermonuclear Experimental Reactor (ITER) project. Work done for ITER this year has focused on developing the needed information for the Non-site Specific Safety Report (NSSR-2).

  12. Rooting the eukaryote tree by using a derived gene fusion.

    PubMed

    Stechmann, Alexandra; Cavalier-Smith, Thomas

    2002-07-05

    Single-gene trees have failed to locate the root of the eukaryote tree because of systematic biases in sequence evolution. Structural genetic data should yield more reliable insights into deep phylogenetic relationships. We searched major protist groups for the presence or absence of a gene fusion in order to locate the root of the eukaryote tree. In striking contrast to previous molecular studies, we show that all eukaryote groups ancestrally with two cilia (bikonts) are evolutionarily derived. The root lies between bikonts and opisthokonts (animals, Fungi, Choanozoa). Amoebozoa either diverged even earlier or are sister of bikonts or (less likely) opisthokonts.

  13. Mutation-associated fusion cancer genes in solid tumors.

    PubMed

    Kaye, Frederic J

    2009-06-01

    Chromosomal translocations and fusion oncogenes serve as the ultimate biomarker for clinicians as they show specificity for distinct histopathologic malignancies while simultaneously encoding an etiologic mutation and a therapeutic target. Previously considered a minor mutational event in epithelial solid tumors, new methodologies that do not rely on the detection of macroscopic cytogenetic alterations, as well as access to large series of annotated clinical material, are expanding the inventory of recurrent fusion oncogenes in both common and rare solid epithelial tumors. Unexpectedly, related assays are also revealing a high number of tandem or chimeric transcripts in normal tissues including, in one provocative case, a template for a known fusion oncogene. These observations may force us to reassess long-held views on the definition of a gene. They also raise the possibility that some rearrangements might represent constitutive forms of a physiological chimeric transcript. Defining the chimeric transcriptome in both health (transcription-induced chimerism and intergenic splicing) and disease (mutation-associated fusion oncogenes) will play an increasingly important role in the diagnosis, prognosis, and therapy of patients with cancer.

  14. Gene expression profile of ewing sarcoma cell lines differing in their EWS-FLI1 fusion type.

    PubMed

    Bandrés, Eva; Malumbres, Raquel; Escalada, Alvaro; Cubedo, Elena; González, Iranzu; Honorato, Beatriz; Zarate, Ruth; García-Foncillas, Jesus; de Alava, Enrique

    2005-10-01

    The t(11;22)(q24;q12) translocation is present in up to 95% of Ewing tumor patients and results in the formation of an EWS-FLI-1 fusion gene that encodes a chimeric transcription factor. Many alternative forms of EWS-FLI-1 exist because of variations in the location of the EWS and FLI-1 genomic breakpoints. Previous reports have shown that the type 1 fusion is associated with a significantly better prognosis than the other fusion types. It has been suggested that the observed clinical discrepancies result from different transactivation potentials of the various EWS-FLI-1 fusion proteins. In an attempt to identify genes whose expression levels are differentially modulated by structurally different EWS-FLI-1 transcription factors, we have used microarray technology to interrogate 19,000 sequence genes to compare gene expression profile of type 1 or non-type 1 Ewing sarcoma cell lines. Data analysis showed few qualitative differences on gene expression; expression of only 41 genes (0.215% of possible sequences analyzed) differed significantly between Ewing tumor cell lines carrying EWS-FLI-1 fusion type 1 with respect to those with non-type 1 fusion.

  15. Final Report on The Theory of Fusion Plasmas

    SciTech Connect

    Steven C. Cowley

    2008-06-17

    Report describes theoretical research in the theory of fusion plasmas funded under grant DE-FG02-04ER54737. This includes work on: explosive instabilities, plasma turbulence, Alfven wave cascades, high beta (pressure) tokamaks and magnetic reconnection. These studies have lead to abetter understanding of fusion plasmas and in particular the future behavior of ITER. More than ten young researchers were involved in this research -- some were funded under the grant.

  16. Anaerobically expressed Escherichia coli genes identified by operon fusion techniques.

    PubMed Central

    Choe, M; Reznikoff, W S

    1991-01-01

    Genes that are expressed under anaerobic conditions were identified by operon fusion techniques with a hybrid bacteriophage of lambda and Mu, lambda placMu53, which creates transcriptional fusions to lacZY. Cells were screened for anaerobic expression on XG medium. Nine strains were selected, and the insertion point of the hybrid phage in each strain was mapped on the Escherichia coli chromosome linkage map. The anaerobic and aerobic expression levels of these genes were measured by beta-galactosidase assays in different medium conditions and in the presence of three regulatory mutations (fnr, narL, and rpoN). The anaerobically expressed genes (aeg) located at minute 99 (aeg-99) and 75 (aeg-75) appeared to be partially regulated by fnr, and aeg-93 is tightly regulated by fnr. aeg-60 requires a functional rpoN gene for its anaerobic expression. aeg-46.5 is repressed by narL. aeg-65A and aeg-65C are partially controlled by fnr but only in media containing nitrate or fumarate. aeg-47.5 and aeg-48.5 were found to be anaerobically induced only in rich media. The effects of a narL mutation on aeg-46.5 expression were observed in all medium conditions regardless of the presence or absence of nitrate. This suggests that narL has a regulatory function in the absence of exogenously added nitrate. PMID:1917846

  17. Characterization of nif regulatory genes in Rhodopseudomonas capsulata using lac gene fusions.

    PubMed

    Kranz, R G; Haselkorn, R

    1985-01-01

    Translational fusions of the Escherichia coli lacZYA operon to Rhodopseudomonas capsulata nif genes were obtained by using mini-MudII1734 [Castilho et al., J. Bacteriol. 158 (1984) 488-495] inserts into cloned fragments of R. capsulata DNA. A lac fusion to the nifH gene, which encodes dinitrogenase reductase, was used to classify Nif- mutations occurring in regulatory genes. Nine mutations were unable to activate nifHDK transcription. The nine mutations define four nif regulatory genes. Three of these genes are located on the same R. capsulata 8.4-kb EcoRI fragment. Each is transcribed independently. One of these (complementing mutant J61) is partially homologous with the ntrC gene of Escherichia coli, based on Southern hybridization. The fourth nif regulatory gene (complementing mutants LJ1, AH1 and AH3) is unlinked to the others. Lac fusions to all four regulatory genes were constructed. Each regulatory gene is weakly expressed compared to derepressed nifH and partially repressed in the presence of ammonia.

  18. Clinical Courses of Two Pediatric Patients with Acute Megakaryoblastic Leukemia Harboring the CBFA2T3-GLIS2 Fusion Gene

    PubMed Central

    Ishibashi, Mayu; Yokosuka, Tomoko; Yanagimachi, Masakatsu D.; Iwasaki, Fuminori; Tsujimoto, Shin-ichi; Sasaki, Koji; Takeuchi, Masanobu; Tanoshima, Reo; Kato, Hiromi; Kajiwara, Ryosuke; Tanaka, Fumiko; Goto, Hiroaki; Yokota, Shumpei

    2016-01-01

    Acute megakaryoblastic leukemia (AMKL) in children without Down syndrome (DS) has an extremely poor outcome with 3-year survival of less than 40%, whereas AMKL in children with DS has an excellent survival rate. Recently, a novel recurrent translocation involving CBFA2T3 and GLIS2 was identified in about 30% of children with non-DS AMKL, and the fusion gene was reported as a strong poor prognostic factor in pediatric AMKL. We report the difficult clinical courses of pediatric patients with AMKL harboring the CBFA2T3-GLIS2 fusion gene. PMID:27094503

  19. Protein functional links in Trypanosoma brucei, identified by gene fusion analysis

    PubMed Central

    2011-01-01

    Background Domain or gene fusion analysis is a bioinformatics method for detecting gene fusions in one organism by comparing its genome to that of other organisms. The occurrence of gene fusions suggests that the two original genes that participated in the fusion are functionally linked, i.e. their gene products interact either as part of a multi-subunit protein complex, or in a metabolic pathway. Gene fusion analysis has been used to identify protein functional links in prokaryotes as well as in eukaryotic model organisms, such as yeast and Drosophila. Results In this study we have extended this approach to include a number of recently sequenced protists, four of which are pathogenic, to identify fusion linked proteins in Trypanosoma brucei, the causative agent of African sleeping sickness. We have also examined the evolution of the gene fusion events identified, to determine whether they can be attributed to fusion or fission, by looking at the conservation of the fused genes and of the individual component genes across the major eukaryotic and prokaryotic lineages. We find relatively limited occurrence of gene fusions/fissions within the protist lineages examined. Our results point to two trypanosome-specific gene fissions, which have recently been experimentally confirmed, one fusion involving proteins involved in the same metabolic pathway, as well as two novel putative functional links between fusion-linked protein pairs. Conclusions This is the first study of protein functional links in T. brucei identified by gene fusion analysis. We have used strict thresholds and only discuss results which are highly likely to be genuine and which either have already been or can be experimentally verified. We discuss the possible impact of the identification of these novel putative protein-protein interactions, to the development of new trypanosome therapeutic drugs. PMID:21729286

  20. TMPRSS2-ERG gene fusion in small cell carcinoma of the prostate.

    PubMed

    Guo, Charles C; Dancer, Jane Y; Wang, Yan; Aparicio, Ana; Navone, Nora M; Troncoso, Patricia; Czerniak, Bogdan A

    2011-01-01

    Recent studies have shown that most prostate cancers carry the TMPRSS2-ERG gene fusion. Here we evaluated the TMPRSS2-ERG gene fusion in small cell carcinoma of the prostate (n = 12) in comparison with small cell carcinoma of the urinary bladder (n = 12) and lung (n = 11). Fluorescence in situ hybridization demonstrated rearrangement of the ERG gene in 8 cases of prostatic small cell carcinoma (67%), and the rearrangement was associated with deletion of the 5' ERG gene in 7 cases, but rearrangement of the ERG gene was not present in any small cell carcinoma of the urinary bladder or lung. Next we evaluated the TMPRSS2-ERG gene fusion in nude mouse xenografts that were derived from 2 prostatic small cell carcinomas carrying the TMPRSS2-ERG gene fusion. Two transcripts encoded by the TMPRSS2-ERG gene fusion were detected by reverse transcriptase polymerase chain reaction, and DNA sequencing demonstrated that the 2 transcripts were composed of fusions of exon 1 of the TMPRSS2 gene to exon 4 or 5 of the ERG gene. Our study demonstrates the specific presence of TMPRSS2-ERG gene fusion in prostatic small cell carcinoma, which may be helpful in distinguishing small cell carcinoma of prostatic origin from nonprostatic origins. The high prevalence of the TMPRSS2-ERG gene fusion in prostatic small cell carcinoma as well as adenocarcinoma implies that small cell carcinoma may share a common pathogenic pathway with adenocarcinoma in the prostate.

  1. Structure and expression of the Drosophila ubiquitin-80-amino-acid fusion-protein gene.

    PubMed Central

    Barrio, R; del Arco, A; Cabrera, H L; Arribas, C

    1994-01-01

    In the fruitfly Drosophila, as in all eukaryotes examined so far, some ubiquitin-coding sequences appear fused to unrelated open reading frames. Two of these fusion genes have been previously described (the homologues of UBI1-UBI2 and UBI4 in yeast), and we report here the organization and expression of a third one, the DUb80 gene (the homologue of UBI3 in yeast). This gene encodes a ubiquitin monomer fused to an 80-amino-acid extension which is homologous with the ribosomal protein encoded by the UB13 gene. The 5' regulatory region of DUb80 shares common features with another ubiquitin fusion gene, DUb52, and with the ribosomal protein genes of Drosophila, Xenopus and mouse. We also find helix-loop-helix protein-binding sequences (E-boxes). The DUb80 gene is transcribed to a 0.9 kb mRNA which is particularly abundant under conditions of high protein synthesis, such as in ovaries and exponentially growing cells. Images Figure 3 Figure 4 PMID:8068011

  2. Fusion Safety Program Annual Report, Fiscal Year 1996

    SciTech Connect

    Longhurst, G.R.; Anderl, R.A.; Cadwallader, L.C.

    1996-12-01

    This report summarizes the major activities of the Fusion Safety Program in FY 1996. The Idaho National Engineering Laboratory (INEL) is the designated lead laboratory, and Lockheed Martin Idaho Technologies Company is the prime contractor for this program. The Fusion Safety Program was initiated in 1979. The objective is to perform research and develop data needed to ensure safety in fusion facilities. Activities include experiments, analysis, code development and application, and other forms of research. These activities are conducted at the INEL, at other DOE laboratories, and at other institutions. Among the technical areas covered in this report are tritium safety, chemical reactions and activation product release, risk assessment failure rate database development, and safety code development and application to fusion safety issues. Most of this work has been done in support of the International Thermonuclear Experimental Reactor (ITER). Work done for ITER this year has focused on developing the needed information for the Non- Site- Specific Safety Report (NSSR-1). A final area of activity described is development of the new DOE Technical Standards for Safety of Magnetic Fusion Facilities.

  3. The yeast ubiquitin genes: a family of natural gene fusions.

    PubMed Central

    Ozkaynak, E; Finley, D; Solomon, M J; Varshavsky, A

    1987-01-01

    Ubiquitin is a 76-residue protein highly conserved among eukaryotes. Conjugation of ubiquitin to intracellular proteins mediates their selective degradation in vivo. We describe a family of four ubiquitin-coding loci in the yeast Saccharomyces cerevisiae. UB11, UB12 and UB13 encode hybrid proteins in which ubiquitin is fused to unrelated ('tail') amino acid sequences. The ubiquitin coding elements of UB11 and UB12 are interrupted at identical positions by non-homologous introns. UB11 and UB12 encode identical 52-residue tails, whereas UB13 encodes a different 76-residue tail. The tail amino acid sequences are highly conserved between yeast and mammals. Each tail contains a putative metal-binding, nucleic acid-binding domain of the form Cys-X2-4-Cys-X2-15-Cys-X2-4-Cys, suggesting that these proteins may function by binding to DNA. The fourth gene, UB14, encodes a polyubiquitin precursor protein containing five ubiquitin repeats in a head-to-tail, spacerless arrangement. All four ubiquitin genes are expressed in exponentially growing cells, while in stationary-phase cells the expression of UB11 and UB12 is repressed. The UB14 gene, which is strongly inducible by starvation, high temperatures and other stresses, contains in its upstream region strong homologies to the consensus 'heat shock box' nucleotide sequence. Elsewhere we show that the essential function of the UB14 gene is to provide ubiquitin to cells under stress. Images Fig. 1. Fig. 7. PMID:3038523

  4. Contribution to Fusion Materials Semiannual Report

    SciTech Connect

    Marian, J; Meier, W

    2012-02-24

    The objectives of this work are the following: (1) The application of micro and mesoscale modeling techniques to study dislocation properties in ferritic and W-based materials; and (2) The development of computational models and tools to study damage accumulation in >1 dpa (fusion-like) conditions, both for Fe and W-based alloys. The high-temperature strength of structural ferritic alloys (ferritic/martensitic steels, ODS steels, bcc refractory alloys) hinges on the thermal stability of second phase particles and their interactions with dislocations. Irradiation damage can modify the structure and stability of both the particles and dislocations, particularly by the introduction of gas atoms, point defects and point defect clusters. The three aspects of materials strength that we are studying are: (a) Computation of dislocation mobility functions (stress-velocity relations) as a function of temperature and dislocation character. This will be done via molecular dynamics (MD) simulations of single dislocation motion under applied shear stress. This is a fundamental input to dislocation dynamics (DD) simulations and also provides fundamental insights into the high-temperature plastic behavior of ferritic materials. (b) Simulations of dislocation-obstacle interactions using MD and DD. This subtask includes simulating the effect on dislocation glide of precipitates (e.g., {alpha}' Cr precipitates), ODS particles, and irradiation induced defect clusters (e.g. voids, dislocation loops, etc.). (c) Implementation of this information (dislocation mobilities and dislocation-defect interaction rules) into DD codes that will allow us to study plasticity of single crystals Fe alloys under relevant irradiation conditions.

  5. Recurrent and pathological gene fusions in breast cancer: current advances in genomic discovery and clinical implications.

    PubMed

    Veeraraghavan, Jamunarani; Ma, Jiacheng; Hu, Yiheng; Wang, Xiao-Song

    2016-07-01

    Gene fusions have long been considered principally as the oncogenic events of hematologic malignancies, but have recently gained wide attention in solid tumors due to several milestone discoveries and the advancement of deep sequencing technologies. With the progress in deep sequencing studies of breast cancer transcriptomes and genomes, the discovery of recurrent and pathological gene fusions in breast cancer is on the focus. Recently, driven by new deep sequencing studies, several recurrent or pathological gene fusions have been identified in breast cancer, including ESR1-CCDC170, SEC16A-NOTCH1, SEC22B-NOTCH2, and ESR1-YAP1 etc. More important, most of these gene fusions are preferentially identified in the more aggressive breast cancers, such as luminal B, basal-like, or endocrine-resistant breast cancer, suggesting recurrent gene fusions as additional key driver events in these tumors other than the known drivers such as the estrogen receptor. In this paper, we have comprehensively summarized the newly identified recurrent or pathological gene fusion events in breast cancer, reviewed the contributions of new genomic and deep sequencing technologies to new fusion discovery and the integrative bioinformatics tools to analyze these data, highlighted the biological relevance and clinical implications of these fusion discoveries, and discussed future directions of gene fusion research in breast cancer.

  6. Colorimetric TMPRSS2-ERG Gene Fusion Detection in Prostate Cancer Urinary Samples via Recombinase Polymerase Amplification.

    PubMed

    Koo, Kevin M; Wee, Eugene J H; Trau, Matt

    2016-01-01

    TMPRSS2 (Exon 1)-ERG (Exon 4) is the most frequent gene fusion event in prostate cancer (PC), and is highly PC-specific unlike the current serum prostate specific antigen (PSA) biomarker. However, TMPRSS2-ERG levels are currently measured with quantitative reverse-transcription PCR (RT-qPCR) which is time-consuming and requires costly equipment, thus limiting its use in clinical diagnostics. Herein, we report a novel rapid, cost-efficient and minimal-equipment assay named "FusBLU" for detecting TMPRSS2-ERG gene fusions from urine. TMPRSS2-ERG mRNA was amplified by isothermal reverse transcription-recombinase polymerase amplification (RT-RPA), magnetically-isolated, and detected through horseradish peroxidase (HRP)-catalyzed colorimetric reaction. FusBLU was specific for TMPRSS2-ERG mRNA with a low visual detection limit of 10(5) copies. We also demonstrated assay readout versatility on 3 potentially useful platforms. The colorimetric readout was detectable by naked eye for a quick yes/no evaluation of gene fusion presence. On the other hand, a more quantitative TMPRSS2-ERG detection was achievable by absorbance/electrochemical measurements. FusBLU was successfully applied to 12 urinary samples and results were validated by gold-standard RT-qPCR. We also showed that sediment RNA was likely the main source of TMPRSS2-ERG mRNA in urinary samples. We believe that our assay is a potential clinical screening tool for PC and could also have wide applications for other disease-related fusion genes.

  7. Colorimetric TMPRSS2-ERG Gene Fusion Detection in Prostate Cancer Urinary Samples via Recombinase Polymerase Amplification

    PubMed Central

    Koo, Kevin M.; Wee, Eugene J.H.; Trau, Matt

    2016-01-01

    TMPRSS2 (Exon 1)-ERG (Exon 4) is the most frequent gene fusion event in prostate cancer (PC), and is highly PC-specific unlike the current serum prostate specific antigen (PSA) biomarker. However, TMPRSS2-ERG levels are currently measured with quantitative reverse-transcription PCR (RT-qPCR) which is time-consuming and requires costly equipment, thus limiting its use in clinical diagnostics. Herein, we report a novel rapid, cost-efficient and minimal-equipment assay named “FusBLU” for detecting TMPRSS2-ERG gene fusions from urine. TMPRSS2-ERG mRNA was amplified by isothermal reverse transcription-recombinase polymerase amplification (RT-RPA), magnetically-isolated, and detected through horseradish peroxidase (HRP)-catalyzed colorimetric reaction. FusBLU was specific for TMPRSS2-ERG mRNA with a low visual detection limit of 105 copies. We also demonstrated assay readout versatility on 3 potentially useful platforms. The colorimetric readout was detectable by naked eye for a quick yes/no evaluation of gene fusion presence. On the other hand, a more quantitative TMPRSS2-ERG detection was achievable by absorbance/electrochemical measurements. FusBLU was successfully applied to 12 urinary samples and results were validated by gold-standard RT-qPCR. We also showed that sediment RNA was likely the main source of TMPRSS2-ERG mRNA in urinary samples. We believe that our assay is a potential clinical screening tool for PC and could also have wide applications for other disease-related fusion genes. PMID:27375789

  8. High-speed biosensing strategy for non-invasive profiling of multiple cancer fusion genes in urine.

    PubMed

    Koo, Kevin M; Wee, Eugene J H; Trau, Matt

    2017-03-15

    Aberrant chromosal rearrangements, such as the multiple variants of TMPRSS2:ERG fusion gene mutations in prostate cancer (PCa), are promising diagnostic and prognostic biomarkers due to their specific expression in cancerous tissue only. Additionally, TMPRSS2:ERG variants are detectable in urine to provide non-invasive PCa diagnostic sampling as an attractive surrogate for needle biopsies. Therefore, rapid and simplistic assays for identifying multiple urinary TMPRSS2:ERG variants are potentially useful to aid in early cancer detection, immediate patient risk stratification, and prompt personalized treatment. However, current strategies for simultaneous detection of multiple gene fusions are limited by tedious and prolonged experimental protocols, thus limiting their use as rapid clinical screening tools. Herein, we report a simple and rapid gene fusion strategy which expliots the specificity of DNA ligase and the speed of isothermal amplification to simultaneously detect multiple fusion gene RNAs within a short sample-to-answer timeframe of 60min. The method has a low detection limit of 2 amol (1000 copies), and was successfully applied for non-invasive fusion gene profiling in patient urine samples with subsequent validation by a PCR-based gold standard approach.

  9. Identification of target genes of synovial sarcoma-associated fusion oncoprotein using human pluripotent stem cells

    SciTech Connect

    Hayakawa, Kazuo; Ikeya, Makoto; Fukuta, Makoto; Woltjen, Knut; Tamaki, Sakura; Takahara, Naoko; Kato, Tomohisa; Sato, Shingo; Otsuka, Takanobu; Toguchida, Junya

    2013-03-22

    Highlights: ► We tried to identify targets of synovial sarcoma (SS)-associated SYT–SSX fusion gene. ► We established pluripotent stem cell (PSC) lines with inducible SYT–SSX gene. ► SYT–SSX responsive genes were identified by the induction of SYT–SSX in PSC. ► SS-related genes were selected from database by in silico analyses. ► 51 genes were finally identified among SS-related genes as targets of SYT–SSX in PSC. -- Abstract: Synovial sarcoma (SS) is a malignant soft tissue tumor harboring chromosomal translocation t(X; 18)(p11.2; q11.2), which produces SS-specific fusion gene, SYT–SSX. Although precise function of SYT–SSX remains to be investigated, accumulating evidences suggest its role in gene regulation via epigenetic mechanisms, and the product of SYT–SSX target genes may serve as biomarkers of SS. Lack of knowledge about the cell-of-origin of SS, however, has placed obstacle in the way of target identification. Here we report a novel approach to identify SYT–SSX2 target genes using human pluripotent stem cells (hPSCs) containing a doxycycline-inducible SYT–SSX2 gene. SYT–SSX2 was efficiently induced both at mRNA and protein levels within three hours after doxycycline administration, while no morphological change of hPSCs was observed until 24 h. Serial microarray analyses identified genes of which the expression level changed more than twofold within 24 h. Surprisingly, the majority (297/312, 95.2%) were up-regulated genes and a result inconsistent with the current concept of SYT–SSX as a transcriptional repressor. Comparing these genes with SS-related genes which were selected by a series of in silico analyses, 49 and 2 genes were finally identified as candidates of up- and down-regulated target of SYT–SSX, respectively. Association of these genes with SYT–SSX in SS cells was confirmed by knockdown experiments. Expression profiles of SS-related genes in hPSCs and human mesenchymal stem cells (hMSCs) were strikingly

  10. Inhibition of protein translation by the DISC1-Boymaw fusion gene from a Scottish family with major psychiatric disorders

    PubMed Central

    Ji, Baohu; Higa, Kerin K.; Kim, Minjung; Zhou, Lynn; Young, Jared W.; Geyer, Mark A.; Zhou, Xianjin

    2014-01-01

    The t(1; 11) translocation appears to be the causal genetic lesion with 70% penetrance for schizophrenia, major depression and other psychiatric disorders in a Scottish family. Molecular studies identified the disruption of the disrupted-in-schizophrenia 1 (DISC1) gene by chromosome translocation at chromosome 1q42. Our previous studies, however, revealed that the translocation also disrupted another gene, Boymaw (also termed DISC1FP1), on chromosome 11. After translocation, two fusion genes [the DISC1-Boymaw (DB7) and the Boymaw-DISC1 (BD13)] are generated between the DISC1 and Boymaw genes. In the present study, we report that expression of the DB7 fusion gene inhibits both intracellular NADH oxidoreductase activities and protein translation. We generated humanized DISC1-Boymaw mice with gene targeting to examine the in vivo functions of the fusion genes. Consistent with the in vitro studies on the DB7 fusion gene, protein translation activity is decreased in the hippocampus and in cultured primary neurons from the brains of the humanized mice. Expression of Gad67, Nmdar1 and Psd95 proteins are also reduced. The humanized mice display prolonged and increased responses to the NMDA receptor antagonist, ketamine, on various mouse genetic backgrounds. Abnormal information processing of acoustic startle and depressive-like behaviors are also observed. In addition, the humanized mice display abnormal erythropoiesis, which was reported to associate with depression in humans. Expression of the DB7 fusion gene may reduce protein translation to impair brain functions and thereby contribute to the pathogenesis of major psychiatric disorders. PMID:24908665

  11. Wnt signaling induces transcription, spatial proximity, and translocation of fusion gene partners in human hematopoietic cells.

    PubMed

    Ugarte, Giorgia D; Vargas, Macarena F; Medina, Matías A; León, Pablo; Necuñir, David; Elorza, Alvaro A; Gutiérrez, Soraya E; Moon, Randall T; Loyola, Alejandra; De Ferrari, Giancarlo V

    2015-10-08

    Chromosomal translocations are frequently associated with a wide variety of cancers, particularly hematologic malignancies. A recurrent chromosomal abnormality in acute myeloid leukemia is the reciprocal translocation t(8;21) that fuses RUNX1 and ETO genes. We report here that Wnt/β-catenin signaling increases the expression of ETO and RUNX1 genes in human hematopoietic progenitors. We found that β-catenin is rapidly recruited into RNA polymerase II transcription factories (RNAPII-Ser5) and that ETO and RUNX1 genes are brought into close spatial proximity upon Wnt3a induction. Notably, long-term treatment of cells with Wnt3a induces the generation a frequent RUNX1-ETO translocation event. Thus, Wnt/β-catenin signaling induces transcription and translocation of RUNX1 and ETO fusion gene partners, opening a novel window to understand the onset/development of leukemia.

  12. Adenoid cystic carcinoma: emerging role of translocations and gene fusions

    PubMed Central

    Wysocki, Piotr T.; Izumchenko, Evgeny; Meir, Juliet; Ha, Patrick K.; Sidransky, David; Brait, Mariana

    2016-01-01

    Adenoid cystic carcinoma (ACC), the second most common salivary gland malignancy, is notorious for poor prognosis, which reflects the propensity of ACC to progress to clinically advanced metastatic disease. Due to high long-term mortality and lack of effective systemic treatment, the slow-growing but aggressive ACC poses a particular challenge in head and neck oncology. Despite the advancements in cancer genomics, up until recently relatively few genetic alterations critical to the ACC development have been recognized. Although the specific chromosomal translocations resulting in MYB-NFIB fusions provide insight into the ACC pathogenesis and represent attractive diagnostic and therapeutic targets, their clinical significance is unclear, and a substantial subset of ACCs do not harbor the MYB-NFIB translocation. Strategies based on detection of newly described genetic events (such as MYB activating super-enhancer translocations and alterations affecting another member of MYB transcription factor family-MYBL1) offer new hope for improved risk assessment, therapeutic intervention and tumor surveillance. However, the impact of these approaches is still limited by an incomplete understanding of the ACC biology, and the manner by which these alterations initiate and drive ACC remains to be delineated. This manuscript summarizes the current status of gene fusions and other driver genetic alterations in ACC pathogenesis and discusses new therapeutic strategies stemming from the current research. PMID:27533466

  13. Fusion breeder studies program: Final report

    SciTech Connect

    Berwald, D.H.

    1986-10-17

    This report is an assessment of technology related to hybrid reactors, especially the Fission-suppressed hybrid. A description of a typical fission-suppressed reactor is given. The economic advantages of the use of a hybrid reactor as part of a fuel cycle center are discussed at length. The inherent safety advantages of the hybrid reactor are analyzed. The report concludes with a proposed timetable for research and development. (JDH)

  14. Fusion Safety Program annual report, fiscal year 1992

    SciTech Connect

    Holland, D.F.; Cadwallader, L.C.; Herring, J.S.; Longhurst, G.R.; McCarthy, K.A.; Merrill, B.J.; Piet, S.J.

    1993-01-01

    This report summarizes the major activities of the Fusion Safety Program in fiscal year 1992. The Idaho National Engineering Laboratory (INEL) is the designated lead laboratory and EG G Idaho, Inc. is the prime contractor for this program. The Fusion Safety Program was initiated in 1979. Activities are conducted at the INEL and in participating organizations including the Westinghouse Hanford Company at the Hanford Engineering Development Laboratory, the Massachusetts Institute of Technology, and the University of Wisconsin. The technical areas covered in the report include tritium safety, activation product release, reactions involving beryllium, reactions involving lithium breeding materials, safety of fusion magnet systems, plasma disruptions, risk assessment failure rate data base, and computer code development for reactor transients. Also included in the report is a summary of the safety and environmental studies performed by the INEL for the Tokamak Physics Experiments and the Tokamak Fusion Test Reactor, the safety analysis for the International Thermonuclear Experimental Reactor design, and the technical support for the ARIES commercial reactor design study.

  15. Fusion Safety Program annual report, fiscal year 1992

    SciTech Connect

    Holland, D.F.; Cadwallader, L.C.; Herring, J.S.; Longhurst, G.R.; McCarthy, K.A.; Merrill, B.J.; Piet, S.J.

    1993-01-01

    This report summarizes the major activities of the Fusion Safety Program in fiscal year 1992. The Idaho National Engineering Laboratory (INEL) is the designated lead laboratory and EG&G Idaho, Inc. is the prime contractor for this program. The Fusion Safety Program was initiated in 1979. Activities are conducted at the INEL and in participating organizations including the Westinghouse Hanford Company at the Hanford Engineering Development Laboratory, the Massachusetts Institute of Technology, and the University of Wisconsin. The technical areas covered in the report include tritium safety, activation product release, reactions involving beryllium, reactions involving lithium breeding materials, safety of fusion magnet systems, plasma disruptions, risk assessment failure rate data base, and computer code development for reactor transients. Also included in the report is a summary of the safety and environmental studies performed by the INEL for the Tokamak Physics Experiments and the Tokamak Fusion Test Reactor, the safety analysis for the International Thermonuclear Experimental Reactor design, and the technical support for the ARIES commercial reactor design study.

  16. Isolation of gene fusions (soi::lacZ) inducible by oxidative stress in Escherichia coli.

    PubMed Central

    Kogoma, T; Farr, S B; Joyce, K M; Natvig, D O

    1988-01-01

    Mu dX phage was used to isolate three gene fusions to the lacZ gene (soi::lacZ; soi for superoxide radical inducible) that were induced by treatment with superoxide radical anion generators such as paraquat and plumbagin. The induction of beta-galactosidase in these fusion strains with the superoxide radical generating agents required aerobic metabolism. Hyperoxygenation (i.e., bubbling of cultures with oxygen gas) also induced the fusions. On the other hand, hydrogen peroxide did not induce the fusions at concentrations that are known to invoke an adaptive response. Introduction of oxyR, htpR, or recA mutations did not affect the induction. Two of the fusion strains exhibited increased sensitivity to paraquat but not to hydrogen peroxide. The third fusion strain showed no increased sensitivity to either agent. All three fusions were located in the 45- to 61-min region of the Escherichia coli chromosome. PMID:2838846

  17. Tandem duplication producing a novel oncogenic BRAF fusion gene defines the majority of pilocytic astrocytomas

    PubMed Central

    Jones, David T. W.; Kocialkowski, Sylvia; Liu, Lu; Pearson, Danita M.; Bäcklund, L. Magnus; Ichimura, Koichi; Collins, V. Peter

    2008-01-01

    Brain tumours are the commonest solid tumours of childhood, and pilocytic astrocytomas (PAs) are the most common central nervous system tumour in 5-19 year-olds. Little is known about the genetic alterations underlying their development. Here we describe a tandem duplication of ∼2Mb at 7q34 occurring in 66% of pilocytic astrocytomas. This rearrangement, which was not observed in a series of 244 higher-grade astrocytomas, results in an in-frame fusion gene incorporating the kinase domain of the BRAF oncogene. We further show that the resulting fusion protein has constitutive BRAF kinase activity, and is able to transform NIH3T3 cells. This is the first report of BRAF activation through rearrangement as a frequent feature in a sporadic tumor. The frequency and specificity of this change underline its potential both as a therapeutic target and a diagnostic tool. PMID:18974108

  18. Identification of novel fusion genes with 28S ribosomal DNA in hematologic malignancies.

    PubMed

    Kobayashi, Satoru; Taki, Tomohiko; Nagoshi, Hisao; Chinen, Yoshiaki; Yokokawa, Yuichi; Kanegane, Hirokazu; Matsumoto, Yosuke; Kuroda, Junya; Horiike, Shigeo; Nishida, Kazuhiro; Taniwaki, Masafumi

    2014-04-01

    Fusion genes are frequently observed in hematologic malignancies and soft tissue sarcomas, and are usually associated with chromosome abnormalities. Many of these fusion genes create in-frame fusion transcripts that result in the production of fusion proteins, and some of which aid tumorigenesis. These fusion proteins are often associated with disease phenotype and clinical outcome, and act as markers for minimal residual disease and indicators of therapeutic targets. Here, we identified the 28S ribosomal DNA (RN28S1) gene as a novel fusion partner of the B-cell leukemia/lymphoma 11B gene (BCL11B), the immunoglobulin κ variable 3-20 gene (IGKV3-20) and the component of oligomeric Golgi complex 1 gene (COG1) in hematologic malignancies. The RN28S1-BCL11B fusion transcript was identified in a case with mixed-lineage (T/myeloid) acute leukemia having t(6;14)(q25;q32) by cDNA bubble PCR using BCL11B primers; however, the gene fused to BCL11B on 14q32 was not on 6q25. IGKV3-20-RN28S1 and COG1-RN28S1 fusion transcripts were identified in the Burkitt lymphoma cell line HBL-5, and the multiple myeloma cell line KMS-18. RN28S1 would not translate, and the breakpoints in partner genes of RN28S1 were within the coding exons, suggesting that disruption of fusion partners by fusion to RN28S1 is the possible mechanism of tumorigenesis. Although further analysis is needed to elucidate the mechanism(s) through which these RN28S1-related fusions play roles in tumorigenesis, our findings provide important insights into the role of rDNA function in human genomic architecture and tumorigenesis.

  19. ZNF384-related fusion genes define a subgroup of childhood B-cell precursor acute lymphoblastic leukemia with a characteristic immunotype

    PubMed Central

    Hirabayashi, Shinsuke; Ohki, Kentaro; Nakabayashi, Kazuhiko; Ichikawa, Hitoshi; Momozawa, Yukihide; Okamura, Kohji; Yaguchi, Akinori; Terada, Kazuki; Saito, Yuya; Yoshimi, Ai; Ogata-Kawata, Hiroko; Sakamoto, Hiromi; Kato, Motohiro; Fujimura, Junya; Hino, Moeko; Kinoshita, Akitoshi; Kakuda, Harumi; Kurosawa, Hidemitsu; Kato, Keisuke; Kajiwara, Ryosuke; Moriwaki, Koichi; Morimoto, Tsuyoshi; Nakamura, Kozue; Noguchi, Yasushi; Osumi, Tomoo; Sakashita, Kazuo; Takita, Junko; Yuza, Yuki; Matsuda, Koich; Yoshida, Teruhiko; Matsumoto, Kenji; Hata, Kenichiro; Kubo, Michiaki; Matsubara, Yoichi; Fukushima, Takashi; Koh, Katsuyoshi; Manabe, Atsushi; Ohara, Akira; Kiyokawa, Nobutaka

    2017-01-01

    Fusion genes involving ZNF384 have recently been identified in B-cell precursor acute lymphoblastic leukemia, and 7 fusion partners have been reported. We further characterized this type of fusion gene by whole transcriptome sequencing and/or polymerase chain reaction. In addition to previously reported genes, we identified BMP2K as a novel fusion partner for ZNF384. Including the EP300-ZNF384 that we reported recently, the total frequency of ZNF384-related fusion genes was 4.1% in 291 B-cell precursor acute lymphoblastic leukemia patients enrolled in a single clinical trial, and TCF3-ZNF384 was the most recurrent, with a frequency of 2.4%. The characteristic immunophenotype of weak CD10 and aberrant CD13 and/or CD33 expression was revealed to be a common feature of the leukemic cells harboring ZNF384-related fusion genes. The signature gene expression profile in TCF3-ZNF384-positive patients was enriched in hematopoietic stem cell features and related to that of EP300-ZNF384-positive patients, but was significantly distinct from that of TCF3-PBX1-positive and ZNF384-fusion-negative patients. However, clinical features of TCF3-ZNF384-positive patients are markedly different from those of EP300-ZNF384-positive patients, exhibiting higher cell counts and a younger age at presentation. TCF3-ZNF384-positive patients revealed a significantly poorer steroid response and a higher frequency of relapse, and the additional activating mutations in RAS signaling pathway genes were detected by whole exome analysis in some of the cases. Our observations indicate that ZNF384-related fusion genes consist of a distinct subgroup of B-cell precursor acute lymphoblastic leukemia with a characteristic immunophenotype, while the clinical features depend on the functional properties of individual fusion partners. PMID:27634205

  20. Inertial Fusion Program. Progress report, July 1-December 31, 1979

    SciTech Connect

    Skoberne, F.

    1981-10-01

    Progress in the development of high-energy short-pulse CO/sub 2/ laser systems for fusion research is reported. Improvements in the Los Alamos National Laboratory eight-beam Helios system are described. These improvements increased the reliability of the laser and permitted the firing of 290 shots, most of which delivered energies of approximately 8 kJ to the target. Modifications to Gemini are outlined, including the installation of a new target-insertion mechanism. The redirection of the Antares program is discussed in detail, which will achieve a total energy of approximatey 40 kJ with two beams. This redirection will bring Antares on-line almost two years earlier than was possible with the full six-beam system, although at a lower energy. Experiments with isentropically imploded Sirius-B targets are discussed, and x-ray radiation-loss data from gold microballoons are presented, which show that these results are essentially identical with those obtained at glass-laser wavelengths. Significant progress in characterizing laser fusion targets is reported. New processes for fabricating glass miroballoon x-ray diagnostic targets, the application of high-quality metallic coatings, and the deposition of thick plastic coatings are described. Results in the development of x-ray diagnostics are reported, and research in the Los Alamos heavy-ion fusion program is summarized. Results of investigations of phase-conjugation research of gaseous saturable absorbers and of the use of alkali-halide crystals in a new class of saturable absorbers are summarized. New containment-vessel concepts for Inertial Confinement Fusion reactors are discussed, and results of a scoping study of four fusion-fission hybrid concepts are presented.

  1. An Efficient Method for Identifying Gene Fusions by Targeted RNA Sequencing from Fresh Frozen and FFPE Samples.

    PubMed

    Scolnick, Jonathan A; Dimon, Michelle; Wang, I-Ching; Huelga, Stephanie C; Amorese, Douglas A

    2015-01-01

    Fusion genes are known to be key drivers of tumor growth in several types of cancer. Traditionally, detecting fusion genes has been a difficult task based on fluorescent in situ hybridization to detect chromosomal abnormalities. More recently, RNA sequencing has enabled an increased pace of fusion gene identification. However, RNA-Seq is inefficient for the identification of fusion genes due to the high number of sequencing reads needed to detect the small number of fusion transcripts present in cells of interest. Here we describe a method, Single Primer Enrichment Technology (SPET), for targeted RNA sequencing that is customizable to any target genes, is simple to use, and efficiently detects gene fusions. Using SPET to target 5701 exons of 401 known cancer fusion genes for sequencing, we were able to identify known and previously unreported gene fusions from both fresh-frozen and formalin-fixed paraffin-embedded (FFPE) tissue RNA in both normal tissue and cancer cells.

  2. An Efficient Method for Identifying Gene Fusions by Targeted RNA Sequencing from Fresh Frozen and FFPE Samples

    PubMed Central

    Scolnick, Jonathan A.; Dimon, Michelle; Wang, I-Ching; Huelga, Stephanie C.; Amorese, Douglas A.

    2015-01-01

    Fusion genes are known to be key drivers of tumor growth in several types of cancer. Traditionally, detecting fusion genes has been a difficult task based on fluorescent in situ hybridization to detect chromosomal abnormalities. More recently, RNA sequencing has enabled an increased pace of fusion gene identification. However, RNA-Seq is inefficient for the identification of fusion genes due to the high number of sequencing reads needed to detect the small number of fusion transcripts present in cells of interest. Here we describe a method, Single Primer Enrichment Technology (SPET), for targeted RNA sequencing that is customizable to any target genes, is simple to use, and efficiently detects gene fusions. Using SPET to target 5701 exons of 401 known cancer fusion genes for sequencing, we were able to identify known and previously unreported gene fusions from both fresh-frozen and formalin-fixed paraffin-embedded (FFPE) tissue RNA in both normal tissue and cancer cells. PMID:26132974

  3. Identification of the merR gene of R100 by using mer-lac gene and operon fusions.

    PubMed Central

    Foster, T J; Brown, N L

    1985-01-01

    Transcriptional (operon) and translational (gene) fusions between the R100 merR gene and lacZ were constructed in vitro in a pBR322 plasmid carrying the mer genes derived from plasmid R100. The translational fusions were oriented in the opposite direction to and divergently from the merTCAD genes. This shows that the reading frame previously thought to be merR was incorrect. Expression of the gene fusion was repressed in trans by a compatible plasmid carrying the R100 merR+ gene, as was a similarly oriented transcriptional fusion. In contrast, expression of beta-galactosidase by the lac fragment located at the same site but in the opposite orientation was at a lower level and was not repressed by merR+. Images PMID:2993235

  4. Inertial Confinement Fusion Annual Report 1999

    SciTech Connect

    Kauffman, Robert L.

    2001-07-01

    The ICF Program has undergone a significant change in 1999 with the decommissioning of the Nova laser and the transfer of much of the experimental program to the OMEGA laser at the University of Rochester. The Nova laser ended operations with the final experiment conducted on May 27, 1999. This marked the end to one of DOE's most successful experimental facilities. Since its commissioning in 1985, Nova performed 13,424 experiments supporting ICF, Defense Sciences, high-power laser research, and basic science research. At the time of its commissioning, Nova was the world's most powerful laser. Its early experiments demonstrated that 3ω light could produce high-drive, low-preheat environment required for indirect-drive ICE. In the early 1990s, the technical program on Nova for indirect drive ignition was defined by the Nova technical contract established by National Academy Review of ICF in 1990. Successful completion of this research program contributed significantly to the recommendation by the ICF Advisory Committee in 1995 to proceed with the construction of the National Ignition Facility? Nova experiments also demonstrated the utility of high-powered lasers for studying the physics of interest to Defense Sciences. Now, high-powered lasers along with pulsed-power machines are the principal facilities for studying high energy density science in DOE's Stockpile Stewardship Program (SSP). In 1997, one beam of Nova was converted to a short pulsed beam producing a petawatt of power in subpicosecond pulses. The petawatt beam was used for pioneering research in short-pulse laser-matter interactions relevant to fast ignitor ICF and short pulsed x-ray, electron, and particle production for use as probes. Nova is being disassembled and the space is being used to support NIF construction. Nova components are being distributed to a number of other laser laboratories around the world for reuse as determined by DOE. This report summarizes the research performed by the ICF

  5. Inertial fusion program. Progress report, July 1-December 31, 1978

    SciTech Connect

    Perkins, R.B.

    1980-11-01

    Progress at Los Alamos Scientific Laboratory (LASL) in the development of high-energy short-pulse CO/sub 2/ laser systems for fusion research is reported. Improvements to LASL's two-beam system, Gemini, are outlined and experimental results are discussed. Our eight-beam system, Helios, was fired successfully on target for the first time, and became the world's most powerful gas laser for laser fusion studies. Work on Antares, our 100- to 200-TW target irradiation system, is summarized, indicating that design work and building construction are 70 and 48% complete, respectively. A baseline design for automatic centering of laser beams onto the various relay mirrors and the optical design of the Antares front end are discussed. The results of various fusion reactor studies are summarized, as well as investigations of synthetic-fuel production through application of fusion energy to hydrogen production by thermochemical water splitting. Studies on increased efficiency of energy extraction in CO/sub 2/ lasers and on lifetimes of cryogenic pellets in a reactor environment are summarized, as well as the results of studies on pellet injection, tracking, and beam synchronization.

  6. Studying Gene Expression: Database Searches and Promoter Fusions to Investigate Transcriptional Regulation in Bacteria†

    PubMed Central

    Martinez-Vaz, Betsy M.; Makarevitch, Irina; Stensland, Shane

    2010-01-01

    A laboratory project was designed to illustrate how to search biological databases and utilize the information provided by these resources to investigate transcriptional regulation in Escherichia coli. The students searched several databases (NCBI Genomes, RegulonDB and EcoCyc) to learn about gene function, regulation, and the organization of transcriptional units. A fluorometer and GFP promoter fusions were used to obtain fluorescence data and measure changes in transcriptional activity. The class designed and performed experiments to investigate the regulation of genes necessary for biosynthesis of amino acids and how expression is affected by environmental signals and transcriptional regulators. Assessment data showed that this activity enhanced students’ knowledge of databases, reporter genes and transcriptional regulation. PMID:23653697

  7. TBL1XR1/TP63: a novel recurrent gene fusion in B-cell non-Hodgkin lymphoma | Office of Cancer Genomics

    Cancer.gov

    Recently, the landscape of single base mutations in diffuse large B-cell lymphoma (DLBCL) was described. Here we report the discovery of a gene fusion between TBL1XR1 and TP63, the only recurrent somatic novel gene fusion identified in our analysis of transcriptome data from 96 DLBCL cases. Based on this cohort and a further 157 DLBCL cases analyzed by FISH, the incidence in de novo germinal center B cell-like (GCB) DLBCL is 5% (6 of 115).

  8. Fusion

    NASA Astrophysics Data System (ADS)

    Herman, Robin

    1990-10-01

    The book abounds with fascinating anecdotes about fusion's rocky path: the spurious claim by Argentine dictator Juan Peron in 1951 that his country had built a working fusion reactor, the rush by the United States to drop secrecy and publicize its fusion work as a propaganda offensive after the Russian success with Sputnik; the fortune Penthouse magazine publisher Bob Guccione sank into an unconventional fusion device, the skepticism that met an assertion by two University of Utah chemists in 1989 that they had created "cold fusion" in a bottle. Aimed at a general audience, the book describes the scientific basis of controlled fusion--the fusing of atomic nuclei, under conditions hotter than the sun, to release energy. Using personal recollections of scientists involved, it traces the history of this little-known international race that began during the Cold War in secret laboratories in the United States, Great Britain and the Soviet Union, and evolved into an astonishingly open collaboration between East and West.

  9. Inertial Fusion Program. Progress report, January-December 1980

    SciTech Connect

    Not Available

    1982-05-01

    This report summarizes research and development effort in support of the Inertial Confinement Fusion program, including absorption measurements with an integrating sphere, generation of high CO/sub 2/-laser harmonics in the backscattered light from laser plasmas, and the effects of hydrogen target contamination on the hot-electron temperature and transport. The development of new diagnostics is outlined and measurements taken with a proximity-focused x-ray streak camera are presented. High gain in phase conjugation using germanium was demonstrated, data were obtained on retropulse isolation by plasmas generated from metal shutters, damage thresholds for copper mirrors at high fluences were characterized, and phase conjugation in the ultraviolet was demonstrated. Significant progress in the characterization of targets, new techniques in target coating, and important advances in the development of low-density, small-cell-size plastic foam that permit highly accurate machining to any desired shape are presented. The results of various fusion reactor system studies are summarized.

  10. Mirror fusion. Quarterly report, April-June 1981

    SciTech Connect

    Not Available

    1981-09-11

    The information in each Quarterly is presented in the same sequence as in the Field Work Package Proposal and Authorization System (WPAS) submissions prepared for the U.S. Department of Energy; the main sections are Applied Plasma Physics, Confinement Systems, Development and Technology, and Mirror Fusion Test Facility (Planning and Projects). On occasion, we shall include information pertaining to the LLNL role as Lead Laboratory for the Open Systems Mirror Fusion Program. Each of these sections is introduced by an overall statement of the goals and purposes of the groups reporting in it. As appropriate within each section, statements of the goals of individual programs and projects are followed by articles containing summaries of significant recent activity and descriptive text.

  11. Post-entrapment genome engineering: first exon size does not affect the expression of fusion transcripts generated by gene entrapment.

    PubMed

    Osipovich, Anna B; Singh, Aparna; Ruley, H Earl

    2005-03-01

    Gene trap mutagenesis in mouse embryonic stem cells has been widely used for genome-wide studies of mammalian gene function. However, while large numbers of genes can be disrupted, individual mutations may suffer from limitations due to the structure and/or placement of targeting vector. To extend the utility of gene trap mutagenesis, replaceable 3' [or poly(A)] gene trap vectors were developed that permit sequences inserted in individual entrapment clones to be engineered by Cre-mediated recombination. 3' traps incorporating different drug resistance genes could be readily exchanged, simply by selecting for the drug-resistance gene of the replacement vector. By substituting different 3' traps, we show that otherwise identical fusion genes containing a large first exon (804 nt) are not expressed at appreciably lower levels than genes expressing small first exons (384 and 151 nt). Thus, size appears to have less effect on the expression and processing of first exons than has been reported for internal exons. Finally, a retroviral poly(A) trap (consisting of a RNA polymerase II promoter, a neomycin-resistance gene, and 5'-splice site) typically produced mutagenized clones in which vector sequences spliced to the 3'-terminal exons of cellular transcription units, suggesting strong selection for fusion transcripts that evade nonsense-mediated decay. The efficient exchange of poly(A) traps should greatly extend the utility of mutant libraries generated by gene entrapment and provides new strategies to study the rules that govern the expression of exons inserted throughout the genome.

  12. Construction of a dual-tag system for gene expression, protein affinity purification and fusion protein processing.

    PubMed

    Motejadded, Hassan; Altenbuchner, Josef

    2009-04-01

    An E. coli vector system was constructed which allows the expression of fusion genes via a L: -rhamnose-inducible promotor. The corresponding fusion proteins consist of the maltose-binding protein and a His-tag sequence for affinity purification, the Saccharomyces cerevisiae Smt3 protein for protein processing by proteolytic cleavage and the protein of interest. The Smt3 gene was codon-optimized for expression in E. coli. In a second rhamnose-inducible vector, the S. cerevisiae Ulp1 protease gene for processing Smt3 fusion proteins was fused in the same way to maltose-binding protein and His-tag sequence but without the Smt3 gene. The enhanced green fluorescent protein (eGFP) was used as reporter and protein of interest. Both fusion proteins (MalE-6xHis-Smt3-eGFP and MalE-6xHis-Ulp1) were efficiently produced in E. coli and separately purified by amylose resin. After proteolytic cleavage the products were applied to a Ni-NTA column to remove protease and tags. Pure eGFP protein was obtained in the flow-through of the column in a yield of around 35% of the crude cell extract.

  13. Inertial Confinement Fusion: Quarterly report, April-June 1996

    SciTech Connect

    Correll, D.

    1996-06-01

    The lead article, `Ion-beam propagation in a low-density reactor chamber for heavy-ion inertial fusion` (p. 89), explores the ability of heavy-ion beams to be adequately transported and focused in an IFE reactor. The next article, `Efficient production and applications of 2- to 10-keV x rays by laser-heated underdense radiators` (p. 96), explores the ability of the NIF to produce sufficient high-energy x rays for diagnostic backlighting, target preheating, or uniform irradiation of large test objects for Nuclear Weapons Effects Testing. For capsule implosion experiments, the increasing energies and distances involved in the NIF compared to Nova require the development of new diagnostics methods. The article `Fusion reaction-rate measurements--Nova and NIF` (p. 115) first reviews the use of time-resolved neutron measurements on Nova to monitor fusion burn histories and then explores the limitations of that technique, principally Doppler broadening, for the proposed NIF. It also explores the use of gamma rays on Nova, thereby providing a proof-of-principle for using gamma rays for monitoring fusion burn histories on the NIF. The articles `The energetics of gas-filled hohlraums` (p. 110) and `Measurements of laser- speckle-induced perturbations in laser-driven foils` (p. 123) report measurements on Nova of two important aspects of implosion experiments. The first characterizes the amount of energy lost from a hohlraum by stimulated Brillouin and Raman scattering as a function of gas fill and laser-beam uniformity. The second of these articles shows that the growth of density nonuniformities implanted on smooth capsule surfaces by laser speckle can be correlated with the effects of physical surface roughness. The article `Laser-tissue interaction modeling with the LATIS computer program` (p. 103) explores the use of modeling to enhance the effectiveness--maximize desired effects and minimize collateral damage--of lasers for medical purposes.

  14. Construction and Expression of Sugar Kinase Transcriptional Gene Fusions by Using the Sinorhizobium meliloti ORFeome▿

    PubMed Central

    Humann, Jodi L.; Schroeder, Brenda K.; Mortimer, Michael W.; House, Brent L.; Yurgel, Svetlana N.; Maloney, Scott C.; Ward, Kristel L.; Fallquist, Heather M.; Ziemkiewicz, Hope T.; Kahn, Michael L.

    2008-01-01

    The Sinorhizobium meliloti ORFeome project cloned 6,314 open reading frames (ORFs) into a modified Gateway entry vector system from which the ORFs could be transferred to destination vectors in vivo via bacterial conjugation. In this work, a reporter gene destination vector, pMK2030, was constructed and used to generate ORF-specific transcriptional fusions to β-glucuronidase (gusA) and green fluorescent protein (gfp) reporter genes. A total of 6,290 ORFs were successfully transferred from the entry vector library into pMK2030. To demonstrate the utility of this system, reporter plasmids corresponding to 30 annotated sugar kinase genes were integrated into the S. meliloti SM1021 and/or SM8530 genome. Expression of these genes was measured using a high-throughput β-glucuronidase assay to track expression on nine different carbon sources. Six ORFs integrated into SM1021 and SM8530 had different basal levels of expression in the two strains. The annotated activities of three other sugar kinases were also confirmed. PMID:18791020

  15. Emergence of FGFR family gene fusions as therapeutic targets in a wide spectrum of solid tumours.

    PubMed

    Parker, Brittany C; Engels, Manon; Annala, Matti; Zhang, Wei

    2014-01-01

    The emergence of fibroblast growth factor receptor (FGFR) family fusions across diverse cancers has brought attention to FGFR-derived cancer therapies. The discovery of the first recurrent FGFR fusion in glioblastoma was followed by discoveries of FGFR fusions in bladder, lung, breast, thyroid, oral, and prostate cancers. Drug targeting of FGFR fusions has shown promising results and should soon be translating into clinical trials. FGFR fusions form as a result of various mechanisms – predominantly deletion for FGFR1, translocation for FGFR2, and tandem duplication for FGFR3. The ability to exploit the unique targetability of FGFR fusions proves that FGFR-derived therapies could have a promising future in cancer therapeutics. Drug targeting of fusion genes has proven to be an extremely effective therapeutic approach for cancers such as the recurrent BCR–ABL1 fusion in chronic myeloid leukaemia. The recent discovery of recurrent FGFR family fusions in several cancer types has brought to attention the unique therapeutic potential for FGFR-positive patients. Understanding the diverse mechanisms of FGFR fusion formation and their oncogenic potential will shed light on the impact of FGFR-derived therapy in the future.

  16. Highly Specific Targeting of the TMPRSS2/ERG Fusion Gene in Prostate Cancer Using Liposomal Nanotechnology

    DTIC Science & Technology

    2012-06-01

    time due to elimination by reticuloendothelial system. To increase stability and blood circulation half- life coating nanoparticles with polymers such...ERG fusion gene in prostate cancer using liposomal nanotechnology PRINCIPAL INVESTIGATOR: Bulent Ozpolat, M.D., Ph.D...fusion gene in prostate cancer using liposomal nanotechnology 5b. GRANT NUMBER W81XWH-09-1-0385 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) 5d

  17. Gene flow despite complex Robertsonian fusions among rock-wallaby (Petrogale) species

    PubMed Central

    Potter, Sally; Moritz, Craig; Eldridge, Mark D. B.

    2015-01-01

    Complex Robertsonian rearrangements, with shared arms in different fusions, are expected to prevent gene flow between hybrids through missegregation during meiosis. Here, we estimate gene flow between recently diverged and chromosomally diverse rock-wallabies (Petrogale) to test for this form of chromosomal speciation. Contrary to expectations, we observe relatively high admixture among species with complex fusions. Our results reinforce the need to consider alternative roles of chromosome change, together with genic divergence, in driving speciation. PMID:26445985

  18. In vivo topological analysis of Ste2, a yeast plasma membrane protein, by using beta-lactamase gene fusions.

    PubMed Central

    Cartwright, C P; Tipper, D J

    1991-01-01

    Gene fusions were constructed between Ste2, the receptor for the Saccharomyces cerevisiae alpha-factor, and beta la, the secreted form of beta-lactamase encoded by the bla gene of pBR322. The Ste2 and beta la components were linked by a processing fragment (P) from the yeast killer preprotoxin containing a C-terminal lysine-arginine site for cleavage by the Golgi-associated Kex2 protease. Ste2 is predicted to have a rhodopsinlike topology, with an external N terminus and seven transmembrane segments. Fusions to three of the four Ste2 domains predicted to be external resulted in beta la secretion from yeast cells. A fusion at a site just preceding the first transmembrane segment was an exception; the product was cell associated, indicating that the first 44 residues of Ste2 are insufficient to direct secretion of beta la; translocation of this domain presumably requires the downstream transmembrane segment. Expression of fusions located in two domains predicted to be cytoplasmic failed to result in beta la secretion. Following insertion of the preprotoxin signal peptide (S) between the Ste2 and P components of these cytoplasmic fusions, secretion of beta la activity occurred, which is consistent with inversion of the orientation of the beta la reporter. Conversely, insertion of S between Ste2 and P in an external fusion sharply reduced beta la secretion. Complementary information about both cytoplasmic and external domains of Ste2 was therefore provided, and most aspects of the predicted topology were confirmed. The steady-state levels of beta la detected were low, presumably because of efficient degradation of the fusions in the secretory pathway; levels, however, were easily detectable. This method should be valuable in the analysis of in vivo topologies of both homologous and foreign plasma membrane proteins expressed in yeast cells. Images PMID:2017168

  19. Acute myeloid leukemia fusion proteins deregulate genes involved in stem cell maintenance and DNA repair

    PubMed Central

    Alcalay, Myriam; Meani, Natalia; Gelmetti, Vania; Fantozzi, Anna; Fagioli, Marta; Orleth, Annette; Riganelli, Daniela; Sebastiani, Carla; Cappelli, Enrico; Casciari, Cristina; Sciurpi, Maria Teresa; Mariano, Angela Rosa; Minardi, Simone Paolo; Luzi, Lucilla; Muller, Heiko; Di Fiore, Pier Paolo; Frosina, Guido; Pelicci, Pier Giuseppe

    2003-01-01

    Acute myelogenous leukemias (AMLs) are genetically heterogeneous and characterized by chromosomal rearrangements that produce fusion proteins with aberrant transcriptional regulatory activities. Expression of AML fusion proteins in transgenic mice increases the risk of myeloid leukemias, suggesting that they induce a preleukemic state. The underlying molecular and biological mechanisms are, however, unknown. To address this issue, we performed a systematic analysis of fusion protein transcriptional targets. We expressed AML1/ETO, PML/RAR, and PLZF/RAR in U937 hemopoietic precursor cells and measured global gene expression using oligonucleotide chips. We identified 1,555 genes regulated concordantly by at least two fusion proteins that were further validated in patient samples and finally classified according to available functional information. Strikingly, we found that AML fusion proteins induce genes involved in the maintenance of the stem cell phenotype and repress DNA repair genes, mainly of the base excision repair pathway. Functional studies confirmed that ectopic expression of fusion proteins constitutively activates pathways leading to increased stem cell renewal (e.g., the Jagged1/Notch pathway) and provokes accumulation of DNA damage. We propose that expansion of the stem cell compartment and induction of a mutator phenotype are relevant features underlying the leukemic potential of AML-associated fusion proteins. PMID:14660751

  20. Fusion Energy Division annual progress report, period ending December 31, 1988

    SciTech Connect

    Sheffield, J.; Berry, L.A.; Saltmarsh, M.J.

    1990-02-01

    This report discusses the following topics on fusion research: toroidal confinement activities; atomic physics and plasma diagnostics development; fusion theory and computation; plasma technology; superconducting magnet development; advanced systems program; fusion materials research; neutron transport; and management services, quality assurance, and safety.

  1. Fusion Energy Division annual progress report, period ending December 31, 1989

    SciTech Connect

    Sheffield, J.; Baker, C.C.; Saltmarsh, M.J.

    1991-07-01

    The Fusion Program of Oak Ridge National Laboratory (ORNL) carries out research in most areas of magnetic confinement fusion. The program is directed toward the development of fusion as an energy source and is a strong and vital component of both the US fusion program and the international fusion community. Issued as the annual progress report of the ORNL Fusion Energy Division, this report also contains information from components of the Fusion Program that are carried out by other ORNL organizations (about 15% of the program effort). The areas addressed by the Fusion Program and discussed in this report include the following: Experimental and theoretical research on magnetic confinement concepts, engineering and physics of existing and planned devices, including remote handling, development and testing of diagnostic tools and techniques in support of experiments, assembly and distribution to the fusion community of databases on atomic physics and radiation effects, development and testing of technologies for heating and fueling fusion plasmas, development and testing of superconducting magnets for containing fusion plasmas, development and testing of materials for fusion devices, and exploration of opportunities to apply the unique skills, technology, and techniques developed in the course of this work to other areas. Highlights from program activities are included in this report.

  2. NUP98 gene fusions and hematopoietic malignancies: common themes and new biologic insights

    PubMed Central

    Gough, Sheryl M.; Slape, Christopher I.

    2011-01-01

    Structural chromosomal rearrangements of the Nucleoporin 98 gene (NUP98), primarily balanced translocations and inversions, are associated with a wide array of hematopoietic malignancies. NUP98 is known to be fused to at least 28 different partner genes in patients with hematopoietic malignancies, including acute myeloid leukemia, chronic myeloid leukemia in blast crisis, myelodysplastic syndrome, acute lymphoblastic leukemia, and bilineage/biphenotypic leukemia. NUP98 gene fusions typically encode a fusion protein that retains the amino terminus of NUP98; in this context, it is important to note that several recent studies have demonstrated that the amino-terminal portion of NUP98 exhibits transcription activation potential. Approximately half of the NUP98 fusion partners encode homeodomain proteins, and at least 5 NUP98 fusions involve known histone-modifying genes. Several of the NUP98 fusions, including NUP98-homeobox (HOX)A9, NUP98-HOXD13, and NUP98-JARID1A, have been used to generate animal models of both lymphoid and myeloid malignancy; these models typically up-regulate HOXA cluster genes, including HOXA5, HOXA7, HOXA9, and HOXA10. In addition, several of the NUP98 fusion proteins have been shown to inhibit differentiation of hematopoietic precursors and to increase self-renewal of hematopoietic stem or progenitor cells, providing a potential mechanism for malignant transformation. PMID:21948299

  3. A rapid and efficient newly established method to detect COL1A1-PDGFB gene fusion in dermatofibrosarcoma protuberans

    SciTech Connect

    Yokoyama, Yoko; Shimizu, Akira; Okada, Etsuko; Ishikawa, Osamu; Motegi, Sei-ichiro

    2012-08-24

    Highlights: Black-Right-Pointing-Pointer We developed new method to rapidly identify COL1A1-PDGFB fusion in DFSP. Black-Right-Pointing-Pointer New PCR method using a single primer pair detected COL1A1-PDGFB fusion in DFSP. Black-Right-Pointing-Pointer This is the first report of DFSP with a novel COL1A1 breakpoint in exon 5. -- Abstract: The detection of fusion transcripts of the collagen type 1{alpha}1 (COL1A1) and platelet-derived growth factor-BB (PDGFB) genes by genetic analysis has recognized as a reliable and valuable molecular tool for the diagnosis of dermatofibrosarcoma protuberans (DFSP). To detect the COL1A1-PDGFB fusion, almost previous reports performed reverse transcription polymerase chain reaction (RT-PCR) using multiplex forward primers from COL1A1. However, it has possible technical difficulties with respect to the handling of multiple primers and reagents in the procedure. The objective of this study is to establish a rapid, easy, and efficient one-step method of PCR using only a single primer pair to detect the fusion transcripts of the COL1A1 and PDGFB in DFSP. To validate new method, we compared the results of RT-PCR in five patients of DFSP between the previous method using multiplex primers and our established one-step RT-PCR using a single primer pair. In all cases of DFSP, the COL1A1-PDGFB fusion was detected by both previous method and newly established one-step PCR. Importantly, we detected a novel COL1A1 breakpoint in exon 5. The newly developed method is valuable to rapidly identify COL1A1-PDGFB fusion transcripts in DFSP.

  4. Fusion Energy Division: Annual progress report, period ending December 31, 1987

    SciTech Connect

    Morgan, O.B. Jr.; Berry, L.A.; Sheffield, J.

    1988-11-01

    The Fusion Program of Oak Ridge National Laboratory (ORNL), a major part of the national fusion program, carries out research in nearly all areas of magnetic fusion. Collaboration among staff from ORNL, Martin Marietta Energy Systems, Inc., private industry, the academic community, and other fusion laboratories, in the United States and abroad, is directed toward the development of fusion as an energy source. This report documents the program's achievements during 1987. Issued as the annual progress report of the ORNL Fusion Energy Division, it also contains information from components of the Fusion Program that are external to the division (about 15% of the program effort). The areas addressed by the Fusion Program include the following: experimental and theoretical research on magnetic confinement concepts, engineering and physics of existing and planned devices, development and testing of diagnostic tools and techniques in support of experiments, assembly and distribution to the fusion community of databases on atomic physics and radiation effects, development and testing of technologies for heating and fueling fusion plasmas, development and testing of superconducting magnets for containing fusion plasmas, and development and testing of materials for fusion devices. Highlights from program activities are included in this report. 126 figs., 15 tabs.

  5. Monitoring of gene expression in bacteria during infections using an adaptable set of bioluminescent, fluorescent and colorigenic fusion vectors.

    PubMed

    Uliczka, Frank; Pisano, Fabio; Kochut, Annika; Opitz, Wiebke; Herbst, Katharina; Stolz, Tatjana; Dersch, Petra

    2011-01-01

    A family of versatile promoter-probe plasmids for gene expression analysis was developed based on a modular expression plasmid system (pZ). The vectors contain different replicons with exchangeable antibiotic cassettes to allow compatibility and expression analysis on a low-, midi- and high-copy number basis. Suicide vector variants also permit chromosomal integration of the reporter fusion and stable vector derivatives can be used for in vivo or in situ expression studies under non-selective conditions. Transcriptional and translational fusions to the reporter genes gfp(mut3.1), amCyan, dsRed2, luxCDABE, phoA or lacZ can be constructed, and presence of identical multiple cloning sites in the vector system facilitates the interchange of promoters or reporter genes between the plasmids of the series. The promoter of the constitutively expressed gapA gene of Escherichia coli was included to obtain fluorescent and bioluminescent expression constructs. A combination of the plasmids allows simultaneous detection and gene expression analysis in individual bacteria, e.g. in bacterial communities or during mouse infections. To test our vector system, we analyzed and quantified expression of Yersinia pseudotuberculosis virulence genes under laboratory conditions, in association with cells and during the infection process.

  6. Report of the Fusion Energy Sciences Advisory Committee. Panel on Integrated Simulation and Optimization of Magnetic Fusion Systems

    SciTech Connect

    Dahlburg, Jill; Corones, James; Batchelor, Donald; Bramley, Randall; Greenwald, Martin; Jardin, Stephen; Krasheninnikov, Sergei; Laub, Alan; Leboeuf, Jean-Noel; Lindl, John; Lokke, William; Rosenbluth, Marshall; Ross, David; Schnack, Dalton

    2002-11-01

    Fusion is potentially an inexhaustible energy source whose exploitation requires a basic understanding of high-temperature plasmas. The development of a science-based predictive capability for fusion-relevant plasmas is a challenge central to fusion energy science, in which numerical modeling has played a vital role for more than four decades. A combination of the very wide range in temporal and spatial scales, extreme anisotropy, the importance of geometric detail, and the requirement of causality which makes it impossible to parallelize over time, makes this problem one of the most challenging in computational physics. Sophisticated computational models are under development for many individual features of magnetically confined plasmas and increases in the scope and reliability of feasible simulations have been enabled by increased scientific understanding and improvements in computer technology. However, full predictive modeling of fusion plasmas will require qualitative improvements and innovations to enable cross coupling of a wider variety of physical processes and to allow solution over a larger range of space and time scales. The exponential growth of computer speed, coupled with the high cost of large-scale experimental facilities, makes an integrated fusion simulation initiative a timely and cost-effective opportunity. Worldwide progress in laboratory fusion experiments provides the basis for a recent FESAC recommendation to proceed with a burning plasma experiment (see FESAC Review of Burning Plasma Physics Report, September 2001). Such an experiment, at the frontier of the physics of complex systems, would be a huge step in establishing the potential of magnetic fusion energy to contribute to the world’s energy security. An integrated simulation capability would dramatically enhance the utilization of such a facility and lead to optimization of toroidal fusion plasmas in general. This science-based predictive capability, which was cited in the FESAC

  7. Repair welding of fusion reactor components. Final technical report

    SciTech Connect

    Chin, B.A.; Wang, C.A.

    1997-09-30

    The exposure of metallic materials, such as structural components of the first wall and blanket of a fusion reactor, to neutron irradiation will induce changes in both the material composition and microstructure. Along with these changes can come a corresponding deterioration in mechanical properties resulting in premature failure. It is, therefore, essential to expect that the repair and replacement of the degraded components will be necessary. Such repairs may require the joining of irradiated materials through the use of fusion welding processes. The present ITER (International Thermonuclear Experimental Reactor) conceptual design is anticipated to have about 5 km of longitudinal welds and ten thousand pipe butt welds in the blanket structure. A recent study by Buende et al. predict that a failure is most likely to occur in a weld. The study is based on data from other large structures, particularly nuclear reactors. The data used also appear to be consistent with the operating experience of the Fast Flux Test Facility (FFTF). This reactor has a fuel pin area comparable with the area of the ITER first wall and has experienced one unanticipated fuel pin failure after two years of operation. The repair of irradiated structures using fusion welding will be difficult due to the entrapped helium. Due to its extremely low solubility in metals, helium will diffuse and agglomerate to form helium bubbles after being trapped at point defects, dislocations, and grain boundaries. Welding of neutron-irradiated type 304 stainless steels has been reported with varying degree of heat-affected zone cracking (HAZ). The objectives of this study were to determine the threshold helium concentrations required to cause HAZ cracking and to investigate techniques that might be used to eliminate the HAZ cracking in welding of helium-containing materials.

  8. Fission-Fusion Neutron Source Progress Report Sept 30, 2009

    SciTech Connect

    Chapline, G F; Daffin, F; Clark, R

    2010-02-19

    In this report the authors describe the progress made in FY09 in evaluating the feasibility of a new concept for using the DT fusion reaction to produce intense pulses of 14 MeV neutrons. In this new scheme the heating of the DT is accomplished using fission fragments rather than ion beams as in conventional magnet confinement fusion schemes or lasers in inertial confinement schemes. As a source of fission fragments they propose using a dust reactor concept introduced some time ago by one of us (RC). An attractive feature of this approach is that there is no need for a large auxiliary power source to heat the DT plasma to the point where self-sustaining fusion become possible. Their scheme does require pulsed magnetic fields, but generating these fields requires only a modest power source. The dust reactor that they propose using for their neutron source would use micron-sized UC pellets suspended in a vacuum as the reactor fuel. Surrounding the fuel with a moderator such as heavy water (D{sub 2}O) would allow the reactor to operate as a thermal reactor and require only modest amounts of HEU. The scheme for using fission fragments to generate intense pulses of 14 MeV neutrons is based on the fission fragment rocket idea. In the fission fragment rocket scheme it was contemplated that the fission fragments produced in a low density reactor core could be guided out of the reactor by large magnetic fields used to form a 'rocket exhaust'. Their adaptation of this idea for the purposes of making a neutron source involves using the fission fragments escaping from one side of a tandem magnet mirror to heat DT gas confined in the adjacent magnetic trap.

  9. Midline lumbar fusion using cortical bone trajectory screws. Preliminary report

    PubMed Central

    Bielecki, Mateusz; Prokopienko, Marek; Nowak, Arkadiusz; Czernicki, Tomasz; Marchel, Andrzej

    2016-01-01

    Introduction Midline lumbar fusion (MIDLF) using cortical bone trajectory is an alternative method of transpedicular spinal fusion for degenerative disease. The new entry points’ location and screwdriving direction allow the approach-related morbidity to be reduced. Aim To present our preliminary experience with the MIDLF technique on the first 5 patients with lumbar degenerative disease and with follow-up of at least 6 months. Material and methods Retrospective analysis was performed on the first 5 patients with foraminal (4) or central (1) stenosis operated on between December 2014 and February 2015. Three patients were fused at L4–L5 and two at the L5–S1 level. Results No intra- or post-operative complications occurred with this approach. An improvement regarding the leading symptom in the early postoperative period (sciatica 4/4, claudication 1/1) was achieved in all patients. The mean improvements in the visual analogue scale for low back and leg pain were 2.2 and 4.8 respectively. The mean Oswestry Disability Index scores were 52% (range: 16–82%) before surgery and 33% (range: 12–56%) at 3-month follow-up (mean improvement 19%). At the most recent follow-up, 4 patients reported the maintenance of the satisfactory result. The early standing and follow-up X-rays showed satisfactory screw placement in all patients. Conclusions In our initial experience, the MIDLF technique seems to be an encouraging alternative to traditional transpedicular trajectory screws when short level lumbar fusion is needed. Nevertheless, longer observations on larger groups of patients are needed to reliably evaluate the safety of the method and the sustainability of the results. PMID:27829938

  10. Recurrent Fusion Genes in Gastric Cancer: CLDN18-ARHGAP26 Induces Loss of Epithelial Integrity.

    PubMed

    Yao, Fei; Kausalya, Jaya P; Sia, Yee Yen; Teo, Audrey S M; Lee, Wah Heng; Ong, Alicia G M; Zhang, Zhenshui; Tan, Joanna H J; Li, Guoliang; Bertrand, Denis; Liu, Xingliang; Poh, Huay Mei; Guan, Peiyong; Zhu, Feng; Pathiraja, Thushangi Nadeera; Ariyaratne, Pramila N; Rao, Jaideepraj; Woo, Xing Yi; Cai, Shaojiang; Mulawadi, Fabianus H; Poh, Wan Ting; Veeravalli, Lavanya; Chan, Chee Seng; Lim, Seong Soo; Leong, See Ting; Neo, Say Chuan; Choi, Poh Sum D; Chew, Elaine G Y; Nagarajan, Niranjan; Jacques, Pierre-Étienne; So, Jimmy B Y; Ruan, Xiaoan; Yeoh, Khay Guan; Tan, Patrick; Sung, Wing-Kin; Hunziker, Walter; Ruan, Yijun; Hillmer, Axel M

    2015-07-14

    Genome rearrangements, a hallmark of cancer, can result in gene fusions with oncogenic properties. Using DNA paired-end-tag (DNA-PET) whole-genome sequencing, we analyzed 15 gastric cancers (GCs) from Southeast Asians. Rearrangements were enriched in open chromatin and shaped by chromatin structure. We identified seven rearrangement hot spots and 136 gene fusions. In three out of 100 GC cases, we found recurrent fusions between CLDN18, a tight junction gene, and ARHGAP26, a gene encoding a RHOA inhibitor. Epithelial cell lines expressing CLDN18-ARHGAP26 displayed a dramatic loss of epithelial phenotype and long protrusions indicative of epithelial-mesenchymal transition (EMT). Fusion-positive cell lines showed impaired barrier properties, reduced cell-cell and cell-extracellular matrix adhesion, retarded wound healing, and inhibition of RHOA. Gain of invasion was seen in cancer cell lines expressing the fusion. Thus, CLDN18-ARHGAP26 mediates epithelial disintegration, possibly leading to stomach H(+) leakage, and the fusion might contribute to invasiveness once a cell is transformed.

  11. A protein disulfide isomerase gene fusion expression system that increases the extracellular productivity of Bacillus brevis.

    PubMed

    Kajino, T; Ohto, C; Muramatsu, M; Obata, S; Udaka, S; Yamada, Y; Takahashi, H

    2000-02-01

    We have developed a versatile Bacillus brevis expression and secretion system based on the use of fungal protein disulfide isomerase (PDI) as a gene fusion partner. Fusion with PDI increased the extracellular production of heterologous proteins (light chain of immunoglobulin G, 8-fold; geranylgeranyl pyrophosphate synthase, 12-fold). Linkage to PDI prevented the aggregation of the secreted proteins, resulting in high-level accumulation of fusion proteins in soluble and biologically active forms. We also show that the disulfide isomerase activity of PDI in a fusion protein is responsible for the suppression of the aggregation of the protein with intradisulfide, whereas aggregation of the protein without intradisulfide was prevented even when the protein was fused to a mutant PDI whose two active sites were disrupted, suggesting that another PDI function, such as chaperone-like activity, synergistically prevented the aggregation of heterologous proteins in the PDI fusion expression system.

  12. Fusion Energy Division progress report, 1 January 1990--31 December 1991

    SciTech Connect

    Sheffield, J.; Baker, C.C.; Saltmarsh, M.J.

    1994-03-01

    The Fusion Program of the Oak Ridge National Laboratory (ORNL), a major part of the national fusion program, encompasses nearly all areas of magnetic fusion research. The program is directed toward the development of fusion as an economical and environmentally attractive energy source for the future. The program involves staff from ORNL, Martin Marietta Energy systems, Inc., private industry, the academic community, and other fusion laboratories, in the US and abroad. Achievements resulting from this collaboration are documented in this report, which is issued as the progress report of the ORNL Fusion Energy Division; it also contains information from components for the Fusion Program that are external to the division (about 15% of the program effort). The areas addressed by the Fusion Program include the following: experimental and theoretical research on magnetic confinement concepts; engineering and physics of existing and planned devices, including remote handling; development and testing of diagnostic tools and techniques in support of experiments; assembly and distribution to the fusion community of databases on atomic physics and radiation effects; development and testing of technologies for heating and fueling fusion plasmas; development and testing of superconducting magnets for containing fusion plasmas; development and testing of materials for fusion devices; and exploration of opportunities to apply the unique skills, technology, and techniques developed in the course of this work to other areas (about 15% of the Division`s activities). Highlights from program activities during 1990 and 1991 are presented.

  13. Model year 2010 Ford Fusion Level-1 testing report.

    SciTech Connect

    Rask, E.; Bocci, D.; Duoba, M.; Lohse-Busch, H.; Energy Systems

    2010-11-23

    As a part of the US Department of Energy's Advanced Vehicle Testing Activity (AVTA), a model year 2010 Ford Fusion was procured by eTec (Phoenix, AZ) and sent to ANL's Advanced Powertrain Research Facility for the purposes of vehicle-level testing in support of the Advanced Vehicle Testing Activity. Data was acquired during testing using non-intrusive sensors, vehicle network information, and facilities equipment (emissions and dynamometer). Standard drive cycles, performance cycles, steady-state cycles, and A/C usage cycles were conducted. Much of this data is openly available for download in ANL's Downloadable Dynamometer Database. The major results are shown in this report. Given the benchmark nature of this assessment, the majority of the testing was done over standard regulatory cycles and sought to obtain a general overview of how the vehicle performs. These cycles include the US FTP cycle (Urban) and Highway Fuel Economy Test cycle as well as the US06, a more aggressive supplemental regulatory cycle. Data collection for this testing was kept at a fairly high level and includes emissions and fuel measurements from an exhaust emissions bench, high-voltage and accessory current/voltage from a DC power analyzer, and CAN bus data such as engine speed, engine load, and electric machine operation. The following sections will seek to explain some of the basic operating characteristics of the MY2010 Fusion and provide insight into unique features of its operation and design.

  14. Inertial fusion program. Progress report, January 1-June 30, 1978

    SciTech Connect

    Skoberne, F.

    1980-05-01

    Studies and experiments aimed at investigating the possibility of restoring wavefront quality in optical systems through phase conjugation are summarized, and work that could lead to the development of highly damage-resistant isolators is discussed. The effects of various parameters on pulse-energy uniformity and of multipass extraction on laser efficiency are reported. Results of equation-of-state, shock propagation, multiburst simulation, and opacity measurements are discussed. Target designs are described that should provide a smooth transition from the exploding-pusher regime of experiments to that of isentropic compression. Progress in target fabrication techniques toward creating a 20-times-liquid-density target are outlined, and efforts that led to the extension of our neutron detection capability to levels of less than 10/sup 3/ n are summarized. The results of various studies of laser fusion application, e.g., for producing ultrahigh-temperature process heat or hydrogen from water decomposition are presented, as well as investigations of fusion-fission hybrids for the production of /sup 233/U from /sup 232/Th.

  15. Fission-Fusion Neutron Source Progress Report July 31, 2009

    SciTech Connect

    Chapline, G; Daffin, F; Clarke, R

    2010-02-19

    In this report the authors describe progress in evaluating the feasibility of a novel concept for producing intense pulses of 14 MeV neutrons using the DT fusion reaction. In this new scheme the heating of the DT is accomplished using fission fragments rather than ion beams as in conventional magnet fusion schemes or lasers in ICF schemes. This has the great advantage that there is no need for any large auxiliary power source. The scheme does require large magnetic fields, but generating these fields, e.g. with superconducting magnets, requires only a modest power source. As a source of fission fragments they propose using a dusty reactor concept introduced some time ago by one of us (RC). The version of the dusty reactor that they propose using for our neutron source would operate as a thermal neutron reactor and use highly enriched uranium in the form of micron sized pellets of UC. Our scheme for using the fission fragments to produce intense pulses of 14 MeV neutrons is based on the fission fragment rocket idea. In the fission fragment rocket scheme it was contemplated that the fission fragments produced in a low density reactor core would then be guided out of the reactor by large magnetic fields. A simple version of this idea would be to use the fission fragments escaping from one side of a tandem magnet mirror to heat DT gas confined in the adjacent magnetic trap.

  16. Use of gene fusions to determine the orientation of gene phoA on the Escherichia coli chromosome.

    PubMed Central

    Sarthy, A; Michaelis, S; Beckwith, J

    1981-01-01

    We present genetic evidence which demonstrates that the phoA gene is transcribed in the clockwise direction on the Escherichia coli chromosome, in contrast to an earlier proposal. Our conclusion is based on analysis of various genetic fusions between the lac operon and the phoA gene. PMID:7007316

  17. EP300-ZNF384 fusion gene product up-regulates GATA3 gene expression and induces hematopoietic stem cell gene expression signature in B-cell precursor acute lymphoblastic leukemia cells.

    PubMed

    Yaguchi, Akinori; Ishibashi, Takeshi; Terada, Kazuki; Ueno-Yokohata, Hitomi; Saito, Yuya; Fujimura, Junya; Shimizu, Toshiaki; Ohki, Kentaro; Manabe, Atsushi; Kiyokawa, Nobutaka

    2017-04-04

    ZNF384-related fusion genes are associated with a distinct subgroup of B-cell precursor acute lymphoblastic leukemias in childhood, with a frequency of approximately 3-4%. We previously identified a novel EP300-ZNF384 fusion gene. Patients with the ZNF384-related fusion gene exhibit a hematopoietic stem cell (HSC) gene expression signature and characteristic immunophenotype with negative or low expression of CD10 and aberrant expression of myeloid antigens, such as CD33 and CD13. However, the molecular basis of this pathogenesis remains completely unknown. In the present study, we examined the biological effects of EP300-ZNF384 expression induced by retrovirus-mediated gene transduction in an REH B-cell precursor acute lymphoblastic leukemia cell line, and observed the acquisition of the HSC gene expression signature and an up-regulation of GATA3 gene expression, as assessed by microarray analysis. In contrast, the gene expression profile induced by wild-type ZNF384 in REH cells was significantly different from that by EP300-ZNF384 expression. Together with the results of reporter assays, which revealed the enhancement of GATA3-promoter activity by EP300-ZNF384 expression, these findings suggest that EP300-ZNF384 mediates GATA3 gene expression and may be involved in the acquisition of the HSC gene expression signature and characteristic immunophenotype in B-cell precursor acute lymphoblastic leukemia cells.

  18. Discovery and characterization of a novel CCND1/MRCK gene fusion in mantle cell lymphoma.

    PubMed

    Masamha, Chioniso Patience; Albrecht, Todd R; Wagner, Eric J

    2016-03-29

    The t(11;14) translocation resulting in constitutive cyclin D1 expression is an early event in mantle cell lymphoma (MCL) transformation. Patients with a highly proliferative phenotype produce cyclin D1 transcripts with truncated 3'UTRs that evade miRNA regulation. Here, we report the recurrence of a novel gene fusion in MCL cell lines and MCL patient isolates that consists of the full protein coding region of cyclin D1 (CCND1) and a 3'UTR consisting of sequences from both the CCND1 3'UTR and myotonic dystrophy kinase-related Cdc42-binding kinase's (MRCK) intron one. The resulting CCND1/MRCK mRNA is resistant to CCND1-targeted miRNA regulation, and targeting the MRCK region of the chimeric 3'UTR with siRNA results in decreased CCND1 levels.

  19. RNA-Seq mapping and detection of gene fusions with a suffix array algorithm.

    PubMed

    Sakarya, Onur; Breu, Heinz; Radovich, Milan; Chen, Yongzhi; Wang, Yulei N; Barbacioru, Catalin; Utiramerur, Sowmi; Whitley, Penn P; Brockman, Joel P; Vatta, Paolo; Zhang, Zheng; Popescu, Liviu; Muller, Matthew W; Kudlingar, Vidya; Garg, Nriti; Li, Chieh-Yuan; Kong, Benjamin S; Bodeau, John P; Nutter, Robert C; Gu, Jian; Bramlett, Kelli S; Ichikawa, Jeffrey K; Hyland, Fiona C; Siddiqui, Asim S

    2012-01-01

    High-throughput RNA sequencing enables quantification of transcripts (both known and novel), exon/exon junctions and fusions of exons from different genes. Discovery of gene fusions-particularly those expressed with low abundance- is a challenge with short- and medium-length sequencing reads. To address this challenge, we implemented an RNA-Seq mapping pipeline within the LifeScope software. We introduced new features including filter and junction mapping, annotation-aided pairing rescue and accurate mapping quality values. We combined this pipeline with a Suffix Array Spliced Read (SASR) aligner to detect chimeric transcripts. Performing paired-end RNA-Seq of the breast cancer cell line MCF-7 using the SOLiD system, we called 40 gene fusions among over 120,000 splicing junctions. We validated 36 of these 40 fusions with TaqMan assays, of which 25 were expressed in MCF-7 but not the Human Brain Reference. An intra-chromosomal gene fusion involving the estrogen receptor alpha gene ESR1, and another involving the RPS6KB1 (Ribosomal protein S6 kinase beta-1) were recurrently expressed in a number of breast tumor cell lines and a clinical tumor sample.

  20. Development of RNA-FISH Assay for Detection of Oncogenic FGFR3-TACC3 Fusion Genes in FFPE Samples

    PubMed Central

    Kojima, Takahiro; Nishimura, Kouichi; Kandori, Shuya; Kawahara, Takashi; Yoshino, Takayuki; Ueno, Satoshi; Iizumi, Yuichi; Mitsuzuka, Koji; Arai, Yoichi; Tsuruta, Hiroshi; Habuchi, Tomonori; Kobayashi, Takashi; Matsui, Yoshiyuki; Ogawa, Osamu; Sugimoto, Mikio; Kakehi, Yoshiyuki; Nagumo, Yoshiyuki; Tsutsumi, Masakazu; Oikawa, Takehiro; Kikuchi, Koji; Nishiyama, Hiroyuki

    2016-01-01

    Introduction and Objectives Oncogenic FGFR3-TACC3 fusions and FGFR3 mutations are target candidates for small molecule inhibitors in bladder cancer (BC). Because FGFR3 and TACC3 genes are located very closely on chromosome 4p16.3, detection of the fusion by DNA-FISH (fluorescent in situ hybridization) is not a feasible option. In this study, we developed a novel RNA-FISH assay using branched DNA probe to detect FGFR3-TACC3 fusions in formaldehyde-fixed paraffin-embedded (FFPE) human BC samples. Materials and Methods The RNA-FISH assay was developed and validated using a mouse xenograft model with human BC cell lines. Next, we assessed the consistency of the RNA-FISH assay using 104 human BC samples. In this study, primary BC tissues were stored as frozen and FFPE tissues. FGFR3-TACC3 fusions were independently detected in FFPE sections by the RNA-FISH assay and in frozen tissues by RT-PCR. We also analyzed the presence of FGFR3 mutations by targeted sequencing of genomic DNA extracted from deparaffinized FFPE sections. Results FGFR3-TACC3 fusion transcripts were identified by RNA-FISH and RT-PCR in mouse xenograft FFPE tissues using the human BC cell lines RT112 and RT4. These cell lines have been reported to be fusion-positive. Signals for FGFR3-TACC3 fusions by RNA-FISH were positive in 2/60 (3%) of non-muscle-invasive BC (NMIBC) and 2/44 (5%) muscle-invasive BC (MIBC) patients. The results of RT-PCR of all 104 patients were identical to those of RNA-FISH. FGFR3 mutations were detected in 27/60 (45%) NMIBC and 8/44 (18%) MIBC patients. Except for one NMIBC patient, FGFR3 mutation and FGFR3-TACC3 fusion were mutually exclusive. Conclusions We developed an RNA-FISH assay for detection of the FGFR3-TACC3 fusion in FFPE samples of human BC tissues. Screening for not only FGFR3 mutations, but also for FGFR3-TACC3 fusion transcripts has the potential to identify additional patients that can be treated with FGFR inhibitors. PMID:27930669

  1. Geophysical data fusion for subsurface imaging. Final report

    SciTech Connect

    1995-10-01

    This report contains the results of a three year, three-phase project whose long-range goal has been to create a means for the more detailed and accurate definition of the near-surface (0--300 ft) geology beneath a site that had been subjected to environmental pollution. The two major areas of research and development have been: improved geophysical field data acquisition techniques; and analytical tools for providing the total integration (fusion) of all site data. The long-range goal of this project has been to mathematically, integrate the geophysical data that could be derived from multiple sensors with site geologic information and any other type of available site data, to provide a detailed characterization of thin clay layers and geological discontinuities at hazardous waste sites.

  2. ESRRA-C11orf20 Is a Recurrent Gene Fusion in Serous Ovarian Carcinoma

    PubMed Central

    Green, Ann E.; Nielsen, Julie S.; Nelson, Brad H.; Drescher, Charles W.; Brown, Patrick O.

    2011-01-01

    Every year, ovarian cancer kills approximately 14,000 women in the United States and more than 140,000 women worldwide. Most of these deaths are caused by tumors of the serous histological type, which is rarely diagnosed before it has disseminated. By deep paired-end sequencing of mRNA from serous ovarian cancers, followed by deep sequencing of the corresponding genomic region, we identified a recurrent fusion transcript. The fusion transcript joins the 5′ exons of ESRRA, encoding a ligand-independent member of the nuclear-hormone receptor superfamily, to the 3′ exons of C11orf20, a conserved but uncharacterized gene located immediately upstream of ESRRA in the reference genome. To estimate the prevalence of the fusion, we tested 67 cases of serous ovarian cancer by RT-PCR and sequencing and confirmed its presence in 10 of these. Targeted resequencing of the corresponding genomic region from two fusion-positive tumor samples identified a nearly clonal chromosomal rearrangement positioning ESRRA upstream of C11orf20 in one tumor, and evidence of local copy number variation in the ESRRA locus in the second tumor. We hypothesize that the recurrent novel fusion transcript may play a role in pathogenesis of a substantial fraction of serous ovarian cancers and could provide a molecular marker for detection of the cancer. Gene fusions involving adjacent or nearby genes can readily escape detection but may play important roles in the development and progression of cancer. PMID:21949640

  3. Fusion gene and splice variant analyses in liquid biopsies of lung cancer patients

    PubMed Central

    Giménez-Capitán, Ana; Karachaliou, Niki; Pérez-Rosado, Ana; Viteri, Santiago; Morales-Espinosa, Daniela; Rosell, Rafael

    2016-01-01

    Obtaining a biopsy of solid tumors requires invasive procedures that strongly limit patient compliance. In contrast, a blood extraction is safe, can be performed at many time points during the course disease and encourages appropriate therapy modifications, potentially improving the patient’s clinical outcome and quality of life. Fusion of the tyrosine kinase genes anaplastic lymphoma kinase (ALK), C-ROS oncogen 1 (ROS 1), rearranged during transfection (RET) and neurotrophic tyrosine kinase 1 (NTRK1) occur in 1–5% of lung adenocarcinomas and constitute therapeutic targets for tyrosine kinase inhibitors. In addition, a MET splicing variant of exon 14, has been reported in 2–4% of lung adenocarcinoma and recent studies suggests that targeted therapies inhibiting MET signaling would be beneficial for patients with this alteration. In this review, we will summarize the new techniques recently developed to detect ALK, RET, ROS and NTRK1 fusions and MET exon 14 splicing variant in liquid biopsy using plasma, serum, circulating tumor cells (CTCs), platelets and exosomes as starting material. PMID:27826534

  4. Fusion Energy Division progress report, January 1, 1992--December 31, 1994

    SciTech Connect

    Sheffield, J.; Baker, C.C.; Saltmarsh, M.J.; Shannon, T.E.

    1995-09-01

    The report covers all elements of the ORNL Fusion Program, including those implemented outside the division. Non-fusion work within FED, much of which is based on the application of fusion technologies and techniques, is also discussed. The ORNL Fusion Program includes research and development in most areas of magnetic fusion research. The program is directed toward the development of fusion as an energy source and is a strong and vital component of both the US and international fusion efforts. The research discussed in this report includes: experimental and theoretical research on magnetic confinement concepts; engineering and physics of existing and planned devices; development and testing of plasma diagnostic tools and techniques; assembly and distribution of databases on atomic physics and radiation effects; development and testing of technologies for heating and fueling fusion plasmas; and development and testing of materials for fusion devices. The activities involving the use of fusion technologies and expertise for non-fusion applications ranged from semiconductor manufacturing to environmental management.

  5. Inertial Confinement Fusion quarterly report, April--June 1995. Volume 5, No. 3

    SciTech Connect

    1995-12-31

    The ICF Quarterly Reports is published four times each fiscal year by the Inertial Confinement Fusion Program at the Lawrence Livermore National Laboratory. The journal reports selected current research within the ICF Program. Major areas of investigation presented here include fusion target theory and design, target fabrication, target experiments, and laser and optical science and technology.

  6. Inertial confinement fusion target component fabrication and technology development report

    NASA Astrophysics Data System (ADS)

    Steinman, D.

    1994-03-01

    On December 30, 1990, the US Department of Energy entered into a contract with General Atomics (GA) to be the Inertial Confinement Fusion Target Component Fabrication and Technology Development Support contractor. This report documents the technical activities which took place under this contract during the period of October 1, 1992 through September 30, 1993. During this period, GA was assigned 18 tasks in support of the Inertial Confinement Fusion program and its laboratories. These tasks included 'Capabilities Activation' and 'Capabilities Demonstration' to enable us to begin production of glass and composite polymer capsules. Capsule delivery tasks included 'Small Glass Shell Deliveries' and 'Composite Polymer Capsules' for Lawrence Livermore National Laboratory (LLNL) and Los Alamos National Laboratory (LANL). We also were asked to provide direct 'Onsite Support' at LLNL and LANL. We continued planning for the transfer of 'Micromachining Equipment from Rocky Flats' and established 'Target Component Micromachining and Electroplating Facilities' at GA. We fabricated over 1100 films and filters of 11 types for Sandia National Laboratory and provided full-time onsite engineering support for target fabrication and characterization. We initiated development of methods to make targets for the Naval Research Laboratory. We investigated spherical interferometry, built an automated capsule sorter, and developed an apparatus for calorimetric measurement of fuel fill for LLNL. We assisted LANL in the 'Characterization of Opaque b-Layered Targets.' We developed deuterated and UV-opaque polymers for use by the University of Rochester's Laboratory for Laser Energetics (UR/LLE) and devised a triple-orifice droplet generator to demonstrate the controlled-mass nature of the microencapsulation process.

  7. A human ESC model for MLL-AF4 leukemic fusion gene reveals an impaired early hematopoietic-endothelial specification.

    PubMed

    Bueno, Clara; Montes, Rosa; Melen, Gustavo J; Ramos-Mejia, Verónica; Real, Pedro J; Ayllón, Verónica; Sanchez, Laura; Ligero, Gertrudis; Gutierrez-Aranda, Iván; Fernández, Agustín F; Fraga, Mario F; Moreno-Gimeno, Inmaculada; Burks, Deborah; Plaza-Calonge, María del Carmen; Rodríguez-Manzaneque, Juan C; Menendez, Pablo

    2012-06-01

    The MLL-AF4 fusion gene is a hallmark genomic aberration in high-risk acute lymphoblastic leukemia in infants. Although it is well established that MLL-AF4 arises prenatally during human development, its effects on hematopoietic development in utero remain unexplored. We have created a human-specific cellular system to study early hemato-endothelial development in MLL-AF4-expressing human embryonic stem cells (hESCs). Functional studies, clonal analysis and gene expression profiling reveal that expression of MLL-AF4 in hESCs has a phenotypic, functional and gene expression impact. MLL-AF4 acts as a global transcriptional activator and a positive regulator of homeobox gene expression in hESCs. Functionally, MLL-AF4 enhances the specification of hemogenic precursors from hESCs but strongly impairs further hematopoietic commitment in favor of an endothelial cell fate. MLL-AF4 hESCs are transcriptionally primed to differentiate towards hemogenic precursors prone to endothelial maturation, as reflected by the marked upregulation of master genes associated to vascular-endothelial functions and early hematopoiesis. Furthermore, we report that MLL-AF4 expression is not sufficient to transform hESC-derived hematopoietic cells. This work illustrates how hESCs may provide unique insights into human development and further our understanding of how leukemic fusion genes, known to arise prenatally, regulate human embryonic hematopoietic specification.

  8. A human ESC model for MLL-AF4 leukemic fusion gene reveals an impaired early hematopoietic-endothelial specification

    PubMed Central

    Bueno, Clara; Montes, Rosa; Melen, Gustavo J; Ramos-Mejia, Verónica; Real, Pedro J; Ayllón, Verónica; Sanchez, Laura; Ligero, Gertrudis; Gutierrez-Aranda, Iván; Fernández, Agustín F; Fraga, Mario F; Moreno-Gimeno, Inmaculada; Burks, Deborah; del Carmen Plaza-Calonge, María; Rodríguez-Manzaneque, Juan C; Menendez, Pablo

    2012-01-01

    The MLL-AF4 fusion gene is a hallmark genomic aberration in high-risk acute lymphoblastic leukemia in infants. Although it is well established that MLL-AF4 arises prenatally during human development, its effects on hematopoietic development in utero remain unexplored. We have created a human-specific cellular system to study early hemato-endothelial development in MLL-AF4-expressing human embryonic stem cells (hESCs). Functional studies, clonal analysis and gene expression profiling reveal that expression of MLL-AF4 in hESCs has a phenotypic, functional and gene expression impact. MLL-AF4 acts as a global transcriptional activator and a positive regulator of homeobox gene expression in hESCs. Functionally, MLL-AF4 enhances the specification of hemogenic precursors from hESCs but strongly impairs further hematopoietic commitment in favor of an endothelial cell fate. MLL-AF4 hESCs are transcriptionally primed to differentiate towards hemogenic precursors prone to endothelial maturation, as reflected by the marked upregulation of master genes associated to vascular-endothelial functions and early hematopoiesis. Furthermore, we report that MLL-AF4 expression is not sufficient to transform hESC-derived hematopoietic cells. This work illustrates how hESCs may provide unique insights into human development and further our understanding of how leukemic fusion genes, known to arise prenatally, regulate human embryonic hematopoietic specification. PMID:22212479

  9. Post-entrapment genome engineering: First exon size does not affect the expression of fusion transcripts generated by gene entrapment

    PubMed Central

    Osipovich, Anna B.; Singh, Aparna; Ruley, H. Earl

    2005-01-01

    Gene trap mutagenesis in mouse embryonic stem cells has been widely used for genome-wide studies of mammalian gene function. However, while large numbers of genes can be disrupted, individual mutations may suffer from limitations due to the structure and/or placement of targeting vector. To extend the utility of gene trap mutagenesis, replaceable 3′ [or poly(A)] gene trap vectors were developed that permit sequences inserted in individual entrapment clones to be engineered by Cre-mediated recombination. 3′ traps incorporating different drug resistance genes could be readily exchanged, simply by selecting for the drug-resistance gene of the replacement vector. By substituting different 3′ traps, we show that otherwise identical fusion genes containing a large first exon (804 nt) are not expressed at appreciably lower levels than genes expressing small first exons (384 and 151 nt). Thus, size appears to have less effect on the expression and processing of first exons than has been reported for internal exons. Finally, a retroviral poly(A) trap (consisting of a RNA polymerase II promoter, a neomycin-resistance gene, and 5′-splice site) typically produced mutagenized clones in which vector sequences spliced to the 3′-terminal exons of cellular transcription units, suggesting strong selection for fusion transcripts that evade nonsense-mediated decay. The efficient exchange of poly(A) traps should greatly extend the utility of mutant libraries generated by gene entrapment and provides new strategies to study the rules that govern the expression of exons inserted throughout the genome. PMID:15741512

  10. Nuclear fusion occurs during mating in Candida albicans and is dependent on the KAR3 gene.

    PubMed

    Bennett, Richard J; Miller, Mathew G; Chua, Penelope R; Maxon, Mary E; Johnson, Alexander D

    2005-02-01

    It is now well established that mating can occur between diploid a and alpha cells of Candida albicans. There is, however, controversy over when, and with what efficiency, nuclear fusion follows cell fusion to create stable tetraploid a/alpha cells. In this study, we have analysed the mating process between C. albicans strains using both cytological and genetic approaches. Using strains derived from SC5314, we used a number of techniques, including time-lapse microscopy, to demonstrate that efficient nuclear fusion occurs in the zygote before formation of the first daughter cell. Consistent with these observations, zygotes micromanipulated from mating mixes gave rise to mononuclear tetraploid cells, even when no selection for successful mating was applied to them. Mating between different clinical isolates of C. albicans revealed that while all isolates could undergo nuclear fusion, the efficiency of nuclear fusion varied in different crosses. We also show that nuclear fusion in C. albicans requires the Kar3 microtubule motor protein. Deletion of the CaKAR3 gene from both mating partners had little or no effect on zygote formation but reduced the formation of stable tetraploids more than 600-fold, as determined by quantitative mating assays. These findings demonstrate that nuclear fusion is an active process that can occur in C. albicans at high frequency to produce stable, mononucleate mating products.

  11. The distribution of BRAF gene fusions in solid tumors and response to targeted therapy.

    PubMed

    Ross, Jeffrey S; Wang, Kai; Chmielecki, Juliann; Gay, Laurie; Johnson, Adrienne; Chudnovsky, Jacob; Yelensky, Roman; Lipson, Doron; Ali, Siraj M; Elvin, Julia A; Vergilio, Jo-Anne; Roels, Steven; Miller, Vincent A; Nakamura, Brooke N; Gray, Adam; Wong, Michael K; Stephens, Philip J

    2016-02-15

    Although the BRAF V600E base substitution is an approved target for the BRAF inhibitors in melanoma, BRAF gene fusions have not been investigated as anticancer drug targets. In our study, a wide variety of tumors underwent comprehensive genomic profiling for hundreds of known cancer genes using the FoundationOne™ or FoundationOne Heme™ comprehensive genomic profiling assays. BRAF fusions involving the intact in-frame BRAF kinase domain were observed in 55 (0.3%) of 20,573 tumors, across 12 distinct tumor types, including 20 novel BRAF fusions. These comprised 29 unique 5' fusion partners, of which 31% (9) were known and 69% (20) were novel. BRAF fusions included 3% (14/531) of melanomas; 2% (15/701) of gliomas; 1.0% (3/294) of thyroid cancers; 0.3% (3/1,062) pancreatic carcinomas; 0.2% (8/4,013) nonsmall-cell lung cancers and 0.2% (4/2,154) of colorectal cancers, and were enriched in pilocytic (30%) vs. nonpilocytic gliomas (1%; p < 0.0001), Spitzoid (75%) vs. nonSpitzoid melanomas (1%; p = 0.0001), acinar (67%) vs. nonacinar pancreatic cancers (<1%; p < 0.0001) and papillary (3%) vs. nonpapillary thyroid cancers (0%; p < 0.03). Clinical responses to trametinib and sorafenib are presented. In conclusion, BRAF fusions are rare driver alterations in a wide variety of malignant neoplasms, but enriched in Spitzoid melanoma, pilocytic astrocytomas, pancreatic acinar and papillary thyroid cancers.

  12. Fusion technology development. Annual report, October 1, 1994--September 30, 1995

    SciTech Connect

    1996-08-01

    In FY95, the General Atomics (GA) Fusion Group made significant contributions to the technology needs of the magnetic fusion program. The work is reported in the following sections on Fusion Power Plant Studies (Section 2), DiMES (Section 3), SiC Composite Studies (Section 4), Magnetic Probe (Section 5) and RF Technology (Section 6). Meetings attended and publications are listed in their respective sections. The overall objective of GA`s fusion technology research is to develop the technologies necessary for fusion to move successfully from present-day physics experiments to ITER and other next-generation fusion experiments, and ultimately to fusion power plants. To achieve this overall objective, they carry out fusion systems design studies to evaluate the technologies needed for next-step experiments and power plants, and they conduct research to develop basic knowledge about these technologies, including plasma technologies, fusion nuclear technologies, and fusion materials. They continue to be committed to the development of fusion power and its commercialization by US industry.

  13. Fusion technology development annual report, October 1, 1995--September 30, 1996

    SciTech Connect

    1997-03-01

    In FY96, the General Atomics (GA) Fusion Group made significant contributions to the technology needs of the magnetic fusion program. The work is reported in the following sections on Fusion Power Plant Design Studies (Section 2), Plasma Interactive Materials (Section 3), SiC/SiC Composite Material Development (Section 4), Magnetic Diagnostic Probes (Section 5) and RF Technology (Section 6). Meetings attended and publications are listed in their respective sections. The overall objective of GA`s fusion technology research is to develop the technologies necessary for fusion to move successfully from present-day physics experiments to ITER and other next-generation fusion experiments, and ultimately to fusion power plants. To achieve this overall objective, the authors carry out fusion systems design studies to evaluate the technologies needed for next-step experiments and power plants, and they conduct research to develop basic knowledge about these technologies, including plasma technologies, fusion nuclear technologies, and fusion materials. They continue to be committed to the development of fusion power and its commercialization by US industry.

  14. mPlum-IFP 1.4 fluorescent fusion protein may display Förster resonance energy transfer associated properties that can be used for near-infrared based reporter gene imaging.

    PubMed

    Lin, Liang-Ting; Wang, Bo-Sheng; Chen, Jyh-Cheng; Liu, Chi-Hsien; Chou, Chien; Chiu, Shu-Jun; Chang, Wen-Yi; Liu, Ren-Shyan; Allen Chang, C; Lee, Yi-Jang

    2013-12-01

    Bacteriophytochrome infrared fluorescent protein (IFP) has a long emission wavelength that is appropriate for detecting pathophysiological effects via near-infrared (NIR) based imaging. However, the brightness and photostability of IFP are suboptimal, although an exogenous supply of biliverdin (BV) IXα is able to enhance these properties. In this study, we fused a far red mPlum fluorescent protein to IFP 1.4 via a linker deoxyribonucleic acid (DNA) sequence encoding eight amino acids. The brightness of mPlum-IFP 1.4 fusion protein at the IFP emission channel was comparable to that of native IFP 1.4 protein when fusion protein and IFP 1.4 were excited by 543 and 633 nm using confocal microscopy, respectively. Visualization of IFP 1.4 fluorescence by excitation of mPlum in mPlum-IFP 1.4 fusion protein is likely to be associated with Förster resonance energy transfer (FRET). The FRET phenomenon was also predicted by acceptor photobleaching using confocal microscopy. Furthermore, the expression of mPlum-IFP 1.4 fusion protein could be detected in cell culture and in xenograft tumors in the absence of BV using in vivo imaging system, although the BV was still essential for detecting native IFP 1.4. Therefore, this innovative-fluorescent fusion protein would be useful for NIR-based imaging in vitro and in vivo.

  15. Spatiotemporal Changes of Calcitonin Gene-Related Peptide Innervation in Spinal Fusion

    PubMed Central

    Zhou, Xiao-Yi; Xu, Xi-Ming; Wu, Sui-Yi; Wang, Fei; Yang, Yi-Lin; Li, Ming

    2016-01-01

    Few studies have investigated the role calcitonin gene-related peptide (CGRP) plays in the process of spinal fusion. The aim of the present study is to observe the temporal and spatial changes of CGRP induced by experimental fusion surgery in rats and elucidate the role of CGRP in spinal fusion. Male Sprague-Dawley rats were used in the study and the specimens were collected on the 7th, 14th, 21st, and 28th day, respectively. Then, histological and immunohistochemical analysis were applied to evaluate the fusion mass and spatiotemporal changes of CGRP chronologically. The results demonstrated that density of CGRP reached peak on the 21st day after surgery and most of the CGRP expression located surrounding the interface of allograft and fibrous tissue where the cells differentiate into osteoblasts, indicating that CGRP might be involved in the process of bone formation and absorption. PMID:27990431

  16. NAB2-STAT6 fusion gene analysis in two cases of meningeal solitary fibrous tumor/hemangiopericytoma with late distant metastases.

    PubMed

    Nakada, Satoko; Minato, Hiroshi; Takegami, Tsutomu; Kurose, Nozomu; Ikeda, Hiroko; Kobayashi, Masako; Sasagawa, Yasuo; Akai, Takuya; Kato, Takashi; Yamamoto, Norio; Nojima, Takayuki

    2015-10-01

    We present two cases of meningeal solitary fibrous tumor (SFT)/hemangiopericytoma (HPC) with immunohistochemistry of STAT6 and analysis of NAB2-STAT6 fusion genes. Case 1 was a 37-year-old male with a left middle fossa tumor; case 2 was a 68-year-old female with a cerebellar tumor. They showed late metastasis to the lung or bone 8 or 13 years, respectively, after the first surgery. Histology of both primary and metastatic tumors showed a cellular hemangiopericytomatous pattern with nuclear atypia. The primary tumors showed nuclear staining of STAT6, but both metastatic tumors showed nuclear and cytoplasmic STAT6. DNA sequencing revealed two kinds of NAB2-STAT6 fusion genes. One consisted of exon 6 of NAB2, intron 6 of NAB2, and the middle of exon 17 of STAT6 (observed in the primary and metastatic tumors of case 1); the other consisted of exon 6 of NAB2 and the beginning of exon 17 of STAT6 (observed in the metastatic tumor of case 2). The primary tumor of case 2 had both fusion genes. To the best of our knowledge, we are the first to report NAB2-STAT6 fusion gene analysis in primary and metastatic meningeal SFT/HPCs and a case showed different fusion gene status in the metastatic tumor.

  17. Cold fusion verification. Final report for period ending 1989

    SciTech Connect

    North, M.H.; Mastny, G.F.; Wesley, E.J.

    1991-03-01

    The objective of this work to verify and reproduce experimental observations of Cold Nuclear Fusion (CNF), as originally reported in 1989. The method was to start with the original report and add such additional information as became available to build a set of operational electrolytic CNF cells. Verification was to be achieved by first observing cells for neutron production, and for those cells that demonstrated a nuclear effect, careful calorimetric measurements were planned. The authors concluded, after laboratory experience, reading published work, talking with others in the field, and attending conferences, that CNF probably is chimera and will go the way of N-rays and polywater. The neutron detector used for these tests was a completely packaged unit built into a metal suitcase that afforded electrostatic shielding for the detectors and self-contained electronics. It was battery-powered, although it was on charge for most of the long tests. The sensor element consists of He detectors arranged in three independent layers in a solid moderating block. The count from each of the three layers as well as the sum of all the detectors were brought out and recorded separately. The neutron measurements were made with both the neutron detector and the sample tested in a cave made of thick moderating material that surrounded the two units on the sides and bottom.

  18. Inertial confinement fusion. Quarterly report, July--September 1993: Volume 3, No. 4

    SciTech Connect

    Sacks, R.A.; Murphy, P.W.; Schleich, D.P.

    1993-12-31

    This report discusses the following research: Diode-pumped solid- state-laser driver for inertial fusion energy power plants; Longitudinal beam dynamics in heavy ion fusion accelerators; Design of the ion sources for heavy ion fusion; Measurement of electron density in laser-produced plasma with a soft x-ray moire deflectometer; and Analysis of weakly nonlinear three-dimensional Rayleigh-Taylor instability growth.

  19. RELATIVE EXPRESSION AND STABILITY OF A CHROMOSOMALLY INTEGRATED AND PLASMID-BORNE MARKER GENE FUSION IN ENVIRONMENTALLY COMPETENT BACTERIA

    EPA Science Inventory

    A xyIE-iceC transcriptional fusion was created by ligating a DNA fragment harboring the cloned xyIE structural gene from the TOL plasmid of Pseudomonas putida mt-2 into the cloned iceC gene of Pseudomonas syringae Cit7. This fusion construct was integrated into chromosome of Pseu...

  20. ChimerDB 3.0: an enhanced database for fusion genes from cancer transcriptome and literature data mining.

    PubMed

    Lee, Myunggyo; Lee, Kyubum; Yu, Namhee; Jang, Insu; Choi, Ikjung; Kim, Pora; Jang, Ye Eun; Kim, Byounggun; Kim, Sunkyu; Lee, Byungwook; Kang, Jaewoo; Lee, Sanghyuk

    2017-01-04

    Fusion gene is an important class of therapeutic targets and prognostic markers in cancer. ChimerDB is a comprehensive database of fusion genes encompassing analysis of deep sequencing data and manual curations. In this update, the database coverage was enhanced considerably by adding two new modules of The Cancer Genome Atlas (TCGA) RNA-Seq analysis and PubMed abstract mining. ChimerDB 3.0 is composed of three modules of ChimerKB, ChimerPub and ChimerSeq. ChimerKB represents a knowledgebase including 1066 fusion genes with manual curation that were compiled from public resources of fusion genes with experimental evidences. ChimerPub includes 2767 fusion genes obtained from text mining of PubMed abstracts. ChimerSeq module is designed to archive the fusion candidates from deep sequencing data. Importantly, we have analyzed RNA-Seq data of the TCGA project covering 4569 patients in 23 cancer types using two reliable programs of FusionScan and TopHat-Fusion. The new user interface supports diverse search options and graphic representation of fusion gene structure. ChimerDB 3.0 is available at http://ercsb.ewha.ac.kr/fusiongene/.

  1. ChimerDB 3.0: an enhanced database for fusion genes from cancer transcriptome and literature data mining

    PubMed Central

    Lee, Myunggyo; Lee, Kyubum; Yu, Namhee; Jang, Insu; Choi, Ikjung; Kim, Pora; Jang, Ye Eun; Kim, Byounggun; Kim, Sunkyu; Lee, Byungwook; Kang, Jaewoo; Lee, Sanghyuk

    2017-01-01

    Fusion gene is an important class of therapeutic targets and prognostic markers in cancer. ChimerDB is a comprehensive database of fusion genes encompassing analysis of deep sequencing data and manual curations. In this update, the database coverage was enhanced considerably by adding two new modules of The Cancer Genome Atlas (TCGA) RNA-Seq analysis and PubMed abstract mining. ChimerDB 3.0 is composed of three modules of ChimerKB, ChimerPub and ChimerSeq. ChimerKB represents a knowledgebase including 1066 fusion genes with manual curation that were compiled from public resources of fusion genes with experimental evidences. ChimerPub includes 2767 fusion genes obtained from text mining of PubMed abstracts. ChimerSeq module is designed to archive the fusion candidates from deep sequencing data. Importantly, we have analyzed RNA-Seq data of the TCGA project covering 4569 patients in 23 cancer types using two reliable programs of FusionScan and TopHat-Fusion. The new user interface supports diverse search options and graphic representation of fusion gene structure. ChimerDB 3.0 is available at http://ercsb.ewha.ac.kr/fusiongene/. PMID:27899563

  2. Gene fusion vehicles for the analysis of gene expression in Rhizobium meliloti.

    PubMed Central

    Kahn, M L; Timblin, C R

    1984-01-01

    A set of plasmid cloning vehicles was developed to facilitate the construction of gene or operon fusions in Rhizobium meliloti. The vehicles also contain a broad-host-range replicon and could be introduced into bacteria either by transformation or by transduction, using bacteriophage P2. Insertion of foreign DNA into a unique restriction endonuclease cleavage site promotes the synthesis of either the Escherichia coli lactose operon or the kanamycin phosphotransferase gene from transposon Tn5. Expression of the lactose operon could be detected by observing the color of Rhizobium colonies on medium that contained a chromogenic indicator. We also determined the growth conditions that make it possible to select either for or against the expression of the E. coli lactose operon in R. meliloti. Recombinant plasmids were constructed by inserting MboI restriction fragments of R. meliloti DNA into one of the vehicles, pMK353 . Expression of beta-galactosidase by a number of these recombinants was measured in both R. meliloti and E. coli. PMID:6327625

  3. Prostate cancer of transition zone origin lacks TMPRSS2-ERG gene fusion.

    PubMed

    Guo, Charles C; Zuo, Geyan; Cao, Dongdong; Troncoso, Patricia; Czerniak, Bogdan A

    2009-07-01

    Recent studies have shown a unique chromosomal rearrangement that leads to the fusion of 5'-transmembrane protein serine proteinase-2 (TMPRSS2) with the EST-related gene (ERG) in prostate cancer. In this study, we used fluorescence in situ hybridization to evaluate TMPRSS2-ERG gene fusion in prostate cancer of different zonal origins. Radical prostatectomy specimens with multifocal prostate cancer were obtained from 30 patients who were treated at our institution. Two separate tumor foci in each specimen, one in the peripheral zone and the other in the transition zone, were selected for gene fusion analysis. The selected peripheral zone tumor foci had a mean Gleason score of 6.8 (range, 6-7) and a mean tumor volume of 1.2 cm(3) (range, 0.1-4.6 cm(3)). The selected transition zone tumor foci had a mean Gleason score of 6.7 (range, 5-8) and a mean tumor volume of 4.0 cm(3) (range, 0.5-9.0 cm(3)). ERG gene rearrangement was not observed in any transition zone tumors; however, it was found in the peripheral zone tumors in 13 cases (43%). In 10 cases, the rearrangement was associated with the deletion of the 5'-end of ERG. In conclusion, we found that TMPRSS2-ERG gene fusion is associated with the zonal origin of prostate cancer. This gene fusion is prevalent in prostate cancer arising from the peripheral zone, but is lacking in prostate cancer arising from the transition zone.

  4. TEL/AML-1 fusion gene. its frequency and prognostic significance in childhood acute lymphoblastic leukemia.

    PubMed

    Jamil, A; Theil, K S; Kahwash, S; Ruymann, F B; Klopfenstein, K J

    2000-10-15

    TEL gene rearrangement due to the 12;21 chromosome translocation is believed to be the most common molecular genetic abnormality in childhood acute lymphoblastic leukemia (ALL). A study was conducted to investigate the frequency and prognostic significance of TEL/AML-1 fusion gene resulting from a cryptic t(12;21)(p13;q22). Bone marrow samples from 86 patients diagnosed over the past 5 years at Columbus Children's Hospital were analyzed by fluorescence in situ hybridization (FISH) technique for TEL/AML-1 fusion gene, using LSI((R)) DNA probes. The positive cases were analyzed for clinical outcome. Patients in this study received treatment according to Children's Cancer Group (CCG) protocols. Fifteen of the 86 cases (17%) were positive for the fusion gene. All were B-cell lineage and except for one, all were CD10 positive. TEL/AML-1 was not found in any T-cell ALL. The mean overall survival (OS) following diagnosis for the TEL/AML-1-positive group was significantly longer than for the TEL/AML-1-negative group by log-rank = 7.84, P = 0.005. Similarly, the event-free survival (EFS) after remission for the positive group (median 94.5 months) was longer than the negative group (median 57 months) by log-rank = 7.19, P = 0.007. This study confirms that the TEL/AML-1 fusion gene may be the most common genetic event in childhood ALL, occurring in 17% of the patients. It appears restricted to the B-cell lineage. In this study, the presence of a TEL/AML-1 fusion gene was statistically significant in predicting both OS and EFS, indicating a favorable clinical outcome for these patients. Screening for TEL/AML-1 should become routine at diagnosis and a useful biological variable for risk stratification in future clinical trials.

  5. Design and Characterization of Novel Recombinant Listeriolysin O–Protamine Fusion Proteins for Enhanced Gene Delivery

    PubMed Central

    2015-01-01

    To improve the efficiency of gene delivery for effective gene therapy, it is essential that the vector carries functional components that can promote overcoming barriers in various steps leading to the transport of DNA from extracellular to ultimately nuclear compartment. In this study, we designed genetically engineered fusion proteins as a platform to incorporate multiple functionalities in one chimeric protein. Prototypes of such a chimera tested here contain two domains: one that binds to DNA; the other that can facilitate endosomal escape of DNA. The fusion proteins are composed of listeriolysin O (LLO), the endosomolytic pore-forming protein from Listeria monocytogenes, and a 22 amino acid sequence of the DNA-condensing polypeptide protamine (PN), singly or as a pair: LLO-PN and LLO-PNPN. We demonstrate dramatic enhancement of the gene delivery efficiency of protamine-condensed DNA upon incorporation of a small amount of LLO-PN fusion protein and further improvement with LLO-PNPN in vitro using cultured cells. Additionally, the association of anionic liposomes with cationic LLO-PNPN/protamine/DNA complexes, yielding a net negative surface charge, resulted in better in vitro transfection efficiency in the presence of serum. An initial, small set of data in mice indicated that the observed enhancement in gene expression could also be applicable to in vivo gene delivery. This study suggests that incorporation of a recombinant fusion protein with multiple functional components, such as LLO–protamine fusion protein, in a nonviral vector is a promising strategy for various nonviral gene delivery systems. PMID:25521817

  6. Establishment of cells to monitor Microprocessor through fusion genes of microRNA and GFP.

    PubMed

    Tsutsui, Motomu; Hasegawa, Hitoki; Adachi, Koichi; Miyata, Maiko; Huang, Peng; Ishiguro, Naoki; Hamaguchi, Michinari; Iwamoto, Takashi

    2008-08-08

    Microprocessor, the complex of Drosha and DGCR8, promotes the processing of primary microRNA to precursor microRNA, which is a crucial step for microRNA maturation. So far, no convenient assay systems have been developed for observing this step in vivo. Here we report the establishment of highly sensitive cellular systems where we can visually monitor the function of Microprocessor. During a series of screening of transfectants with fusion genes of the EGFP cDNA and primary microRNA genes, we have obtained certain cell lines where introduction of siRNA against DGCR8 or Drosha strikingly augments GFP signals. In contrast, these cells have not responded to Dicer siRNA; thus they have a unique character that GFP signals should be negatively and specifically correlated to the action of Microprocessor among biogenesis of microRNA. These cell lines can be useful tools for real-time analysis of Microprocessor action in vivo and identifying its novel modulators.

  7. Massachusetts Institute of Technology Plasma Fusion Center 1992--1993 report to the President

    SciTech Connect

    Not Available

    1993-07-01

    This report discusses research being conducted at MIT`s plasma fusion center. Some of the areas covered are: plasma diagnostics; rf plasma heating; gyrotron research; treatment of solid waste by arc plasma; divertor experiments; tokamak studies; and plasma and fusion theory.

  8. Particle-beam-fusion progress report, July 1979 through December 1979

    SciTech Connect

    Not Available

    1981-01-01

    The following chapters are included in this semi-annual progress report: (1) fusion target studies, (2) target experiments, (3) particle-beam source developments, (4) particle beam experiments, (5) pulsed power, (6) pulsed power applications, and (7) electron beam fusion accelerator project. (MOW)

  9. Fusion Energy Division annual progress report period ending December 31, 1986

    SciTech Connect

    Morgan, O.B. Jr.; Berry, L.A.; Sheffield, J.

    1987-10-01

    This annual report on fusion energy discusses the progress on work in the following main topics: toroidal confinement experiments; atomic physics and plasma diagnostics development; plasma theory and computing; plasma-materials interactions; plasma technology; superconducting magnet development; fusion engineering design center; materials research and development; and neutron transport. (LSP)

  10. The brain expressed x-linked gene 1 (Bex1) regulates myoblast fusion

    PubMed Central

    Yue, Feng; Kuang, Shihuan

    2015-01-01

    Skeletal muscle development (myogenesis) is a complex but precisely orchestrated process involving spatiotemporal regulation of the proliferation, differentiation and fusion of myogenic progenitor cells (myoblasts). Here we identify brain expressed x-linked gene 1 (Bex1) as a transient, developmentally regulated gene involved in myoblast fusion. Bex1 expression is undetectable in adult muscles or in quiescent muscle stem cells (satellite cells). During embryonic myogenesis, however, Bex1 is robustly expressed by myogenin+ differentiating myoblasts, but not by Pax7+ proliferating myoblasts. Interestingly, Bex1 is initially localized in the cytoplasm and then translocates into the nucleus. During adult muscle regeneration, Bex1 is highly expressed in newly regenerated myofibers and the expression is rapidly downregulated during maturation. Consistently, in cultured myoblasts, Bex1 is not expressed at the proliferation stage but transiently expressed upon induction of myogenic differentiation, following a similar cytoplasm to nucleus translocation pattern as seen in vivo. Using gain- and loss-of-function studies, we found that overexpression of Bex1 promotes the fusion of primary myoblasts without affecting myogenic differentiation and myogenin expression. Conversely, Bex1 knockout myoblasts exhibit obvious fusion defects, even though they express normal levels of myogenin and differentiate normally. These results elucidate a novel role of Bex1 in myogenesis through regulating myoblast fusion. PMID:26586200

  11. Characterization of the genomic features and expressed fusion genes in micropapillary carcinomas of the breast

    PubMed Central

    Natrajan, Rachael; Wilkerson, Paul M; Marchiò, Caterina; Piscuoglio, Salvatore; Ng, Charlotte KY; Wai, Patty; Lambros, Maryou B; Samartzis, Eleftherios P; Dedes, Konstantin J; Frankum, Jessica; Bajrami, Ilirjana; Kopec, Alicja; Mackay, Alan; A'hern, Roger; Fenwick, Kerry; Kozarewa, Iwanka; Hakas, Jarle; Mitsopoulos, Costas; Hardisson, David; Lord, Christopher J; Kumar-Sinha, Chandan; Ashworth, Alan; Weigelt, Britta; Sapino, Anna; Chinnaiyan, Arul M; Maher, Christopher A; Reis-Filho, Jorge S

    2014-01-01

    Micropapillary carcinoma (MPC) is a rare histological special type of breast cancer, characterized by an aggressive clinical behaviour and a pattern of copy number aberrations (CNAs) distinct from that of grade- and oestrogen receptor (ER)-matched invasive carcinomas of no special type (IC-NSTs). The aims of this study were to determine whether MPCs are underpinned by a recurrent fusion gene(s) or mutations in 273 genes recurrently mutated in breast cancer. Sixteen MPCs were subjected to microarray-based comparative genomic hybridization (aCGH) analysis and Sequenom OncoCarta mutation analysis. Eight and five MPCs were subjected to targeted capture and RNA sequencing, respectively. aCGH analysis confirmed our previous observations about the repertoire of CNAs of MPCs. Sequencing analysis revealed a spectrum of mutations similar to those of luminal B IC-NSTs, and recurrent mutations affecting mitogen-activated protein kinase family genes and NBPF10. RNA-sequencing analysis identified 17 high-confidence fusion genes, eight of which were validated and two of which were in-frame. No recurrent fusions were identified in an independent series of MPCs and IC-NSTs. Forced expression of in-frame fusion genes (SLC2A1–FAF1 and BCAS4–AURKA) resulted in increased viability of breast cancer cells. In addition, genomic disruption of CDK12 caused by out-of-frame rearrangements was found in one MPC and in 13% of HER2-positive breast cancers, identified through a re-analysis of publicly available massively parallel sequencing data. In vitro analyses revealed that CDK12 gene disruption results in sensitivity to PARP inhibition, and forced expression of wild-type CDK12 in a CDK12-null cell line model resulted in relative resistance to PARP inhibition. Our findings demonstrate that MPCs are neither defined by highly recurrent mutations in the 273 genes tested, nor underpinned by a recurrent fusion gene. Although seemingly private genetic events, some of the fusion transcripts found

  12. A gene fusion at a homeobox locus: alterations in leaf shape and implications for morphological evolution.

    PubMed Central

    Chen, J J; Janssen, B J; Williams, A; Sinha, N

    1997-01-01

    Compound leaves are seen in many angiosperm genera and are thought to be either fundamentally different from simple leaves or elaborations of simple leaves. The knotted1-like homeobox (knox) genes are known to regulate plant development. When overexpressed in homologous or heterologous species, this family of genes can cause changes in leaf morphology, including excessive leaf compounding in tomato. We describe here an instance of a spontaneously arisen fusion between a gene encoding a metabolic enzyme and a homeodomain protein. We show that the fusion results in overexpression of the homeodomain protein and a change in morphology that approximates the changes caused by overexpression of the same gene under the control of the cauliflower mosaic virus 35S promoter in transgenic plants. Exon-shuffling events can account for the modularity of proteins. If the shuffled exons are associated with altered promoters, changes in gene expression patterns can result. Our results show that gene fusions of this nature can cause changes in expression patterns that lead to altered morphology. We suggest that such phenomena may have played a role in the evolution of form. PMID:9286107

  13. Integrated genomic analyses identify frequent gene fusion events and VHL inactivation in gastrointestinal stromal tumors

    PubMed Central

    Sun, Choong-Hyun; Park, Inho; Lee, Seungmook; Kwon, Jekeun; Do, Ingu; Hong, Min Eui; Van Vrancken, Michael; Lee, Jeeyun; Park, Joon Oh; Cho, Jeonghee; Kim, Kyoung-Mee; Sohn, Tae Sung

    2016-01-01

    Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. We sequenced nine exomes and transcriptomes, and two genomes of GISTs for integrated analyses. We detected 306 somatic variants in nine GISTs and recurrent protein-altering mutations in 29 genes. Transcriptome sequencing revealed 328 gene fusions, and the most frequently involved fusion events were associated with IGF2 fused to several partner genes including CCND1, FUS, and LASP1. We additionally identified three recurrent read-through fusion transcripts: POLA2-CDC42EP2, C8orf42-FBXO25, and STX16-NPEPL1. Notably, we found intragenic deletions in one of three exons of the VHL gene and increased mRNAs of VEGF, PDGF-β, and IGF-1/2 in 56% of GISTs, suggesting a mechanistic link between VHL inactivation and overexpression of hypoxia-inducible factor target genes in the absence of hypoxia. We also identified copy number gain and increased mRNA expression of AMACR, CRIM1, SKP2, and CACNA1E. Mapping of copy number and gene expression results to the KEGG pathways revealed activation of the JAK-STAT pathway in small intestinal GISTs and the MAPK pathway in wild-type GISTs. These observations will allow us to determine the genetic basis of GISTs and will facilitate further investigation to develop new therapeutic options. PMID:25987131

  14. Integrated genomic analyses identify frequent gene fusion events and VHL inactivation in gastrointestinal stromal tumors.

    PubMed

    Kang, Guhyun; Yun, Hongseok; Sun, Choong-Hyun; Park, Inho; Lee, Seungmook; Kwon, Jekeun; Do, Ingu; Hong, Min Eui; Van Vrancken, Michael; Lee, Jeeyun; Park, Joon Oh; Cho, Jeonghee; Kim, Kyoung-Mee; Sohn, Tae Sung

    2016-02-09

    Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. We sequenced nine exomes and transcriptomes, and two genomes of GISTs for integrated analyses. We detected 306 somatic variants in nine GISTs and recurrent protein-altering mutations in 29 genes. Transcriptome sequencing revealed 328 gene fusions, and the most frequently involved fusion events were associated with IGF2 fused to several partner genes including CCND1, FUS, and LASP1. We additionally identified three recurrent read-through fusion transcripts: POLA2-CDC42EP2, C8orf42-FBXO25, and STX16-NPEPL1. Notably, we found intragenic deletions in one of three exons of the VHL gene and increased mRNAs of VEGF, PDGF-β, and IGF-1/2 in 56% of GISTs, suggesting a mechanistic link between VHL inactivation and overexpression of hypoxia-inducible factor target genes in the absence of hypoxia. We also identified copy number gain and increased mRNA expression of AMACR, CRIM1, SKP2, and CACNA1E. Mapping of copy number and gene expression results to the KEGG pathways revealed activation of the JAK-STAT pathway in small intestinal GISTs and the MAPK pathway in wild-type GISTs. These observations will allow us to determine the genetic basis of GISTs and will facilitate further investigation to develop new therapeutic options.

  15. TMPRSS2-ERG Fusion Gene Expression in Prostate Tumor Cells and Its Clinical and Biological Significance in Prostate Cancer Progression

    PubMed Central

    St. John, Jason; Powell, Katelyn; Conley-LaComb, M. Katie; Chinni, Sreenivasa R.

    2012-01-01

    TMPRSS2-Ets gene fusions were identified in prostate cancers where the promoter of transmembrane protease, serine 2 (TMPRSS2) fused with coding sequence of the erythroblastosis virus E26 (Ets) gene family members. TMPRSS2 is an androgen responsive transmembrane serine protease. Ets family members are oncogenic transcription factors that contain a highly conserved Ets DNA binding domain and an N-terminal regulatory domain. Fusion of these gene results in androgen dependent transcription of Ets factor in prostate tumor cells. The ERG is the most common fusion partner with TMPRSS2 promoter in prostate cancer patients. The high prevalence of these gene fusions, in particular TMPRSS2-ERG, makes them attractive as potential diagnostic and prognostic indicators, as well as making them a potential target for tailored therapies. This review focuses on the clinical and biological significance of TMPRSS2-ERG fusions and their role in PC development and progression. PMID:23264855

  16. Acute leukemias of different lineages have similar MLL gene fusions encoding related chimeric proteins resulting from chromosomal translocation

    SciTech Connect

    Corral, J.; Forster, A.; Thompson, S.; Rabbitts, T.H. ); Lampert, F. ); Kaneko, Y. ); Slater, R.; Kroes, W.G. ); Van Der Schoot, C.E. ); Ludwig, W.D. ); Karpas, A. ); Pocock, C.; Cotter, F. )

    1993-09-15

    The MLL gene, on human chromosome 11q23, undergoes chromosomal translocation in acute leukemias, resulting in gene fusion with AF4 (chromosome 4) and ENL (chromosome 19). The authors report here translocation of MLL with nine different chromosomes and two paracentric chromosome 11 deletions in early B cell, B- or T-cell lineage, or nonlymphocytic acute leukemias. The mRNA translocation junction from 22t(4;11) patients, including six adult leukemias, and nine t(11;19) tumors reveals a remarkable conservation of breakpoints within MLL, AF4, or ENL genes, irrespective of tumor phenotype. Typically, the breakpoints are upstream of the zinc-finger region of MLL, and deletion of this region can accompany translocation, supporting the der(11) chromosome as the important component in leukemogenesis. Partial sequence of a fusion between MLL and the AFX1 gene from chromosome X shows the latter to be rich in Ser/Pro codons, like the ENL mRNA. These data suggest that the heterogeneous 11q23 abnormalities might cause attachment of Ser/Pro-rich segments to the NH[sub 2] terminus of MLL, lacking the zinc-finger region, and that translocation occurs in early hematopoietic cells, before commitment to distinct lineages. 36 refs., 2 figs.

  17. Functional characterization of the 5' flanking region of human ubiquitin fusion degradation 1 like gene (UFD1L).

    PubMed

    Amati, Francesca; Conti, Emanuela; Botta, Annalisa; Amicucci, Paola; Dallapiccola, Bruno; Novelli, Giuseppe

    2002-06-01

    UFD1L (Ubiquitin Fusion Degradation 1 Like) gene encodes for a component of a multi-complex involved in the degradation of ubiquitin fusion proteins. The gene maps on chromosome 22q11, in a region commonly deleted in severe congenital disorders such as DiGeorge (DGS) and velo-cardio-facial (VCFS) syndromes. UFD1L is a single copy gene ubiquitously expressed in high levels in the pharyngeal pouches and fourth branchial arch artery during development. To understand the regulation of UFD1L expression we performed a functional analysis of its 5' regulatory region. 5'-RACE and primer extension analyses revealed the presence of different transcription start sites in adult and fetal tissues. UFD1L 5' flanking region contains a TATA-box motif and is also very GC-rich with a CpG island encompassing exon 1. Transcriptional activity of this region was examined by transfection experiments of promoter-GFP reporter gene constructs in a human epithelial cell line. These experiments revealed the importance of the region between -17 and -463 nt which contains the TATA-box. EMSA assay resulted in the detection of five functional consensus sequences respectively for the transcription complex TFIID and for the transcription factors AP-1 (one site), AP-2 (one) and Sp1 (two).

  18. Fusion reactor materials. Semiannual progress report for period ending September 30, 1993

    SciTech Connect

    Rowcliffe, A.F.; Burn, G.L.; Knee`, S.S.; Dowker, C.L.

    1994-02-01

    This is the fifteenth in a series of semiannual technical progress reports on fusion reactor materials. This report combines research and development activities which were previously reported separately in the following progress reports: Alloy Development for Irradiation Performance; Damage Analysis and Fundamental Studies; Special purpose Materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials programs being conducted in support of the Magnetic Fusion Energy Program of the U.S. Department of Energy. The Fusion Reactor Materials Program is a national effort involving several national laboratories, universities, and industries. The purpose of this series of reports is to provide a working technical record for the use of the program participants, and to provide a means of communicating the efforts of materials scientists to the rest of the fusion community, both nationally and worldwide.

  19. NTRK gene fusions as novel targets of cancer therapy across multiple tumour types

    PubMed Central

    Sartore-Bianchi, Andrea; Siena, Salvatore

    2016-01-01

    The tropomyosin receptor kinase (Trk) receptor family comprises 3 transmembrane proteins referred to as Trk A, B and C (TrkA, TrkB and TrkC) receptors that are encoded by the NTRK1, NTRK2 and NTRK3 genes, respectively. These receptor tyrosine kinases are expressed in human neuronal tissue and play an essential role in the physiology of development and function of the nervous system through activation by neurotrophins. Gene fusions involving NTRK genes lead to transcription of chimeric Trk proteins with constitutively activated or overexpressed kinase function conferring oncogenic potential. These genetic abnormalities have recently emerged as targets for cancer therapy, because novel compounds have been developed that are selective inhibitors of the constitutively active rearranged proteins. Developments in this field are being aided by next generation sequencing methods as tools for unbiased gene fusions discovery. In this article, we review the role of NTRK gene fusions across several tumour histologies, and the promises and challenges of targeting such genetic alterations for cancer therapy. PMID:27843590

  20. Isolation and characterization of Escherichia coli strains containing new gene fusions (soi::lacZ) inducible by superoxide radicals.

    PubMed Central

    Mito, S; Zhang, Q M; Yonei, S

    1993-01-01

    Gene fusions in Escherichia coli that showed increased beta-galactosidase expression in response to treatment with a superoxide radical (O2-) generator, methyl viologen (MV), were obtained. These fusions were constructed by using a Mud(Ap lac) phage to insert the lactose structural genes randomly into the E. coli chromosome. Ampicillin-resistant colonies were screened for increased expression of beta-galactosidase on X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) plates containing MV at 1.25 micrograms/ml. Other O2- generators, menadione and plumbagin, also induced beta-galactosidase activity in these fusion strains. The induction by these drugs occurred only under aerobic conditions. Hyperoxygenation also elicited an induction of the fusions. On the other hand, no significant induction was observed with hydrogen peroxide and cumene hydroperoxide. The induction of these fusions by MV was not dependent on the peroxide stress control mediated by the oxyR gene or on the recA-dependent SOS system. These fusions were named soi (superoxide inducible)::lacZ. The induction of beta-galactosidase was significantly reduced by introducing a soxS::Tn10 locus into the fusion strains, indicating that the soi genes are members of the soxRS regulon. Five of the fusions were located in 6 to 26 min of the E. coli genetic map, while three fusions were located in 26 to 36 min, indicating that these fusions are not related to genes already known to be inducible by O2- under the control of soxRS. At least five mutants containing the soi::lacZ fusion were more sensitive to MV and menadione than the wild-type strain, suggesting that the products of these soi genes play an important role in protection against oxidative stress. PMID:8386722

  1. Gene fusion analysis of membrane protein topology: a direct comparison of alkaline phosphatase and beta-lactamase fusions.

    PubMed Central

    Prinz, W A; Beckwith, J

    1994-01-01

    To compare two approaches to analyzing membrane protein topology, a number of alkaline phosphatase fusions to membrane proteins were converted to beta-lactamase fusions. While some alkaline phosphatase fusions near the N terminus of cytoplasmic loops of membrane proteins have anomalously high levels of activity, the equivalent beta-lactamase fusions do not. This disparity may reflect differences in the folding of beta-lactamase and alkaline phosphatase in the cytoplasm. PMID:7929016

  2. Recurrent MET fusion genes represent a drug target in pediatric glioblastoma.

    PubMed

    2016-11-01

    Pediatric glioblastoma is one of the most common and most deadly brain tumors in childhood. Using an integrative genetic analysis of 53 pediatric glioblastomas and five in vitro model systems, we identified previously unidentified gene fusions involving the MET oncogene in ∼10% of cases. These MET fusions activated mitogen-activated protein kinase (MAPK) signaling and, in cooperation with lesions compromising cell cycle regulation, induced aggressive glial tumors in vivo. MET inhibitors suppressed MET tumor growth in xenograft models. Finally, we treated a pediatric patient bearing a MET-fusion-expressing glioblastoma with the targeted inhibitor crizotinib. This therapy led to substantial tumor shrinkage and associated relief of symptoms, but new treatment-resistant lesions appeared, indicating that combination therapies are likely necessary to achieve a durable clinical response.

  3. Two novel imatinib-responsive PDGFRA fusion genes in chronic eosinophilic leukaemia.

    PubMed

    Curtis, Claire E; Grand, Francis H; Musto, Pellegrino; Clark, Andrew; Murphy, John; Perla, Gianni; Minervini, Maria M; Stewart, Janet; Reiter, Andreas; Cross, Nicholas C P

    2007-07-01

    We identified two patients with a t(2;4)(p24;q12) and a t(4;12)(q2?3;p1?2), respectively, in association with BCR-ABL and FIP1L1-PDGFRA negative chronic eosinophilic leukaemia. Molecular analysis revealed a novel STRN-PDGFRA fusion for the t(2;4) and ETV6-PDGFRA for the t(4;12). The fusions were confirmed by specific amplification of the genomic breakpoints, reverse transcription polymerase chain reaction and fluorescence in situ hybridisation. Both patients were treated with imatinib and, following a rapid haematological response, achieved cytogenetic remission and a major molecular response. In conclusion, PDGFRA fuses to diverse partner genes in myeloid disorders. Identification of these fusions is important as they are particularly sensitive to imatinib.

  4. A Search for Gene Fusions/Translocations in Breast Cancer

    DTIC Science & Technology

    2012-10-01

    pseudogenes derived from AURKA (kidney samples), RHOB (colon samples), and HMGB1 ( myeloproliferative neoplasms [MPNs]) (Figure 3A, top). Interestingly...key clusters are labeled with their corresponding parental gene symbols. MPN, myeloproliferative neoplasms . See also Table S6.transcripts in samples

  5. Fusion of the TEL gene on 12p13 to the AML1 gene on 21q22 in acute lymphoblastic leukemia

    SciTech Connect

    Barker, G.F.; Golub, T.R.; Gilliland, D.G.; Bohlander, S.K.; Rowley, J.D.; Heibert, S.W.; Raimondi, S.C.; Ward, D.C.; Bray-Ward, P.; Morgan, E.

    1995-05-23

    Chromosomal rearrangements involving band 12p13 are found in a wide variety of human leukemias but are particularly common in childhood acute lymphoblastic leukemia. The genes involved in these rearrangements, however, have not been identified. We now report the cloning of a t(12;21) translocation breakpoint involving 12p13 and 21q22 in two cases of childhood pre-B acute lymphoblastic leukemia, in which t(12;21) rearrangements were not initially apparent. The consequence of the translocation is fusion of the helix-loop-helix domain of TEL, an ETS-like putative transcription factor, to the DNA-binding and transactivation domains of the transcription factor AML1. These data show that TEL, previously shown to be fused to the platelet-derived growth factor receptor {beta} in chronic myelomonocytic leukemia, can be implicated in the pathogenesis of leukemia through its fusion to either a receptor tyrosine kinase or a transcription factor. The TEL-AML1 fusion also indicates that translocations affecting the AML1 gene can be associated with lymphoid, as well as myeloid, malignancy. 23 refs., 5 figs.

  6. Report of the heavy-ion fusion task group

    SciTech Connect

    Sawyer, G.A.; Booth, L.A.; Henderson, D.B.; Jameson, R.A.; Kindel, J.M.; Knapp, E.A.; Pollock, R.; Talbert, W.L.; Thode, L.E.; Williams, J.M.

    1980-02-01

    An assessment of heavy-ion fusion has been completed. Energetic heavy ions, for example 10-GeV uranium, provided by an rf linac or an induction linac, are used as alternatives to laser light to drive inertial confinement fusion pellets. The assessment has covered accelerator technology, transport of heavy-ion beams, target interaction physics, civilian power issues, and military applications. It is concluded that particle accelerators promise to be efficient pellet drivers, but that there are formidable technical problems to be solved. It is recommended that a moderate level research program on heavy-ion fusion be pursued and that LASL should continue to work on critical issues in accelerator development, beam transport, reactor systems studies, and target physics over the next few years.

  7. Laser fusion driven breeder design study. Final report

    SciTech Connect

    Berwald, D.H.; Massey, J.V.

    1980-12-01

    The results of the Laser Fusion Breeder Design Study are given. This information primarily relates to the conceptual design of an inertial confinement fusion (ICF) breeder reactor (or fusion-fission hybrid) based upon the HYLIFE liquid metal wall protection concept developed at Lawrence Livermore National Laboratory. The blanket design for this breeder is optimized to both reduce fissions and maximize the production of fissile fuel for subsequent use in conventional light water reactors (LWRs). When the suppressed fission blanket is compared with its fast fission counterparts, a minimal fission rate in the blanket results in a unique reactor safety advantage for this concept with respect to reduced radioactive inventory and reduced fission product decay afterheat in the event of a loss-of-coolant-accident.

  8. FY-2013 FES (Fusion Energy Sciences) Joint Research Target Report

    SciTech Connect

    Fenstermacher, M. E.; Garofalo, A. M.; Gerhardt, S. P.; Hubbard, A.; Maingi, R.; Whyte, D.

    2013-09-30

    investigated. The research will strengthen the basis for extrapolation of stationary regimes which combine high energy confinement with good particle and impurity control, to ITER and other future fusion facilities for which avoidance of large ELMs is a critical issue. Data from the Alcator C-Mod tokamak (MIT), DIII-D tokamak (General Atomics), and NSTX spherical tokamak (PPPL) contribute to this report. Experiments specifically motivated by this research target were conducted on DIII-D, with a national team of researchers from GA, LLNL, PPPL, MIT and ORNL contributing. Both the Alcator C-Mod and NSTX-U teams contributed analysis of previously collected data, as those two facilities did not operate in FY2013. Within each of the three research groups, members from both the host institutions and collaborating institutions made critical contributions. Highlights from these research activities are provided, with additional details.

  9. Dissection of left iliac artery during anterior lumbar interspace fusion: Report of a case.

    PubMed

    Fischer, Uwe M; Davies, Mark G; El Sayed, Hosam

    2015-04-01

    Vascular injury is an uncommon complication of spine surgery. Among the different approaches, anterior lumbar interbody fusion has increased potential for vascular injuries, since the great vessels and their branches overly the disc spaces to be operated on, and retraction of these vessels is necessary to gain adequate surgical exposure. The reported incidence for anterior lumbar interbody fusion-associated vascular injuries ranges from 0% to 18.1%, with venous laceration as the most common type. We report a case of anterior lumbar interbody fusion-associated left common iliac artery dissection leading to delayed acute limb ischemia developing in early post-operative period.

  10. Selection against Robertsonian fusions involving housekeeping genes in the house mouse: integrating data from gene expression arrays and chromosome evolution.

    PubMed

    Ruiz-Herrera, Aurora; Farré, Marta; Ponsà, Montserrat; Robinson, Terence J

    2010-11-01

    Monobrachial homology resulting from Robertsonian (Rb) fusions is thought to contribute to chromosomal speciation through underdominance. Given the karyotypic diversity characterizing wild house mouse populations [Mus musculus domesticus, (MMU)], variation that results almost exclusively from Rb fusions (diploid numbers range from 22 to 40) and possibly whole arm reciprocal translocations (WARTs), this organism represents an excellent model for testing hypotheses of chromosomal evolution. Previous studies of chromosome size and recombination rates have failed to explain the bias for certain chromosomes to be involved more frequently than others in these rearrangements. Here, we show that the pericentromeric region of one such chromosome, MMU19, which is infrequently encountered as a fusion partner in wild populations, is significantly enriched for housekeeping genes when compared to other chromosomes in the genome. These data suggest that there is selection against breakpoints in the pericentromeric region and provide new insights into factors that constrain chromosomal reorganizations in house mice. Given the anticipated increase in vertebrate whole genome sequences, the examination of gene content and expression profiles of the pericentromeric regions of other mammalian lineages characterized by Rb fusions (i.e., other rodents, bats, and bovids, among others) is both achievable and crucial to developing broadly applicable models of chromosome evolution.

  11. Origin and Evolution of a Chimeric Fusion Gene in Drosophila subobscura, D. madeirensis and D. guanche

    PubMed Central

    Jones, Corbin D.; Custer, Andrew W.; Begun, David J.

    2005-01-01

    An understanding of the mutational and evolutionary mechanisms underlying the emergence of novel genes is critical to studies of phenotypic and genomic evolution. Here we describe a new example of a recently formed chimeric fusion gene that occurs in Drosophila guanche, D. madeirensis, and D. subobscura. This new gene, which we name Adh-Twain, resulted from an Adh mRNA that retrotransposed into the Gapdh-like gene, CG9010. Adh-Twain is transcribed; its 5′ promoters and transcription patterns appear similar to those of CG9010. Population genetic and phylogenetic analyses suggest that the amino acid sequence of Adh-Twain evolved rapidly via directional selection shortly after it arose. Its more recent history, however, is characterized by slower evolution consistent with increasing functional constraints. We present a model for the origin of this new gene and discuss genetic and evolutionary factors affecting the evolution of new genes and functions. PMID:15781692

  12. The relationship of TMPRSS2-ERG gene fusion between primary and metastatic prostate cancers

    PubMed Central

    Guo, Charles C.; Wang, Yan; Xiao, Li; Troncoso, Patricia; Czerniak, Bogdan A.

    2013-01-01

    Recent studies have revealed the presence of TMPRSS2-ERG gene fusion in both primary and metastatic prostatic cancers (PCAs). However, the relationship between primary and corresponding metastatic PCAs with respect to the status of this gene fusion remains unclear. Using fluorescence in situ hybridization, we evaluated the rearrangement of the ERG gene in the radical prostatectomy (RP) specimens and corresponding lymph node metastases from 19 patients with PCA. The mean age of the patients was 61 years and the median Gleason score in the RP specimens was 7 (4+3). PCA was unifocal in 6 cases and multifocal in 13 cases, including 10 with 2 foci and 3 with 3 foci. In the primary PCAs, rearrangement of the ERG gene was observed in 13 cases and associated with deletion of the 5’ ERG gene in 8 cases. In the metastases, the ERG rearrangement was present in 10 cases and associated with deletion of the 5’ ERG gene in 6 cases. In unifocal PCAs, the status of the ERG rearrangement was concordant between the primary PCA and metastasis in 5 of 6 cases. In multifocal PCA, despite a significant interfocal discordance, the status of the ERG rearrangement was concordant between the index (largest) primary tumor focus and metastasis in all 13 cases. Our study demonstrates a close relationship of the TMPRSS2-ERG gene fusion status between primary and metastatic PCA. The concordance of the ERG gene rearrangement status between the index primary tumor focus and metastasis suggests that metastasis most likely arises from the index tumor focus in multifocal PCA. PMID:21937078

  13. The relationship of TMPRSS2-ERG gene fusion between primary and metastatic prostate cancers.

    PubMed

    Guo, Charles C; Wang, Yan; Xiao, Li; Troncoso, Patricia; Czerniak, Bogdan A

    2012-05-01

    Recent studies have revealed the presence of TMPRSS2-ERG gene fusion in both primary and metastatic prostate cancers. However, the relationship between primary and corresponding metastatic prostate cancers with respect to the status of this gene fusion remains unclear. Using fluorescence in situ hybridization, we evaluated the rearrangement of the ERG gene in the radical prostatectomy specimens and corresponding lymph node metastases from 19 patients with prostate cancer. The mean age of the patients was 61 years, and the median Gleason score in the radical prostatectomy specimens was 7 (4 + 3). Prostate cancer was unifocal in 6 cases and multifocal in 13 cases, including 10 with 2 foci and 3 with 3 foci. In the primary prostate cancers, rearrangement of the ERG gene was observed in 13 cases and associated with deletion of the 5' ERG gene in 8 cases. In the metastases, the ERG rearrangement was present in 10 cases and associated with deletion of the 5' ERG gene in 6 cases. In unifocal prostate cancers, the status of the ERG rearrangement was concordant between the primary prostate cancer and metastasis in 5 of 6 cases. In multifocal prostate cancer, despite a significant interfocal discordance, the status of the ERG rearrangement was concordant between the index (largest) primary tumor focus and metastasis in all 13 cases. Our study demonstrates a close relationship of the TMPRSS2-ERG gene fusion status between primary and metastatic prostate cancer. The concordance of the ERG gene rearrangement status between the index primary tumor focus and metastasis suggests that metastasis most likely arises from the index tumor focus in multifocal prostate cancer.

  14. Gene expression, single nucleotide variant and fusion transcript discovery in archival material from breast tumors.

    PubMed

    Norton, Nadine; Sun, Zhifu; Asmann, Yan W; Serie, Daniel J; Necela, Brian M; Bhagwate, Aditya; Jen, Jin; Eckloff, Bruce W; Kalari, Krishna R; Thompson, Kevin J; Carr, Jennifer M; Kachergus, Jennifer M; Geiger, Xochiquetzal J; Perez, Edith A; Thompson, E Aubrey

    2013-01-01

    Advantages of RNA-Seq over array based platforms are quantitative gene expression and discovery of expressed single nucleotide variants (eSNVs) and fusion transcripts from a single platform, but the sensitivity for each of these characteristics is unknown. We measured gene expression in a set of manually degraded RNAs, nine pairs of matched fresh-frozen, and FFPE RNA isolated from breast tumor with the hybridization based, NanoString nCounter (226 gene panel) and with whole transcriptome RNA-Seq using RiboZeroGold ScriptSeq V2 library preparation kits. We performed correlation analyses of gene expression between samples and across platforms. We then specifically assessed whole transcriptome expression of lincRNA and discovery of eSNVs and fusion transcripts in the FFPE RNA-Seq data. For gene expression in the manually degraded samples, we observed Pearson correlations of >0.94 and >0.80 with NanoString and ScriptSeq protocols, respectively. Gene expression data for matched fresh-frozen and FFPE samples yielded mean Pearson correlations of 0.874 and 0.783 for NanoString (226 genes) and ScriptSeq whole transcriptome protocols respectively, p<2x10(-16). Specifically for lincRNAs, we observed superb Pearson correlation (0.988) between matched fresh-frozen and FFPE pairs. FFPE samples across NanoString and RNA-Seq platforms gave a mean Pearson correlation of 0.838. In FFPE libraries, we detected 53.4% of high confidence SNVs and 24% of high confidence fusion transcripts. Sensitivity of fusion transcript detection was not overcome by an increase in depth of sequencing up to 3-fold (increase from ~56 to ~159 million reads). Both NanoString and ScriptSeq RNA-Seq technologies yield reliable gene expression data for degraded and FFPE material. The high degree of correlation between NanoString and RNA-Seq platforms suggests discovery based whole transcriptome studies from FFPE material will produce reliable expression data. The RiboZeroGold ScriptSeq protocol performed

  15. Gene Fusion Analysis in the Battle against the African Endemic Sleeping Sickness

    PubMed Central

    Trimpalis, Philip; Koumandou, Vassiliki Lila; Pliakou, Evangelia; Anagnou, Nicholas P.; Kossida, Sophia

    2013-01-01

    The protozoan Trypanosoma brucei causes African Trypanosomiasis or sleeping sickness in humans, which can be lethal if untreated. Most available pharmacological treatments for the disease have severe side-effects. The purpose of this analysis was to detect novel protein-protein interactions (PPIs), vital for the parasite, which could lead to the development of drugs against this disease to block the specific interactions. In this work, the Domain Fusion Analysis (Rosetta Stone method) was used to identify novel PPIs, by comparing T. brucei to 19 organisms covering all major lineages of the tree of life. Overall, 49 possible protein-protein interactions were detected, and classified based on (a) statistical significance (BLAST e-value, domain length etc.), (b) their involvement in crucial metabolic pathways, and (c) their evolutionary history, particularly focusing on whether a protein pair is split in T. brucei and fused in the human host. We also evaluated fusion events including hypothetical proteins, and suggest a possible molecular function or involvement in a certain biological process. This work has produced valuable results which could be further studied through structural biology or other experimental approaches so as to validate the protein-protein interactions proposed here. The evolutionary analysis of the proteins involved showed that, gene fusion or gene fission events can happen in all organisms, while some protein domains are more prone to fusion and fission events and present complex evolutionary patterns. PMID:23874788

  16. SFM: A novel sequence-based fusion method for disease genes identification and prioritization.

    PubMed

    Yousef, Abdulaziz; Moghadam Charkari, Nasrollah

    2015-10-21

    The identification of disease genes from human genome is of great importance to improve diagnosis and treatment of disease. Several machine learning methods have been introduced to identify disease genes. However, these methods mostly differ in the prior knowledge used to construct the feature vector for each instance (gene), the ways of selecting negative data (non-disease genes) where there is no investigational approach to find them and the classification methods used to make the final decision. In this work, a novel Sequence-based fusion method (SFM) is proposed to identify disease genes. In this regard, unlike existing methods, instead of using a noisy and incomplete prior-knowledge, the amino acid sequence of the proteins which is universal data has been carried out to present the genes (proteins) into four different feature vectors. To select more likely negative data from candidate genes, the intersection set of four negative sets which are generated using distance approach is considered. Then, Decision Tree (C4.5) has been applied as a fusion method to combine the results of four independent state-of the-art predictors based on support vector machine (SVM) algorithm, and to make the final decision. The experimental results of the proposed method have been evaluated by some standard measures. The results indicate the precision, recall and F-measure of 82.6%, 85.6% and 84, respectively. These results confirm the efficiency and validity of the proposed method.

  17. Adenoviral-Mediated Imaging of Gene Transfer Using a Somatostatin Receptor-Cytosine Deaminase Fusion Protein

    PubMed Central

    Lears, Kimberly A.; Parry, Jesse J.; Andrews, Rebecca; Nguyen, Kim; Wadas, Thaddeus J.; Rogers, Buck E.

    2015-01-01

    Suicide gene therapy is a process by which cells are administered a gene that encodes a protein capable of converting a nontoxic prodrug into an active toxin. Cytosine deaminase (CD) has been widely investigated as a means of suicide gene therapy due to the enzyme’s ability to convert the prodrug 5-fluorocytosine (5-FC) into the toxic compound 5-fluorouracil (5-FU). However, the extent of gene transfer is a limiting factor in predicting therapeutic outcome. The ability to monitor gene transfer, non-invasively, would strengthen the efficiency of therapy. In this regard, we have constructed and evaluated a replication-deficient adenovirus (Ad) containing the human somatostatin receptor subtype 2 (SSTR2) fused with a C-terminal yeast CD gene for the non-invasive monitoring of gene transfer and therapy. The resulting Ad (AdSSTR2-yCD) was evaluated in vitro in breast cancer cells to determine the function of the fusion protein. These studies demonstrated that the both the SSTR2 and yCD were functional in binding assays, conversion assays, and cytotoxicity assays. In vivo studies similarly demonstrated the functionality using conversion assays, biodistribution studies, and small animal positron-emission tomography (PET) imaging studies. In conclusion, the fusion protein has been validated as useful for the non-invasive imaging of yCD expression and will be evaluated in the future for monitoring yCD-based therapy. PMID:25837665

  18. Adenoviral-mediated imaging of gene transfer using a somatostatin receptor-cytosine deaminase fusion protein.

    PubMed

    Lears, K A; Parry, J J; Andrews, R; Nguyen, K; Wadas, T J; Rogers, B E

    2015-03-01

    Suicide gene therapy is a process by which cells are administered a gene that encodes a protein capable of converting a nontoxic prodrug into an active toxin. Cytosine deaminase (CD) has been widely investigated as a means of suicide gene therapy owing to the enzyme's ability to convert the prodrug 5-fluorocytosine (5-FC) into the toxic compound 5-fluorouracil (5-FU). However, the extent of gene transfer is a limiting factor in predicting therapeutic outcome. The ability to monitor gene transfer, non-invasively, would strengthen the efficiency of therapy. In this regard, we have constructed and evaluated a replication-deficient adenovirus (Ad) containing the human somatostatin receptor subtype 2 (SSTR2) fused with a C-terminal yeast CD gene for the non-invasive monitoring of gene transfer and therapy. The resulting Ad (AdSSTR2-yCD) was evaluated in vitro in breast cancer cells to determine the function of the fusion protein. These studies demonstrated that both the SSTR2 and yCD were functional in binding assays, conversion assays and cytotoxicity assays. In vivo studies similarly demonstrated the functionality using conversion assays, biodistribution studies and small animal positron-emission tomography (PET) imaging studies. In conclusion, the fusion protein has been validated as useful for the non-invasive imaging of yCD expression and will be evaluated in the future for monitoring yCD-based therapy.

  19. Final report on the Magnetized Target Fusion Collaboration

    SciTech Connect

    John Slough

    2009-09-08

    Nuclear fusion has the potential to satisfy the prodigious power that the world will demand in the future, but it has yet to be harnessed as a practical energy source. The entry of fusion as a viable, competitive source of power has been stymied by the challenge of finding an economical way to provide for the confinement and heating of the plasma fuel. It is the contention here that a simpler path to fusion can be achieved by creating fusion conditions in a different regime at small scale (~ a few cm). One such program now under study, referred to as Magnetized Target Fusion (MTF), is directed at obtaining fusion in this high energy density regime by rapidly compressing a compact toroidal plasmoid commonly referred to as a Field Reversed Configuration (FRC). To make fusion practical at this smaller scale, an efficient method for compressing the FRC to fusion gain conditions is required. In one variant of MTF a conducting metal shell is imploded electrically. This radially compresses and heats the FRC plasmoid to fusion conditions. The closed magnetic field in the target plasmoid suppresses the thermal transport to the confining shell, thus lowering the imploding power needed to compress the target. The undertaking to be described in this proposal is to provide a suitable target FRC, as well as a simple and robust method for inserting and stopping the FRC within the imploding liner. The timescale for testing and development can be rapidly accelerated by taking advantage of a new facility funded by the Department of Energy. At this facility, two inductive plasma accelerators (IPA) were constructed and tested. Recent experiments with these IPAs have demonstrated the ability to rapidly form, accelerate and merge two hypervelocity FRCs into a compression chamber. The resultant FRC that was formed was hot (T&ion ~ 400 eV), stationary, and stable with a configuration lifetime several times that necessary for the MTF liner experiments. The accelerator length was less than

  20. Inferring orthologous gene regulatory networks using interspecies data fusion

    PubMed Central

    Penfold, Christopher A.; Millar, Jonathan B. A.; Wild, David L.

    2015-01-01

    Motivation: The ability to jointly learn gene regulatory networks (GRNs) in, or leverage GRNs between related species would allow the vast amount of legacy data obtained in model organisms to inform the GRNs of more complex, or economically or medically relevant counterparts. Examples include transferring information from Arabidopsis thaliana into related crop species for food security purposes, or from mice into humans for medical applications. Here we develop two related Bayesian approaches to network inference that allow GRNs to be jointly inferred in, or leveraged between, several related species: in one framework, network information is directly propagated between species; in the second hierarchical approach, network information is propagated via an unobserved ‘hypernetwork’. In both frameworks, information about network similarity is captured via graph kernels, with the networks additionally informed by species-specific time series gene expression data, when available, using Gaussian processes to model the dynamics of gene expression. Results: Results on in silico benchmarks demonstrate that joint inference, and leveraging of known networks between species, offers better accuracy than standalone inference. The direct propagation of network information via the non-hierarchical framework is more appropriate when there are relatively few species, while the hierarchical approach is better suited when there are many species. Both methods are robust to small amounts of mislabelling of orthologues. Finally, the use of Saccharomyces cerevisiae data and networks to inform inference of networks in the budding yeast Schizosaccharomyces pombe predicts a novel role in cell cycle regulation for Gas1 (SPAC19B12.02c), a 1,3-beta-glucanosyltransferase. Availability and implementation: MATLAB code is available from http://go.warwick.ac.uk/systemsbiology/software/. Contact: d.l.wild@warwick.ac.uk Supplementary information: Supplementary data are available at Bioinformatics

  1. The role of FLI-1-EWS, a fusion gene reciprocal to EWS-FLI-1, in Ewing sarcoma.

    PubMed

    Elzi, David J; Song, Meihua; Houghton, Peter J; Chen, Yidong; Shiio, Yuzuru

    2015-11-01

    Ewing sarcoma is a cancer of bone and soft tissue in children that is characterized by a chromosomal translocation involving EWS and an Ets family transcription factor, most commonly FLI-1. The EWS-FLI-1 fusion oncogene is widely believed to play a central role in Ewing sarcoma. The EWS-FLI-1 gene product regulates the expression of a number of genes important for cancer progression, can transform mouse cells such as NIH3T3 and C3H10T1/2, and is necessary for proliferation and tumorigenicity of Ewing sarcoma cells, suggesting that EWS-FLI-1 is the causative oncogene. However, a variety of evidence also suggest that EWS-FLI-1 alone cannot fully explain the Ewing sarcomagenesis. Here we report that FLI-1-EWS, a fusion gene reciprocal to EWS-FLI-1, is frequently expressed in Ewing sarcoma. We present evidence suggesting that endogenous FLI-1-EWS is required for Ewing sarcoma growth and that FLI-1-EWS cooperates with EWS-FLI-1 in human mesenchymal stem cells, putative cells of origin of Ewing sarcoma, through abrogation of the proliferation arrest induced by EWS- FLI-1.

  2. Technical Letter Report - Preliminary Assessment of NDE Methods on Inspection of HDPE Butt Fusion Piping Joints for Lack of Fusion

    SciTech Connect

    Crawford, Susan L.; Cumblidge, Stephen E.; Doctor, Steven R.; Hall, Thomas E.; Anderson, Michael T.

    2008-05-29

    was conducted. Millimeter (mm) waves were also used to inspect these assemblies. Fluor and NDE Innovations, Inc. conducted TOFD inspections using their commercially available equipment on all 24 specimens. These NDE inspection results were reviewed and several of the specimens were selected for destructive evaluation using a microtome to slice small blocks of blank and fusion joint material. This interim report provides a status/summary of the work that has been conducted to date. In the areas selected for destructive testing where there were strong acoustic responses, LOF was verified. In areas where there were no NDE responses, no LOF was found. It needs to be noted that only a small amount of material has been destructively characterized at this point and further work is planned to determine if these trends hold up. Some of the material from three of the assemblies was sent off for mechanical testing but the results were not available to be included in this status report. The initial work shows that at least some of the LOF is providing NDE responses that have been verified through destructive testing. Thus, there is promise that a volumetric examination can be conducted on HDPE butt fusion joints. The future work will lead to quantifying what various NDE methods can detect, what they miss, and what they incorrectly characterize as defective.

  3. CONFERENCE REPORT: Summary of the 8th IAEA Technical Meeting on Fusion Power Plant Safety

    NASA Astrophysics Data System (ADS)

    Girard, J. Ph.; Gulden, W.; Kolbasov, B.; Louzeiro-Malaquias, A.-J.; Petti, D.; Rodriguez-Rodrigo, L.

    2008-01-01

    Reports were presented covering a selection of topics on the safety of fusion power plants. These included a review on licensing studies developed for ITER site preparation surveying common and non-common issues (i.e. site dependent) as lessons to a broader approach for fusion power plant safety. Several fusion power plant models, spanning from accessible technology to more advanced-materials based concepts, were discussed. On the topic related to fusion-specific technology, safety studies were reported on different concepts of breeding blanket modules, tritium handling and auxiliary systems under normal and accident scenarios' operation. The testing of power plant relevant technology in ITER was also assessed in terms of normal operation and accident scenarios, and occupational doses and radioactive releases under these testings have been determined. Other specific safety issues for fusion have also been discussed such as availability and reliability of fusion power plants, dust and tritium inventories and component failure databases. This study reveals that the environmental impact of fusion power plants can be minimized through a proper selection of low activation materials and using recycling technology helping to reduce waste volume and potentially open the route for its reutilization for the nuclear sector or even its clearance into the commercial circuit. Computational codes for fusion safety have been presented in support of the many studies reported. The on-going work on establishing validation approaches aiming at improving the prediction capability of fusion codes has been supported by experimental results and new directions for development have been identified. Fusion standards are not available and fission experience is mostly used as the framework basis for licensing and target design for safe operation and occupational and environmental constraints. It has been argued that fusion can benefit if a specific fusion approach is implemented, in particular

  4. SUPPRESSOR OF FRIGIDA (SUF4) Supports Gamete Fusion via Regulating Arabidopsis EC1 Gene Expression.

    PubMed

    Resentini, Francesca; Cyprys, Philipp; Steffen, Joshua G; Alter, Svenja; Morandini, Piero; Mizzotti, Chiara; Lloyd, Alan; Drews, Gary N; Dresselhaus, Thomas; Colombo, Lucia; Sprunck, Stefanie; Masiero, Simona

    2017-01-01

    The EGG CELL1 (EC1) gene family of Arabidopsis (Arabidopsis thaliana) comprises five members that are specifically expressed in the egg cell and redundantly control gamete fusion during double fertilization. We investigated the activity of all five EC1 promoters in promoter-deletion studies and identified SUF4 (SUPPRESSOR OF FRIGIDA4), a C2H2 transcription factor, as a direct regulator of the EC1 gene expression. In particular, we demonstrated that SUF4 binds to all five Arabidopsis EC1 promoters, thus regulating their expression. The down-regulation of SUF4 in homozygous suf4-1 ovules results in reduced EC1 expression and delayed sperm fusion, which can be rescued by expressing SUF4-β-glucuronidase under the control of the SUF4 promoter. To identify more gene products able to regulate EC1 expression together with SUF4, we performed coexpression studies that led to the identification of MOM1 (MORPHEUS' MOLECULE1), a component of a silencing mechanism that is independent of DNA methylation marks. In mom1-3 ovules, both SUF4 and EC1 genes are down-regulated, and EC1 genes show higher levels of histone 3 lysine-9 acetylation, suggesting that MOM1 contributes to the regulation of SUF4 and EC1 gene expression.

  5. A malignant inflammatory myofibroblastic tumor of the hypopharynx harboring the 3a/b variants of the EML4-ALK fusion gene

    PubMed Central

    Muscarella, Lucia Anna; Rossi, Giulio; Trombetta, Domenico; La Torre, Annamaria; Di Candia, Leonarda; Mengoli, Maria Cecilia; Sparaneo, Angelo; Fazio, Vito Michele; Graziano, Paolo

    2017-01-01

    Inflammatory myofibroblastic tumors (IMT) in the head and neck region are rare neoplasms that generally mimic benign/low-grade neoplasms. Overexpression of anaplastic lymphoma kinase (ALK) has been reported in 50% of IMT cases, secondary to ALK activation by structural rearrangements in the ALK gene, which results in a fusion protein with echinoderm microtubule associated protein like 4 (EML4) in ~20% of cases. The present study describes a case of a 74-year-old woman with a malignant IMT in the right posterior hypopharynx harboring a previously unreported chromosomal rearrangement resulting in EML4 and ALK gene fusion. Strong ALK immunoreactivity was observed in neoplastic cells, while fluorescent in situ hybridization combined with fluorescent fragment analysis and direct sequencing identified the first case of the 3a/b variants of the EML4-ALK fusion gene in IMT. The results of the current study highlight the uncommon occurrence of ALK-positive IMT in the head/neck region and demonstrate the importance of integrating different molecular methodologies to identify unequivocal gene fusion characterization. PMID:28356934

  6. Paediatric and adult soft tissue sarcomas with NTRK1 gene fusions: a subset of spindle cell sarcomas unified by a prominent myopericytic/haemangiopericytic pattern.

    PubMed

    Haller, Florian; Knopf, Jasmin; Ackermann, Anne; Bieg, Matthias; Kleinheinz, Kortine; Schlesner, Matthias; Moskalev, Evgeny A; Will, Rainer; Satir, Ali Abdel; Abdelmagid, Ibtihalat E; Giedl, Johannes; Carbon, Roman; Rompel, Oliver; Hartmann, Arndt; Wiemann, Stefan; Metzler, Markus; Agaimy, Abbas

    2016-04-01

    Neoplasms with a myopericytomatous pattern represent a morphological spectrum of lesions encompassing myopericytoma of the skin and soft tissue, angioleiomyoma, myofibromatosis/infantile haemangiopericytoma and putative neoplasms reported as malignant myopericytoma. Lack of reproducible phenotypic and genetic features of malignant myopericytic neoplasms have prevented the establishment of myopericytic sarcoma as an acceptable diagnostic category. Following detection of a LMNA-NTRK1 gene fusion in an index case of paediatric haemangiopericytoma-like sarcoma by combined whole-genome and RNA sequencing, we identified three additional sarcomas harbouring NTRK1 gene fusions, termed 'spindle cell sarcoma, NOS with myo/haemangiopericytic growth pattern'. The patients were two children aged 11 months and 2 years and two adults aged 51 and 80 years. While the tumours of the adults were strikingly myopericytoma-like, but with clear-cut atypical features, the paediatric cases were more akin to infantile myofibromatosis/haemangiopericytoma. All cases contained numerous thick-walled dysplastic-like vessels with segmental or diffuse nodular myxohyaline myo-intimal proliferations of smooth muscle actin-positive cells, occasionally associated with thrombosis. Immunohistochemistry showed variable expression of smooth muscle actin and CD34, but other mesenchymal markers, including STAT6, were negative. This study showed a novel variant of myo/haemangiopericytic sarcoma with recurrent NTRK1 gene fusions. Given the recent introduction of a novel therapeutic approach targeting NTRK fusion-positive neoplasms, recognition of this rare but likely under-reported sarcoma variant is strongly encouraged.

  7. Improved Controls for Fusion RF Systems. Final technical report

    SciTech Connect

    Casey, Jeffrey A.

    2011-11-08

    We have addressed the specific requirements for the integrated systems controlling an array of klystrons used for Lower Hybrid Current Drive (LHCD). The immediate goal for our design was to modernize the transmitter protection system (TPS) for LHCD on the Alcator C-Mod tokamak at the MIT Plasma Science and Fusion Center (MIT-PSFC). Working with the Alcator C-Mod team, we have upgraded the design of these controls to retrofit for improvements in performance and safety, as well as to facilitate the upcoming expansion from 12 to 16 klystrons. The longer range goals to generalize the designs in such a way that they will be of benefit to other programs within the international fusion effort was met by designing a system which was flexible enough to address all the MIT system requirements, and modular enough to adapt to a large variety of other requirements with minimal reconfiguration.

  8. [Construction of the plant expression vector with hepatitis a capsid protein fusion gene and genetic transformation of Citrus. Sinensis Osbeck].

    PubMed

    Hu, Rong; Wei, Hong; Chen, Shan-Chun; He, Yong-Rui

    2004-07-01

    The use of edible plants for the production and delivery of vaccine proteins could provide an economical alternative to fermentation systems. The construction of the plant expression vector pBI121-A was reported, which contained a fusion gene encoding hepatitis A capsid proteins. The gene was located between the left and right Ti border sequences under the control of CaMV35S promoter. The vector was identified via PCR and restriction enzyme analysis and was introduced into Agrobacterium tumerifacience LBA4404. The transgenic Citrus plants were produced by Agrobacterium-mediated transformation of epicotyl segments.13 putatively transformed plants through the kanamycin selection were micrografted onto the seedlings. The presence and integration of the transgene had been verified by PCR analysis. The result showed that five transformants were integrated and the transformation efficiency was 4.1%.

  9. Fusion Plasma Theory: Task 1, Magnetic confinement Fusion Plasma Theory. Annual progress report, November 16, 1992--November 15, 1993

    SciTech Connect

    Callen, J.D.

    1993-10-01

    The research performed under this grant during the current year has concentrated on few tokamak plasma confinement issues: applications of our new Chapman-Enskog-like approach for developing hybrid fluid/kinetic descriptions of tokamak plasmas; multi-faceted studies as part of our development of a new interacting island paradigm for the tokamak equilibrium`` and transport; investigations of the resolution power of BES and ECE diagnostics for measuring core plasma fluctuations; and studies of net transport in the presence of fluctuating surfaces. Recent progress and publications in these areas, and in the management of the NERSC node and the fusion theory workstations are summarized briefly in this report.

  10. Kinase impact assessment in the landscape of fusion genes that retain kinase domains: a pan-cancer study.

    PubMed

    Kim, Pora; Jia, Peilin; Zhao, Zhongming

    2016-12-24

    Assessing the impact of kinase in gene fusion is essential for both identifying driver fusion genes (FGs) and developing molecular targeted therapies. Kinase domain retention is a crucial factor in kinase fusion genes (KFGs), but such a systematic investigation has not been done yet. To this end, we analyzed kinase domain retention (KDR) status in chimeric protein sequences of 914 KFGs covering 312 kinases across 13 major cancer types. Based on 171 kinase domain-retained KFGs including 101 kinases, we studied their recurrence, kinase groups, fusion partners, exon-based expression depth, short DNA motifs around the break points and networks. Our results, such as more KDR than 5'-kinase fusion genes, combinatorial effects between 3'-KDR kinases and their 5'-partners and a signal transduction-specific DNA sequence motif in the break point intronic sequences, supported positive selection on 3'-kinase fusion genes in cancer. We introduced a degree-of-frequency (DoF) score to measure the possible number of KFGs of a kinase. Interestingly, kinases with high DoF scores tended to undergo strong gene expression alteration at the break points. Furthermore, our KDR gene fusion network analysis revealed six of the seven kinases with the highest DoF scores (ALK, BRAF, MET, NTRK1, NTRK3 and RET) were all observed in thyroid carcinoma. Finally, we summarized common features of 'effective' (highly recurrent) kinases in gene fusions such as expression alteration at break point, redundant usage in multiple cancer types and 3'-location tendency. Collectively, our findings are useful for prioritizing driver kinases and FGs and provided insights into KFGs' clinical implications.

  11. Inertial Confinement Fusion quarterly report, January--March 1995. Volume 5, No. 2

    SciTech Connect

    1995-09-01

    The ICF quarterly report is published by the Inertial Confinement Fusion Program at the Lawrence Livermore National Laboratory. Topics included this quarter include: the role of the National Ignition Facility in the development of Inertial Confinement Fusion, laser-plasma interactions in large gas-filled hohlraums, evolution of solid-state induction modulators for a heavy-ion recirculator, the National Ignition Facility project, and terminal-level relaxation in Nd-doped laser material.

  12. CONFERENCE REPORT: 11th EU-US Transport Task Force workshop on transport in fusion plasmas

    NASA Astrophysics Data System (ADS)

    Connor, J. W.; Angioni, C.; Diamond, P. H.; Hammett, G. W.; Hidalgo, C.; Loarte, A.; Mantica, P.

    2007-04-01

    This report summarizes the contributions presented at the 11th EU-US Transport Task Force workshop on transport in fusion plasmas, held in Marseilles, France, 4-7 September, 20068The present workshop: http://www-fusion-magnetique.cea.fr/ttf2006.. There were sessions on momentum transport, multi-scale physics, electron transport, particle transport and transport in the scrape-off layer.

  13. The rationale of vectored gene-fusion vaccines against cancer: evolving strategies and latest evidence

    PubMed Central

    Ragonnaud, Emeline

    2013-01-01

    The development of vaccines that target tumor antigens in cancer has proven difficult. A major reason for this is that T cells specific for tumor self-antigens and neoantigens are eliminated or inactivated through mechanisms of tolerance. Antigen fusion strategies which increase the ability of vaccines to stimulate T cells that have escaped tolerance mechanisms, may have a particular potential as immunotherapies. This review highlights antigen fusion strategies that have been successful in stimulating the induction of T-cell immunity against cancer and counteracting tumor-associated tolerance. In preclinical studies, these strategies have shown to improve the potency of vectored vaccines through fusion of tumor antigen to proteins or protein domains that increase CD4+ T-cell help, CD8+ T-cell responses or both the CD4+ and CD8+ T-cell responses. However, in clinical trials such strategies seem to be less efficient when provided as a DNA vaccine. The first clinical trial using a viral vectored fusion-gene vaccine is expected to be tested as a partner in a heterologous prime-boost regimen directed against cervical cancer. PMID:24757514

  14. [Analysis of DEK-CAN fusion gene expression in acute myeloid leukemia patients with 6; 9 chromosome translocation].

    PubMed

    Wang, Ya-Lun; Wang, Tong; Xu, Feng; Gang, Yan; Wang, Jie

    2006-04-01

    This study was aimed to explore the relationship of 6; 9 chromosome translocation with DEK-CAN fusion gene expression in patients with acute myeloid leukemia (AML) and its clinical significance. Chromosome specimens were prepared by routine method after short-term culture of bone marrow cells; karyotype analysis was performed by R banding technique; the expression of fusion gene DEK-CAN was analyzed by RT-nested-PCR in mononuclear cells of bone marrow or peripheral blood of 4 AML patients, for 3 patients received allo-BMT out of 4 patients the dynamic follow-up was performed. The results indicated that t (6; 9) (p23; q34) was confirmed by chromosome karyotype analysis in the four AML patients. The DEK-CAN fusion gene was found during in all four de novo, relapsed and CR patients (100%). And the expression of DEK-CAN fusion gene enhanced apparently in de novo and relapsed patients, and weakened in CR patient. DEK-CAN mRNA was found in the three patients during 1-24 months after allo-BMT. Clinical data showed 2 patients relapsed and died after CR for 1-24 months; the other two patients received allo-BMT got CR and still survive. It is concluded that DEK-CAN fusion gene is the molecular basis in pathogenesis of AML. The detection of DEK-CAN fusion gene is significant for diagnosis of AML, evaluation of curative effect, and predication of prognosis.

  15. Origin of the plant Tm-1-like gene via two independent horizontal transfer events and one gene fusion event

    PubMed Central

    Yang, Zefeng; Liu, Li; Fang, Huimin; Li, Pengcheng; Xu, Shuhui; Cao, Wei; Xu, Chenwu; Huang, Jinling; Zhou, Yong

    2016-01-01

    The Tomato mosaic virus (ToMV) resistance gene Tm-1 encodes a direct inhibitor of ToMV RNA replication to protect tomato from infection. The plant Tm-1-like (Tm-1L) protein is predicted to contain an uncharacterized N-terminal UPF0261 domain and a C-terminal TIM-barrel signal transduction (TBST) domain. Homologous searches revealed that proteins containing both of these two domains are mainly present in charophyte green algae and land plants but absent from glaucophytes, red algae and chlorophyte green algae. Although Tm-1 homologs are widely present in bacteria, archaea and fungi, UPF0261- and TBST-domain-containing proteins are generally encoded by different genes in these linages. A co-evolution analysis also suggested a putative interaction between UPF0261- and TBST-domain-containing proteins. Phylogenetic analyses based on homologs of these two domains revealed that plants have acquired UPF0261- and TBST-domain-encoding genes through two independent horizontal gene transfer (HGT) events before the origin of land plants from charophytes. Subsequently, gene fusion occurred between these two horizontally acquired genes and resulted in the origin of the Tm-1L gene in streptophytes. Our results demonstrate a novel evolutionary mechanism through which the recipient organism may acquire genes with functional interaction through two different HGT events and further fuse them into one functional gene. PMID:27647002

  16. Ewing Sarcoma With ERG Gene Rearrangements: A Molecular Study Focusing on the Prevalence of FUS-ERG and Common Pitfalls in Detecting EWSR1-ERG Fusions by FISH

    PubMed Central

    Chen, Sonja; Deniz, Kemal; Sung, Yun-Shao; Zhang, Lei; Dry, Sarah; Antonescu, Cristina R.

    2016-01-01

    The genetics of Ewing sarcoma (ES) are characterized by a canonical fusion involving EWSR1 gene and a member of the ETS family of transcription factors, such as FLI1 and ERG. In fact, ERG gene rearrangements represent the second most common molecular alteration, with EWSR1-ERG being identified in 5–10% of cases, while only a handful of reports document a FUS-ERG fusion. In this study, we focus on ES with ERG gene abnormalities, specifically to investigate the prevalence and clinicopathologic features of FUS-ERG fusions in a large cohort of small blue round cell tumors (SBRCTs) and compare to the eight reported FUS-positive ES. Among the 85 SBRCTs tested, seven (8.2%) cases harbored FUS gene rearrangements; six fused to ERG and one with FEV. During this investigation we came across a number of ERG-rearranged ES lacking both EWSR1 and FUS abnormalities by FISH. In one case, RNA sequencing identified an EWSR1-ERG transcript despite the negative EWSR1 rearrangements by FISH. Additional 3-color FISH fusion assay demonstrated the fusion of EWSR1 and ERG signals in all four cases negative for break-apart EWSR1 FISH. These results emphasize a potential pitfall of relying on EWSR1 FISH assay alone for diagnosis of ES. In cases with classic morphology and/or strong CD99 and ERG immunoreactivity, additional molecular testing should be applied, such as ERG FISH or RT-PCR/next generation sequencing, for a more definitive diagnosis. Although our study group is small, there were no differences noted between the clinical, morphologic features and immunoprofile of the different subsets of ERG-rearranged SBRCTs. PMID:26690869

  17. Membrane fusion inducers, chloroquine and spermidine increase lipoplex-mediated gene transfection

    SciTech Connect

    Wong-Baeza, Carlos; Bustos, Israel; Serna, Manuel; Tescucano, Alonso; Alcantara-Farfan, Veronica; Ibanez, Miguel; Montanez, Cecilia; Wong, Carlos; Baeza, Isabel

    2010-05-28

    Gene transfection into mammalian cells can be achieved with viral and non-viral vectors. Non-viral vectors, such as cationic lipids that form lipoplexes with DNA, are safer and more stable than viral vectors, but their transfection efficiencies are lower. Here we describe that the simultaneous treatment with a membrane fusion inducer (chlorpromazine or procainamide) plus the lysosomotropic agent chloroquine increases lipoplex-mediated gene transfection in human (HEK293 and C-33 A) and rat (PC12) cell lines (up to 9.2-fold), as well as in situ in BALB/c mice spleens and livers (up to 6-fold); and that the polyamine spermidine increases lipoplex-mediated gene transfection and expression in cell cultures. The use of these four drugs provides a novel, safe and relatively inexpensive way to considerably increase lipoplex-mediated gene transfection efficiency.

  18. Deep RNA sequencing analysis of readthrough gene fusions in human prostate adenocarcinoma and reference samples

    PubMed Central

    2011-01-01

    Background Readthrough fusions across adjacent genes in the genome, or transcription-induced chimeras (TICs), have been estimated using expressed sequence tag (EST) libraries to involve 4-6% of all genes. Deep transcriptional sequencing (RNA-Seq) now makes it possible to study the occurrence and expression levels of TICs in individual samples across the genome. Methods We performed single-end RNA-Seq on three human prostate adenocarcinoma samples and their corresponding normal tissues, as well as brain and universal reference samples. We developed two bioinformatics methods to specifically identify TIC events: a targeted alignment method using artificial exon-exon junctions within 200,000 bp from adjacent genes, and genomic alignment allowing splicing within individual reads. We performed further experimental verification and characterization of selected TIC and fusion events using quantitative RT-PCR and comparative genomic hybridization microarrays. Results Targeted alignment against artificial exon-exon junctions yielded 339 distinct TIC events, including 32 gene pairs with multiple isoforms. The false discovery rate was estimated to be 1.5%. Spliced alignment to the genome was less sensitive, finding only 18% of those found by targeted alignment in 33-nt reads and 59% of those in 50-nt reads. However, spliced alignment revealed 30 cases of TICs with intervening exons, in addition to distant inversions, scrambled genes, and translocations. Our findings increase the catalog of observed TIC gene pairs by 66%. We verified 6 of 6 predicted TICs in all prostate samples, and 2 of 5 predicted novel distant gene fusions, both private events among 54 prostate tumor samples tested. Expression of TICs correlates with that of the upstream gene, which can explain the prostate-specific pattern of some TIC events and the restriction of the SLC45A3-ELK4 e4-e2 TIC to ERG-negative prostate samples, as confirmed in 20 matched prostate tumor and normal samples and 9 lung cancer

  19. Report of the Integrated Program Planning Activity for the DOE Fusion Energy Sciences Program

    SciTech Connect

    2000-12-01

    This report of the Integrated Program Planning Activity (IPPA) has been prepared in response to a recommendation by the Secretary of Energy Advisory Board that, ''Given the complex nature of the fusion effort, an integrated program planning process is an absolute necessity.'' We, therefore, undertook this activity in order to integrate the various elements of the program, to improve communication and performance accountability across the program, and to show the inter-connectedness and inter-dependency of the diverse parts of the national fusion energy sciences program. This report is based on the September 1999 Fusion Energy Sciences Advisory Committee's (FESAC) report ''Priorities and Balance within the Fusion Energy Sciences Program''. In its December 5,2000, letter to the Director of the Office of Science, the FESAC has reaffirmed the validity of the September 1999 report and stated that the IPPA presents a framework and process to guide the achievement of the 5-year goals listed in the 1999 report. The National Research Council's (NRC) Fusion Assessment Committee draft final report ''An Assessment of the Department of Energy's Office of Fusion Energy Sciences Program'', reviewing the quality of the science in the program, was made available after the IPPA report had been completed. The IPPA report is, nevertheless, consistent with the recommendations in the NRC report. In addition to program goals and the related 5-year, 10-year, and 15-year objectives, this report elaborates on the scientific issues associated with each of these objectives. The report also makes clear the relationships among the various program elements, and cites these relationships as the reason why integrated program planning is essential. In particular, while focusing on the science conducted by the program, the report addresses the important balances between the science and energy goals of the program, between the MFE and IFE approaches, and between the domestic and international aspects

  20. Analysis of API2-MALT1 fusion, trisomies, and immunoglobulin VH genes in pulmonary mucosa-associated lymphoid tissue lymphoma.

    PubMed

    Xia, Hongjing; Nakayama, Takahisa; Sakuma, Hidenori; Yamada, Seiji; Sato, Fumihiko; Takino, Hisashi; Okabe, Mitsukuni; Fujiyoshi, Yukio; Hattori, Hideo; Inagaki, Hiroshi

    2011-09-01

    Pulmonary mucosa-associated lymphoid tissue lymphoma is unique in that chronic inflammation is rare and that API2-MALT1 fusion, resulting from t(11;18)(q21;q21), occurs frequently. In this study, we examined 20 cases for API2-MALT1 fusion using the multiplex reverse-transcription polymerase chain reaction and looked for trisomy 3, trisomy 18, and abnormalities of MALT1 and IGH genes using fluorescence in situ hybridization. In addition, we analyzed VH genes by subcloning of the monoclonal polymerase chain reaction products. Of 20 cases studied, we detected gene abnormalities in 16: API2-MALT1 fusion in 9, trisomy 3 in 5, trisomy 18 in 4, MALT1 abnormality in 13, and IGH abnormality in 1. MALT1 gene abnormalities were concordant with API2-MALT1 fusion or trisomy 18. One case showed API2-MALT1 fusion and trisomy 3. On detection of API2-MALT1 fusion and trisomies, we were able to divide our cases into 3 groups, API2-MALT1 positive, trisomy positive, and no detectable gene abnormality, suggesting that tumor development had processed along different genetic pathways. All 20 cases were analyzed for VH genes. Most of the VH genes selected by the lymphomas belonged to the VH3 family, but there was no restriction to any particular VH fragment. Of interest, VH genes were unmutated in 7 cases, suggesting that T-cell-independent extrafollicular B-cell maturation may be important in the development of this lymphoma. In addition, both mutated and unmutated tumor cases were found to carry the API2-MALT1 fusion and trisomy 3. This observation suggests that these gene abnormalities may occur in microenvironments found before or outside of follicular germinal centers.

  1. Anti-colorectal cancer effect of interleukin-2 and interferon-β fusion gene driven by carcinoembryonic antigen promoter

    PubMed Central

    Wang, Yan; Wang, Mengchun; Li, Yan

    2016-01-01

    This study was designed to investigate the antitumor effects of combined interleukin-2/interferon-β-based gene therapy in colorectal cancer. Transfection of the fusion gene expression plasmid induced significant apoptosis of Lovo cells. Additionally, the fusion gene exhibited strong inhibitory activity against tumor growth and apoptosis when being injected into the nude mice implanted with human colon cancer cells. Furthermore, the tail-vein injection showed a more notable effect than direct injection into tumor. These results suggest that the combined interleukin-2/interferon-β-based gene therapy with the carcinoembryonic antigen promoter might be an effective antitumor strategy. PMID:27313471

  2. Ethanol utilization regulatory protein: profile alignments give no evidence of origin through aldehyde and alcohol dehydrogenase gene fusion.

    PubMed Central

    Nicholas, H. B.; Persson, B.; Jörnvall, H.; Hempel, J.

    1995-01-01

    The suggestion that the ethanol regulatory protein from Aspergillus has its evolutionary origin in a gene fusion between aldehyde and alcohol dehydrogenase genes (Hawkins AR, Lamb HK, Radford A, Moore JD, 1994, Gene 146:145-158) has been tested by profile analysis with aldehyde and alcohol dehydrogenase family profiles. We show that the degree and kind of similarity observed between these profiles and the ethanol regulatory protein sequence is that expected from random sequences of the same composition. This level of similarity fails to support the suggested gene fusion. PMID:8580855

  3. Fusion reactor materials: Semiannual progress report for the period ending March 31, 1987

    SciTech Connect

    none,

    1987-09-01

    This is the second in a series of semiannual technical progress reports on fusion reactor materials. This report combines research and development activities in the following areas: (1) Alloy Development for Irradiation Performance; (2) Damage Analysis and Fundamental Studies; and (3) Special Purpose Materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials program being conducted in support of the Magnetic Fusion Energy Program of the US Department of Energy. Separate analytics were prepared for the reports in this volume.

  4. Fusion Materials Semiannual Progress Report for Period Ending December 31, 1998

    SciTech Connect

    Rowcliff, A.F.; Burn, G.

    1999-04-01

    This is the twenty-fifth in a series of semiannual technical progress reports on fusion materials. This report combines the full spectrum of research and development activities on both metallic and non-metallic materials with primary emphasis on the effects of the neutronic and chemical environment on the properties and performance of materials for in-vessel components. This effort forms one element of the materials program being conducted in support of the Fusion Energy Sciences Program of the U.S. Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately.

  5. Fusion Materials Semiannual Progress Report for the Period Ending June 30, 1999

    SciTech Connect

    Rowcliffe, A.F.

    1999-09-01

    This is the twenty-sixth in a series of semiannual technical progress reports on fusion materials. This report combines the full spectrum of research and development activities on both metallic and non-metallic materials with primary emphasis on the effects of the neutronic and chemical environment on the properties and performance of materials for in-vessel components. This effort forms one element of the materials program being conducted in support of the Fusion Energy Sciences Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and its reported separately.

  6. Z-inertial fusion energy: power plant final report FY 2006.

    SciTech Connect

    Anderson, Mark; Kulcinski, Gerald; Zhao, Haihua; Cipiti, Benjamin B.; Olson, Craig Lee; Sierra, Dannelle P.; Meier, Wayne; McConnell, Paul E.; Ghiaasiaan, M. (Georgia Institute of Technology, Atlanta, GA); Kern, Brian (Georgia Institute of Technology, Atlanta, GA); Tajima, Yu (University of California, Los Angeles, CA); Campen, Chistopher (University of California, Berkeley, CA); Sketchley, Tomas (University of California, Los Angeles, CA); Moir, R (Lawrence Livermore National Laboratories); Bardet, Philippe M. (University of California, Berkeley, CA); Durbin, Samuel; Morrow, Charles W.; Vigil, Virginia L (University of Wisconsin, Madison, WI); Modesto-Beato, Marcos A.; Franklin, James Kenneth; Smith, James Dean; Ying, Alice; Cook, Jason T.; Schmitz, Lothar (University of California, Los Angeles, CA); Abdel-Khalik, S. (Georgia Institute of Technology, Atlanta, GA); Farnum, Cathy Ottinger; Abdou, Mohamed A.; Bonazza, Riccardo; Rodriguez, Salvador B.; Sridharan, Kumar (University of Wisconsin, Madison, WI); Rochau, Gary Eugene; Gudmundson, Jesse; Peterson, Per F.; Marriott, Ed; Oakley, Jason

    2006-10-01

    This report summarizes the work conducted for the Z-inertial fusion energy (Z-IFE) late start Laboratory Directed Research Project. A major area of focus was on creating a roadmap to a z-pinch driven fusion power plant. The roadmap ties ZIFE into the Global Nuclear Energy Partnership (GNEP) initiative through the use of high energy fusion neutrons to burn the actinides of spent fuel waste. Transmutation presents a near term use for Z-IFE technology and will aid in paving the path to fusion energy. The work this year continued to develop the science and engineering needed to support the Z-IFE roadmap. This included plant system and driver cost estimates, recyclable transmission line studies, flibe characterization, reaction chamber design, and shock mitigation techniques.

  7. Innovative approaches to inertial confinement fusion reactors: Final report

    SciTech Connect

    Bourque, R.F.; Schultz, K.R.

    1986-11-01

    Three areas of innovative approaches to inertial confinement fusion (ICF) reactor design are given. First, issues pertaining to the Cascade reactor concept are discussed. Then, several innovative concepts are presented which attempt to directly recover the blast energy from a fusion target. Finally, the Turbostar concept for direct recovery of that energy is evaluated. The Cascade issues discussed are combustion of the carbon granules in the event of air ingress, the use of alternate granule materials, and the effect of changes in carbon flow on details of the heat exchanger. Carbon combustion turns out to be a minor problem. Four ICF innovative concepts were considered: a turbine with ablating surfaces, a liquid piston system, a wave generator, and a resonating pump. In the final analysis, none show any real promise. The Turbostar concept of direct recovery is a very interesting idea and appeared technically viable. However, it shows no efficiency gain or any decrease in capital cost compared to reactors with conventional thermal conversion systems. Attempts to improve it by placing a close-in lithium sphere around the target to increase gas generation increased efficiency only slightly. It is concluded that these direct conversion techniques require thermalization of the x-ray and debris energy, and are Carnot limited. They therefore offer no advantage over existing and proposed methods of thermal energy conversion or direct electrical conversion.

  8. Protein interaction maps for complete genomes based on gene fusion events

    NASA Astrophysics Data System (ADS)

    Enright, Anton J.; Iliopoulos, Ioannis; Kyrpides, Nikos C.; Ouzounis, Christos A.

    1999-11-01

    A large-scale effort to measure, detect and analyse protein-protein interactions using experimental methods is under way. These include biochemistry such as co-immunoprecipitation or crosslinking, molecular biology such as the two-hybrid system or phage display, and genetics such as unlinked noncomplementing mutant detection. Using the two-hybrid system, an international effort to analyse the complete yeast genome is in progress. Evidently, all these approaches are tedious, labour intensive and inaccurate. From a computational perspective, the question is how can we predict that two proteins interact from structure or sequence alone. Here we present a method that identifies gene-fusion events in complete genomes, solely based on sequence comparison. Because there must be selective pressure for certain genes to be fused over the course of evolution, we are able to predict functional associations of proteins. We show that 215 genes or proteins in the complete genomes of Escherichia coli, Haemophilus influenzae and Methanococcus jannaschii are involved in 64 unique fusion events. The approach is general, and can be applied even to genes of unknown function.

  9. Xp11.2 translocation renal cell carcinoma with NONO-TFE3 gene fusion: morphology, prognosis, and potential pitfall in detecting TFE3 gene rearrangement.

    PubMed

    Xia, Qiu-Yuan; Wang, Zhe; Chen, Ni; Gan, Hua-Lei; Teng, Xiao-Dong; Shi, Shan-Shan; Wang, Xuan; Wei, Xue; Ye, Sheng-Bing; Li, Rui; Ma, Heng-Hui; Lu, Zhen-Feng; Zhou, Xiao-Jun; Rao, Qiu

    2017-03-01

    Xp11 translocation renal cell carcinomas are characterized by several different translocations involving the TFE3 gene. Tumors with different specific gene fusions may have different clinicopathological manifestations. Fewer than 10 renal cell carcinoma cases with NONO-TFE3 have been described. Here we examined eight additional cases of this rare tumor using clinicopathological, immunohistochemical, and molecular analyses. The male-to-female ratio of our study cohort was 1:1, and the median age was 30 years. The most distinctive feature of the tumors was that they exhibited glandular/tubular or papillary architecture that was lined with small-to-medium cuboidal to high columnar cells with indistinct cell borders and an abundantly clear or flocculent eosinophilic cytoplasm. The nuclei were oriented toward the luminal surface and were round and uniform in shape, which resulted in the appearance of secretory endometrioid subnuclear vacuolization. The distinct glandular/tubular or papillary architecture was often accompanied by sheets of epithelial cells that presented a biphasic pattern. Immunohistochemically, all eight cases demonstrated moderate (2+) or strong (3+) positive staining for TFE3, CD10, RCC marker, and PAX-8. None of the tumors were immunoreactive for CK7, Cathepsin K, Melan-A, HMB45, Ksp-cadherin, Vimentin, CA9, 34βE12 or CD117. NONO-TFE3 fusion transcripts were identified in six cases by RT-PCR. All eight cases showed equivocal split signals with a distance of nearly 2 signal diameters and sometimes had false-negative results. Furthermore, we developed a fluorescence in situ hybridization (FISH) assay to serve as an adjunct diagnostic tool for the detection of the NONO-TFE3 fusion gene and used this method to detect the fusion gene in all eight cases. Long-term follow-up (range, 10-102 months) was available for 7 patients. All 7 patients were alive with no evidence of recurrent disease or disease progression after their initial resection. This report

  10. Development of GFP fusions for examination of the effects of the space environment on gene expression in Escherichia coli

    NASA Astrophysics Data System (ADS)

    Mancinelli, R.; Fahlen, T.

    The goal of the In situ Space Gene Expression on Nano-satillites (ISGEN) program is to be ready to fly technology that can support a fully automated experiment to quantify changes in model organisms in situ in low earth orbit in a free flyer platform in less than two years. A straightforward gene expression assay that meets the ISGEN flight objective for testing flight hardware as well as return data regarding the effects of microgravity on gene expression has been developed. Escherichia coli K-12, a bacterium that exhibits changes in its growth pattern when flown in micro-gravity on the Space Shuttle, was used. The scientific objective of this work is to determine if there is a discernable change in metabolic and stress pathway gene expression due to growth in the space environment. To that end, we have linked the green fluorescent protein (GFP) reporter gfp to phoP, a gene that responds to extracellular Mg2+ levels, and pykF, a gene involved in the glycolytic pathway that responds to changes in intracellular pyruvate. These genes respond to the metabolic needs of the cell and may be altered in the micro-gravity environment. E. coli cells containing a plasmid encoding the phoP-gfp-mut3 reporter construct were grown with or without MgSO_4. The effect of the added MgSO_4 is the repression of the expression of GFP. This is the expected result if GFP expression were under the control of a magnesium-regulated promoter such as phoP. Consistent with the negative feedback loop, we observe repression of GFP production in cells containing our pykF-gfp plasmid construct, when grown in the presence of excess glucose. Thus, the pykF-gfp fusion functions as a glucose sensor.

  11. A unique RPW8-encoding class of genes that originated in early land plants and evolved through domain fission, fusion, and duplication

    PubMed Central

    Zhong, Yan; Cheng, Zong-Ming (Max)

    2016-01-01

    Duplication, lateral gene transfer, domain fusion/fission and de novo domain creation play a key role in formation of initial common ancestral protein. Abundant protein diversities are produced by domain rearrangements, including fusions, fissions, duplications, and terminal domain losses. In this report, we explored the origin of the RPW8 domain and examined the domain rearrangements that have driven the evolution of RPW8-encoding genes in land plants. The RPW8 domain first emerged in the early land plant, Physcomitrella patens, and it likely originated de novo from a non-coding sequence or domain divergence after duplication. It was then incorporated into the NBS-LRR protein to create a main sub-class of RPW8-encoding genes, the RPW8-NBS-encoding genes. They evolved by a series of genetic events of domain fissions, fusions, and duplications. Many species-specific duplication events and tandemly duplicated clusters clearly demonstrated that species-specific and tandem duplications played important roles in expansion of RPW8-encoding genes, especially in gymnosperms and species of the Rosaceae. RPW8 domains with greater Ka/Ks values than those of the NBS domains indicated that they evolved faster than the NBS domains in RPW8-NBSs. PMID:27678195

  12. Fusion materials semiannual progress report for period ending December 31, 1999

    SciTech Connect

    Burn, G.

    2000-03-01

    This is the twenty-seventh in a series of semiannual technical progress reports on fusion materials. This report combines the full spectrum of research and development activities on both metallic and non-metallic materials with primary emphasis on the effects of the neutronic and chemical environment on the properties and performance of materials for in-vessel components.

  13. Fusion materials semiannual progress report for the period ending March 31, 1995

    SciTech Connect

    1995-07-01

    This is the eighteenth in a series of semiannual technical progress reports on fusion materials. This report combines research and development activities which were previously reported separately in the following progress reports: {sm_bullet} Alloy Development for Irradiation Performance. {sm_bullet} Damage Analysis and Fundamental Studies. {sm_bullet} Special Purpose Materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials programs being conducted in support of the Magnetic Fusion Energy Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. The Fusion Materials Program is a national effort involving several national laboratories, universities, and industries. The purpose of this series of reports is to provide a working technical record for the use of the program participants, and to provide a means of communicating the efforts of materials scientists to the rest of the fusion community, both nationally and worldwide. This report has been compiled and edited under the guidance of A.F. Rowcliffe by Gabrielle Burn, Oak Ridge National Laboratory. Their efforts, and the efforts of the many persons who made technical contributions, are gratefully acknowledged.

  14. Construction and characterization of Escherichia coli polA-lacZ gene fusions.

    PubMed Central

    Ward, D F; Murray, N E

    1980-01-01

    The promoter of the polA gene of Escherichia coli K-12 was fused to the lacZ gene by selecting deletions within a lambda lacZ polA transducing phage. Four fusions, deleting varying amounts of the polA gene, were characterized. The polA promoter was found to be approximately 3% as active as the fully induced lac promoter. This figure is compatible with the normal intracellular level of deoxyribonucleic acid polymerase I. No evidence was found for outogenous regulation of transcription from the polA promoter. Expression from this promoter was influenced by neither recA nor mitomycin C, but uvrD and uvrE mutations reduced expression slightly. Images PMID:6445899

  15. Fusion in the Era of Burning Plasma Studies: Workforce Planning for 2004 to 2014. Final report to FESA C

    SciTech Connect

    none,

    2004-03-29

    This report has been prepared in response to Dr. R. Orbach’s request of the Fusion Energy Sciences Advisory Committee (FESAC) to “address the issue of workforce development in the U.S. fusion program.” The report addresses three key questions: what is the current status of the fusion science, technology, and engineering workforce; what is the workforce that will be needed and when it will be needed to ensure that the U.S. is an effective partner in ITER and to enable the U.S. to successfully carry out the fusion program; and, what can be done to ensure a qualified, diversified, and sufficiently large workforce and a pipeline to maintain that workforce? In addressing the charge, the Panel considers a workforce that allows for a vigorous national program of fusion energy research that includes participation in magnetic fusion (ITER) and inertial fusion (NIF) burning plasma experiments.

  16. Isolation of ara-lac gene fusions in Salmonella typhimurium LT2 by using transducing bacteriophage Mu d (Apr lac).

    PubMed

    Lee, J H; Heffernan, L; Wilcox, G

    1980-09-01

    A specialized Mu transducing phage containing a gene encoding ampicillin resistance and the lac structural genes without the lac promotor [Mu d(apr lac)] has been constructed and used to create gene fusions in Escherichia coli (M. J. Cadadaban and S. N. Cohen, Proc. Natl. Acad. Sci. U.S.A. 76:4530--4533, 1979). Transposition of the Mu d(Apr lac) phage to chromosomal sites can result in lac expression being controlled by a chromosomal promoter. We have constructed an Escherichia coli K-12 strain in which the Mu d(Apr lac) phage is integrated into an F factor. The F+::Mu d(Apr lac) was then transferred by conjugation into a Salmonella typhimurium strain that was sensitive to L-arabinose. Strains containing gene fusions were selected as L-arabinose-resistant colonies after partial induction of the phage. Two classes of ara-lac fusion strains were isolated: (i) araC-lac fusions in which the expression of beta-galactosidase synthesis was constitutuve and not inducible by L-arabinose; and ((ii) fusion of the lac genes to the ara structural genes in which the expression of beta-galatosidase synthesis was induced 263-fold by L-arabinose.

  17. A Preliminary Report on the CO2 Laser for Lumbar Fusion: Safety, Efficacy and Technical Considerations.

    PubMed

    Villavicencio, Alan T; Burneikiene, Sigita; Babuska, Jason M; Nelson, Ewell L; Mason, Alexander; Rajpal, Sharad

    2015-04-01

    The purpose of this study was to evaluate potential technical advantages of the CO2 laser technology in mini-open transforaminal lumbar interbody fusion (TLIF) surgeries and report our preliminary clinical data on the safety and clinical outcomes. There is currently no literature discussing the recently redeveloped CO2 laser technology application for lumbar fusion. Safety and clinical outcomes were compared between two groups: 24 patients that underwent CO2 laser-assisted one-level TLIF surgeries and 30 patients that underwent standard one-level TLIF surgeries without the laser. There were no neural thermal injuries or other intraoperative laser-related complications encountered in this cohort of patients. At a mean follow-up of 17.4 months, significantly reduced lower back pain scores (P=0.013) were reported in the laser-assisted patient group compared to a standard fusion patient group. Lower extremity radicular pain intensity scores were similar in both groups. Laser-assisted TLIF surgeries showed a tendency (P = 0.07) of shorter operative times that was not statistically significant. Based on this preliminary clinical report, the safety of the CO2 laser device for lumbar fusion surgeries was assessed. There were no neural thermal injuries or other intraoperative laser-related complications encountered in this cohort of patients. Further investigation of CO2 laser-assisted lumbar fusion procedures is warranted in order to evaluate its effect on clinical outcomes.

  18. Fusion materials semiannual progress report for the period ending June 30, 1998

    SciTech Connect

    Burn, G.

    1998-09-01

    This is the twenty-fourth in a series of semiannual technical progress reports on fusion materials. This report combines the full spectrum of research and development activities on both metallic and non-metallic materials with primary emphasis on the effects of the neutronic and chemical environment on the properties and performance of materials for in-vessel components. This effort forms one element of the materials program being conducted in support of the Fusion Energy Sciences Program of the US Department of Energy. Selected papers have been indexed separately for inclusion in the Energy Science and Technology Database.

  19. Fusion reactor materials semiannual progress report for the period ending March 31, 1993

    SciTech Connect

    Not Available

    1993-07-01

    This is the fourteenth in a series of semiannual technical progress reports on fusion reactor materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials programs being conducted in support of the Magnetic Fusion Energy Program of the US Depart of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. Separate abstracts were prepared for each individual section.

  20. [Cloning of CTB-PROIN fusion gene and its expression in Escherichia coli].

    PubMed

    Chen, Li; Ouyang, Feng-Xiu; Qian, Bing-Jun; Ren, Hong; Wang, Qiang; Jiang, Qing-Wu; Wang, Yu-Jiong; Liu, Jing-Bo; Liang, Wan-Qi; Zhang, Da-Bing

    2005-03-01

    A fusion gene CTB-PROIN, in which Proinsulin gene was fused to the 3' end of CTB gene by a hinge peptide 'GPGP', was constructed and cloned into pET-30a(+) to obtain a prokaryotic expression vector pETCPI. Subsequently the recombinant plasmid pETCPI was transformed into E. coli stain BL21 (DE3). After induced by IPTG, the expression product was analyzed by sodium dodecyl sulphate-polyacrylamide gel (15%) electrophoresis (SDS-PAGE), and its result indicated that the recombinant protein CTB-PROIN was expressed and accumulated as inclusion bodies. The recombinant CTB-PROIN protein accumulated to the level of 25% of total bacterial proteins. After inclusion bodies was denaturalized and refolded in vitro, significant assembly of monomers had occurred, and the recombinant protein represented assembled pentamers. The results of western blotting analysis also demonstrated that the fusion protein could be recognized by the anti-CT and anti-insulin antibody, respectively. In addition, the result of the CTB-PROIN-GM1 binding assay, that the protein could bind to monosialoganglioside specifically, showed it possesed biological activity in vitro. These results provided the possibility of developing a cheaper and more efficient oral vaccine for type I diabetes using such constructs.

  1. Fusion reactor materials: Semiannual progress report for period ending September 30, 1987

    SciTech Connect

    none,

    1988-03-01

    This is the third in a series of semiannual technical progress reports on fusion reactor materials. This report combines research and development activities which were previously reported separately in the following technical progress reports: Alloy Development for Irradiation Performances; Damage Analysis and Fundamental Studies; Special Purpose Materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials program being conducted in support of the Magnetic Fusion Energy Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. The Fusion Reactor Materials Program is a national effort involving several national laboratories, universities, and industries. The purpose of this series of reports is to provide a working technical record for the use of the program participants, and to provide a means of communicating the efforts of materials scientists to the rest of the fusion community, both nationally and worldwide.

  2. Fusion reactor materials semiannual progress report for period ending September 30, 1990

    SciTech Connect

    Not Available

    1991-04-01

    This is the ninth in series of semiannual technical progress reports on fusion reactor materials. This report combines research and development activities which were previously reported separately in the following technical progress reports: Alloy Development of Irradiation Performance; Damage Analysis and Fundamental Studies; and Special Purpose Materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials program being conducted in support of the Magnetic Fusion Energy Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. The Fusion Reactor Materials Program is a national effort involving several national laboratories, universities, and industries. The purpose of this series of reports is to provide a working technical record for the use of the program participants, and to provide a means of communicating the efforts of materials scientists to the rest of the fusion community, both nationally and worldwide.

  3. Fusion reactor materials semiannual progress report for the period ending September 30, 1989

    SciTech Connect

    none,

    1989-01-01

    This is the seventh in a series of semiannual technical progress reports on fusion reactor materials. This report combines research and development activities which were previously reported separately in the following technical progress reports: alloy development for irradiation performance, damage analysis and fundamental studies, and special purpose materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials program being conducted in support of the Magnetic Fusion Energy Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. The Fusion Reactor Materials Program is a national effort involving several national laboratories, universities, and industries. The purpose of this series of reports is to provide a working technical record for the use of the program participants, and to provide a means of communicating the efforts of materials scientists to the rest of the fusion community, both nationally and worldwide.

  4. Fusion Reactor Materials semiannual progress report for the period ending March 31, 1992

    SciTech Connect

    Not Available

    1992-07-01

    This is the twelfth in a series of semiannual technical progress reports on fusion reactor materials. This report combines research and development activities which were previously reported separately in the following progress reports: Alloy Development for Irradiation Performance; Damage Analysis and Fundamental Studies; and Special Purpose Materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials programs being conducted in support of the Magnetic Fusion Energy Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. The Fusion Reactor Materials Program is a national effort involving several national laboratories, universities, and industries. The purpose of this series of reports is to provide a working technical record for the use of the program participants, and to provide a means of communicating the efforts of materials scientists to the rest of the fusion community, both nationally and worldwide.

  5. Fusion reactor materials semiannual progress report for the period ending March 31, 1991

    SciTech Connect

    none,

    1991-07-01

    This is the tenth in a series of semiannual technical progress reports on fusion reactor materials. This report combines research and development activities which were previously reported separately in the following progress reports: alloy development for irradiation performance; damage analysis and fundamental studies; special purpose materials. These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials program being conducted in support of the Magnetic Fusion Energy Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. The Fusion Reactor Materials Program is a national effort involving several national laboratories, universities, and industries. The purpose of this series of reports is to provide a working technical record for the use of program participants, and to provide a means of communicating the efforts of materials scientists to the test of the fusion community, both nationally and worldwide.

  6. Fusion of the Escherichia coli lac genes to the ara promoter: a general technique using bacteriophage Mu-1 insertions.

    PubMed

    Casadaban, M J

    1975-03-01

    The lac genes were fused to the ara promoter by means of phage phi 80 translocations of the lac and ara genes to att80. Homology for a crossover between the nonhomologous ara and lac operons was provided by mu insertions. Selection for recombinants within the mu insertions generated strains that had the ara promoter on one side of a mu insertion and the lac genes on the other side. ara-lac fusions were obtained from these strains by deleting the mu insertion. These fusions extend the techniques available for studies on the lac operon to studies on the ara operon. It should be possible to fuse other operons by this method.

  7. Pandemic H1N1 influenza A directly induces a robust and acute inflammatory gene signature in primary human bronchial epithelial cells downstream of membrane fusion.

    PubMed

    Paquette, Stéphane G; Banner, David; Chi, Le Thi Bao; Leόn, Alberto J; Xu, Luoling; Ran, Longsi; Huang, Stephen S H; Farooqui, Amber; Kelvin, David J; Kelvin, Alyson A

    2014-01-05

    Pandemic H1N1 influenza A (H1N1pdm) elicits stronger pulmonary inflammation than previously circulating seasonal H1N1 influenza A (sH1N1), yet mechanisms of inflammatory activation in respiratory epithelial cells during H1N1pdm infection are unclear. We investigated host responses to H1N1pdm/sH1N1 infection and virus entry mechanisms in primary human bronchial epithelial cells in vitro. H1N1pdm infection rapidly initiated a robust inflammatory gene signature (3 h post-infection) not elicited by sH1N1 infection. Protein secretion inhibition had no effect on gene induction. Infection with membrane fusion deficient H1N1pdm failed to induce robust inflammatory gene expression which was rescued with restoration of fusion ability, suggesting H1N1pdm directly triggered the inflammatory signature downstream of membrane fusion. Investigation of intra-virion components revealed H1N1pdm viral RNA (vRNA) triggered a stronger inflammatory phenotype than sH1N1 vRNA. Thus, our study is first to report H1N1pdm induces greater inflammatory gene expression than sH1N1 in vitro due to direct virus-epithelial cell interaction.

  8. MODELLING THERMODYNAMICS OF ALLOYS FOR FUSION APPLICATION . Semi annual report for the Fusion Program

    SciTech Connect

    Caro, J A

    2007-07-31

    This research has two main objectives: (1) The development of computational tools to evaluate alloy properties, using the information contained in thermodynamic functions. We aim at improving the ability of classical potentials to account for complex alloy behavior; and (2) The application of these tools to predict properties of alloys under irradiation. Atomistic simulations of alloys at the empirical level face the challenge of correctly modeling basic thermodynamic properties. In the periods reported previously we develop a methodology to generalize many-body classic potentials to incorporate complex formation energy curves. Application to Fe-Cr allows us to predict the implications of the ab initio results of formation energy on the phase diagram of this alloy and to get a detailed insight into the processes leading to precipitation of {alpha}{prime} phase under irradiation. In particular in this period we report on the consequences of the negative heat of formation at low Cr composition on the short range order SRO existing in the {alpha} phase. We elaborate a simple description of SRO on a two phase alloy and compare the predictions with experiments. We provide a key to rationalize a diversity of experiments on SRO versus annealing time or irradiation dose.

  9. Prokaryotic ancestry and gene fusion of a dual localized peroxiredoxin in malaria parasites

    PubMed Central

    Djuika, Carine F.; Huerta-Cepas, Jaime; Przyborski, Jude M.; Deil, Sophia; Sanchez, Cecilia P.; Doerks, Tobias; Bork, Peer; Lanzer, Michael; Deponte, Marcel

    2015-01-01

    Horizontal gene transfer has emerged as a crucial driving force for the evolution of eukaryotes. This also includes Plasmodium falciparum and related economically and clinically relevant apicomplexan parasites, whose rather small genomes have been shaped not only by natural selection in different host populations but also by horizontal gene transfer following endosymbiosis. However, there is rather little reliable data on horizontal gene transfer between animal hosts or bacteria and apicomplexan parasites. Here we show that apicomplexan homologues of peroxiredoxin 5 (Prx5) have a prokaryotic ancestry and therefore represent a special subclass of Prx5 isoforms in eukaryotes. Using two different immunobiochemical approaches, we found that the P. falciparum Prx5 homologue is dually localized to the parasite plastid and cytosol. This dual localization is reflected by a modular Plasmodium-specific gene architecture consisting of two exons. Despite the plastid localization, our phylogenetic analyses contradict an acquisition by secondary endosymbiosis and support a gene fusion event following a horizontal prokaryote-to-eukaryote gene transfer in early apicomplexans. The results provide unexpected insights into the evolution of apicomplexan parasites as well as the molecular evolution of peroxiredoxins, an important family of ubiquitous, usually highly concentrated thiol-dependent hydroperoxidases that exert functions as detoxifying enzymes, redox sensors and chaperones. PMID:28357258

  10. Definition of the ovalbumin gene promoter by transfer of an ovalglobin fusion gene into cultured cells.

    PubMed Central

    Knoll, B J; Zarucki-Schulz, T; Dean, D C; O'Malley, B W

    1983-01-01

    In order to study the initiation of transcription from the ovalbumin gene promoter, we constructed a hybrid gene (ovalglobin) in which 753 bps of ovalbumin gene 5'-flanking sequence were joined to the chicken adult beta-globin gene. When transfected into HeLa S3 cells, ovalglobin gene transcription initiated at the ovalbumin gene cap site, as measured by S1 nuclease and primer extension analysis. Deletion of 5'-flanking sequences to position -95 had little effect on transcription; deletion to -77 reduced transcription to about 20% of the wild type level and deletion to -48 reduced the level to about 2%. A deletion to -24, removing the sequence TATATAT, abolished transcription entirely. Hormonal regulation of the ovalglobin gene was observed when primary oviduct cells were used as recipients for DNA transfection. Under these conditions, addition of progesterone increased the level of ovalglobin transcripts to more than 10 times the uninduced level. Images PMID:6314256

  11. Spatial and temporal analysis of gene expression during growth and fusion of the mouse facial prominences.

    PubMed

    Feng, Weiguo; Leach, Sonia M; Tipney, Hannah; Phang, Tzulip; Geraci, Mark; Spritz, Richard A; Hunter, Lawrence E; Williams, Trevor

    2009-12-16

    Orofacial malformations resulting from genetic and/or environmental causes are frequent human birth defects yet their etiology is often unclear because of insufficient information concerning the molecular, cellular and morphogenetic processes responsible for normal facial development. We have, therefore, derived a comprehensive expression dataset for mouse orofacial development, interrogating three distinct regions - the mandibular, maxillary and frontonasal prominences. To capture the dynamic changes in the transcriptome during face formation, we sampled five time points between E10.5-E12.5, spanning the developmental period from establishment of the prominences to their fusion to form the mature facial platform. Seven independent biological replicates were used for each sample ensuring robustness and quality of the dataset. Here, we provide a general overview of the dataset, characterizing aspects of gene expression changes at both the spatial and temporal level. Considerable coordinate regulation occurs across the three prominences during this period of facial growth and morphogenesis, with a switch from expression of genes involved in cell proliferation to those associated with differentiation. An accompanying shift in the expression of polycomb and trithorax genes presumably maintains appropriate patterns of gene expression in precursor or differentiated cells, respectively. Superimposed on the many coordinated changes are prominence-specific differences in the expression of genes encoding transcription factors, extracellular matrix components, and signaling molecules. Thus, the elaboration of each prominence will be driven by particular combinations of transcription factors coupled with specific cell:cell and cell:matrix interactions. The dataset also reveals several prominence-specific genes not previously associated with orofacial development, a subset of which we externally validate. Several of these latter genes are components of bidirectional

  12. Spatial and Temporal Analysis of Gene Expression during Growth and Fusion of the Mouse Facial Prominences

    PubMed Central

    Feng, Weiguo; Leach, Sonia M.; Tipney, Hannah; Phang, Tzulip; Geraci, Mark; Spritz, Richard A.; Hunter, Lawrence E.; Williams, Trevor

    2009-01-01

    Orofacial malformations resulting from genetic and/or environmental causes are frequent human birth defects yet their etiology is often unclear because of insufficient information concerning the molecular, cellular and morphogenetic processes responsible for normal facial development. We have, therefore, derived a comprehensive expression dataset for mouse orofacial development, interrogating three distinct regions – the mandibular, maxillary and frontonasal prominences. To capture the dynamic changes in the transcriptome during face formation, we sampled five time points between E10.5–E12.5, spanning the developmental period from establishment of the prominences to their fusion to form the mature facial platform. Seven independent biological replicates were used for each sample ensuring robustness and quality of the dataset. Here, we provide a general overview of the dataset, characterizing aspects of gene expression changes at both the spatial and temporal level. Considerable coordinate regulation occurs across the three prominences during this period of facial growth and morphogenesis, with a switch from expression of genes involved in cell proliferation to those associated with differentiation. An accompanying shift in the expression of polycomb and trithorax genes presumably maintains appropriate patterns of gene expression in precursor or differentiated cells, respectively. Superimposed on the many coordinated changes are prominence-specific differences in the expression of genes encoding transcription factors, extracellular matrix components, and signaling molecules. Thus, the elaboration of each prominence will be driven by particular combinations of transcription factors coupled with specific cell:cell and cell:matrix interactions. The dataset also reveals several prominence-specific genes not previously associated with orofacial development, a subset of which we externally validate. Several of these latter genes are components of bidirectional

  13. Institute for Fusion Studies, Final Technical Report, December 1, 1995 - February 29, 2004

    SciTech Connect

    Dr. James Van Dam

    2005-02-14

    During the 2001-2003 grant period, Institute for Fusion Studies (IFS) scientist made notable progress in a number of research areas. This report summarizes the work that has been accomplished in the following areas: (1) Magnetohydrodynamics; (2) Burning plasma and energetic particle physics; (3) Turbulent transport; (4) Computational physics; (5) Fundamental Theory; (6) Innovative confinement concepts; and (7) Plasma applications.

  14. Fusion materials semiannual progress report for the period ending December 31, 1997

    SciTech Connect

    Burn, G.

    1998-03-01

    This is the twenty-third in a series of semiannual technical progress reports on fusion materials. This report combines the full spectrum of research and development activities on both metallic and non-metallic materials with primary emphasis on the effects of the neutronic and chemical environment on the properties and performance of materials for in-vessel components. This effort forms one element of the materials program being conducted in support of the Fusion Energy Sciences Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. The Fusion Materials Program is a national effort involving several national laboratories, universities, and industries. A large fraction of this work, particularly in relation to fission reactor experiments, is carried out collaboratively with their partners in Japan, Russia, and the European Union. The purpose of this series of reports is to provide a working technical record for the use of the program participants, and to provide a means of communicating the efforts of materials scientists to the rest of the fusion community, both nationally and worldwide. Selected papers have been indexed separately for inclusion in the Energy Science and Technology Database.

  15. CONFERENCE REPORT: 13th EU-US Transport Task Force Workshop on transport in fusion plasmas

    NASA Astrophysics Data System (ADS)

    Connor, J. W.; Fasoli, A.; Hidalgo, C.; Kirk, A.; Naulin, V.; Peeters, A. G.; Tala, T.

    2009-04-01

    This report summarizes the contributions presented at the 13th EU-US Transport Task Force Workshop on transport in fusion plasmas, held in Copenhagen, Denmark, 1-4 September 2008. There were sessions on core heat and particle transport; core and edge momentum transport; edge and scrape-off-layer transport and MHD and fast particle interaction with transport.

  16. On the origin of protein synthesis factors: a gene duplication/fusion model.

    PubMed

    Cousineau, B; Leclerc, F; Cedergren, R

    1997-12-01

    Sequence similarity has given rise to the proposal that IF-2, EF-G, and EF-Tu are related through a common ancestor. We evaluate this proposition and whether the relationship can be extended to other factors of protein synthesis. Analysis of amino acid sequence similarity gives statistical support for an evolutionary affiliation among IF-1, IF-2, IF-3, EF-Tu, EF-Ts, and EF-G and suggests further that this association is a result of gene duplication/fusion events. In support of this mechanism, the three-dimensional structures of IF-3, EF-Tu, and EF-G display a predictable domain structure and overall conformational similarity. The model that we propose consists of three consecutives duplication/fusion events which would have taken place before the divergence of the three superkingdoms: eubacteria, archaea, and eukaryotes. The root of this protein superfamily tree would be an ancestor of the modern IF-1 gene sequence. The repeated fundamental motif of this protein superfamily is a small RNA binding domain composed of two alpha-helices packed along side of an antiparallel beta-sheet.

  17. Expression pattern of the RAR alpha-PML fusion gene in acute promyelocytic leukemia.

    PubMed Central

    Alcalay, M; Zangrilli, D; Fagioli, M; Pandolfi, P P; Mencarelli, A; Lo Coco, F; Biondi, A; Grignani, F; Pelicci, P G

    1992-01-01

    Two chimeric genes, PML-RAR alpha and RAR alpha-PML, are formed as a consequence of the acute promyelocytic leukemia (APL)-specific reciprocal translocation of chromosomes 15 and 17 [t(15;17)]. PML-RAR alpha is expressed as a fusion protein. We investigated the organization and expression pattern of the RAR alpha-PML gene in a series of APL patients representative of the molecular heterogeneity of the t(15;17) and found (i) two types of RAR alpha-PML mRNA junctions (RAR alpha exon 2/PML exon 4 or RAR alpha exon 2/PML exon 7) that maintain the RAR alpha and PML longest open reading frames aligned and are the result of chromosome 15 breaking at two different sites; and (ii) 10 different RAR alpha-PML fusion transcripts that differ for the assembly of their PML coding exons. A RAR alpha-PML transcript was present in most, but not all, APL patients. Images PMID:1317574

  18. Horizontal gene transfers and cell fusions in microbiology, immunology and oncology (Review).

    PubMed

    Sinkovics, Joseph G

    2009-09-01

    Evolving young genomes of archaea, prokaryota and unicellular eukaryota were wide open for the acceptance of alien genomic sequences, which they often preserved and vertically transferred to their descendants throughout three billion years of evolution. Established complex large genomes, although seeded with ancestral retroelements, have come to regulate strictly their integrity. However, intruding retroelements, especially the descendents of Ty3/Gypsy, the chromoviruses, continue to find their ways into even the most established genomes. The simian and hominoid-Homo genomes preserved and accommodated a large number of endogenous retroviral genomic segments. These retroelements may mature into exogenous retroviruses, or into functional new genes. Phages and viruses have been instrumental in incorporating and transferring host cell genes. These events profoundly influenced and altered the course of evolution. Horizontal (lateral) gene transfers (HGT) overwhelmed the genomes of the ancient protocells and the evolving unicellular microorganisms, actually leading to their Cambrian explosion. While the rigidly organized genomes of multicellular organisms increasingly resist H/LGT, de-differentiated cells assuming the metabolism of their onto- or phylogenetic ancestors, open up widely to the practice of H/LGT by direct transfer, or to transfers mediated by viruses, or by cell fusions. This activity is intensified in malignantly transformed cells, thus rendering these subjects receptive to therapy with oncolytic viruses and with viral vectors of tumor-suppressive or immunogenic genetic materials. Naturally formed hybrids of dendritic and tumor cells are often tolerogenic, whereas laboratory products of these unisons may be immunogenic in the hosts of origin. As human breast cancer stem cells are induced by a treacherous class of CD8+ T cells to undergo epithelial to mesenchymal (ETM) transition and to yield to malignant transformation by the omnipresent proto

  19. Phase 1 report on sensor technology, data fusion and data interpretation for site characterization

    SciTech Connect

    Beckerman, M.

    1991-10-01

    In this report we discuss sensor technology, data fusion and data interpretation approaches of possible maximal usefulness for subsurface imaging and characterization of land-fill waste sites. Two sensor technologies, terrain conductivity using electromagnetic induction and ground penetrating radar, are described and the literature on the subject is reviewed. We identify the maximum entropy stochastic method as one providing a rigorously justifiable framework for fusing the sensor data, briefly summarize work done by us in this area, and examine some of the outstanding issues with regard to data fusion and interpretation. 25 refs., 17 figs.

  20. Bilateral Supernumerary Deciduous Maxillary Lateral Incisors with Fusion: Report of a Rare Case

    PubMed Central

    Ghaderi, Faezeh; Rafiee, Azade

    2016-01-01

    Dental anomaly in number, size and shape might be due to excessive activation of dental lamina during the morpho-differentiation stage. Fusion is one of the most unusual and rare anomalies of shape of the teeth. It frequently involves the supernumerary teeth resulting in a challenging differential diagnosis with gemination. Dental anomalies may result in different problems such as delayed eruption and crowding; thus, early diagnosis is required for effective intervention and proper in-time treatment. The case reported here is a 4-year-old girl with bilateral supernumerary primary maxillary lateral incisors associated with fusion between primary maxillary left lateral incisor and supernumerary lateral tooth. PMID:26966712

  1. Use of gene fusions to study outer membrane protein localization in Escherichia coli.

    PubMed Central

    Silhavy, T J; Shuman, H A; Beckwith, J; Schwartz, M

    1977-01-01

    Escherichia coli strains have been isolated that produce hybrid proteins comprised of an NH2-terminal sequence from the lamB gene product (an outer membrane protein) and a major portion of the COOH-terminal sequence of beta-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23; a cytoplasmic protein). These proteins exhibit beta-galactosidase activity. One such strain, pop 3105, produces a hybrid protein containing very little of the lamB gene protein; the protein is found in the cytoplasm. The protein found in a second strain, pop 3186, contains much more of the lamB gene protein; a substantial fraction of the beta-galactosidase activity is found in the outer membrane, probably facing outward. These results indicate that information necessary to direct the lamB gene product to its outer membrane location is located within the lamB gene itself. The properties of such fusion strains open up the prospect of a precise genetic analysis of the genetic components involved in protein transport. Images PMID:414221

  2. Variations for susceptibilities to ultraviolet induced cellular inactivation and gene segregation among protoplast fusion hybrids of Candida albicans.

    PubMed

    Sarachek, A; Henderson, L A

    1988-01-01

    Hybrids of the naturally diploid, asexual and opportunistically pathogenic yeast, Candida albicans, can be obtained artificially by protoplast fusion. Evidence is presented that gene conversion and reciprocal recombination contribute to ultraviolet (UV)-induced segregations of heterozygous markers from both diploid and hybrid strains, and that hybrids also segregate through induced chromosome loss. Heterozygous diploid strains independently derived from the same wild-type diploid stock were alike in post-UV survival and segregational responses, and the organization of a four gene linkage group identified in diploids from the segregant products of reciprocal recombinations was transmitted intact to all hybrids from fusions between diploids of isogenic or nonisogenic backgrounds. However, hybrids arising independently from a given fusion cross differed significantly from each other in post-UV survival, absolute ability to segregate some parental markers, frequency of gene segregation, and proclivities for each of the three mechanisms of gene segregation. The bearings of the genetic backgrounds of parental strains and of growth temperatures during hybrid formation on each of these variables are described. The findings emphasize that awareness of the intrinsic heterogeneities of fusion hybrids is essential for reliable application of the protoplast fusion procedure to genetic analysis of C. albicans.

  3. Methods of economic analysis applied to fusion research. Fourth annual report

    SciTech Connect

    Hazelrigg, Jr, G A

    1980-12-31

    The current study reported here has involved three separate tasks. The first task deals with the development of expected utility analysis techniques for economic evaluation of fusion research. A decision analytic model is developed for the incorporation of market uncertainties, as well as technological uncertainties in an economic evaluation of long-range energy research. The model is applied to the case of fusion research. The second task deals with the potential effects of long-range energy RD and D on fossil fuel prices. ECON's previous fossil fuel price model is extended to incorporate a dynamic demand function. The dynamic demand function supports price fluctuations such as those observed in the marketplace. The third task examines alternative uses of fusion technologies, specifically superconducting technologies and first wall materials to determine the potential for alternative, nonfusion use of these technologies. In both cases, numerous alternative uses are found.

  4. Establishment of an Institute for Fusion Studies. Technical progress report, November 1, 1994--October 31, 1995

    SciTech Connect

    1995-07-01

    The Institute for Fusion Studies is a national center for theoretical fusion plasma physics research. Its purposes are to (1) conduct research on theoretical questions concerning the achievement of controlled fusion energy by means of magnetic confinement--including both fundamental problems of long-range significance, as well as shorter-term issues; (2) serve as a national and international center for information exchange by hosting exchange visits, conferences, and workshops; and (3) train students and postdoctoral research personnel for the fusion energy program and plasma physics research areas. During FY 1995, a number of significant scientific advances were achieved at the IFS, both in long-range fundamental problems as well as in near-term strategic issues, consistent with the Institute`s mandate. Examples of these achievements include, for example, tokamak edge physics, analytical and computational studies of ion-temperature-gradient-driven turbulent transport, alpha-particle-excited toroidal Alfven eigenmode nonlinear behavior, sophisticated simulations for the Numerical Tokamak Project, and a variety of non-tokamak and non-fusion basic plasma physics applications. Many of these projects were done in collaboration with scientists from other institutions. Research discoveries are briefly described in this report.

  5. Dysregulated Glycoprotein B-Mediated Cell-Cell Fusion Disrupts Varicella-Zoster Virus and Host Gene Transcription during Infection.

    PubMed

    Oliver, Stefan L; Yang, Edward; Arvin, Ann M

    2017-01-01

    The highly conserved herpesvirus glycoprotein complex gB/gH-gL mediates membrane fusion during virion entry and cell-cell fusion. Varicella-zoster virus (VZV) characteristically forms multinucleated cells, or syncytia, during the infection of human tissues, but little is known about this process. The cytoplasmic domain of VZV gB (gBcyt) has been implicated in cell-cell fusion regulation because a gB[Y881F] substitution causes hyperfusion. gBcyt regulation is necessary for VZV pathogenesis, as the hyperfusogenic mutant gB[Y881F] is severely attenuated in human skin xenografts. In this study, gBcyt-regulated fusion was investigated by comparing melanoma cells infected with wild-type-like VZV or hyperfusogenic mutants. The gB[Y881F] mutant exhibited dramatically accelerated syncytium formation in melanoma cells caused by fusion of infected cells with many uninfected cells, increased cytoskeleton reorganization, and rapid displacement of nuclei to dense central structures compared to pOka using live-cell confocal microscopy. VZV and human transcriptomes were concurrently investigated using whole transcriptome sequencing (RNA-seq) to identify viral and cellular responses induced when gBcyt regulation was disrupted by the gB[Y881F] substitution. The expression of four vital VZV genes, ORF61 and the genes for glycoproteins gC, gE, and gI, was significantly reduced at 36 h postinfection for the hyperfusogenic mutants. Importantly, hierarchical clustering demonstrated an association of differential gene expression with dysregulated gBcyt-mediated fusion. A subset of Ras GTPase genes linked to membrane remodeling were upregulated in cells infected with the hyperfusogenic mutants. These data implicate gBcyt in the regulation of gB fusion function that, if unmodulated, triggers cellular processes leading to hyperfusion that attenuates VZV infection.

  6. Cold fusion, Alchemist's dream

    SciTech Connect

    Clayton, E.D.

    1989-09-01

    In this report the following topics relating to cold fusion are discussed: muon catalysed cold fusion; piezonuclear fusion; sundry explanations pertaining to cold fusion; cosmic ray muon catalysed cold fusion; vibrational mechanisms in excited states of D{sub 2} molecules; barrier penetration probabilities within the hydrogenated metal lattice/piezonuclear fusion; branching ratios of D{sub 2} fusion at low energies; fusion of deuterons into {sup 4}He; secondary D+T fusion within the hydrogenated metal lattice; {sup 3}He to {sup 4}He ratio within the metal lattice; shock induced fusion; and anomalously high isotopic ratios of {sup 3}He/{sup 4}He.

  7. Use of hupS::lacZ gene fusion to study regulation of hydrogenase expression in Rhodobacter capsulatus: stimulation by H2.

    PubMed Central

    Colbeau, A; Vignais, P M

    1992-01-01

    The Escherichia coli beta-galactosidase enzyme was used as a reporter molecule for genetic fusions in Rhodobacter capsulatus. DNA fragments that were from the upstream region of the hydrogenase structural operon hupSLM and contained 5' hupS sequences were fused in frame to a promoterless lacZ gene, yielding fusion proteins comprising the putative signal sequence and the first 22 amino acids of the HupS protein joined to the eight amino acid of beta-galactosidase. We demonstrate the usefulness of the hupS::lacZ fusion in monitoring regulation of hydrogenase gene expression. The activities of plasmid-determined beta-galactosidase and chromosome-encoded hydrogenase changed in parallel in response to various growth conditions (light or dark, aerobiosis or anaerobiosis, and presence or absence of ammonia or of H2), showing that changes in hydrogenase activity were due to changes in enzyme synthesis. Molecular hydrogen stimulated hydrogenase synthesis in dark, aerobic cultures and in illuminated, anaerobic cultures. Analysis of hupS::lacZ expression in various mutants indicated that neither the hydrogenase structural genes nor NifR4 (sigma 54) was essential for hydrogen regulation of hydrogenase synthesis. PMID:1624420

  8. Inertial confinement fusion quarterly report, October-December 1996

    SciTech Connect

    Hammer, J.

    1997-01-01

    The articles in this issue report progress on: Supernova Hydrodynamics Experiments on the Nova Laser; Characterization of Laser-Driven Shock Waves Using Interferometry; Absolute Equation of State Measurements of Compressed Liquid Deuterium Using Nova; Low-Density-Foam Shells; Tetrahedral Hohlraums; The Rosseland Mean Opacity of a Composite Material at High Temperatures.

  9. Identification and assembly of V genes as idiotype-specific DNA fusion vaccines in multiple myeloma.

    PubMed

    Sahota, Surinder S; Townsend, Mark; Stevenson, Freda K

    2005-01-01

    Tumor-specific markers are important in identifying and tracking malignant cells. In this regard, functionally rearranged immunoglobulin variable (V) region genes in B-cell tumors fulfill and extend these criteria. V genes provide signature motifs in tumor cells and can delineate critical features of the clonal history of the cell of origin. They also define a tumor-specific antigen, which can be targeted for immunotherapy. Our focus has been on using novel DNA fusion vaccines to induce antitumor immunity. Here, we describe in detail the methods for identifying tumor-derived V genes at the nucleotide level in the malignant plasma cells of multiple myeloma. We further present the methodology for assembly of tumor V genes as single-chain variable region fragments (scFv), fused in frame with an immunopotentiating nontoxic bacterial sequence, Fragment C (FrC) of tetanus toxin. These scFv.FrC DNA vaccines provide protection in myeloma models and are currently in clinical trials. The vaccines are patient specific and can be rapidly assembled for clinical use.

  10. Analysis of the fusion protein gene of the porcine rubulavirus LPMV: comparative analysis of paramyxovirus F proteins.

    PubMed

    Berg, M; Bergvall, A C; Svenda, M; Sundqvist, A; Moreno-López, J; Linné, T

    1997-01-01

    Complementary DNA clones representing the fusion (F) protein gene of the porcine rubulavirus LPMV were isolated and sequenced. The F gene was found to be 1,845 nucleotides long containing one long open reading frame capable of encoding a protein of 541 amino acids. The cleavage motif for F0 into F1 and F2 is His-Arg-Lys-Lys-Arg. A sequence comparison and a phylogenetic analysis was performed in order to identify possible functional domains of paramyxovirus fusion proteins and also to classify the porcine rubulavirus. The F gene of LPMV is most closely related to the human mumps virus and simian virus type 5 F genes, and is therefore classified into the rubulavirus genus. A coding region for a small hydrophobic protein was however not found between the F and hemagglutinin-neuraminidase (HN) genes as previously found in both SV5 and mumps.

  11. Long Term Non-Invasive Imaging of Embryonic Stem Cells Using Reporter Genes

    PubMed Central

    Sun, Ning; Lee, Andrew; Wu, Joseph C.

    2013-01-01

    Development of non-invasive and accurate methods to track cell fate following delivery will greatly expedite transition of embryonic stem (ES) cell therapy to the clinic. Here we describe a protocol for the in vivo monitoring of stem cell survival, proliferation, and migration using reporter genes. We established stable ES cell lines constitutively expressing double fusion (DF; enhanced green fluorescent protein and firefly luciferase) or triple fusion (TF; monomeric red fluorescent protein, firefly luciferase, and herpes simplex virus thymidine kinase) reporter genes using lentiviral transduction. We used fluorescence activated cell sorting to purify these populations in vitro, bioluminescence imaging and positron emission tomography imaging to track them in vivo, and fluorescence immunostaining to confirm the results ex vivo. Unlike other methods of cell tracking such as iron particle and radionuclide labeling, reporter genes are inherited genetically and can be used to monitor cell proliferation and survival for the lifetime of transplanted cells and their progeny. PMID:19617890

  12. Functional screening of antibiotic resistance genes from human gut microbiota reveals a novel gene fusion.

    PubMed

    Cheng, Gong; Hu, Yongfei; Yin, Yeshi; Yang, Xi; Xiang, Chunsheng; Wang, Baohong; Chen, Yanfei; Yang, Fengling; Lei, Fang; Wu, Na; Lu, Na; Li, Jing; Chen, Quanze; Li, Lanjuan; Zhu, Baoli

    2012-11-01

    The human gut microbiota has a high density of bacteria that are considered a reservoir for antibiotic resistance genes (ARGs). In this study, one fosmid metagenomic library generated from the gut microbiota of four healthy humans was used to screen for ARGs against seven antibiotics. Eight new ARGs were obtained: one against amoxicillin, six against d-cycloserine, and one against kanamycin. The new amoxicillin resistance gene encodes a protein with 53% identity to a class D β-lactamase from Riemerella anatipestifer RA-GD. The six new d-cycloserine resistance genes encode proteins with 73-81% identity to known d-alanine-d-alanine ligases. The new kanamycin resistance gene encodes a protein of 274 amino acids with an N-terminus (amino acids 1-189) that has 42% identity to the 6'-aminoglycoside acetyltransferase [AAC(6')] from Enterococcus hirae and a C-terminus (amino acids 190-274) with 35% identity to a hypothetical protein from Clostridiales sp. SSC/2. A functional study on the novel kanamycin resistance gene showed that only the N-terminus conferred kanamycin resistance. Our results showed that functional metagenomics is a useful tool for the identification of new ARGs.

  13. Ionic solid hydrogen fuel: Experimental investigation of cluster-impact fusion. Final report 1 Jul-30 Sep 90

    SciTech Connect

    Bae, Y.K.; Lorents, D.C.; Young, S.; Staider, K.

    1991-01-24

    This report discusses the investigation of a new novel fusion method: the use of deuterium-containing clusters for igniting fusion (cluster-impact fusion, CIF). If CIF becomes practical, the energy density that can be achieved with the DT fuel is 3.4 X 10 to the 14 power J/kg, which is eight orders of magnitude larger than the energy density of LO{sub X}/H{sub 2} (1,600,000 j/kg). For missions with specific impulse less than 100,000 s, the CIF rocket performance will be essentially identical to that of antimatter. Under the current project, we have successfully built and demonstrated a new cluster-impact fusion facility by extending the existing hydrogen cluster facility. We are confident that the new fusion method discovered at Brookhaven is very promising; thus, we are ready to investigate means of increasing the fusion yield to a practical level for rocket propulsion applications.

  14. Genetic diversity of fusion gene (ORF 117), an analogue of vaccinia virus A27L gene of capripox virus isolates.

    PubMed

    Dashprakash, M; Venkatesan, Gnanavel; Ramakrishnan, Muthannan Andavar; Muthuchelvan, Dhanavelu; Sankar, Muthu; Pandey, Awadh Bihari; Mondal, Bimelendu

    2015-04-01

    The fusion gene (ORF 117) sequences of twelve (n = 12) capripox virus isolates namely sheeppox (SPPV) and goatpox (GTPV) viruses from India were demonstrated for their genetic and phylogenetic relationship among them. All the isolates were confirmed for their identity by routine PCR before targeting ORF 117 gene for sequence analysis. The designed primers specifically amplified ORF 117 gene as 447 bp fragment from total genomic DNA extracted from all the isolates. Sequence analysis revealed a significant percentage of identity among GTPV, SPPV and between them at both nucleotide and amino acid levels. The topology of the phylogenetic tree revealed that three distinct clusters corresponding to SPPV, GTPV and lumpy skin disease virus was formed. However, SPPV Pune/08 and SPPV Roumanian Fanar isolates were clustered into GTPV group as these two isolates showed a 100 and 99.3 % identity with GTPV isolates of India at nt and aa levels, respectively. Protein secondary structure and 3D view was predicted and found that it has high antigenic index and surface probability with low hydrophobicity, and it can be targeted for expression and its evaluation to explore its diagnostic potential in epidemiological investigation in future.

  15. Codon-Optimized NADH Oxidase Gene Expression and Gene Fusion with Glycerol Dehydrogenase for Bienzyme System with Cofactor Regeneration

    PubMed Central

    Zhou, Qiang; Wang, Shizhen

    2015-01-01

    NADH oxidases (NOXs) play an important role in maintaining balance of NAD+/NADH by catalyzing cofactors regeneration. The expression of nox gene from Lactobacillus brevis in Escherichia coli BL21 (BL21 (DE3)) was studied. Two strategies, the high AT-content in the region adjacent to the initiation codon and codon usage of the whole gene sequence consistent with the host, obtained the NOX activity of 59.9 U/mg and 73.3 U/mg (crude enzyme), with enhanced expression level of 2.0 and 2.5-folds, respectively. Purified NOX activity was 213.8 U/mg. Gene fusion of glycerol dehydrogenase (GDH) and NOX formed bifuctional multi-enzymes for bioconversion of glycerol coupled with coenzyme regeneration. Kinetic parameters of the GDH-NOX for each substrate, glycerol and NADH, were calculated as Vmax(Glycerol) 20 μM/min, Km(Glycerol) 19.4 mM, Vmax (NADH) 12.5 μM/min and Km (NADH) 51.3 μM, respectively, which indicated the potential application of GDH-NOX for quick glycerol analysis and dioxyacetone biosynthesis. PMID:26115038

  16. Effects of an adenoviral vector containing a suicide gene fusion on growth characteristics of breast cancer cells.

    PubMed

    Kong, Heng; Liu, Chunli; Zhu, Ting; Huang, Zonghai; Yang, Liucheng; Li, Qiang

    2014-12-01

    The herpes simplex virus thymidine kinase/ganciclovir (HSV‑TK/GCV) and the cytosine deaminase/5‑fluorocytosine (CD/5‑FC) systems have been widely applied in suicide gene therapy for cancer. Although suicide gene therapy has been successfully used in vitro and in vivo studies, the number of studies on the effects of recombinant adenoviruses (Ads) containing suicide genes on target cancer cells is limited. The aim of this study was to examine whether recombinant Ads containing the CD/TK fusion gene affect cell proliferation of breast cancer cells in vitro. In the present study, we explored the use of a recombinant adenoviral vector to deliver the CD/TK fusion gene to the breast cancer cell line MCF‑7. We found that the recombinant adenoviral vector efficiently infected MCF‑7 cells. Western blot analysis revealed that CD and TK proteins are expressed in the infected cells. The infected breast cancer cells did not show any significant changes in morphology, ultrastructure, cell growth, and cell‑cycle distribution compared to the uninfected cells. This study revealed that the Ad‑vascular endothelial growth factor promoter (VEGFp)‑CD/TK vector is non‑toxic to MCF‑7 cells at the appropriate titer. Our results indicate that it is feasible to use a recombinant adenoviral vector containing the CD/TK fusion gene in suicide gene therapy to target breast cancer cells.

  17. Antitumor activities of the targeted multi-tyrosine kinase inhibitor lenvatinib (E7080) against RET gene fusion-driven tumor models.

    PubMed

    Okamoto, Kiyoshi; Kodama, Kotaro; Takase, Kazuma; Sugi, Naoko Hata; Yamamoto, Yuji; Iwata, Masao; Tsuruoka, Akihiko

    2013-10-28

    RET gene fusions are recurrent oncogenes identified in thyroid and lung carcinomas. Lenvatinib is a multi-tyrosine kinase inhibitor currently under evaluation in several clinical trials. Here we evaluated lenvatinib in RET gene fusion-driven preclinical models. In cellular assays, lenvatinib inhibited auto-phosphorylation of KIF5B-RET, CCDC6-RET, and NcoA4-RET. Lenvatinib suppressed the growth of CCDC6-RET human thyroid and lung cancer cell lines, and as well, suppressed anchorage-independent growth and tumorigenicity of RET gene fusion-transformed NIH3T3 cells. These results demonstrate that lenvatinib can exert antitumor activity against RET gene fusion-driven tumor models by inhibiting oncogenic RET gene fusion signaling.

  18. Inducible expression of a fusion gene encoding two proteinase inhibitors leads to insect and pathogen resistance in transgenic rice.

    PubMed

    Quilis, Jordi; López-García, Belén; Meynard, Donaldo; Guiderdoni, Emmanuel; San Segundo, Blanca

    2014-04-01

    Plant proteinase inhibitors (PIs) are considered as candidates for increased insect resistance in transgenic plants. Insect adaptation to PI ingestion might, however, compromise the benefits received by transgenic expression of PIs. In this study, the maize proteinase inhibitor (MPI), an inhibitor of insect serine proteinases, and the potato carboxypeptidase inhibitor (PCI) were fused into a single open reading frame and introduced into rice plants. The two PIs were linked using either the processing site of the Bacillus thuringiensis Cry1B precursor protein or the 2A sequence from the foot-and-mouth disease virus (FMDV). Expression of each fusion gene was driven by the wound- and pathogen-inducible mpi promoter. The mpi-pci fusion gene was stably inherited for at least three generations with no penalty on plant phenotype. An important reduction in larval weight of Chilo suppressalis fed on mpi-pci rice, compared with larvae fed on wild-type plants, was observed. Expression of the mpi-pci fusion gene confers resistance to C. suppressalis (striped stem borer), one of the most important insect pest of rice. The mpi-pci expression systems described may represent a suitable strategy for insect pest control, better than strategies based on the use of single PI genes, by preventing insect adaptive responses. The rice plants expressing the mpi-pci fusion gene also showed enhanced resistance to infection by the fungus Magnaporthe oryzae, the causal agent of the rice blast disease. Our results illustrate the usefulness of the inducible expression of the mpi-pci fusion gene for dual resistance against insects and pathogens in rice plants.

  19. Drosophila Erect wing (Ewg) controls mitochondrial fusion during muscle growth and maintenance by regulation of the Opa1-like gene.

    PubMed

    Rai, Mamta; Katti, Prasanna; Nongthomba, Upendra

    2014-01-01

    Mitochondrial biogenesis and morphological changes are associated with tissue-specific functional demand, but the factors and pathways that regulate these processes have not been completely identified. A lack of mitochondrial fusion has been implicated in various developmental and pathological defects. The spatiotemporal regulation of mitochondrial fusion in a tissue such as muscle is not well understood. Here, we show in Drosophila indirect flight muscles (IFMs) that the nuclear-encoded mitochondrial inner membrane fusion gene, Opa1-like, is regulated in a spatiotemporal fashion by the transcription factor/co-activator Erect wing (Ewg). In IFMs null for Ewg, mitochondria undergo mitophagy and/or autophagy accompanied by reduced mitochondrial functioning and muscle degeneration. By following the dynamics of mitochondrial growth and shape in IFMs, we found that mitochondria grow extensively and fuse during late pupal development to form the large tubular mitochondria. Our evidence shows that Ewg expression during early IFM development is sufficient to upregulate Opa1-like, which itself is a requisite for both late pupal mitochondrial fusion and muscle maintenance. Concomitantly, by knocking down Opa1-like during early muscle development, we show that it is important for mitochondrial fusion, muscle differentiation and muscle organization. However, knocking down Opa1-like, after the expression window of Ewg did not cause mitochondrial or muscle defects. This study identifies a mechanism by which mitochondrial fusion is regulated spatiotemporally by Ewg through Opa1-like during IFM differentiation and growth.

  20. Annual report of the Summit Members' Working Group on Controlled Thermonuclear Fusion (Fusin Working Group (FWG))

    SciTech Connect

    none,

    1987-04-01

    The Summit Members' Working Group on Controlled Thermonuclear Fusion (Fusion Working Group (FWG)) was established in 1983 in response to the Declaration of the Heads of State and Government at the Versailles Economic Summit meeting of 1982, and in response to the subsequent report of the Working Group in Technology, Growth and Employment (TGE) as endorsed at the Williamsburg Summit meeting, 1983. This document contains the complete written record of each of the three FWG meetings which include the minutes, lists of attendees, agendas, statements, and summary conclusions as well as the full reports of the Technical Working Party. In addition, there is a pertinent exchange of correspondence between FWG members on the role of the Technical Working Party and a requested background paper on the modalities associated with a possible future ETR project.

  1. Fusion development and technology. Technical progress report, October 15, 1990--October 14, 1993

    SciTech Connect

    Montgomery, D.B.

    1992-06-01

    This report discusses the following: superconducting magnet technology; high field superconductors; advanced magnetic system and divertor development; poloidal field coils; gyrotron development; commercial reactor studies--aries; ITER physics: alpha physics and alcator R&D for ITER; lower hybrid current drive and heating in the ITER device; ITER superconducting PF scenario and magnet analysis; ITER systems studies; and safety, environmental and economic factors in fusion development.

  2. Inertial Confinement Fusion quarterly report, October--December 1994. Volume 5, No. 1

    SciTech Connect

    1995-09-01

    The ICF quarterly report is published by the Inertial Confinement Fusion Program at the Lawrence Livermore National Laboratory. Topics included in this issue include: system description and initial performance results for beamlet, design and performance of the beamlet amplifiers and optical switch, beamlet pulse-generation and wavefront-control system, large-aperture, high- damage-threshold optics for beamlet, beamlet pulsed power system, beamlet laser diagnostics, and beam propagation and frequency conversion modeling for the beamlet laser.

  3. Inertial confinement fusion quarterly report, October--December 1992. Volume 3, No. 1

    SciTech Connect

    Dixit, S.N.

    1992-12-31

    This report contains papers on the following topics: The Beamlet Front End: Prototype of a new pulse generation system;imaging biological objects with x-ray lasers; coherent XUV generation via high-order harmonic generation in rare gases; theory of high-order harmonic generation; two-dimensional computer simulations of ultra- intense, short-pulse laser-plasma interactions; neutron detectors for measuring the fusion burn history of ICF targets; the recirculator; and lasnex evolves to exploit computer industry advances.

  4. Low-density hydrocarbon foams for laser fusion targets: Progress report, 1987

    SciTech Connect

    Haendler, B.L.; Buckley, S.R.; Chen, C.; Cook, A.R.; Cook, R.C.; Hair, L.M.; Kong, F.M.; Kramer, H.D.; Letts, S.A.; Overturf, G.E. III

    1988-06-01

    This report describes progress made in the development of direct-drive hydrocarbon foam targets for laser inertial confinement fusion during 1987. The foam materials are polystyrene, resorcinol-formaldehyde, carbonized resorcinol-formaldehyde, and cellulose acetate. The processes for making the foams, their properties, characterization techniques, and the relationship of their properties to target specifications are presented. Progress in the creation and testing of prototype targets is also described.

  5. Overexpression of Recombinant Human Teriparatide, rhPTH (1–34) in Escherichia coli : An Innovative Gene Fusion Approach

    PubMed Central

    Bakhtiari, Nahid; Amini Bayat, Zahra; Sagharidouz, Sepideh; Vaez, Mohsen

    2017-01-01

    Background: Parathyroid hormone is an 84-amino acid peptide secreted by the parathyroid glands. Its physiological role is maintenance of normal serum calcium level and bone remodeling. Biological activity of this hormone is related to N-terminal 1–34 amino acids. The recombinant form of hormone (1–34) has been approved for treatment of osteoporosis from 2002. In this study, a novel fusion partner has been developed for preparation of high yield recombinant 1–34 amino acids of hPTH. Methods: Novel nucleotide cassette designed encoding a chimeric fusion protein comprising of a fusion partner consisting of a His-tag in N-terminal, 53 amino acids belong to Escherichia coli (E. coli) β-galactosidase (LacZ) gene, a linker sequence for increasing of expression and protection of target peptide structure from fusion tag effect, an Enteropeptidase cleavage site, rhPTH (1–34) gene fragment. Optimized fusion gene was synthesized and ligated into pET-28a vector under control of T7 promoter, and then transformed in E. coli (DH5α) cells. Positive clones containing this gene were double digested with NcoI and-BamHI and also approved by sequencing. Gene overexpression was observed in SDS-PAGE after induction with 0.2 mM IPTG. Confirmation of gene expression was performed by western blotting using anti-His-tag antibody conjugated with peroxidase. Results: By this fusion gene design approach, we achieved a high level expression of the rhPTH, where it represented at least 43.7% of the total protein as determined by SDS-PAGE and confirmed by western blotting. Conclusion: In addition to high level expression of the designed gene in this work, specific amino acid sequence of bacterial β-galactosidase was selected as major part of carrier tag for protection of this hormone as important step of recombinant rhPTH with relevant isoelectronic point (pI). This innovation resulted in recombinant production of hPTH very well and the gene construct could be applied as a pattern for

  6. Pandemic H1N1 influenza A directly induces a robust and acute inflammatory gene signature in primary human bronchial epithelial cells downstream of membrane fusion

    SciTech Connect

    Paquette, Stéphane G.; Banner, David; Chi, Le Thi Bao; Leon, Alberto J.; Xu, Luoling; Ran, Longsi; Huang, Stephen S.H.; Farooqui, Amber; and others

    2014-01-05

    Pandemic H1N1 influenza A (H1N1pdm) elicits stronger pulmonary inflammation than previously circulating seasonal H1N1 influenza A (sH1N1), yet mechanisms of inflammatory activation in respiratory epithelial cells during H1N1pdm infection are unclear. We investigated host responses to H1N1pdm/sH1N1 infection and virus entry mechanisms in primary human bronchial epithelial cells in vitro. H1N1pdm infection rapidly initiated a robust inflammatory gene signature (3 h post-infection) not elicited by sH1N1 infection. Protein secretion inhibition had no effect on gene induction. Infection with membrane fusion deficient H1N1pdm failed to induce robust inflammatory gene expression which was rescued with restoration of fusion ability, suggesting H1N1pdm directly triggered the inflammatory signature downstream of membrane fusion. Investigation of intra-virion components revealed H1N1pdm viral RNA (vRNA) triggered a stronger inflammatory phenotype than sH1N1 vRNA. Thus, our study is first to report H1N1pdm induces greater inflammatory gene expression than sH1N1 in vitro due to direct virus–epithelial cell interaction. - Highlights: • We investigated H1N1pdm/sH1N1 infection in primary epithelial cells. • H1N1pdm directly initiated a robust inflammatory gene signature, sH1N1 did not. • H1N1pdm viral RNA triggered a stronger response than sH1N1. • H1N1pdm induces greater response due to direct virus–cell interaction. • These results have potential to impact vaccine and therapeutic development.

  7. LAMTOR1-PRKCD and NUMA1-SFMBT1 fusion genes identified by RNA sequencing in aneurysmal benign fibrous histiocytoma with t(3;11)(p21;q13).

    PubMed

    Panagopoulos, Ioannis; Gorunova, Ludmila; Bjerkehagen, Bodil; Lobmaier, Ingvild; Heim, Sverre

    2015-11-01

    RNA sequencing of an aneurysmal benign fibrous histiocytoma with the karyotype 46,XY,t(3;11)(p21;q13),del(6)(p23)[17]/46,XY[2] showed that the t(3;11) generated two fusion genes: LAMTOR1-PRKCD and NUMA1-SFMBT1. RT-PCR together with Sanger sequencing verified the presence of fusion transcripts from both fusion genes. In the LAMTOR1-PRKCD fusion, the part of the PRKCD gene coding for the catalytic domain of the serine/threonine kinase is under control of the LAMTOR1 promoter. In the NUMA1-SFMBT1 fusion, the part of the SFMBT1 gene coding for two of four malignant brain tumor domains and the sterile alpha motif domain is controlled by the NUMA1 promoter. The data support a neoplastic genesis of aneurysmal benign fibrous histiocytoma and indicate a pathogenetic role for LAMTOR1-PRKCD and NUMA1-SFMBT1.

  8. EWS-Oct-4B, an alternative EWS-Oct-4 fusion gene, is a potent oncogene linked to human epithelial tumours

    PubMed Central

    Kim, S; Lim, B; Kim, J

    2010-01-01

    Background: Characterisation of EWS-Oct-4 translocation fusion product in bone and soft-tissue tumours revealed a chimeric gene resulting from an in-frame fusion between EWS (Ewing's sarcoma gene) exons 1–6 and Oct-4 exons 1–4. Recently, an alternative form of the fusion protein between the EWS and Oct-4 genes, named EWS-Oct-4B, was reported in two types of epithelial tumours, a hidradenoma of the skin and a mucoepidermoid carcinoma of the salivary glands. As the N-terminal and POU domains of the EWS-Oct-4 and EWS-Oct-4B proteins are not structurally identical, we decided to investigate the functional consequences of the EWS-Oct-4B fusion. Methods: In this report, we have characterised the EWS-Oct-4B fusion protein. To investigate how the EWS-Oct-4B protein contributes to tumourigenesis in human cancers, we analysed its DNA-binding activity, subcellular localisation, transcriptional activation behaviour, and oncogenic properties. Results: We found that this new chimeric gene encodes a nuclear protein that binds DNA with the same sequence specificity as the parental Oct-4 protein or the fusion EWS-Oct-4 protein. We show that the nuclear localisation signal of EWS-Oct-4B is dependent on the POU DNA-binding domain, and we identified a cluster of basic amino acids, 269RKRKR273, in the POU domain that specifically mediates the nuclear localisation of EWS-Oct-4B. Comparison of the properties of EWS-Oct-4B and EWS-Oct-4 indicated that EWS-Oct-4B is a less-potent transcriptional activator of a reporter construct carrying the Oct-4-binding sites. Deletion analysis of the functional domains of EWS-Oct-4B revealed that the EWS N-terminal domain (NTD)B, POU, and C-terminal domain (CTD) are necessary for its full transactivation potential. Despite its reduced activity as a transcriptional activator, EWS-Oct-4B regulated the expression of fgf-4 (fibroblast growth factor-4) and nanog, which are potent mitogens, as well as of Oct-4 downstream target genes, the promoters of

  9. Code development incorporating environmental, safety, and economic aspects of fusion reactors (FY 89--91). Final report

    SciTech Connect

    Ho, S.K.; Fowler, T.K.; Holdren, J.P.

    1991-11-01

    This report discusses the following aspects of Fusion reactors.: Activation Analysis; Tritium Inventory; Environmental and Safety Indices and Their Graphical Representation; Probabilistic Risk Assessment (PRA) and Decision Analysis; Plasma Burn Control -- Application to ITER; and Other Applications.

  10. Laser-Fusion Studies at NRL. A Report to AEC for the Period July 1973 to June 1974,

    DTIC Science & Technology

    This report summarizes work done at NRL on laser fusion related problems during FY 1974. Included are sections related to laser development , and to experimental and theoretical studies of laser plasma interaction.

  11. Laser-Fusion Studies at NRL. A Report to ERDA for the Period July 1974 to June 1975.

    DTIC Science & Technology

    This report summarizes work done at NRL on laser fusion related to problems during FY 1975. Included are sections related to laser development and to experimental studies of laser plasma interaction. (Author)

  12. Long-Term Endurance Exercise in Humans Stimulates Cell Fusion of Myoblasts along with Fusogenic Endogenous Retroviral Genes In Vivo

    PubMed Central

    Suhr, Frank; Konou, Thierry M.; Tappe, Kim A.; Toigo, Marco; Jung, Hans H.; Henke, Christine; Steigleder, Ruth; Strissel, Pamela L.; Huebner, Hanna; Beckmann, Matthias W.; van der Keylen, Piet; Schoser, Benedikt; Schiffer, Thorsten; Frese, Laura; Bloch, Wilhelm; Strick, Reiner

    2015-01-01

    Myogenesis is defined as growth, differentiation and repair of muscles where cell fusion of myoblasts to multinucleated myofibers is one major characteristic. Other cell fusion events in humans are found with bone resorbing osteoclasts and placental syncytiotrophoblasts. No unifying gene regulation for natural cell fusions has been found. We analyzed skeletal muscle biopsies of competitive cyclists for muscle-specific attributes and expression of human endogenous retrovirus (ERV) envelope genes due to their involvement in cell fusion of osteoclasts and syncytiotrophoblasts. Comparing muscle biopsies from post- with the pre-competitive seasons a significant 2.25-fold increase of myonuclei/mm fiber, a 2.38-fold decrease of fiber area/nucleus and a 3.1-fold decrease of satellite cells (SCs) occurred. We propose that during the pre-competitive season SC proliferation occurred following with increased cell fusion during the competitive season. Expression of twenty-two envelope genes of muscle biopsies demonstrated a significant increase of putative muscle-cell fusogenic genes Syncytin-1 and Syncytin-3, but also for the non-fusogenic erv3. Immunohistochemistry analyses showed that Syncytin-1 mainly localized to the sarcolemma of myofibers positive for myosin heavy-chain isotypes. Cellular receptors SLC1A4 and SLC1A5 of Syncytin-1 showed significant decrease of expression in post-competitive muscles compared with the pre-competitive season, but only SLC1A4 protein expression localized throughout the myofiber. Erv3 protein was strongly expressed throughout the myofiber, whereas envK1-7 localized to SC nuclei and myonuclei. Syncytin-1 transcription factors, PPARγ and RXRα, showed no protein expression in the myofiber, whereas the pCREB-Ser133 activator of Syncytin-1 was enriched to SC nuclei and myonuclei. Syncytin-1, Syncytin-3, SLC1A4 and PAX7 gene regulations along with MyoD1 and myogenin were verified during proliferating or actively-fusing human primary myoblast cell

  13. Long-Term Endurance Exercise in Humans Stimulates Cell Fusion of Myoblasts along with Fusogenic Endogenous Retroviral Genes In Vivo.

    PubMed

    Frese, Sebastian; Ruebner, Matthias; Suhr, Frank; Konou, Thierry M; Tappe, Kim A; Toigo, Marco; Jung, Hans H; Henke, Christine; Steigleder, Ruth; Strissel, Pamela L; Huebner, Hanna; Beckmann, Matthias W; van der Keylen, Piet; Schoser, Benedikt; Schiffer, Thorsten; Frese, Laura; Bloch, Wilhelm; Strick, Reiner

    2015-01-01

    Myogenesis is defined as growth, differentiation and repair of muscles where cell fusion of myoblasts to multinucleated myofibers is one major characteristic. Other cell fusion events in humans are found with bone resorbing osteoclasts and placental syncytiotrophoblasts. No unifying gene regulation for natural cell fusions has been found. We analyzed skeletal muscle biopsies of competitive cyclists for muscle-specific attributes and expression of human endogenous retrovirus (ERV) envelope genes due to their involvement in cell fusion of osteoclasts and syncytiotrophoblasts. Comparing muscle biopsies from post- with the pre-competitive seasons a significant 2.25-fold increase of myonuclei/mm fiber, a 2.38-fold decrease of fiber area/nucleus and a 3.1-fold decrease of satellite cells (SCs) occurred. We propose that during the pre-competitive season SC proliferation occurred following with increased cell fusion during the competitive season. Expression of twenty-two envelope genes of muscle biopsies demonstrated a significant increase of putative muscle-cell fusogenic genes Syncytin-1 and Syncytin-3, but also for the non-fusogenic erv3. Immunohistochemistry analyses showed that Syncytin-1 mainly localized to the sarcolemma of myofibers positive for myosin heavy-chain isotypes. Cellular receptors SLC1A4 and SLC1A5 of Syncytin-1 showed significant decrease of expression in post-competitive muscles compared with the pre-competitive season, but only SLC1A4 protein expression localized throughout the myofiber. Erv3 protein was strongly expressed throughout the myofiber, whereas envK1-7 localized to SC nuclei and myonuclei. Syncytin-1 transcription factors, PPARγ and RXRα, showed no protein expression in the myofiber, whereas the pCREB-Ser133 activator of Syncytin-1 was enriched to SC nuclei and myonuclei. Syncytin-1, Syncytin-3, SLC1A4 and PAX7 gene regulations along with MyoD1 and myogenin were verified during proliferating or actively-fusing human primary myoblast cell

  14. A novel type of EWS-CHOP fusion gene in myxoid liposarcoma

    SciTech Connect

    Matsui, Yoshito . E-mail: ymatsui@sb4.so-net.ne.jp; Ueda, Takafumi; Kubo, Takahiro; Hasegawa, Tadashi; Tomita, Yasuhiko; Okamoto, Mina; Myoui, Akira; Kakunaga, Shigeki; Yasui, Natsuo; Yoshikawa, Hideki

    2006-09-22

    The cytogenetic hallmark of myxoid type and round cell type liposarcoma consists of reciprocal translocation of t(12;16)(q13;p11) and t(12;22)(q13;q12), which results in fusion of TLS/FUS and CHOP, and EWS and CHOP, respectively. Nine structural variations of the TLS/FUS-CHOP chimeric transcript have been reported, however, only two types of EWS-CHOP have been described. We describe here a case of myxoid liposarcoma containing a novel EWS-CHOP chimeric transcript and identified the breakpoint occurring in intron 13 of EWS. Reverse transcription-polymerase chain reaction and direct sequence showed that exon 13 of EWS was in-frame fused to exon 2 of CHOP. Genomic analysis revealed that the breaks were located in intron 13 of EWS and intron 1 of CHOP.

  15. Microwave generation for magnetic fusion energy applications. Progress report, September 15, 1991--July 15, 1992

    SciTech Connect

    Antonsen, T.M. Jr.; Destler, W.W.; Granatstein, V.L.; Levush, B.

    1992-01-01

    This progress report encompasses work on two separate projects, both related to developing sources for electron cyclotron resonance heating of magnetic fusion plasmas. The report is therefore divided into two parts as follows: Free electron laser with small period wigglers; and theory and modeling of high frequency, high power gryotron operation. Task A is experimental and eventually aims at developing continuously tunable, cw sources for ECRH with power per unit as high as 5 megawatts. Task B provides gryotron theory and modeling in support of the gryotron development programs at MIT and Varian.

  16. Fusion reactor materials: Semiannual progress report for period ending September 30, 1986

    SciTech Connect

    none,

    1987-09-01

    These activities are concerned principally with the effects of the neutronic and chemical environment on the properties and performance of reactor materials; together they form one element of the overall materials program being conducted in support of the Magnetic Fusion Energy Program of the US Department of Energy. The major areas of concern covered in this report are irradiation facilities, test matrices, and experimental methods; dosimetry, damage parameters and activation calculations; materials engineering and design requirements; radiation effects; development of structural alloys; solid breeding materials; ceramics and superconducting magnet materials. There are 61 reports cataloged separately. (LSP)

  17. Differential transactivation by orphan nuclear receptor NOR1 and its fusion gene product EWS/NOR1: possible involvement of poly(ADP-ribose) polymerase I, PARP-1.

    PubMed

    Ohkura, Naganari; Nagamura, Yuko; Tsukada, Toshihiko

    2008-10-15

    In extraskeletal myxoid chondrosarcoma, a chromosomal translocation creates a gene fusion between EWS and an orphan nuclear receptor, NOR1. The resulting fusion protein EWS/NOR1 has been believed to lead to malignant transformation by functioning as a transactivator for NOR1-target genes. By comparing the gene expression profiles of NOR1- and EWS/NOR1-overexpressing cells, we found that they largely shared up-regulated genes, but no significant correlation was observed with respect to the transactivation levels of each gene. In addition, the proteins associated with NOR1 and EWS/NOR1 were mostly the same in these cells. The results suggest that these proteins differentially transactivate overlapping target genes through a similar transcriptional machinery. To clarify the mechanisms underlying the transcriptional divergence between NOR1 and EWS/NOR1, we searched for alternatively associated proteins, and identified poly(ADP-ribose) polymerase I (PARP-1) as an NOR1-specific binding protein. Consistent with its binding properties, PARP-1 acted as a transcriptional repressor of NOR1, but not EWS/NOR1, in a luciferase reporter assay employing PARP-1(-/-) fibroblasts. Interestingly, suppressive activity of PARP-1 was observed in a DNA response element-specific manner, and in a subtype-specific manner toward the NR4A family (Nur77, Nurr1, and NOR1), suggesting that PARP-1 plays a role in the diversity of transcriptional regulation mediated by the NR4A family in normal cells. Altogether, our findings suggest that NOR1 and EWS/NOR1 regulate overlapping target genes differently by utilizing associated proteins, including PARP-1; and that EWS/NOR1 may acquire oncogenic activities by avoiding (or gaining) transcription factor-specific modulation by the associated proteins.

  18. Modified mariner transposons for random inducible-expression insertions and transcriptional reporter fusion insertions in Bacillus subtilis.

    PubMed

    Pozsgai, Eric R; Blair, Kris M; Kearns, Daniel B

    2012-02-01

    Transposons are mobile genetic elements bounded by insertion sequences that are recognized by a specific mobilizing transposase enzyme. The transposase may mobilize not only the insertion sequences but also intervening DNA. mariner is a particularly efficient transposon for the random chromosomal integration of genes and insertional mutagenesis. Here, we modify an existing mariner transposon, TnYLB, such that it can easily be genetically manipulated and introduced into Bacillus subtilis. We generate a series of three new mariner derivatives that mobilize spectinomycin, chloramphenicol, and kanamycin antibiotic resistance cassettes. Furthermore, we generate a series of transposons with a strong, outward-oriented, optionally isopropyl-β-D-thiogalactopyranoside (IPTG)-inducible promoter for the random overexpression of neighboring genes and a series of transposons with a promoterless lacZ gene for the random generation of transcriptional reporter fusions. We note that the modification of the base transposon is not restricted to B. subtilis and should be applicable to any mariner-compatible host organism, provided that in vitro mutagenesis or an in vivo species-specific delivery vector is employed.

  19. The TITAN Reversed-Field Pinch fusion reactor study: Scoping phase report

    SciTech Connect

    Not Available

    1987-01-01

    The TITAN research program is a multi-institutional effort to determine the potential of the Reversed-Field Pinch (RFP) magnetic fusion concept as a compact, high-power-density, and ''attractive'' fusion energy system from economic (cost of electricity, COE), environmental, and operational viewpoints. In particular, a high neutron wall loading design (18 MW/m/sup 2/) has been chosen as the reference case in order to quantify the issue of engineering practicality, to determine the physics requirements and plasma operating mode, to assess significant benefits of compact systems, and to illuminate the main drawbacks. The program has been divided into two phases, each roughly one year in length: the Scoping Phase and the Design Phase. During the scoping phase, the TITAN design team has defined the parameter space for a high mass power density (MPD) RFP reactor, and explored a variety of approaches to the design of major subsystems. Two major design approaches consistent with high MPD and low COE, the lithium-vanadium blanket design and aqueous loop-in-pool design, have been selected for more detailed engineering evaluation in the design phase. The program has retained a balance in its approach to investigating high MPD systems. On the one hand, parametric investigations of both subsystems and overall system performance are carried out. On the other hand, more detailed analysis and engineering design and integration are performed, appropriate to determining the technical feasibility of the high MPD approach to RFP fusion reactors. This report describes the work of the scoping phase activities of the TITAN program. A synopsis of the principal technical findings and a brief description of the TITAN multiple-design approach is given. The individual chapters on Plasma Physics and Engineering, Parameter Systems Studies, Divertor, Reactor Engineering, and Fusion Power Core Engineering have been cataloged separately.

  20. Accelerator and Fusion Research Division annual report, October 1981-September 1982. Fiscal year 1982

    SciTech Connect

    Johnson, R.K.; Bouret, C.

    1983-05-01

    This report covers the activities of LBL's Accelerator and Fusion Research Division (AFRD) during 1982. In nuclear physics, the Uranium Beams Improvement Project was concluded early in the year, and experimentation to exploit the new capabilities began in earnest. Technical improvement of the Bevalac during the year centered on a heavy-ion radiofrequency quadrupole (RFQ) as part of the local injector upgrade, and we collaborated in studies of high-energy heavy-ion collision facilities. The Division continued its collaboration with Fermilab to design a beam-cooling system for the Tevatron I proton-antiprotron collider and to engineer the needed cooling components for the antiproton. The high-field magnet program set yet another record for field strength in an accelerator-type dipole magnet (9.2 T at 1.8 K). The Division developed the design for the Advanced Light Source (ALS), a 1.3-GeV electron storage ring designed explicitly (with low beam emittance and 12 long straight sections) to generate high-brilliance synchrotron light from insertion devices. The Division's Magnetic Fusion Energy group continued to support major experiments at the Princeton Plasma Physics Laboratory, the Lawrence Livermore National Laboratory (LLNL), and General Atomic Co. by developing positive-ion-based neutral-beam injectors. Progress was made toward converting our major source-test facility into a long-pulse national facility, the Neutral Beam Engineering Test Facility, which was completed on schedule and within budget in 1983. Heavy Ion Fusion research focused on planning, theoretical studies, and beam-transport experiments leading toward a High Temperature Experiment - a major test of this promising backup approach to fusion energy.

  1. THE GENERAL ATOMICS FUSION THEORY PROGRAM ANNUAL REPORT FOR GRANT YEAR 2004

    SciTech Connect

    PROJECT STAFF

    2004-12-01

    The dual objective of the fusion theory program at General Atomics (GA) is to significantly advance our scientific understanding of the physics of fusion plasmas and to support the DIII-D and other tokamak experiments. The program plan is aimed at contributing significantly to the Fusion Energy Science and the Tokamak Concept Improvement goals of the Office of Fusion Energy Sciences (OFES).

  2. Fusion materials semiannual progress report for the period ending December 31, 1996

    SciTech Connect

    1997-04-01

    This is the twenty-first in a series of semiannual technical progress reports on fusion materials. This report combines the full spectrum of research and development activities on both metallic and non-metallic materials with primary emphasis on the effects of the neutronic and chemical environment on the properties and performance of materials for in-vessel components. This effort forms one element of the materials program being conducted in support of the Fusion Energy Sciences Program of the US Department of Energy. The other major element of the program is concerned with the interactions between reactor materials and the plasma and is reported separately. The report covers the following topics: vanadium alloys; silicon carbide composite materials; ferritic/martensitic steels; copper alloys and high heat flux materials; austenitic stainless steels; insulating ceramics and optical materials; solid breeding materials; radiation effects, mechanistic studies and experimental methods; dosimetry, damage parameters, and activation calculations; materials engineering and design requirements; and irradiation facilities, test matrices, and experimental methods.

  3. Importance of NAB2-STAT6 Fusion in the Diagnosis of Pancreatic Solitary Fibrous Tumor with Hamartoma-Like Features: A Case Report and Review of the Literature

    PubMed Central

    Tanaka, Kei; Kishimoto, Takashi; Ohtsuka, Masayuki; Nakatani, Yukio; Miyazaki, Masaru

    2015-01-01

    We report a case of pancreatic hamartoma-like solitary fibrous tumor which was differentiated from pancreatic hamartoma with the detection of NAB2-STAT6 fusion, a specific mutation for solitary fibrous tumors. A pancreatic well-demarcated solid nodule, 21 × 17 mm, of 82-year-old man was surgically enucleated. Microscopic findings were close to a pancreatic hamartoma that consisted of sparsely distributed pancreatic ducts and acini in heavily collagenized fibrous stroma. Neither islet nor peripheral nerve existed in the tumor. The fibroblastic cells in the stroma were immune-positive for CD34, CD99, and bcl-2. But these expressions were not decisive in the differentiation between solitary fibrous tumor and pancreatic hamartoma, because CD34 was positive for both tumors, and CD99 and bcl-2 expressions were not elucidated in the previous cases of pancreatic hamartomas. Thus, we evaluated NAB2-STAT6 fusion. The fibroblastic cells were positive for STAT6 and sequencing analysis revealed the gene fusion between NAB2 exon 4 and STAT6 exon 2, with which the final diagnos is of solitary fibrous tumor was achieved. In conclusion, detection of NAB2-STAT6 fusion has a great diagnostic value for pancreatic solitary fibrous tumors with hamartoma-like features. PMID:26425382

  4. A Naturally Occurring rev1-vpu Fusion Gene Does Not Confer a Fitness Advantage to HIV-1

    PubMed Central

    Langer, Simon M.; Hopfensperger, Kristina; Iyer, Shilpa S.; Kreider, Edward F.; Learn, Gerald H.; Lee, Lan-Hui; Hahn, Beatrice H.; Sauter, Daniel

    2015-01-01

    Background Pandemic strains of HIV-1 (group M) encode a total of nine structural (gag, pol, env), regulatory (rev, tat) and accessory (vif, vpr, vpu, nef) genes. However, some subtype A and C viruses exhibit an unusual gene arrangement in which the first exon of rev (rev1) and the vpu gene are placed in the same open reading frame. Although this rev1-vpu gene fusion is present in a considerable fraction of HIV-1 strains, its functional significance is unknown. Results Examining infectious molecular clones (IMCs) of HIV-1 that encode the rev1-vpu polymorphism, we show that a fusion protein is expressed in infected cells. Due to the splicing pattern of viral mRNA, however, these same IMCs also express a regular Vpu protein, which is produced at much higher levels. To investigate the function of the fusion gene, we characterized isogenic IMC pairs differing only in their ability to express a Rev1-Vpu protein. Analysis in transfected HEK293T and infected CD4+ T cells showed that all of these viruses were equally active in known Vpu functions, such as down-modulation of CD4 or counteraction of tetherin. Furthermore, the polymorphism did not affect Vpu-mediated inhibition of NF-кB activation or Rev-dependent nuclear export of incompletely spliced viral mRNAs. There was also no evidence for enhanced replication of Rev1-Vpu expressing viruses in primary PBMCs or ex vivo infected human lymphoid tissues. Finally, the frequency of HIV-1 quasispecies members that encoded a rev1-vpu fusion gene did not change in HIV-1 infected individuals over time. Conclusions Expression of a rev1-vpu fusion gene does not affect regular Rev and Vpu functions or alter HIV-1 replication in primary target cells. Since there is no evidence for increased replication fitness of rev1-vpu encoding viruses, this polymorphism likely emerged in the context of other mutations within and/or outside the rev1-vpu intergenic region, and may have a neutral phenotype. PMID:26554585

  5. Final Report: Establishment of an Institute for Fusion Studies, June 1, 1980 - March 1, 1998

    SciTech Connect

    Hazeltine, Richard D.

    1999-12-03

    The mission of the Institute for Fusion Studies has been to serve as a national center for theoretical fusion and plasma physics research. As an independent scientific group of critical size, its objectives were to conduct research on fundamental phenomena important to fusion; to serve as a center for fusion theory exchange activities with other countries; to exchange scientific developments with other academic disciplines; and to train students and postdoctoral fellows in fusion and plasma physics research.

  6. Cloning and characterization of the gene encoding human NPL4, a protein interacting with the ubiquitin fusion-degradation protein (UFD1L).

    PubMed

    Botta, A; Tandoi, C; Fini, G; Calabrese, G; Dallapiccola, B; Novelli, G

    2001-09-05

    The ubiquitin fusion-degradation gene (UFD1L) encodes the human homologue of the yeast ubiquitin fusion-degradation 1 protein, an essential component of the ubiquitin-dependent proteolytic turnover and mRNA processing. Although the UFD1L gene has been mapped in the region commonly deleted in patients with DiGeorge syndrome (DGS)/velocardiofacial syndrome (VCFS), correlation between its haploinsufficiency and the phenotype has not yet been established. The only functional data available about mammalian Ufd1p is the ability to form a complex with the rat Npl4 protein, a component of the nuclear pore complex. In this paper we report the cloning and molecular characterization of the human NPL4 gene. This gene encodes for a protein 96% homologous to the rat Npl4, and 44 and 34% homologous to the C. elegans and S. cerevisiae Npl4 gene products, respectively. Fluorescence in situ hybridization experiments on human metaphases localized the NPL4 gene on the most telomeric region of chromosome 17q. Northern blots analysis on foetal and adult human tissues revealed a major approximately 4.5 kb transcript most abundant in heart, brain, kidney and skeletal muscle. In order to test a potential relationship between nuclear transport defects and some aspect of the DGS/VCFS phenotype, we also exclude the presence of mutations in the NPL4 coding sequence in a subset of patients with DGS/VCFS and no detectable 22q11 deletion or mutations at the UFD1L locus.

  7. Bony fusion of the maxilla and mandible as a sequelae of noma: A rare case report

    PubMed Central

    Awasthi, Ujjwala Rastogi; Mody, Bharat M.; Suma, Gundareddy N.; Garg, Shruti

    2015-01-01

    Noma is a gangrenous disease of the orofacial region that leads to severe facial tissue destruction and is a significant cause of death among children. With the advent of modern antibiotics and improved nutrition, children with noma may survive into adulthood, but must face the challenge of undergoing repair of the sequelae of noma. This report describes a case of bony fusion of the maxilla and mandible in a 28-year-old female patient, which was a sequelae of a childhood case of noma. PMID:26389063

  8. Heavy Ion Fusion Accelerator Research (HIFAR) year-end report, April 1, 1989--September 30, 1989

    SciTech Connect

    Not Available

    1989-12-01

    This report contains the following topics on heavy ion fusion: MBE-4 drifting beam quadrupole operating range; transverse emittance growth in MBE-4; an improved ion source for MBE-4; drifting beam studies on MBE-4; 2-MV injector; improvements in lifetime of the C{sup +} source; injector control system; Maxwell spark gap test update; ILSE cosine 2{theta} quadrupole magnet development; electrostatic quadrupole prototype development activity; induction accelerator cell development; effect of a spread in beamlet currents on longitudinal stability; and heavy ion linac driver analysis.

  9. Fusion of splicing factor genes PSF and NonO (p54nrb) to the TFE3 gene in papillary renal cell carcinoma.

    PubMed

    Clark, J; Lu, Y J; Sidhar, S K; Parker, C; Gill, S; Smedley, D; Hamoudi, R; Linehan, W M; Shipley, J; Cooper, C S

    1997-10-01

    We demonstrate that the cytogenetically defined translocation t(X;1)(p11.2;p34) observed in papillary renal cell carcinomas results in the fusion of the splicing factor gene PSF located at 1p34 to the TFE3 helix-loop-helix transcription factor gene at Xp11.2. In addition we define an X chromosome inversion inv(X)(p11.2;q12) that results in the fusion of the NonO (p54nrb) gene to TFE3. NonO (p54nrb), the human homologue of the Drosophila gene NonAdiss which controls the male courtship song, is closely related to PSF and also believed to be involved in RNA splicing. In each case the rearrangement results in the fusion of almost the entire splicing factor protein to the TFE3 DNA-binding domain. These observations suggest the possibility of intriguing links between the processes of RNA splicing, DNA transcription and oncogenesis.

  10. Regulation of fos-lacZ fusion gene expression in primary mouse epidermal keratinocytes isolated from transgenic mice.

    PubMed Central

    Bollag, W B; Xiong, Y; Ducote, J; Harmon, C S

    1994-01-01

    The expression of a fos-lacZ fusion gene was studied in primary mouse epidermal keratinocytes obtained from transgenic mice. This gene construct contains the entire upstream regulatory sequence of c-fos, and expression of the endogenous and fusion gene was shown by Northern analysis to correlate upon induction with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). Using a chromogenic substrate of beta-galactosidase, we also demonstrated that expression of the fusion gene product, like that of Fos, was localized to the cell nucleus. In addition, we showed that epidermal keratinocytes responded to dialysed fetal bovine serum (FBS), TPA and high-calcium medium with enhanced Fos-lacZ expression and an inhibition of proliferation. The time course of induction of Fos-lacZ expression was similar for dialysed FBS and TPA, with a peak approximately 2 h after exposure. Exposure for approximately 24 h to an elevated extracellular calcium concentration was required to elicit an increase in Fos-lacZ expression. The lack of an immediate effect of raising medium calcium levels on Fos-lacZ expression contrasted with the rapidity of its effect on DNA synthesis, which was significantly inhibited within 6-8 h. In addition, we found that the protein kinase C inhibitor Ro 31-7549 blocked Fos-lacZ expression induced by TPA but had little or no effect on that elicited by high calcium levels. Thus, although our results indicate that the fos gene product may be involved in mediating epidermal keratinocyte growth arrest in response to differentiative agents such as FBS, TPA and high medium calcium levels, the exact role of this gene product remains unclear. Images Figure 1 Figure 2 PMID:8198544

  11. Regulation of fos-lacZ fusion gene expression in primary mouse epidermal keratinocytes isolated from transgenic mice.

    PubMed

    Bollag, W B; Xiong, Y; Ducote, J; Harmon, C S

    1994-05-15

    The expression of a fos-lacZ fusion gene was studied in primary mouse epidermal keratinocytes obtained from transgenic mice. This gene construct contains the entire upstream regulatory sequence of c-fos, and expression of the endogenous and fusion gene was shown by Northern analysis to correlate upon induction with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). Using a chromogenic substrate of beta-galactosidase, we also demonstrated that expression of the fusion gene product, like that of Fos, was localized to the cell nucleus. In addition, we showed that epidermal keratinocytes responded to dialysed fetal bovine serum (FBS), TPA and high-calcium medium with enhanced Fos-lacZ expression and an inhibition of proliferation. The time course of induction of Fos-lacZ expression was similar for dialysed FBS and TPA, with a peak approximately 2 h after exposure. Exposure for approximately 24 h to an elevated extracellular calcium concentration was required to elicit an increase in Fos-lacZ expression. The lack of an immediate effect of raising medium calcium levels on Fos-lacZ expression contrasted with the rapidity of its effect on DNA synthesis, which was significantly inhibited within 6-8 h. In addition, we found that the protein kinase C inhibitor Ro 31-7549 blocked Fos-lacZ expression induced by TPA but had little or no effect on that elicited by high calcium levels. Thus, although our results indicate that the fos gene product may be involved in mediating epidermal keratinocyte growth arrest in response to differentiative agents such as FBS, TPA and high medium calcium levels, the exact role of this gene product remains unclear.

  12. Role of starvation genes in the survival of deep subsurface bacterial communities. Final report

    SciTech Connect

    Matin, A.; Schmidt, T.; Caldwell, D.

    1998-11-01

    The investigation dealt with several aspects of subsurface bacterial survival and their nature. Mutants of Pseudomonas putida, a common environmental bacterium with counterparts in the subsurface, were isolated by transposon mutagenesis. These mutants were highly sensitive to starvation stress. Reporter gene fusions also showed that these genes were starvation genes since they were induced several fold when the cultures were started. Since the regulatory religions (promoters) of starvation genes are of interest in bioremediation and in experiments designed to understand the roles of starvation genes in the maintenance of microbial community structure, the promoter of one of these genes (pstarv1, contained in strain MK107) was characterized in detail. As a preliminary to these studies, the growth characteristics of Pseudomonas putida MK1 and MK107 were compared for cells growing in batch cultures or as an attached monolayer in microstat cultures.

  13. Increased gene copy number of ERG on chromosome 21 but not TMPRSS2–ERG fusion predicts outcome in prostatic adenocarcinomas

    PubMed Central

    Toubaji, Antoun; Albadine, Roula; Meeker, Alan K; Isaacs, William B; Lotan, Tamara; Haffner, Michael C; Chaux, Alcides; Epstein, Jonathan I; Han, Misop; Walsh, Patrick C; Partin, Alan W; De Marzo, Angelo M; Platz, Elizabeth A; Netto, George J

    2012-01-01

    The role of TMPRSS2–ERG gene fusion in prostate cancer prognostication remains controversial. We evaluated the prognostic role of TMPRSS2–ERG fusion using fluorescence in situ hybridization analysis in a case–control study nested in The Johns Hopkins retropubic radical prostatectomy cohort. In all, 10 tissue microarrays containing paired tumors and normal tissues obtained from 172 cases (recurrence) and 172 controls (non-recurrence) matched on pathological grade, stage, race/ethnicity, and age at the time of surgery were analyzed. All radical prostatectomies were performed at our institution between 1993 and 2004. Recurrence was defined as biochemical recurrence, development of clinical evidence of metastasis, or death from prostate carcinoma. Each tissue microarray spot was scored for the presence of TMPRSS2–ERG gene fusion and for ERG gene copy number gains. The odds ratio of recurrence and 95% confidence intervals were estimated from conditional logistic regression. Although the percentage of cases with fusion was slightly lower in cases than in controls (50 vs 57%), the difference was not statistically significant (P=0.20). The presence of fusion due to either deletion or split event was not associated with recurrence. Similarly, the presence of duplicated ERG deletion, duplicated ERG split, or ERG gene copy number gain with a single ERG fusion was not associated with recurrence. ERG gene polysomy without fusion was significantly associated with recurrence (odds ratio 2.0, 95% confidence interval 1.17–3.42). In summary, TMPRSS2–ERG fusion was not prognostic for recurrence after retropubic radical prostatectomy for clinically localized prostate cancer, although men with ERG gene copy number gain without fusion were twice more likely to recur. PMID:21743434

  14. Structure and expression of the Drosophila ubiquitin-52-amino-acid fusion-protein gene.

    PubMed Central

    Cabrera, H L; Barrio, R; Arribas, C

    1992-01-01

    Ubiquitin belongs to a multigene family. In Drosophila two members of this family have been previously described. We report here the organization and expression of a third member, the DUb52 gene, isolated by screening a Drosophila melanogaster genomic library. This gene encodes an ubiquitin monomer fused to a 52-amino acid extension protein. There are no introns interrupting the coding sequence. Recently, it has been described that this extension encodes a ribosomal protein in Saccharomyces, Dictyostelium, and Arabidopsis. The present results show that the 5' regulatory region of DUb52 shares common features with the ribosomal protein genes of Drosophila, Xenopus and mouse, including GC- and pyrimidine-rich regions. Moreover, sequences similar to the consensus Ribo-box in Neurospora crassa have been identified. Furthermore, a sequence has been found that is similar to the binding site for the TFIIIA distal element factor from Xenopus laevis. The DUb52 gene is transcribed to a 0.9 kb mRNA that is expressed constitutively throughout development and is particularly abundant in ovaries. In addition, the DUb52 gene has been found to be preferentially transcribed in exponentially growing Drosophila cells. Images Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. PMID:1381584

  15. Plant expansins in bacteria and fungi: evolution by horizontal gene transfer and independent domain fusion.

    PubMed

    Nikolaidis, Nikolas; Doran, Nicole; Cosgrove, Daniel J

    2014-02-01

    Horizontal gene transfer (HGT) has been described as a common mechanism of transferring genetic material between prokaryotes, whereas genetic transfers from eukaryotes to prokaryotes have been rarely documented. Here we report a rare case of HGT in which plant expansin genes that code for plant cell-wall loosening proteins were transferred from plants to bacteria, fungi, and amoebozoa. In several cases, the species in which the expansin gene was found is either in intimate association with plants or is a known plant pathogen. Our analyses suggest that at least two independent genetic transfers occurred from plants to bacteria and fungi. These events were followed by multiple HGT events within bacteria and fungi. We have also observed that in bacteria expansin genes have been independently fused to DNA fragments that code for an endoglucanase domain or for a carbohydrate binding module, pointing to functional convergence at the molecular level. Furthermore, the functional similarities between microbial expansins and their plant xenologs suggest that these proteins mediate microbial-plant interactions by altering the plant cell wall and therefore may provide adaptive advantages to these species. The evolution of these nonplant expansins represents a unique case in which bacteria and fungi have found innovative and adaptive ways to interact with and infect plants by acquiring genes from their host. This evolutionary paradigm suggests that despite their low frequency such HGT events may have significantly contributed to the evolution of prokaryotic and eukaryotic species.

  16. Schinzel-Giedion syndrome: report of splenopancreatic fusion and proposed diagnostic criteria.

    PubMed

    Lehman, Anna M; McFadden, Deborah; Pugash, Denise; Sangha, Karan; Gibson, William T; Patel, Millan S

    2008-05-15

    We report on the 46th patient with Schinzel-Giedion syndrome (SGS) and the first observation of splenopancreatic fusion in this syndrome. In the antenatal period, a male fetus was found to have bilateral hydronephrosis. Postnatally, in keeping with a diagnosis of SGS, there were large fontanelles, ocular hypertelorism, a wide, broad forehead, midface retraction, a short, upturned nose, macroglossia, and a short neck. Other anomalies included cardiac defects, widened and dense long bone cortices, cerebral ventriculomegaly, and abnormal fundi. Splenopancreatic fusion, usually encountered in trisomy 13, was found on autopsy. Schinzel-Giedion syndrome is likely a monogenic condition for which neither the heritability pattern nor pathogenesis has yet been determined. A clinical diagnosis may be made by identifying the facial phenotype, including prominent forehead, midface retraction, and short, upturned nose, plus one of either of the two other major distinguishing features: typical skeletal abnormalities or hydronephrosis. Typical skeletal anomalies include a sclerotic skull base, wide supraoccipital-exoccipital synchondrosis, increased cortical density or thickness, and broad ribs. Other highly supportive features include neuroepithelial tumors (found in 17%), hypertrichosis, and brain abnormalities. Severe developmental delay and poor survival are constant features in reported patients.

  17. EML4-ALK fusion gene and efficacy of an ALK kinase inhibitor in lung cancer

    PubMed Central

    Koivunen, Jussi P.; Mermel, Craig; Zejnullahu, Kreshnik; Murphy, Carly; Lifshits, Eugene; Holmes, Alison J.; Choi, Hwan Geun; Kim, Jhingook; Chiang, Derek; Thomas, Roman; Lee, Jinseon; Richards, William G.; Sugarbaker, David J.; Ducko, Christopher; Lindeman, Neal; Marcoux, J. Paul; Engelman, Jeffrey A.; Gray, Nathanael S.; Lee, Charles; Meyerson, Matthew; Jänne, Pasi A.

    2011-01-01

    Purpose The EML4-ALK fusion gene has been detected in ~7% of Japanese non-small cell lung cancers (NSCLC). We determined the frequency of EML4-ALK in Caucasian NSCLCs and in NSCLC cell lines. We also determined whether TAE684, a specific ALK kinase inhibitor, would inhibit the growth of EML4-ALK containing cell lines in vitro and in vivo. Experimental Design We screened 305 primary NSCLCs (both US (n=138) and Korean (n=167) patients) and 83 NSCLC cell lines using RT-PCR and by exon array analyses. We evaluated the efficacy of TAE684 against NSCLC cell lines in vitro and in vivo. Results We detected 4 different variants, including two novel variants, of EML4-ALK using RT-PCR in 8/305 tumors (3%) and in 3/83 (3.6%) NSCLC cell lines. All EML4-ALK containing tumors and cell lines were adenocarcinomas. EML4-ALK was detected more frequently in NSCLC patients who were never or light (< 10 pack years) cigarette smokers compared to current/former smokers (6% vs. 1%; p=0.049). TAE684 inhibited the growth of 1 of 3 (H3122) EML4-ALK containing cell lines in vitro and in vivo, inhibited Akt phosphorylation and caused apoptosis. In another EML4-ALK cell line, DFCI032, TAE684 was ineffective due to co-activation of EGFR and ERBB2. The combination of TAE684 and CL-387,785 (EGFR/ERBB2 kinase inhibitor), inhibited growth and Akt phosphorylation and led to apoptosis in the DFCI032 cell line. Conclusions EML4-ALK is found in the minority of NSCLCs. ALK kinase inhibitors alone or in combination may nevertheless be clinically effective treatments for NSCLC patients whose tumors contain EML4-ALK. PMID:18594010

  18. Targeted endostatin-cytosine deaminase fusion gene therapy plus 5-fluorocytosine suppresses ovarian tumor growth.

    PubMed

    Sher, Y-P; Chang, C-M; Juo, C-G; Chen, C-T; Hsu, J L; Lin, C-Y; Han, Z; Shiah, S-G; Hung, M-C

    2013-02-28

    There are currently no effective therapies for cancer patients with advanced ovarian cancer, therefore developing an efficient and safe strategy is urgent. To ensure cancer-specific targeting, efficient delivery, and efficacy, we developed an ovarian cancer-specific construct (Survivin-VISA-hEndoyCD) composed of the cancer specific promoter survivin in a transgene amplification vector (VISA; VP16-GAL4-WPRE integrated systemic amplifier) to express a secreted human endostatin-yeast cytosine deaminase fusion protein (hEndoyCD) for advanced ovarian cancer treatment. hEndoyCD contains an endostatin domain that has tumor-targeting ability for anti-angiogenesis and a cytosine deaminase domain that converts the prodrug 5-fluorocytosine (5-FC) into the chemotherapeutic drug, 5-fluorouracil. Survivin-VISA-hEndoyCD was found to be highly specific, selectively express secreted hEndoyCD from ovarian cancer cells, and induce cancer-cell killing in vitro and in vivo in the presence of 5-FC without affecting normal cells. In addition, Survivin-VISA-hEndoyCD plus 5-FC showed strong synergistic effects in combination with cisplatin in ovarian cancer cell lines. Intraperitoneal (i.p.) treatment with Survivin-VISA-hEndoyCD coupled with liposome attenuated tumor growth and prolonged survival in mice bearing advanced ovarian tumors. Importantly, there was virtually no severe toxicity when hEndoyCD is expressed by Survivin-VISA plus 5-FC compared with CMV plus 5-FC. Thus, the current study demonstrates an effective cancer-targeted gene therapy that is worthy of development in clinical trials for treating advanced ovarian cancer.

  19. [Construction of eukaryotic recombinant vector and expression in COS7 cell of LipL32-HlyX fusion gene from Leptospira serovar Lai].

    PubMed

    Huang, Bi; Bao, Lang; Zhong, Qi; Zhang, Huidong; Zhang, Ying

    2009-04-01

    This study was conducted to construct eukaryotic recombinant vector of LipL32-HlyX fusion gene from Leptospira serovar Lai and express it in mammalian cell. Both of LipL32 gene and HlyX gene were amplified from Leptospira strain O17 genomic DNA by PCR. Then with the two genes as template, LipL32-HlyX fusion gene was obtained by SOE PCR (gene splicing by overlap extension PCR). The fusion gene was then cloned into pcDNA3.1 by restriction nuclease digestion. Having been transformed into E. coli DH5alpha, the recombiant plasmid was identified by restriction nuclease digestion, PCR analysis and sequencing. The recombinant plasmid was then transfected into COS7 cell whose expression was detected by RT-PCR and Western blotting analysis. RT-PCR amplified a fragment about 2000 bp and Western blotting analysis found a specific band about 75 KD which was consistent with the expected fusion protein size. In conclusion, the successful construction of eukaryotic recombinant vector containing LipL32-HlyX fusion gene and the effective expression in mammalian have laid a foundation for the application of Leptospira DNA vaccine.

  20. Characterization of foot-and-mouth disease virus gene products with antisera against bacterially synthesized fusion proteins

    SciTech Connect

    Strebel, K.; Beck, E.; Strohmaier, K.; Schaller, H.

    1986-03-01

    Defined segments of the cloned foot-and-mouth disease virus genome corresponding to all parts of the coding region were expressed in Escherichia coli as fusions to the N-terminal part of the MS2-polymerase gene under the control of the inducible lambdaPL promoter. All constructs yielded large amounts of proteins, which were purified and used to raise sequence-specific antisera in rabbits. These antisera were used to identify the corresponding viral gene products in /sup 35/S-labeled extracts from foot-and-mouth disease virus-infected BHK cells. This allowed us to locate unequivocally all mature foot-and-mouth disease virus gene products in the nucleotide sequence, to identify precursor-product relationships, and to detect several foot-and mouth disease virus gene products not previously identified in vivo or in vitro.

  1. Dominant negative selection of vaccinia virus using a thymidine kinase/thymidylate kinase fusion gene and the prodrug azidothymidine

    SciTech Connect

    Holzer, Georg W. . E-mail: falknef@baxter.com

    2005-07-05

    The Escherichia coli thymidine kinase/thymidylate kinase (tk/tmk) fusion gene encodes an enzyme that efficiently converts the prodrug 3'-azido-2',3'-dideoxythymidine (AZT) into its toxic triphosphate derivative, a substance which stops DNA chain elongation. Integration of this marker gene into vaccinia virus that normally is not inhibited by AZT allowed the establishment of a powerful selection procedure for recombinant viruses. In contrast to the conventional vaccinia thymidine kinase (tk) selection that is performed in tk-negative cell lines, AZT selection can be performed in normal (tk-positive) cell lines. The technique is especially useful for the generation of replication-deficient vaccinia viruses and may also be used for gene knock-out studies of essential vaccinia genes.

  2. Incidental finding of two rare developmental anomalies: Fusion and dilaceration: A case report and literature review

    PubMed Central

    Sultan, Nishat

    2015-01-01

    A number of developmental anomalies of morphology are there. However, as compared to the more common oral diseases like caries or periodontal problems, they account for a relatively lower number. When present, they may pose various problems of esthetic, function, malocclusion, or possible disposition to other oral problems. Hence, though rare, their timely diagnosis is very vital in proper treatment planning to avoid unseen complications during extractions, endodontic or orthodontic treatment. The present case is of a patient reporting with two very rare developmental anomalies, that is, fusion and root dilaceration, in contralateral sides of the same patient. To the knowledge of the author, reportedly it is the first such case. The terminologies, etiology, and epidemiology of both these anomalies are also discussed. PMID:26604610

  3. Overexpression of HMGA2-LPP fusion transcripts promotes expression of the {alpha} 2 type XI collagen gene

    SciTech Connect

    Kubo, Takahiro; Matsui, Yoshito . E-mail: ymatsui@sb4.so-net.ne.jp; Goto, Tomohiro; Yukata, Kiminori; Yasui, Natsuo

    2006-02-10

    In a subset of human lipomas, a specific t (3; 12) chromosome translocation gives rise to HMGA2-LPP fusion protein, containing the amino (N)-terminal DNA binding domains of HMGA2 fused to the carboxyl (C)-terminal LIM domains of LPP. In addition to its role in adipogenesis, several observations suggest that HMGA2-LPP is linked to chondrogenesis. Here, we analyzed whether HMGA2-LPP promotes chondrogenic differentiation, a marker of which is transactivation of the {alpha} 2 type XI collagen gene (Col11a2). Real-time PCR analysis showed that HMGA2-LPP and COL11A2 were co-expressed. Luciferase assay demonstrated that either of HMGA2-LPP, wild-type HMGA2 or the N-terminal HMGA2 transactivated the Col11a2 promoter in HeLa cells, while the C-terminal LPP did not. RT-PCR analysis revealed that HMGA2-LPP transcripts in lipomas with the fusion were 591-fold of full-length HMGA2 transcripts in lipomas without the fusion. These results indicate that in vivo overexpression of HMGA2-LPP promotes chondrogenesis by upregulating cartilage-specific collagen gene expression through the N-terminal DNA binding domains.

  4. Cold fusion research

    SciTech Connect

    1989-11-01

    I am pleased to forward to you the Final Report of the Cold Fusion Panel. This report reviews the current status of cold fusion and includes major chapters on Calorimetry and Excess Heat, Fusion Products and Materials Characterization. In addition, the report makes a number of conclusions and recommendations, as requested by the Secretary of Energy.

  5. Making genes green: creating green fluorescent protein (GFP) fusions with blunt-end PCR products.

    PubMed

    Lo, W; Rodgers, W; Hughes, T

    1998-07-01

    The jellyfish green fluorescent protein (GFP) has proven to be a useful tool in protein localization and trafficking studies. Fused to GFP, a protein of interest can be visualized and tracked in vivo through fluorescence microscopy. However, the process of making these fusion proteins is often tedious and painstaking. Here, we describe a simple and quick method for creating GFP fusion proteins using blunt-end PCR product ligation.

  6. The Chicken Gene Nomenclature Committee report.

    PubMed

    Burt, David W; Carrë, Wilfrid; Fell, Mark; Law, Andy S; Antin, Parker B; Maglott, Donna R; Weber, Janet A; Schmidt, Carl J; Burgess, Shane C; McCarthy, Fiona M

    2009-07-14

    Comparative genomics is an essential component of the post-genomic era. The chicken genome is the first avian genome to be sequenced and it will serve as a model for other avian species. Moreover, due to its unique evolutionary niche, the chicken genome can be used to understand evolution of functional elements and gene regulation in mammalian species. However comparative biology both within avian species and within amniotes is hampered due to the difficulty of recognising functional orthologs. This problem is compounded as different databases and sequence repositories proliferate and the names they assign to functional elements proliferate along with them. Currently, genes can be published under more than one name and one name sometimes refers to unrelated genes. Standardized gene nomenclature is necessary to facilitate communication between scientists and genomic resources. Moreover, it is important that this nomenclature be based on existing nomenclature efforts where possible to truly facilitate studies between different species. We report here the formation of the Chicken Gene Nomenclature Committee (CGNC), an international and centralized effort to provide standardized nomenclature for chicken genes. The CGNC works in conjunction with public resources such as NCBI and Ensembl and in consultation with existing nomenclature committees for human and mouse. The CGNC will develop standardized nomenclature in consultation with the research community and relies on the support of the research community to ensure that the nomenclature facilitates comparative and genomic studies.

  7. Comprehensive therapy of a fusion between a mandibular lateral incisor and supernumerary tooth: case report.

    PubMed

    Onçag, Ozant; Candan, Umit; Arikan, Fatih

    2005-08-01

    The term fusion is used to define a developmental anomaly characterised by the union of two adjacent teeth. In the case reported here, clinical and radiographic examinations suggested a unilateral fusion between the mandibular left permanent incisor and a super-numerary tooth. Radiographs showed that the fused teeth had two distinct pulp chambers and canals. A diagnosis of chronic periapical abscess of the supernumerary tooth was made. Before root canal therapy, a periodontal surgical procedure was performed to section the central incisor and its fused supernumerary. Also, odontoplasty was performed on the roots, to establish an anatomy consistent with a normal central incisor. Later, the chronic apical abscess on the supernumerary tooth was instrumented chemo-mechanically, root canal filling was performed and an anterior composite resin restoration was placed. The patient was evaluated for one year after root canal therapy. The tooth was asymptomatic, not exhibiting any pathological root resorption or alveolar resorption, and the anterior composite restoration was intact. Instead of extracting the supernumerary tooth, the application of endodontic, periodontal, and restorative procedures proved to be an alternative treatment.

  8. Heavy ion fusion accelerator research (HIFAR) year-end report, April 1, 1987-September 30, 1987

    SciTech Connect

    Not Available

    1987-12-01

    The basic objective of the Heavy Ion Fusion Accelerator Research (HIFAR) program is to access the suitabilty of heavy ion accelerators as iginiters for Inertial Confinement Fusion (ICF). A specific accerelator techonolgy, the induction linac, has been studied at the Lawerence Berkeley Laboratory and has reached the point at which its viability for ICF applications can be assessed over the next few years. The HIFAR program addresses the generation of high-power, high-brightness beams of heavy ions, the understanding of the scaling laws in this novel physics regime, and the vadidation of new accelerator strategies, to cut costs. The papers in this report that address these goals are: MBE-4 mechanical progress, alignment of MBE-4, a compact energy analyzer for MBE-4, Cs/sup +/ injector modeling with the EGUN code, an improved emittance scanning system for HIFAR, 2-MV injector, carbon arc source development, beam combining in ILSE, emittance growth due to transverse beam combining in ILSE - particle simulation results, achromatic beam combiner for ILSE, additional elements for beam merging, quadrupole magnet design for ILSE, and waveforms and longitudinal beam-parameters for ILSE.

  9. Investigation of electromagnetic launcher behavior for impact fusion. Annual report, July 1, 1983-May 1, 1984

    SciTech Connect

    Thio, Y.C.

    1984-06-01

    A program to develop an ultrahigh velocity accelerator (SUVAC), based on the electromagnetic railgun accelerator concept and sponsored by the US Department of Energy, has been initiated at Westinghouse R and D Center. The program involves the construction over a 4-year period (July 1983 to June 1987) of a multi-stage railgun accelerator which has the potential of accelerating a 1-g projectile to about 30 km/s (Mach 100). The scientific objective of the program is to use the accelerator so built as the experimental apparatus to investigate the potential technical problems of accelerating macroparticles to velocity presently thought to be required to produce impact fusion. The program is part of a joint program with the University of Washington to develop the scientific and technological basis to achieve controlled thermonuclear fusion by hypervelocity impact. This report summarizes the progress made in the first year of the program. It covers work done for the period July 1, 1983 to May 1, 1984.

  10. The EWSR1/NR4A3 fusion protein of extraskeletal myxoid chondrosarcoma activates the PPARG nuclear receptor gene.

    PubMed

    Filion, C; Motoi, T; Olshen, A B; Laé, M; Emnett, R J; Gutmann, D H; Perry, A; Ladanyi, M; Labelle, Y

    2009-01-01

    The NR4A3 nuclear receptor is implicated in the development of extraskeletal myxoid chondrosarcoma (EMC), primitive sarcoma unrelated to conventional chondrosarcomas, through a specific fusion with EWSR1 resulting in an aberrant fusion protein that is thought to disrupt the transcriptional regulation of specific target genes. We performed an expression microarray analysis of EMC tumours expressing the EWSR1/NR4A3 fusion protein, comparing their expression profiles to those of other sarcoma types. We thereby identified a set of genes significantly overexpressed in EMC relative to other sarcomas, including PPARG and NDRG2. Western blot or immunohistochemical analyses confirm that PPARG and NDRG2 are expressed in tumours positive for EWSR1/NR4A3. Bioinformatic analysis identified a DNA response element for EWSR1/NR4A3 in the PPARG promoter, and band-shift experiments and transient transfections indicate that EWSR1/NR4A3 can activate transcription through this element. Western blots further show that an isoform of the native NR4A3 receptor lacking the C-terminal domain is very highly expressed in tumours positive for EWSR1/NR4A3, and co-transfections of this isoform along with EWSR1/NR4A3 indicate that it may negatively regulate the activity of the fusion protein on the PPARG promoter. These results suggest that the overall expression of PPARG in EMC may be regulated in part by the balance between EWSR1/NR4A3 and NR4A3, and that PPARG may play a crucial role in the development of these tumours. The specific up-regulation of PPARG by EWSR1/NR4A3 may also have potential therapeutic implications.

  11. EDITORIAL: Special issue: overview reports from the Fusion Energy Conference (FEC) (Daejeon, South Korea, 2010) Special issue: overview reports from the Fusion Energy Conference (FEC) (Daejeon, South Korea, 2010)

    NASA Astrophysics Data System (ADS)

    Thomas, Paul

    2011-09-01

    The group of 27 papers published in this special issue of Nuclear Fusion aims to monitor the worldwide progress made in the period 2008-2010 in the field of thermonuclear fusion. Of these papers, 22 are based on overview reports presented at the 23rd Fusion Energy Conference (FEC 2010) and five are summary reports. The conference was hosted by the Republic of Korea and organized by the IAEA in cooperation with the National Fusion Research Institute and the Daejeon Metropolitan City. It took place in Daejeon on 11-16 October 2010. The overviews presented at the conference have been rewritten and extended for the purpose of this special issue and submitted to the standard double-referee peer-review of Nuclear Fusion. The articles are placed in the following sequence: Conference summaries of the sessions devoted to: Tokamak and stellarator experiments, experimental divertor physics and plasma wall interaction experiments, stability experiments and waves and fast particles; ITER activities, fusion technology, safety and economics; Magnetic confinement theory and modelling; Inertial confinement fusion; Innovative confinement concepts, operational scenarios and confinement. Overview articles, presented in programme order, are as follows: Tokamaks Overview of KSTAR initial experiments; Recent progress in RF heating and long-pulse experiments on EAST; Overview of JET results; DIII-D contributions toward the scientific basis for sustained burning plasmas; Overview of JT-60U results toward the resolution of key physics and engineering issues in ITER and JT-60SA; Overview of physics results from NSTX; Overview of ASDEX Upgrade results; Overview of physics results from MAST; Contribution of Tore Supra in preparation of ITER; Overview of FTU results; Overview of experimental results on the HL-2A tokamak; Progress and scientific results in the TCV tokamak; Overview of the JT-60SA project; Recent results of the T-10 tokamak; The reconstruction and research progress of the TEXT

  12. NAB2-STAT6 Gene Fusion in Meningeal Hemangiopericytoma and Solitary Fibrous Tumor.

    PubMed

    Fritchie, Karen J; Jin, Long; Rubin, Brian P; Burger, Peter C; Jenkins, Sarah M; Barthelmeß, Sarah; Moskalev, Evgeny A; Haller, Florian; Oliveira, Andre M; Giannini, Caterina

    2016-03-01

    Meningeal solitary fibrous tumor (SFT) and hemangiopericytoma (HPC) are considered to be distinct entities in the WHO Classification of CNS Tumours (2007). They harbor NAB2-STAT6 fusions similar to their soft tissue counterparts, supporting the view that they are part of a tumor continuum. We examined 30 meningeal-based tumors originally diagnosed as either SFT or HPC. These showed a spectrum of morphologic features and were diagnosed as SFTs, malignant SFTs, HPCs, or tumors with "intermediate" features. All of the tumors showed nuclear expression of STAT6. SFTs consistently expressed diffuse CD34, while HPCs and intermediate tumors had heterogeneous staining. NAB2-STAT6 fusions were identified in 20 cases, including 7 with exon 4-exon 3, 9 with exon 6-exon 17, and 4 with exon 6-exon 18 fusions. NAB2 exon 4-STAT6 exon 3 fusion correlated with classic SFT morphology and older age and showed a trend toward less mitotic activity; there was also a trend toward more aggressive behavior in tumors lacking NAB2 exon 4-STAT6 exon 3. Thus, despite their clinical and morphologic differences, meningeal-based SFTs, HPCs, and tumors with intermediate features, similar to their soft tissue counterparts, form a histopathologic spectrum unified by STAT6 immunoexpression and NAB2-STAT6 fusion.

  13. Protein-Protein Fusion Catalyzed by Sortase A

    PubMed Central

    Levary, David A.; Parthasarathy, Ranganath; Boder, Eric T.; Ackerman, Margaret E.

    2011-01-01

    Chimeric proteins boast widespread use in areas ranging from cell biology to drug delivery. Post-translational protein fusion using the bacterial transpeptidase sortase A provides an attractive alternative when traditional gene fusion fails. We describe use of this enzyme for in vitro protein ligation and report the successful fusion of 10 pairs of protein domains with preserved functionality — demonstrating the robust and facile nature of this reaction. PMID:21494692

  14. Bioluminescent reporters for catabolic gene expression and pollutant bioavailability

    SciTech Connect

    Heitzer, A.; DiGrazia, P.M.; Sayler, G.S. . Center for Environmental Biotechnology); Burlage, R.S. )

    1991-01-01

    The application of visualized catabolic nah-gene expression using a luxCDABE gene fusion provides a valuable method to measure quantitatively and specifically naphthalene and salicylate bioavailability. It has been demonstrated that the physiological state of the test culture together with the intrinsic regulation mechanisms of the naphthalene degradation pathway as well as the physiological aspects of the lux gene fusion have to be taken into account. The method presented provides a high potential for in situ bioprocess monitoring. In addition, the results obtained with immobilized cells provide a basis for the development of biosensors for environmental applications in specific pollutant monitoring in waste streams and soil slurry systems but, as a general method, also for more conventional biotechnological process control. 8 refs., 2 figs., 1 tab.

  15. Visualizing Presynaptic Calcium Dynamics and Vesicle Fusion with a Single Genetically Encoded Reporter at Individual Synapses.

    PubMed

    Jackson, Rachel E; Burrone, Juan

    2016-01-01

    Synaptic transmission depends on the influx of calcium into the presynaptic compartment, which drives neurotransmitter release. Genetically encoded reporters are widely used tools to understand these processes, particularly pHluorin-based reporters that report vesicle exocytosis and endocytosis through pH dependent changes in fluorescence, and genetically encoded calcium indicators (GECIs) that exhibit changes in fluorescence upon binding to calcium. The recent expansion of the color palette of available indicators has made it possible to image multiple probes simultaneously within a cell. We have constructed a single molecule reporter capable of concurrent imaging of both presynaptic calcium influx and exocytosis, by fusion of sypHy, the vesicle associated protein synaptophysin containing a GFP-based pHluorin sensor, with the red-shifted GECI R-GECO1. Due to the fixed stoichiometry of the two probes, the ratio of the two responses can also be measured, providing an all optical correlate of the calcium dependence of release. Here, we have characterized stimulus-evoked sypHy-RGECO responses of hippocampal synapses in vitro, exploring the effects of different stimulus strengths and frequencies as well as variations in external calcium concentrations. By combining live sypHy-RGECO imaging with post hoc fixation and immunofluorescence, we have also investigated correlations between structural and functional properties of synapses.

  16. Visualizing Presynaptic Calcium Dynamics and Vesicle Fusion with a Single Genetically Encoded Reporter at Individual Synapses

    PubMed Central

    Jackson, Rachel E.; Burrone, Juan

    2016-01-01

    Synaptic transmission depends on the influx of calcium into the presynaptic compartment, which drives neurotransmitter release. Genetically encoded reporters are widely used tools to understand these processes, particularly pHluorin-based reporters that report vesicle exocytosis and endocytosis through pH dependent changes in fluorescence, and genetically encoded calcium indicators (GECIs) that exhibit changes in fluorescence upon binding to calcium. The recent expansion of the color palette of available indicators has made it possible to image multiple probes simultaneously within a cell. We have constructed a single molecule reporter capable of concurrent imaging of both presynaptic calcium influx and exocytosis, by fusion of sypHy, the vesicle associated protein synaptophysin containing a GFP-based pHluorin sensor, with the red-shifted GECI R-GECO1. Due to the fixed stoichiometry of the two probes, the ratio of the two responses can also be measured, providing an all optical correlate of the calcium dependence of release. Here, we have characterized stimulus-evoked sypHy-RGECO responses of hippocampal synapses in vitro, exploring the effects of different stimulus strengths and frequencies as well as variations in external calcium concentrations. By combining live sypHy-RGECO imaging with post hoc fixation and immunofluorescence, we have also investigated correlations between structural and functional properties of synapses. PMID:27507942

  17. HSP70 and modified HPV 16 E7 fusion gene without the addition of a signal peptide gene sequence as a candidate therapeutic tumor vaccine.

    PubMed

    Zong, Jinbao; Wang, Changyuan; Wang, Qingyong; Peng, Qinglin; Xu, Yufei; Xie, Xixiu; Xu, Xuemei

    2013-12-01

    Millions of women are currently infected with high-risk human papillomavirus (HPV), which is considered to be a major risk factor for cervical cancer. Thus, it is urgent to develop therapeutic vaccines to eliminate the established infections or HPV-related diseases. In the present study, using the mycobacterium tuberculosis heat shock protein 70 (MtHSP70) gene linked to the modified HPV 16 E7 (mE7) gene, we generated two potential therapeutic HPV DNA vaccines, mE7/MtHSP70 and SigmE7/MtHSP70, the latter was linked to the signal peptide gene sequence of human CD33 at the upstream of the fusion gene. We found that vaccination with the mE7/MtHSP70 DNA vaccine induced a stronger E7-specific CD8+ T cell response and resulted in a more significant therapeutic effect against E7-expressing tumor cells in mice. Our results demonstrated that HSP70 can play a more important role in mE7 and MtHSP70 fusion DNA vaccine without the help of a signal peptide. This may facilitate the use of HSP70 and serve as a significant reference for future study.

  18. A GAL4-HP1 fusion protein targeted near heterochromatin promotes gene silencing.

    PubMed

    Seum, C; Spierer, A; Delattre, M; Pauli, D; Spierer, P

    2000-11-01

    We have constructed a new reporter transgene, Winkelried, equipped with a synthetic binding site for the yeast GAL4 transcriptional activator. The binding site is inserted between the white and lacZ reporter genes, and is flanked by FRT sequences. These elements allow excision of the GAL4 binding site by crossing the transgenic line with an FLP recombinase producing strain. We have generated by X-ray irradiation two independent chromosomal rearrangements, Heidi and Tell, relocating Winkelried next to pericentromeric heterochromatin. These rearrangements induce variegation of both white and lacZ. Variegation of Winkelried in the rearranged transgenic lines responds to the loss and excess of doses of the dominant suppressors of position-effect variegation (PEV) Su(var)3-7 and Su(var)2-5. Winkelried therefore constitutes a unique tool to test the effect on variegation in cis of any factor fused to the GAL4 DNA binding domain. Indeed, a chimeric protein, made of the DNA binding site of GAL4 and of HP1, the modifier of PEV encoded by Su(var)2-5, is shown to enhance variegation of Heidi and Tell. Excision of the binding sites for GAL4 in the variegating rearrangements Heidi and Tell abolishes the modifier effect of the GAL4-HP1 chimera. Therefore, in the Heidi and Tell rearrangements, enhancement of position-effect variegation depends strictly both on the concentration of GAL4-HP1 and on the presence of its binding site in the vicinity of the reporter genes.

  19. Workshop on Accelerators for Heavy Ion Fusion: Summary Report of the Workshop

    SciTech Connect

    Seidl, P.A.; Barnard, J.J.

    2011-04-29

    The Workshop on Accelerators for Heavy Ion Fusion was held at Lawrence Berkeley National Laboratory May 23-26, 2011. The workshop began with plenary sessions to review the state of the art in HIF (heavy ion fusion), followed by parallel working groups, and concluded with a plenary session to review the results. There were five working groups: IFE (inertial fusion energy) targets, RF approach to HIF, induction accelerator approach to HIF, chamber and driver interface, ion sources and injectors.

  20. [Fusion research/tokamak]. Final report, 1 May 1988--30 April 1994

    SciTech Connect

    1994-12-31

    The objectives of the Fusion Research Center Program are: (1) to advance /the transport studies of tokamaks, including the development and maintenance of the Magnetic Fusion Energy Database, and (2) to provide theoretical interpretation, modeling and equilibrium and stability studies for the text-upgrade tokamak. Work is described on five basic categories: (1) magnetic fusion energy database; (2) computational support and numerical modeling; (3) support for TEXT-upgrade and diagnostics; (4) transport studies; and (5) Alfven waves.

  1. Identification and characterization of the fusion transcript, composed of the apterous homolog and a putative protein phosphatase gene, generated by 1.5-Mb interstitial deletion in the vestigial (Vg) mutant of Bombyx mori.

    PubMed

    Fujii, T; Abe, H; Katsuma, S; Shimada, T

    2011-05-01

    The vestigial (Vg) mutant is a Z-linked mutant that causes vestigial wings in the silkworm, Bombyx mori. We have previously reported a 1.5-Mb interstitial deletion on the Z chromosome bearing the Vg mutation (Z(Vg) chromosome). In this study, we found that exons 3-8 of a gene named Bmptp-Z encoding a putative tyrosine-specific protein phosphatase are deleted by the 1.5-Mb interstitial deletion. We found that a gene encoding the Bombyx homolog of Drosophila Apterous (BmAp-A) protein is located 4.5 kb downstream of the distal breakpoint of the 1.5-Mb interstitial deletion. Moreover, an in-frame fusion transcript composed of the 5' part of Bmptp-Z and the 3' part of Bmap-A is generated specific to the Z(Vg) chromosome. Effects of the in-frame fusion transcript on the vestigial phenotype are discussed.

  2. Optimizing Transgene Configuration and Protein Fusions to Maximize Dopamine Production for the Gene Therapy of Parkinson's Disease.

    PubMed

    Stewart, Hannah J; Ralph, G Scott; Fong-Wong, Liang; Strickland, Iain; McCloskey, Laura; Barnes, Lucy; Blount, Ian; Wells, Owen; Truran, Christelle J M; Kingsman, Alan J; Palfi, Stéphane; Mitrophanous, Kyriacos A

    2016-09-01

    Pharmacological dopamine replacement therapies provide the most well-established treatments for Parkinson's disease (PD). However, these long-term treatments can lead to motor complications and off-target effects. ProSavin(®), a lentiviral vector (LV)-based gene therapy approach aimed at restoring local and continuous dopamine production, through delivery of three enzymes in the dopamine biosynthesis pathway, was demonstrated to be safe and well-tolerated in a phase I/II clinical study of patients with advanced PD. Although improvements in motor behaviour were observed, the data indicated that higher levels of dopamine replacement might be required to maximize benefit. We attempted to increase production of dopamine, and its precursor L-Dopa in LV-transduced cells, by optimizing the gene order in the ProSavin expression cassette, and by creating fusions of two or three of the transgenes, using linker sequences. In vitro analysis showed that several gene arrangements provided significantly increased dopamine and/or L-Dopa production compared with ProSavin, and that LV titers and transgene expression were not affected by introducing gene fusions. One vector, equine infectious anemia virus (EIAV)-TCiA, was selected for further characterization and showed significant improvements in dopamine and L-Dopa production compared with ProSavin, in human neuronal cells. Further characterization of EIAV-TCiA demonstrated expression of all three dopamine enzymes in vivo and faithful delivery and integration of the expected gene expression cassette within the genome of target cells, as assessed by Northern and Southern blotting. In conclusion, we have developed a novel LV vector with an increased capacity for L-Dopa and dopamine production compared with the current ProSavin vector. Clinical evaluation of this vector will be performed to assess the benefits in patients with PD.

  3. Analysis of a MULE-cyanide hydratase gene fusion in Verticillium dahliae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genome of the phytopathogenic fungus Verticillium dahliae encodes numerous Class II “cut-and-paste” transposable elements, including those of a small group of MULE transposons. We have previously identified a fusion event between a MULE transposon sequence and sequence encoding a cyanide hydrata...

  4. LLE 1998 annual report, October 1997--September 1998. Inertial fusion program and National Laser Users` Facility program

    SciTech Connect

    1999-01-01

    This report summarizes research at the Laboratory for Laser Energetics (LLE), the operation of the National Laser Users` Facility (NLUF), and programs involving the education of high school, undergraduate, and graduate students for FY98. Research summaries cover: progress in laser fusion; diagnostic development; laser and optical technology; and advanced technology for laser targets.

  5. Code development incorporating environmental, safety, and economic aspects of fusion reactors (FY 92--94). Final report

    SciTech Connect

    Ho, S.K.; Fowler, T.K.; Holdren, J.P.

    1994-11-01

    This is the Final Report for a three-year (FY 92--94) study of the Environmental, Safety, and Economic (ESE) aspects of fusion energy systems, emphasizing development of computerized approaches suitable for incorporation as modules in fusion system design codes. First, as is reported in Section 2, the authors now have operating a simplified but complete environment and safety evaluation code, BESAFE. The first tests of BESAFE as a module of the SUPERCODE, a design optimization systems code at LLNL, are reported in Section 3. Secondly, as reported in Section 4, the authors have maintained a strong effort in developing fast calculational schemes for activation inventory evaluation. In addition to these major accomplishments, considerable progress has been made on research on specific topics as follows. A tritium modeling code TRIDYN was developed in collaboration with the TSTA group at LANL and the Fusion Nuclear Technology group at UCLA. A simplified algorithm has been derived to calculate the transient temperature profiles in the blanket during accidents. The scheme solves iteratively a system of non-linear ordinary differential equations describing about 10 regions of the blanket by preserving energy balance. The authors have studied the physics and engineering aspects of divertor modeling for safety applications. Several modifications in the automation and characterization of environmental and safety indices have been made. They have applied this work to the environmental and safety comparisons of stainless steel with alternative structural materials for fusion reactors. A methodology in decision analysis utilizing influence and decision diagrams has been developed to model fusion reactor design problems. Most of the work during this funding period has been reported in 26 publications including theses, journal publications, conference papers, and technical reports, as listed in Section 11.

  6. Inertial confinement fusion. ICF quarterly report, October 1993--December 1993, Volume 4, Number 1

    SciTech Connect

    Powell, H.T.; Schleich, D.P.; Murphy, P.W.

    1994-05-01

    In the 1990 National Academy of Sciences (NAS) report of its review of the U.S. Inertial Confinement Fusion (ICF) Program, it was recommended that a high priority be placed on completing the Precision Nova Project and its associated experimental campaign. Since fiscal year 1990, the lab has therefore campaigned vigorously on Nova and in its supporting laboratories to develop the Precision Nova capabilities needed to perform the stressful target experiments recommended in the 1990 NAS report. The activities to enable these experiments have been directed at improvements in three areas - the Nova laser, target fabrication capabilities, and target diagnostics. As summarized in the five articles in this report, the Precision Nova improvements have been successfully completed. These improvements have had a positive impact on target performance and on the ability to diagnose the results, as evidenced by the HEP-1 experimental results. The five articles generally concentrate on improvements to the capabilities rather than on the associated target physics experiments. Separate abstracts are included for each paper.

  7. High-level SUMO-mediated fusion expression of ABP-dHC-cecropin A from multiple joined genes in Escherichia coli.

    PubMed

    Zhang, Jiaxin; Movahedi, Ali; Wei, Zhiheng; Sang, Ming; Wu, Xiaolong; Wang, Mengyang; Wei, Hui; Pan, Huixin; Yin, Tongming; Zhuge, Qiang

    2016-09-15

    The antimicrobial peptide ABP-dHC-cecropin A is a small cationic peptide with potent activity against a wide range of bacterial species. Evidence of antifungal activity has also been suggested; however, evaluation of this peptide has been limited due to the low expression of cecropin proteins in Escherichia coli. To improve the expression level of ABP-dHC-cecropin A in E. coli, tandem repeats of the ABP-dHC-cecropin A gene were constructed and expressed as fusion proteins (SUMO-nABP-dHC-cecropin, n = 1, 2, 3, 4) via pSUMO-nABP-dHC-cecropin A vectors (n = 1, 2, 3, 4). Comparison of the expression levels of soluble SUMO-nABP-dHC-cecropin A fusion proteins (n = 1, 2, 3, 4) suggested that BL21 (DE3)/pSUMO-3ABP-dHC-cecropin A is an ideal recombinant strain for ABP-dHC-cecropin A production. Under the selected conditions of cultivation and isopropylthiogalactoside (IPTG) induction, the expression level of ABP-dHC-cecropin A was as high as 65 mg/L, with ∼21.3% of the fusion protein in soluble form. By large-scale fermentation, protein production reached nearly 300 mg/L, which is the highest yield of ABP-dHC-cecropin A reported to date. In antibacterial experiments, the efficacy was approximately the same as that of synthetic ABP-dHC-cecropin A. This method provides a novel and effective means of producing large amounts of ABP-dHC-cecropin A.

  8. Inertial Confinement Fusion quarterly report, January-March 1998, volume 8, number 2

    SciTech Connect

    Kruer, W

    1998-03-31

    The coupling of laser light with plasmas is one of the key physics issues for the use of high-power lasers for inertial fusion, high-energy-density physics, and scientific stockpile stewardship. The coupling physics is extremely rich and challenging, particularly in the large plasmas to be accessed on the National Ignition Facility (NIF). The coupling mechanisms span the gamut from classical inverse bremsstrahlung absorption to a variety of nonlinear optical processes. These include stimulated Raman scattering (SRS) from electron plasma waves, stimulated Brillouin scattering (SBS) from ion sound waves, resonant decay into electron plasma and ion sound waves, and laser beam filamentation. These processes depend on laser intensity and produce effects such as changes in the efficiency and location of the energy deposition or generation of a component of very energetic electrons, which can preheat capsules. Coupling physics issues have an extremely high leverage. The coupling models are clearly very important ingredients for detailed calculations of laser-irradiated target behavior. Improved understanding and models enable a more efficient use of laser facilities, which becomes even more important as these facilities become larger and more expensive. Advances in the understanding also allow a more timely and cost-effective identification of new applications of high-power lasers, such as for generation of high-temperature hohlraums and compact x-ray sources, or for discovery of advanced fusion schemes. Finally, the interaction of intense electromagnetic waves with ionized media is a fundamental topic of interest to numerous areas of applied science and is an excellent test bed for advancing plasma science and computational modeling of complex phenomena. This issue of the ICF Quarterly Report is dedicated to laser--plasma interactions. The eight articles present a cross section of the broad progress in understanding the key interaction issues, such as laser beam bending

  9. An MSC2 Promoter-lacZ Fusion Gene Reveals Zinc-Responsive Changes in Sites of Transcription Initiation That Occur across the Yeast Genome

    PubMed Central

    Wu, Yi-Hsuan; Taggart, Janet; Song, Pamela Xiyao; MacDiarmid, Colin; Eide, David J.

    2016-01-01

    The Msc2 and Zrg17 proteins of Saccharomyces cerevisiae form a complex to transport zinc into the endoplasmic reticulum. ZRG17 is transcriptionally induced in zinc-limited cells by the Zap1 transcription factor. In this report, we show that MSC2 mRNA also increases (~1.5 fold) in zinc-limited cells. The MSC2 gene has two in-frame ATG codons at its 5’ end, ATG1 and ATG2; ATG2 is the predicted initiation codon. When the MSC2 promoter was fused at ATG2 to the lacZ gene, we found that unlike the chromosomal gene this reporter showed a 4-fold decrease in lacZ mRNA in zinc-limited cells. Surprisingly, β-galactosidase activity generated by this fusion gene increased ~7 fold during zinc deficiency suggesting the influence of post-transcriptional factors. Transcription of MSC2ATG2-lacZ was found to start upstream of ATG1 in zinc-replete cells. In zinc-limited cells, transcription initiation shifted to sites just upstream of ATG2. From the results of mutational and polysome profile analyses, we propose the following explanation for these effects. In zinc-replete cells, MSC2ATG2-lacZ mRNA with long 5’ UTRs fold into secondary structures that inhibit translation. In zinc-limited cells, transcripts with shorter unstructured 5’ UTRs are generated that are more efficiently translated. Surprisingly, chromosomal MSC2 did not show start site shifts in response to zinc status and only shorter 5’ UTRs were observed. However, the shifts that occur in the MSC2ATG2-lacZ construct led us to identify significant transcription start site changes affecting the expression of ~3% of all genes. Therefore, zinc status can profoundly alter transcription initiation across the yeast genome. PMID:27657924

  10. Biosensing of BCR/ABL fusion gene using an intensity-interrogation surface plasmon resonance imaging system

    NASA Astrophysics Data System (ADS)

    Wu, Jiangling; Huang, Yu; Bian, Xintong; Li, DanDan; Cheng, Quan; Ding, Shijia

    2016-10-01

    In this work, a custom-made intensity-interrogation surface plasmon resonance imaging (SPRi) system has been developed to directly detect a specific sequence of BCR/ABL fusion gene in chronic myelogenous leukemia (CML). The variation in the reflected light intensity detected from the sensor chip composed of gold islands array is proportional to the change of refractive index due to the selective hybridization of surface-bound DNA probes with target ssDNA. SPRi measurements were performed with different concentrations of synthetic target DNA sequence. The calibration curve of synthetic target sequence shows a good relationship between the concentration of synthetic target and the change of reflected light intensity. The detection limit of this SPRi measurement could approach 10.29 nM. By comparing SPRi images, the target ssDNA and non-complementary DNA sequence are able to be distinguished. This SPRi system has been applied for assay of BCR/ABL fusion gene extracted from real samples. This nucleic acid-based SPRi biosensor therefore offers an alternative high-effective, high-throughput label-free tool for DNA detection in biomedical research and molecular diagnosis.

  11. Synergistic antitumor effect of a human papillomavirus DNA vaccine harboring E6E7 fusion gene and vascular endothelial growth factor receptor 2 gene.

    PubMed

    Gao, Jie; Fan, Lei; Ma, Wei; Xiao, Huan

    2016-09-01

    Human papillomavirus (HPV) has been identified as the primary etiological factor in cervical cancer as well as in subsets of anogenital and oropharyngeal cancers. The two HPV viral oncoproteins, E6 and E7, are uniquely and consistently expressed in all HPV-infected cells and are therefore promising targets for therapeutic vaccination. In order to achieve a synergistic antitumor and anti-angiogenesis effect, we designed and constructed a novel DNA vaccine that can express the HPV 16 E6E7 fusion protein and VEGFR2 in the same reading frame. A series of DNA plasmids encoding E6E7, VEGFR2 and their conjugates were constructed and injected into mice. The resultant humoral and cellular immune responses were detected by ELISA and enzyme-linked immunospot (ELISPOT), respectively. To evaluate the antitumor efficacy of these plasmids, tumor-bearing mice expressing the E6E7 fusion protein were constructed. After injection into the tumor-bearing mouse model, the plasmid harboring the E6E7 fusion gene and VEGFR2 showed stronger inhibition of tumor growth than the plasmid expressing E6E7 or VEGFR2 alone, which indicated that the combination of E6E7 and VEGFR2 could exert a synergistic antitumor effect. These observations emphasize the potential of a synergistic antitumor and anti-angiogenesis strategy using a DNA vaccine, which could be a promising approach for tumor immunotherapy.

  12. IFMIF, International Fusion Materials Irradiation Facility conceptual design activity cost report

    SciTech Connect

    Rennich, M.J.

    1996-12-01

    This report documents the cost estimate for the International Fusion Materials Irradiation Facility (IFMIF) at the completion of the Conceptual Design Activity (CDA). The estimate corresponds to the design documented in the Final IFMIF CDA Report. In order to effectively involve all the collaborating parties in the development of the estimate, a preparatory meeting was held at Oak Ridge National Laboratory in March 1996 to jointly establish guidelines to insure that the estimate was uniformly prepared while still permitting each country to use customary costing techniques. These guidelines are described in Section 4. A preliminary cost estimate was issued in July 1996 based on the results of the Second Design Integration Meeting, May 20--27, 1996 at JAERI, Tokai, Japan. This document served as the basis for the final costing and review efforts culminating in a final review during the Third IFMIF Design Integration Meeting, October 14--25, 1996, ENEA, Frascati, Italy. The present estimate is a baseline cost estimate which does not apply to a specific site. A revised cost estimate will be prepared following the assignment of both the site and all the facility responsibilities.

  13. Region between the canine distemper virus M and F genes modulates virulence by controlling fusion protein expression.

    PubMed

    Anderson, Danielle E; von Messling, Veronika

    2008-11-01

    Morbilliviruses, including measles and canine distemper virus (CDV), are nonsegmented, negative-stranded RNA viruses that cause severe diseases in humans and animals. The transcriptional units in their genomes are separated by untranslated regions (UTRs), which contain essential transcription and translation signals. Due to its increased length, the region between the matrix (M) protein and fusion (F) protein open reading frames is of particular interest. In measles virus, the entire F 5' region is untranslated, while several start codons are found in most other morbilliviruses, resulting in a long F protein signal peptide (Fsp). To characterize the role of this region in morbillivirus pathogenesis, we constructed recombinant CDVs, in which either the M-F UTR was replaced with that between the nucleocapsid (N) and phosphoprotein (P) genes, or 106 Fsp residues were deleted. The Fsp deletion alone had no effect in vitro and in vivo. In contrast, substitution of the UTR was associated with a slight increase in F gene and protein expression. Animals infected with this virus either recovered completely or experienced prolonged disease and death due to neuroinvasion. The combination of both changes resulted in a virus with strongly increased F gene and protein expression and complete attenuation. Taken together, our results provide evidence that the region between the morbillivirus M and F genes modulates virulence through transcriptional control of the F gene expression.

  14. Inertial Confinement Fusion Target Component Fabrication and Technology Development report. Annual report, October 1, 1992--September 30, 1993

    SciTech Connect

    Steinman, D.

    1994-03-01

    On December 30, 1990, the US Department of Energy entered into a contract with General Atomics (GA) to be the Inertial Confinement Fusion Target Component Fabrication and Technology Development Support contractor. This report documents the technical activities which took place under this contract during the period of October 1, 1992 through September 30, 1993. During this period, GA was assigned 18 tasks in support of the Inertial Confinement Fusion program and its laboratories. These tasks included ``Capabilities Activation`` and ``Capabilities Demonstration`` to enable us to begin production of glass and composite polymer capsules. Capsule delivery tasks included ``Small Glass Shell Deliveries`` and ``Composite Polymer Capsules`` for Lawrence Livermore National Laboratory (LLNL) and Los Alamos National Laboratory (LANL). We also were asked to provide direct ``Onsite Support`` at LLNL and LANL. We continued planning for the transfer of ``Micromachining Equipment from Rocky Flats`` and established ``Target Component Micromachining and Electroplating Facilities`` at GA. We fabricated over 1100 films and filters of 11 types for Sandia National Laboratory and provided full-time onsite engineering support for target fabrication and characterization. We initiated development of methods to make targets for the Naval Research Laboratory. We investigated spherical interferometry, built an automated capsule sorter, and developed an apparatus for calorimetric measurement of fuel fill for LLNL. We assisted LANL in the ``Characterization of Opaque b-Layered Targets.`` We developed deuterated and UV-opaque polymers for use by the University of Rochester`s Laboratory for Laser Energetics (UR/LLE) and devised a triple-orifice droplet generator to demonstrate the controlled-mass nature of the microencapsulation process.

  15. Increased expression of the diabetes gene SOX4 reduces insulin secretion by impaired fusion pore expansion

    PubMed Central

    Collins, Stephan C.; Do, Hyun Woong; Hastoy, Benoit; Hugill, Alison; Adam, Julie; Chibalina, Margarita V.; Galvanovskis, Juris; Godazgar, Mahdieh; Lee, Sheena; Goldsworthy, Michelle; Salehi, Albert; Tarasov, Andrei I.; Rosengren, Anders H.; Cox, Roger; Rorsman, Patrik

    2016-01-01

    The transcription factor Sox4 has been proposed to underlie the increased type-2 diabetes risk linked to an intronic SNP in CDKAL1. In a mouse model expressing a mutant form of Sox4, glucose-induced insulin secretion is reduced by 40% despite normal intracellular Ca2+ signalling and depolarization-evoked exocytosis. This paradox is explained by a 4-fold increase in kiss-and-run exocytosis (as determined by single-granule exocytosis measurements), in which the fusion pore connecting the granule lumen to the exterior only expands to a diameter of 2 nm that does not allow the exit of insulin. Microarray analysis indicated that this correlated with an increased expression of the exocytosis-regulating protein Stxbp6. In a large collection of human islet preparations (n=63), STXBP6 expression and GIIS correlated positively and negatively with SOX4 expression, respectively. Overexpression of SOX4 in the human insulin-secreting cell EndoC-βH2 interfered with granule emptying and inhibited hormone release, the latter effect was reversed by silencing of STXBP6. These data suggest that increased SOX4 expression inhibits insulin secretion and increased diabetes risk by upregulation of STXBP6 and an increase in kiss-and-run exocytosis at the expense of full fusion. We propose that pharmacological interventions promoting fusion pore expansion may be effective in diabetes therapy. PMID:26993066

  16. Genes encoding Cher-TPR fusion proteins are predominantly found in gene clusters encoding chemosensory pathways with alternative cellular functions.

    PubMed

    Muñoz-Martínez, Francisco; García-Fontana, Cristina; Rico-Jiménez, Miriam; Alfonso, Carlos; Krell, Tino

    2012-01-01

    Chemosensory pathways correspond to major signal transduction mechanisms and can be classified into the functional families flagellum-mediated taxis, type four pili-mediated taxis or pathways with alternative cellular functions (ACF). CheR methyltransferases are core enzymes in all of these families. CheR proteins fused to tetratricopeptide repeat (TPR) domains have been reported and we present an analysis of this uncharacterized family. We show that CheR-TPRs are widely distributed in GRAM-negative but almost absent from GRAM-positive bacteria. Most strains contain a single CheR-TPR and its abundance does not correlate with the number of chemoreceptors. The TPR domain fused to CheR is comparatively short and frequently composed of 2 repeats. The majority of CheR-TPR genes were found in gene clusters that harbor multidomain response regulators in which the REC domain is fused to different output domains like HK, GGDEF, EAL, HPT, AAA, PAS, GAF, additional REC, HTH, phosphatase or combinations thereof. The response regulator architectures coincide with those reported for the ACF family of pathways. Since the presence of multidomain response regulators is a distinctive feature of this pathway family, we conclude that CheR-TPR proteins form part of ACF type pathways. The diversity of response regulator output domains suggests that the ACF pathways form a superfamily which regroups many different regulatory mechanisms, in which all CheR-TPR proteins appear to participate. In the second part we characterize WspC of Pseudomonas putida, a representative example of CheR-TPR. The affinities of WspC-Pp for S-adenosylmethionine and S-adenosylhomocysteine were comparable to those of prototypal CheR, indicating that WspC-Pp activity is in analogy to prototypal CheRs controlled by product feed-back inhibition. The removal of the TPR domain did not impact significantly on the binding constants and consequently not on the product feed-back inhibition. WspC-Pp was found to be

  17. Gene identification and analysis: an application of neural network-based information fusion

    SciTech Connect

    Matis, S.; Xu, Y.; Shah, M.B.; Mural, R.J.; Einstein, J.R.; Uberbacher, E.C.

    1996-10-01

    Identifying genes within large regions of uncharacterized DNA is a difficult undertaking and is currently the focus of many research efforts. We describe a gene localization and modeling system called GRAIL. GRAIL is a multiple sensor-neural network based system. It localizes genes in anonymous DNA sequence by recognizing gene features related to protein-coding slice sites, and then combines the recognized features using a neural network system. Localized coding regions are then optimally parsed into a gene mode. RNA polymerase II promoters can also be predicted. Through years of extensive testing, GRAIL consistently localizes about 90 percent of coding portions of test genes with a false positive rate of about 10 percent. A number of genes for major genetic diseases have been located through the use of GRAIL, and over 1000 research laboratories worldwide use GRAIL on regular bases for localization of genes on their newly sequenced DNA.

  18. Fusion Energy Division annual progress report period ending December 31, 1983

    SciTech Connect

    Not Available

    1984-09-01

    The Fusion Program carries out work in a number of areas: (1) experimental and theoretical research on two magnetic confinement concepts - the ELMO Bumpy Torus (EBT) and the tokamak, (2) theoretical and engineering studies on a third concept - the stellarator, (3) engineering and physics of present-generation fusion devices, (4) development and testing of diagnostic tools and techniques, (5) development and testing of materials for fusion devices, (6) development and testing of the essential technologies for heating and fueling fusion plasmas, (7) development and testing of the superconducting magnets that will be needed to confine these plasmas, (8) design of future devices, (9) assessment of the environmental impact of fusion energy, and (10) assembly and distribution to the fusion community of data bases on atomic physics and radiation effects. The interactions between these activities and their integration into a unified program are major factors in the success of the individual activities, and the ORNL Fusion Program strives to maintain a balance among these activities that will lead to continued growth.

  19. Establishment of an Institute for Fusion Studies. Technical progress report, November 1, 1991--October 31, 1992

    SciTech Connect

    Hazeltine, R.D.

    1992-07-01

    The Institute for Fusion Studies is a national center for theoretical fusion plasma physics research. Its purposes are: (1) to conduct research on theoretical questions concerning the achievement of controlled fusion energy by means of magnetic confinement--including both fundamental problems of long-range significance, as well as shorter-term issues; (2) to serve as a center for information exchange, nationally and internationally, by hosting exchange visits, conferences, and workshops; (3) and to train students and postdoctoral research personnel for the fusion energy program and plasma physics research areas. The theoretical research results that are obtained by the Institute contribute mainly to the progress of national and international efforts in nuclear fusion research, whose goal is the development of fusion power.as a basic energy source. In addition to its primary focus on fusion physics, the Institute is also involved with research in related fields, such as advanced computing techniques, nonlinear dynamics, plasma astrophysics, and accelerator physics. The work of EFS scientists continued to receive national and international recognition. Numerous invited papers were given during the past year at workshops, conferences, and scientific meetings. Last year IFS scientists published 95 scientific articles in technical journals and monographs.

  20. CONFERENCE REPORT: Summary of the 16th IAEA Technical Meeting on 'Research using Small Fusion Devices'

    NASA Astrophysics Data System (ADS)

    Gribkov, V.; Van Oost, G.; Malaquias, A.; Herrera, J.

    2006-10-01

    Common research topics that are being studied in small, medium and large devices such as H-mode like or improved confinement, turbulence and transport are reported. These included modelling and diagnostic developments for edge and core, to characterize plasma density, temperature, electric potential, plasma flows, turbulence scale, etc. Innovative diagnostic methods were designed and implemented which could be used to develop experiments in small devices (in some cases not possible in large devices due to higher power deposition) to allow a better understanding of plasma edge and core properties. Reports are given addressing research in linear devices that can be used to study particular plasma physics topics relevant for other magnetic confinement devices such as the radial transport and the modelling of self-organized plasma jets involved in spheromak-like plasma formation. Some aspects of the work presented are of interest to the astrophysics community since they are believed to shed light on the basis of the physics of stellar jets. On the dense magnetized plasmas (DMP) topic, the present status of research, operation of new devices, plasma dynamics modelling and diagnostic developments is reported. The main devices presented belong to the class of Z-pinches, mostly plasma foci, and several papers were presented under this topic. The physics of DMP is important both for the main-stream fusion investigations as well as for providing the basis for elaboration of new concepts. New high-current technology introduced in the DMP devices design and construction make these devices nowadays more reliably fitted to various applications and give the possibility to widen the energy range used by them in both directions—to the multi-MJ level facilities and down to miniature plasma focus devices with energy of just a few J.

  1. Urokinase-Targeted Fusion by Oncolytic Sendai Virus Eradicates Orthotopic Glioblastomas by Pronounced Synergy With Interferon-β Gene

    PubMed Central

    Hasegawa, Yuzo; Kinoh, Hiroaki; Iwadate, Yasuo; Onimaru, Mitsuho; Ueda, Yasuji; Harada, Yui; Saito, Satoru; Furuya, Aki; Saegusa, Takashi; Morodomi, Yosuke; Hasegawa, Mamoru; Saito, Shigeyoshi; Aoki, Ichio; Saeki, Naokatsu; Yonemitsu, Yoshikazu

    2010-01-01

    Glioblastoma multiforme (GM), the most frequent primary malignant brain tumor, is highly invasive due to the expression of proteases, including urokinase-type plasminogen activator (uPA). Here, we show the potential of our new and powerful recombinant Sendai virus (rSeV) showing uPA-specific cell-to-cell fusion activity [rSeV/dMFct14 (uPA2), named “BioKnife”] for GM treatment, an effect that was synergistically enhanced by arming BioKnife with the interferon-β (IFN-β) gene. BioKnife killed human GM cell lines efficiently in a uPA-dependent fashion, and this killing was prevented by PA inhibitor-1. Rat gliosarcoma 9L cells expressing both uPA and its functional receptor uPAR (9L-L/R) exhibited high uPA activity on the cellular surface and were highly susceptible to BioKnife. Although parent 9L cells (9L-P) were resistant to BioKnife and to BioKnife expressing IFN-β (BioKnife-IFNβ), cell–cell fusion of 9L-L/R strongly facilitated the expression of IFN-β, and in turn, IFN-β significantly accelerated the fusion activity of BioKnife. A similar synergy was seen in a rat orthotopic brain GM model with 9L-L/R in vivo; therefore, these results suggest that BioKnife-IFNβ may have significant potential to improve the survival of GM patients in a clinical setting. PMID:20606645

  2. Instrumented reduction and monosegmental fusion for Meyerding Grade IV developmental spondylolisthesis: a report of 3 cases.

    PubMed

    Mizuno, Kentaro; Mikami, Yasuo; Nagae, Masateru; Tonomura, Hitoshi; Ikeda, Takumi; Fujiwara, Hiroyoshi; Kubo, Toshikazu

    2014-12-01

    There are numerous reports of treatment methods for spondylolisthesis with a Meyerding Grade of more than III. In high dysplastic spondylosthesis, surgical treatment was selected because there is considered to be a high possibility of low back pain and lower limb neurological symptoms worsening if slippage progresses. Monosegmental lumbar interbody fusion (L5-S1) with a pedicle screw system (PPS) was used to treat three cases of Meyerding Grade IV developmental spondylolisthesis. Patients gave written informed consent. The spondylolisthesis was reduced to Meyerding Grade I and sagittal balance improved in all three cases. In two cases with severe spinal instability, there were no postoperative neurological complications and the course was favorable. However, in one case with little spinal mobility due to vertebral body dysplasia, despite performing sufficient decompression of the nerve root at L5 and slow reduction to avoid placing excessive tension on the nerve root, a transient neurological disorder was observed. A PPS was used to increase the reduction strength and favorable reduction was possible. However, in the case with a long clinical course and the case with poor spinal mobility, since the mobility and plasticity of the nerve root itself may have been reduced, it was considered that reduction should be performed carefully using intraoperative neurological monitoring.

  3. Deep vein thrombosis due to migrated graft bone after posterior lumbosacral interbody fusion. Case report.

    PubMed

    Yoshimoto, Hisashi; Sato, Shigenobu; Nakagawa, Izumi; Hyakumachi, Takahiko; Yanagibashi, Yasushi; Nitta, Fumihito; Masuda, Takeshi

    2007-01-01

    The authors report the case of an 83-year-old woman with refractory sciatica attributable to isthmic spondylolisthesis at L-5. Her symptoms were successfully improved after posterior lumbar interbody fusion (PLIF) at L5-S1; however, notable swelling in her left leg suddenly developed 2 days postoperatively. Anterior migration of a fragment of bone graft was demonstrated on computed tomography scanning, and there was obvious occlusion of the left common iliac vein (CIV) on magnetic resonance venography. Ultrasonography revealed a thrombus in the left CIV at the site of compression. To prevent a pulmonary embolism during manipulation of the affected vein, an inferior vena cava filter was placed just before excision of the migrated bone fragment. The swelling in the patient's leg subsided quickly after the surgery, and she was treated with heparin and warfarin to prevent recurrent deep vein thrombosis (DVT). Six months after the second surgery, complete restoration of blood flow to the left CIV and no recurrence of DVT were demonstrated on magnetic resonance venography. Especially in elderly patients with degenerative disc disease, excessive curettage and impaction of disc materials during the PLIF procedure may cause migration of bone graft fragments. Surgeons should be aware of the possible vascular complications of PLIF.

  4. Alloy development for irradiation performance in fusion reactors. Annual report, September 1979-September 1980

    SciTech Connect

    Harling, O K; Grant, N J

    1980-12-01

    This report summarizes the research and development work performed during the second year of an M.I.T. project directed toward the development of improved structural alloys for the fusion reactor first wall application. Several new alloys have been produced by rapid solidification. Emphasis in alloy design and production has been placed on producing austenitic Type 316SS with fine dispersions of TiC and Al/sub 2/O/sub 3/ particles. Results of mechanical and microstructural tests are presented. A number of neutron irradiations have been initiated on samples fabricated from alloys produced in this project. A dual beam, heavy ion and helium ion, irradiation was completed using several alloys and a range of temperatures, damage rates and total doses. Modeling of irradiation phenomena has been continued with emphasis in the last year upon understanding the effect of recoil resolution on relatively stable second phase particles. Work continued to fully characterize the microstructure of several ZrB/sub 2/ doped stainless steels.

  5. Construction, expression, and characterization of Arabidopsis thaliana 4CL and Arachis hypogaea RS fusion gene 4CL::RS in Escherichia coli.

    PubMed

    Zhang, Erhao; Guo, Xuefeng; Meng, Zhifen; Wang, Jin; Sun, Jia; Yao, Xi; Xun, Hang

    2015-09-01

    Resveratrol is an important antioxidant that confers several beneficial effects on human health. 4-coumarate coenzyme A ligase (4CL) and resveratrol synthase (RS) are key rate-limiting enzymes in the biosynthetic pathway of resveratrol. Using gene fusion technology, the fusion gene, 4CL::RS, was constructed by the 4CL gene from Arabidopsis thaliana and RS gene from Arachis hypogaea. DNAMAN analysis showed that the fusion gene encoded a 964-amino acid protein with an approximate weight of 104.7 kDa and a pI of 5.63. A prokaryotic expression vector containing Nco-I and EcoR-I restriction sites, pET-30a/4CL::RS, was identified by liquid culture bacterial PCR, enzyme digestion, and sequencing, and then used in the induction of expression. Subsequently, a biosynthetic pathway of resveratrol was constructed in Escherichia coli BL21(DE3) that harbored pET-30a/4CL::RS. The recombinant strains were induced to express the fusion protein at 28 °C for 8 h. After bacterial cells were disrupted by hypothermic ultrasonication, the 4CL::RS fusion protein was thoroughly separated from tags using Ni-NTA affinity chromatography, and then detected by SDS-PAGE analysis. When the recombinant strains expressed the fusion protein, the precursor, p-coumaric acid, was converted to resveratrol. In the present study, the final concentration of resveratrol derived from 1 mM p-coumaric acid was 80.524 mg/L, with a 35.28 % (mol/mol) conversion yield.

  6. Functional consequences of a gene duplication and fusion event in an arginine kinase.

    PubMed

    Compaan, Deanne M; Ellington, W Ross

    2003-05-01

    Arginine kinase (AK) from the foot of the razor clam Ensis directus consists of two full-length AK domains, denoted D1 and D2, fused in a single polypeptide chain. The full-length cDNA for Ensis AK was obtained and its deduced amino acid sequence was analyzed in the context of the X-ray crystal structure of a typical, monomeric AK. Both domains of Ensis AK contain most of the residues currently thought to be critical in catalysis, suggesting that both AK domains are catalytically competent. The full-length Ensis AK, a D2-NusA-His-tag fusion protein and a D2-truncated AK (enterokinase cleavage product of the fusion protein) were expressed in Escherichia coli and purified. All recombinant AK constructs displayed high enzyme activity. Attempts at expressing active D1 alone, D2 alone or a D1-NusA-His-tag fusion protein were unsuccessful. The catalytic properties of the active proteins were compared with the corresponding properties of recombinant AK from the horseshoe crab Limulus polyphemus, which is a typical monomeric AK. In contrast to expectations, the kinetic results strongly suggest that Ensis AK has only one active domain, namely D2. The K(cat) values for all Ensis constructs were roughly twice that of typical AKs, indicating higher overall catalytic throughput at the competent active site. Furthermore, both the full-length and truncated D2 Ensis AKs showed no synergism of substrate binding unlike typical AKs. The D2-NusA-His-tag fusion construct actually displayed negative synergism of substrate binding, which means that, in effect, the first substrate bound acts as a competitive inhibitor of the second. The conservation of the structure of the apparently inactive D1 may be related to constraints imposed by structural changes that could potentially impact substrate binding in D2 and/or possibly influence the proper folding of the enzyme during synthesis. Overall, the results from the present study indicate that the AK contiguous dimer from Ensis directus

  7. Fusion energy division annual progress report, period ending December 31, 1980

    SciTech Connect

    Not Available

    1981-11-01

    The ORNL Program encompasses most aspects of magnetic fusion research including research on two magnetic confinement programs (tokamaks and ELMO bumpy tori); the development of the essential technologies for plasma heating, fueling, superconducting magnets, and materials; the development of diagnostics; the development of atomic physics and radiation effect data bases; the assessment of the environmental impact of magnetic fusion; the physics and engineering of present-generation devices; and the design of future devices. The integration of all of these activities into one program is a major factor in the success of each activity. An excellent example of this integration is the extremely successful application of neutral injection heating systems developed at ORNL to tokamaks both in the Fusion Energy Division and at Princeton Plasma Physics Laboratory (PPPL). The goal of the ORNL Fusion Program is to maintain this balance between plasma confinement, technology, and engineering activities.

  8. Fusion driver study. Final technical report, April 1, 1978-March 31, 1980

    SciTech Connect

    Friedman, H.W.

    1980-04-01

    A conceptual design of a multi-megajoule, repetitively pulsed CO/sub 2/ laser system for Inertial Confinement Fusion is presented. System configurations consisting of 50 to 100 kJ modules operating at subatmospheric pressures with multiple pass optical extraction appear feasible with present or near term technology. Overall laser system efficiencies of greater than 10% at repetition rates in excess of 10 Hz are possible with the state-of-the-art pulsed power technology. The synthesis of all the laser subsystems into a specific configuration for a Laser Fusion Driver depends upon the reactor chamber(s) layout, subsystem reliability and restrictions on overall dimensions of the fusion driver. A design is presented which stacks power amplifier modules in series in a large torus with centrally located reactor chamber. Cost estimates of the overall Laser Fusion Driver are also presented.

  9. Accelerator and Fusion Research Division annual report, fiscal year 1980, October 1979-September 1980

    SciTech Connect

    Not Available

    1981-03-01

    Research during October 1979 to September 1980 is summarized. Areas covered include: accelerator operations; positron-electron project; stochastic beam cooling; high-field superconducting magnets; accelerator theory; neutral beam sources; and heavy ion fusion. (GHT)

  10. SECAD-- a Schema-based Environment for Configuring, Analyzing and Documenting Integrated Fusion Simulations. Final report

    SciTech Connect

    Shasharina, Svetlana

    2012-05-23

    SECAD is a project that developed a GUI for running integrated fusion simulations as implemented in FACETS and SWIM SciDAC projects. Using the GUI users can submit simulations locally and remotely and visualize the simulation results.

  11. Internet and web projects for fusion plasma science and education. Final technical report

    SciTech Connect

    Eastman, Timothy E.

    1999-08-30

    The plasma web site at http://www.plasmas.org provides comprehensive coverage of all plasma science and technology with site links worldwide. Prepared to serve the general public, students, educators, researchers, and decision-makers, the site covers basic plasma physics, fusion energy, magnetic confinement fusion, high energy density physics include ICF, space physics and astrophysics, pulsed-power, lighting, waste treatment, plasma technology, plasma theory, simulations and modeling.

  12. Minimally invasive atlantoaxial fusion: cadaveric study and report of 5 clinical cases.

    PubMed

    Srikantha, Umesh; Khanapure, Kiran S; Jagannatha, Aniruddha T; Joshi, Krishna C; Varma, Ravi G; Hegde, Alangar S

    2016-12-01

    OBJECTIVE Minimally invasive techniques are being increasingly used to treat disorders of the cervical spine. They have a potential to reduce the postoperative neck discomfort subsequent to extensive muscle dissection associated with conventional atlantoaxial fusion procedures. The aim of this paper was to elaborate on the technique and results of minimally invasive atlantoaxial fusion. MATERIALS Minimally invasive atlantoaxial fusion was done initially in 4 fresh-frozen cadavers and subsequently in 5 clinical cases. Clinical cases included patients with reducible atlantoaxial instability and undisplaced or minimally displaced odontoid fractures. The surgical technique is illustrated in detail. RESULTS Among the cadaveric specimens, all C-1 lateral mass screws were in the correct position and 2 of the 8 C-2 screws had a vertebral canal breach. Among clinical cases, all C-1 lateral mass screws were in the correct position. Only one C-2 screw had a Grade 2 vertebral canal breach, which was clinically insignificant. None of the patients experienced neurological worsening or implant-related complications at follow-up. Evidence of rib graft fusion or C1-2 joint fusion was successfully demonstrated in 4 cases, and flexion-extension radiographs done at follow-up did not show mobility in any case. CONCLUSIONS Minimally invasive atlantoaxial fusion is a safe and effective alternative to the conventional approach in selected cases. Larger series with direct comparison to the conventional approach will be required to demonstrate clinical benefit presumed to be associated with a minimally invasive approach.

  13. Inertial confinement fusion research and development studies. Final report, October 1979-August 1980

    SciTech Connect

    Bullis, R.; Finkelman, M.; Leng, J.; Luzzi, T.; Ojalvo, I.; Powell, E.; Sedgley, D.

    1980-08-01

    These Inertial Confinement Fusion (ICF) research and development studies were selected for structural, thermal, and vacuum pumping analyses in support of the High Yield Lithium Injection Fusion Energy (HYLIFE) concept development. An additional task provided an outlined program plan for an ICF Engineering Test Facility, using the HYLIFE concept as a model, although the plan is generally applicable to other ICF concepts. The HYLIFE is one promising type of ICF concept which features a falling array of liquid lithium jets. These jets surround the fusion reaction to protect the first structural wall (FSW) of the vacuum chamber by absorbing the fusion energy, and to act as the tritium breeder. The fusion energy source is a deuterium-tritium pellet injected into the chamber every second and driven by laser or heavy ion beams. The studies performed by Grumman have considered the capabilities of specific HYLIFE features to meet life requirements and the requirement to recover to preshot conditions prior to each subsequent shot. The components under investigation were the FSW which restrains the outward motion of the liquid lithium, the nozzle plate which forms the falling jet array, the graphite shield which is in direct top view of the fusion pellet, and the vacuum pumping system. The FSW studies included structural analysis, and definition of an experimental program to validate computer codes describing lithium motion and the resulting impact on the wall.

  14. Positron Emission Tomography Reporter Genes and Reporter Probes: Gene and Cell Therapy Applications

    PubMed Central

    Yaghoubi, Shahriar S.; Campbell, Dean O.; Radu, Caius G.; Czernin, Johannes

    2012-01-01

    Positron emission tomography (PET) imaging reporter genes (IRGs) and PET reporter probes (PRPs) are amongst the most valuable tools for gene and cell therapy. PET IRGs/PRPs can be used to non-invasively monitor all aspects of the kinetics of therapeutic transgenes and cells in all types of living mammals. This technology is generalizable and can allow long-term kinetics monitoring. In gene therapy, PET IRGs/PRPs can be used for whole-body imaging of therapeutic transgene expression, monitoring variations in the magnitude of transgene expression over time. In cell or cellular gene therapy, PET IRGs/PRPs can be used for whole-body monitoring of therapeutic cell locations, quantity at all locations, survival and proliferation over time and also possibly changes in characteristics or function over time. In this review, we have classified PET IRGs/PRPs into two groups based on the source from which they were derived: human or non-human. This classification addresses the important concern of potential immunogenicity in humans, which is important for expansion of PET IRG imaging in clinical trials. We have then discussed the application of this technology in gene/cell therapy and described its use in these fields, including a summary of using PET IRGs/PRPs in gene and cell therapy clinical trials. This review concludes with a discussion of the future direction of PET IRGs/PRPs and recommends cell and gene therapists collaborate with molecular imaging experts early in their investigations to choose a PET IRG/PRP system suitable for progression into clinical trials. PMID:22509201

  15. The t(10;11)(p13;q14) in the U937 cell line results in the fusion of the AF10 gene and CALM, encoding a new member of the AP-3 clathrin assembly protein family.

    PubMed Central

    Dreyling, M H; Martinez-Climent, J A; Zheng, M; Mao, J; Rowley, J D; Bohlander, S K

    1996-01-01

    The translocation t(10;11)(p13;q14) is a recurring chromosomal abnormality that has been observed in patients with acute lymphoblastic leukemia as well as acute myeloid leukemia. We have recently reported that the monocytic cell line U937 has a t(10;11)(p13;q14) translocation. Using a combination of positional cloning and candidate gene approach, we cloned the breakpoint and were able to show that AF10 is fused to a novel gene that we named CALM (Clathrin Assembly Lymphoid Myeloid leukemia gene) located at 11q14. AF10, a putative transcription factor, had recently been cloned as one of the fusion partners of MLL. CALM has a very high homology in its N-terminal third to the murine ap-3 gene which is one of the clathrin assembly proteins. The N-terminal region of ap-3 has been shown to bind to clathrin and to have a high-affinity binding site for phosphoinositols. The identification of the CALM/AF10 fusion gene in the widely used U937 cell line will contribute to our understanding of the malignant phenotype of this line. Images Fig. 1 Fig. 3 PMID:8643484

  16. Evidence that the multifunctional polypeptides of vertebrate and fungal fatty acid synthases have arisen by independent gene fusion events.

    PubMed

    McCarthy, A D; Goldring, J P; Hardie, D G

    1983-10-17

    The enoyl reductase (NADPH binding site) of rabbit mammary fatty acid synthase has been radioactively labelled using pyridoxal phosphate and sodium [3H]borohydride. Using this method we have been able to add this site to the four sites whose location has already been mapped within the multifunctional polypeptide chain of the protein. The results show that the enoyl reductase lies between the 3-oxoacylsynthase and the acyl carrier. This confirms that the active sites occur in a different order on the single multifunctional polypeptide of vertebrate fatty acid synthase and the two multifunctional polypeptides of fungal fatty acid synthase, and suggests that these two systems have arisen by independent gene fusion events.

  17. Osiris and SOMBRERO inertial confinement fusion power plant designs. Volume 1, Executive summary and overview, Final report

    SciTech Connect

    Meier, W.R.; Bieri, R.L.; Monsler, M.J.

    1992-03-01

    Conceptual designs and assessments have been completed for two inertial fusion energy (IFE) electric power plants. The detailed designs and results of the assessment studies are presented in this report. Osiris is a heavy-ion-beam (HIB) driven power plant and SOMBRERO is a Krypton-Fluoride (KrF) laser-driven power plant. Both plants are sized for a net electric power of 1000 MWe.

  18. Postoperative Cervical Haematoma Complicated by Ipsilateral Carotid Thrombosis and Aphasia after Anterior Cervical Fusion: A Case Report

    PubMed Central

    Chin, Kingsley R.; Seale, Jason; Butron, Veronica

    2013-01-01

    Hematoma alone is the most common vascular complication reported after anterior cervical decompression and fusion (ACDF). We present this case to report the occurrence of postoperative cervical hematoma complicated by ipsilateral carotid thrombosis and aphasia after an uncomplicated C4–6 ACDF. This is a case of a 65-year-old woman who underwent revision fusions of the C4-5 and C6-7 levels complicated by postoperative cervical hematoma and carotid thrombosis. The patient's history, clinical examination, imaging findings, and treatment are reported. The revision fusions were performed and deemed routine. Approximately eight hours later 200 mL of blood was evacuated from a postoperative cervical hematoma. The patient became unresponsive and disoriented a few hours after evacuating the hematoma. Computed tomography and magnetic resonance imaging of the brain were normal, but magnetic resonance angiography demonstrated total occlusion of the left carotid artery. Thrombectomy was performed and the patient was discharged without residual deficits. At the latest followup she is fully functional and asymptomatic in her neck. We suggest, after evacuating a cervical hematoma, an evaluation of the carotids be made with MRA or cerebral angiography, as this may demonstrate a clot before the patient develops symptoms. PMID:23533432

  19. Cytotoxicity of HSVtk and hrTNF-alpha fusion genes with IRES in treatment of gastric cancer.

    PubMed

    Zhang, Jian-Hua; Wan, Ming-Xi; Pan, Bo-Rong; Yu, Bing

    2006-04-28

    The efficacy of the suicide gene therapy by using the herpes simplex virus thymidine kinase/ganciclovir (HSVtk/GCV) system for the treatment of cancer is limited because of the insufficient gene transfer and the low killing activity. To enhance the anti-tumor activity, we probed into whether recombinant retroviral expression vector PLXSN expressing both HSVtk and TNF-alpha genes could potentiate the destruction of SGC7901. The pL(tk-TNF-alpha)SN harboring HSVtk and TNF-alpha genes in sequence was constructed with a bicistronic unit including the internal ribosomal entry site, the recombinant retroviruses were transferred into SGC7901 cells by lipofectamine, and pEGFP and Western blot analysis were used to detect the expression of fusion genes in transfected SGC7901 cells, and then apoptosis of the transfected cells were detected by using the TdT-mediated dUTP nick end labeling, flow cytometric analysis and transmission electron microscopy. In vitro study, the transfected gastric cancer cells were maintained in the GCV-contained medium, to assay the cell killing effect and bystander effect. In vivo experiments, retroviral serum plasmids were transfected into tumor-bearing nude mice, to observe the changes of tumor volumes and survival of the mice. In vitro there was no significant difference of cell survival rate between the three groups. However, in vivo results showed that tk/GCV, tk-TNF-alpha/GCV and TNF-alpha could inhibit the tumor growth, and the obvious anti-tumor effect was shown in tk-TNF-alpha/GCV group, and TNF-alpha obviously enhanced the anti-tumor effect in vivo. The pathologic examination showed necrosis of the cancer in the treated groups.

  20. Transgenic Mouse Model Harboring the Transcriptional Fusion Ccl20-Luciferase as a Novel Reporter of Pro-Inflammatory Response

    PubMed Central

    Crispo, Martina; Van Maele, Laurye; Tabareau, Julien; Cayet, Delphine; Errea, Agustina; Ferreira, Ana María; Rumbo, Martin; Sirard, Jean Claude

    2013-01-01

    The chemokine CCL20, the unique ligand of CCR6 functions as an attractant of immune cells. Expression of CCL20 is induced by Toll-like Receptor (TLR) signaling or proinflammatory cytokine stimulation. However CCL20 is also constitutively produced at specific epithelial sites of mucosa. This expression profile is achieved by transcriptional regulation. In the present work we characterized regulatory features of mouse Ccl20 gene. Transcriptional fusions between the mouse Ccl20 promoter and the firefly luciferase (luc) encoding gene were constructed and assessed in in vitro and in vivo assays. We found that liver CCL20 expression and luciferase activity were upregulated by systemic administration of the TLR5 agonist flagellin. Using shRNA and dominant negative form specific for mouse TLR5, we showed that this expression was controlled by TLR5. To address in situ the regulation of gene activity, a transgenic mouse line harboring a functional Ccl20-luc fusion was generated. The luciferase expression was highly concordant with Ccl20 expression in different tissues. Our data indicate that the transgenic mouse model can be used to monitor activation of innate response in vivo. PMID:24265691

  1. Transgenic mouse model harboring the transcriptional fusion ccl20-luciferase as a novel reporter of pro-inflammatory response.

    PubMed

    Crispo, Martina; Van Maele, Laurye; Tabareau, Julien; Cayet, Delphine; Errea, Agustina; Ferreira, Ana María; Rumbo, Martin; Sirard, Jean Claude

    2013-01-01

    The chemokine CCL20, the unique ligand of CCR6 functions as an attractant of immune cells. Expression of CCL20 is induced by Toll-like Receptor (TLR) signaling or proinflammatory cytokine stimulation. However CCL20 is also constitutively produced at specific epithelial sites of mucosa. This expression profile is achieved by transcriptional regulation. In the present work we characterized regulatory features of mouse Ccl20 gene. Transcriptional fusions between the mouse Ccl20 promoter and the firefly luciferase (luc) encoding gene were constructed and assessed in in vitro and in vivo assays. We found that liver CCL20 expression and luciferase activity were upregulated by systemic administration of the TLR5 agonist flagellin. Using shRNA and dominant negative form specific for mouse TLR5, we showed that this expression was controlled by TLR5. To address in situ the regulation of gene activity, a transgenic mouse line harboring a functional Ccl20-luc fusion was generated. The luciferase expression was highly concordant with Ccl20 expression in different tissues. Our data indicate that the transgenic mouse model can be used to monitor activation of innate response in vivo.

  2. FY2014 FES (Fusion Energy Sciences) Theory & Simulation Performance Target, Final Report

    SciTech Connect

    Fu, Guoyong; Budny, Robert; Gorelenkov, Nikolai; Poli, Francesca; Chen, Yang; McClenaghan, Joseph; Lin, Zhihong; Spong, Don; Bass, Eric; Waltz, Ron

    2014-10-14

    We report here the work done for the FY14 OFES Theory Performance Target as given below: "Understanding alpha particle confinement in ITER, the world's first burning plasma experiment, is a key priority for the fusion program. In FY 2014, determine linear instability trends and thresholds of energetic particle-driven shear Alfven eigenmodes in ITER for a range of parameters and profiles using a set of complementary simulation models (gyrokinetic, hybrid, and gyrofluid). Carry out initial nonlinear simulations to assess the effects of the unstable modes on energetic particle transport". In the past year (FY14), a systematic study of the alpha-driven Alfven modes in ITER has been carried out jointly by researchers from six institutions involving seven codes including the transport simulation code TRANSP (R. Budny and F. Poli, PPPL), three gyrokinetic codes: GEM (Y. Chen, Univ. of Colorado), GTC (J. McClenaghan, Z. Lin, UCI), and GYRO (E. Bass, R. Waltz, UCSD/GA), the hybrid code M3D-K (G.Y. Fu, PPPL), the gyro-fluid code TAEFL (D. Spong, ORNL), and the linear kinetic stability code NOVA-K (N. Gorelenkov, PPPL). A range of ITER parameters and profiles are specified by TRANSP simulation of a hybrid scenario case and a steady-state scenario case. Based on the specified ITER equilibria linear stability calculations are done to determine the stability boundary of alpha-driven high-n TAEs using the five initial value codes (GEM, GTC, GYRO, M3D-K, and TAEFL) and the kinetic stability code (NOVA-K). Both the effects of alpha particles and beam ions have been considered. Finally, the effects of the unstable modes on energetic particle transport have been explored using GEM and M3D-K.

  3. Inertial Confinement Fusion. Annual report 10/1/98 through 9/30/99

    SciTech Connect

    Gibson, Jane

    1999-12-01

    General Atomics (GA) has served as the Inertial Confinement Fusion (ICF) Target Component Fabrication and Technology Development Support contractor for the U.S. Department of Energy since December 30, 1990. This report documents the technical activities of the period October 1, 1998 through September 30, 1999. During this period, GA and our partner Schafer Corporation were assigned 17 formal tasks in support of the ICF program and its five laboratories. A portion of the effort on these tasks included providing direct ''Onsite Support'' at Lawrence Livermore National Laboratory (LLNL), Los Alamos National Laboratory (LANL), and Sandia National Laboratory (SNL). We fabricated and delivered over 1790 hohlraum mandrels and numerous other micromachined components to LLNL, LANL, and SNL. We produced more than 1380 glass and plastic target capsules over a wide range of sizes and designs (plus over 300 near target-quality capsules) for LLNL, LANL, SNL, and University of Rochester/Laboratory for Laser Energetic (UR/LLE). We also delivered various target foils and films for Naval Research Lab (NRL) and UWLLE in FY99. We fabricated a device to polish NIF-sized beryllium shells and prepared a laboratory for the safe operation of beryllium polishing activities. This report describes these target fabrication activities and the target fabrication and characterization development activities that made the deliveries possible. During FY99, the GA/Schafer portion of the GA/Schafer-UR/LLE-LANL team effort for design, procurement, installation, and testing of the OMEGA Cryogenic Target System (OCTS) that will field cryogenic targets on OMEGA was completed. All components of the OCTS were procured, fabricated, assembled, tested, and shipped to UR/LLE. Only minor documentation tasks remain to be done in FY00. The ICF program is anticipating experiments at the OMEGA laser and the National Ignition Facility (NIF) which will require targets containing cryogenic layered D2 or deuterium

  4. Enhancing potency of siRNA targeting fusion genes by optimization outside of target sequence

    PubMed Central

    Gavrilov, Kseniya; Seo, Young-Eun; Tietjen, Gregory T.; Cui, Jiajia; Cheng, Christopher J.; Saltzman, W. Mark

    2015-01-01

    Canonical siRNA design algorithms have become remarkably effective at predicting favorable binding regions within a target mRNA, but in some cases (e.g., a fusion junction site) region choice is restricted. In these instances, alternative approaches are necessary to obtain a highly potent silencing molecule. Here we focus on strategies for rational optimization of two siRNAs that target the junction sites of fusion oncogenes BCR-ABL and TMPRSS2-ERG. We demonstrate that modifying the termini of these siRNAs with a terminal G-U wobble pair or a carefully selected pair of terminal asymmetry-enhancing mismatches can result in an increase in potency at low doses. Importantly, we observed that improvements in silencing at the mRNA level do not necessarily translate to reductions in protein level and/or cell death. Decline in protein level is also heavily influenced by targeted protein half-life, and delivery vehicle toxicity can confound measures of cell death due to silencing. Therefore, for BCR-ABL, which has a long protein half-life that is difficult to overcome using siRNA, we also developed a nontoxic transfection vector: poly(lactic-coglycolic acid) nanoparticles that release siRNA over many days. We show that this system can achieve effective killing of leukemic cells. These findings provide insights into the implications of siRNA sequence for potency and suggest strategies for the design of more effective therapeutic siRNA molecules. Furthermore, this work points to the importance of integrating studies of siRNA design and delivery, while heeding and addressing potential limitations such as restricted targetable mRNA regions, long protein half-lives, and nonspecific toxicities. PMID:26627251

  5. Conference Report on the 3rd International Symposium on Lithium Application for Fusion Devices

    NASA Astrophysics Data System (ADS)

    Mazzitelli, G.; Hirooka, Y.; Hu, J. S.; Mirnov, S. V.; Nygren, R.; Shimada, M.; Ono, M.; Tabares, F. L.

    2015-02-01

    The third International Symposium on Lithium Application for Fusion Device (ISLA-2013) was held on 9-11 October 2013 at ENEA Frascati Centre with growing participation and interest from the community working on more general aspect of liquid metal research for fusion energy development. ISLA-2013 has been confirmed to be the largest and the most important meeting dedicated to liquid metal application for the magnetic fusion research. Overall, 45 presentation plus 5 posters were given, representing 28 institutions from 11 countries. The latest experimental results from nine magnetic fusion devices were presented in 16 presentations from NSTX (PPPL, USA), FTU (ENEA, Italy), T-11M (Trinity, RF), T-10 (Kurchatov Institute, RF), TJ-II (CIEMAT, Spain), EAST(ASIPP, China), HT-7 (ASIPP, China), RFX (Padova, Italy), KTM (NNC RK, Kazakhstan). Sessions were devoted to the following: (I) lithium in magnetic confinement experiments (facility overviews), (II) lithium in magnetic confinement experiments (topical issues), (III) special session on liquid lithium technology, (IV) lithium laboratory test stands, (V) Lithium theory/modelling/comments, (VI) innovative lithium applications and (VII) special Session on lithium-safety and lithium handling. There was a wide participation from the fusion technology communities, including IFMIF and TBM communities providing productive exchange with the physics oriented magnetic confinement liquid metal research groups. This international workshop will continue on a biennial basis (alternating with the Plasma-Surface Interactions (PSI) Conference) and the next workshop will be held at CIEMAT, Madrid, Spain, in 2015.

  6. Conference report on the 3rd International Symposium on Lithium Application for Fusion Devices

    SciTech Connect

    Mazzitelli, Guiseppe; Hirooka, Y.; Hu, J. S.; Mirnov, S. V.; Nygren, R.; Shimada, M.; Ono, M.; Tabares, F. L.

    2015-01-14

    The third International Symposium on Lithium Application for Fusion Device (ISLA-2013) was held on 9-11 October 2013 at ENEA Frascati Centre with growing participation and interest from the community working on more general aspect of liquid metal research for fusion energy development. ISLA-2013 has been confirmed to be the largest and the most important meeting dedicated to liquid metal application for the magnetic fusion research. Overall, 45 presentation plus 5 posters were given, representing 28 institutions from 11 countries. The latest experimental results from nine magnetic fusion devices were presented in 16 presentations from NSTX (PPPL, USA), FTU (ENEA, Italy), T-11M (Trinity, RF), T-10 (Kurchatov Institute, RF), TJ-II (CIEMAT, Spain), EAST(ASIPP, China), HT-7 (ASIPP, China), RFX (Padova, Italy), KTM (NNC RK, Kazakhstan). Sessions were devoted to the following: (I) lithium in magnetic confinement experiments (facility overviews), (II) lithium in magnetic confinement experiments (topical issues), (III) special session on liquid lithium technology, (IV) lithium laboratory test stands, (V) Lithium theory/modelling/comments, (VI) innovative lithium applications and (VII) special Session on lithium-safety and lithium handling. There was a wide participation from the fusion technology communities, including IFMIF and TBM communities providing productive exchange with the physics oriented magnetic confinement liquid metal research groups. Furthermore, this international workshop will continue on a biennial basis (alternating with the Plasma-Surface Interactions (PSI) Conference) and the next workshop will be held at CIEMAT, Madrid, Spain, in 2015.

  7. Single-cell RNA-seq reveals activation of unique gene groups as a consequence of stem cell-parenchymal cell fusion.

    PubMed

    Freeman, Brian T; Jung, Jangwook P; Ogle, Brenda M

    2016-03-21

    Fusion of donor mesenchymal stem cells with parenchymal cells of the recipient can occur in the brain, liver, intestine and heart following transplantation. The therapeutic benefit or detriment of resultant hybrids is unknown. Here we sought a global view of phenotypic diversification of mesenchymal stem cell-cardiomyocyte hybrids and associated time course. Using single-cell RNA-seq, we found hybrids consistently increase ribosome components and decrease genes associated with the cell cycle suggesting an increase in protein production and decrease in proliferation to accommodate the fused state. But in the case of most other gene groups, hybrids were individually distinct. In fact, though hybrids can express a transcriptome similar to individual fusion partners, approximately one-third acquired distinct expression profiles in a single day. Some hybrids underwent reprogramming, expressing pluripotency and cardiac precursor genes latent in parental cells and associated with developmental and morphogenic gene groups. Other hybrids expressed genes associated with ontologic cancer sets and two hybrids of separate experimental replicates clustered with breast cancer cells, expressing critical oncogenes and lacking tumor suppressor genes. Rapid transcriptional diversification of this type garners consideration in the context of cellular transplantation to damaged tissues, those with viral infection or other microenvironmental conditions that might promote fusion.

  8. Construction and uses of a new transposable element whose insertion is able to produce gene fusions with the neomycin-phosphotransferase-coding region of Tn903.

    PubMed

    Ratet, P; Richaud, F

    1986-01-01

    We describe the construction of a transposable element derived from the Mu phage that upon insertion is able to create a gene fusion between the region of Tn903 coding for neomycin phosphotransferase (NPT I), which confers resistance to aminoglycosides including kanamycin (KmR), neomycin and G418, and the control elements of the gene where the insertion occurs. A chloramphenicol (Cm) transacetylase gene (cat) that confers resistance to Cm is present in the transposon so that transposition events can be monitored even when no active fusions with the nptI coding region occur. The transposase gene is deleted and, therefore, this transposon is perfectly stable upon insertion. The properties of this new transposable element were studied by obtaining gene fusions between the Escherichia coli L-arabinose operon and 'nptI gene. In some of them the KmR phenotype is induced by arabinose. Insertions of this element in cloned fragments of the T-DNA region of Agrobacterium rhizogenes were also isolated. Some of them confer a KmR phenotype upon its E. coli carriers, which indicates that portions of the T-DNA are expressed in these cells.

  9. Molecular characterization of partial fusion gene and C-terminus extension length of haemagglutinin-neuraminidase gene of recently isolated Newcastle disease virus isolates in Malaysia

    PubMed Central

    2010-01-01

    Background Newcastle disease (ND), caused by Newcastle disease virus (NDV), is a highly contagious disease of birds and has been one of the major causes of economic losses in the poultry industry. Despite routine vaccination programs, sporadic cases have occasionally occurred in the country and remain a constant threat to commercial poultry. Hence, the present study was aimed to characterize NDV isolates obtained from clinical cases in various locations of Malaysia between 2004 and 2007 based on sequence and phylogenetic analysis of partial F gene and C-terminus extension length of HN gene. Results The coding region of eleven NDV isolates fusion (F) gene and carboxyl terminal region of haemagglutinin-neuraminidase (HN) gene including extensions were amplified by reverse transcriptase PCR and directly sequenced. All the isolates have shown to have non-synonymous to synonymous base substitution rate ranging between 0.081 - 0.264 demonstrating presence of negative selection. Analysis based on F gene showed the characterized isolates possess three different types of protease cleavage site motifs; namely 112RRQKRF117, 112RRRKRF117 and 112GRQGRL117 and appear to show maximum identities with isolates in the region such as cockatoo/14698/90 (Indonesia), Ch/2000 (China), local isolate AF2240 indicating the high similarity of isolates circulating in the South East Asian countries. Meanwhile, one of the isolates resembles commonly used lentogenic vaccine strains. On further characterization of the HN gene, Malaysian isolates had C-terminus extensions of 0, 6 and 11 amino acids. Analysis of the phylogenetic tree revealed that the existence of three genetic groups; namely, genotype II, VII and VIII. Conclusions The study concluded that the occurrence of three types of NDV genotypes and presence of varied carboxyl terminus extension lengths among Malaysian isolates incriminated for sporadic cases. PMID:20691110

  10. Research Needs for Magnetic Fusion Energy Sciences. Report of the Research Needs Workshop (ReNeW) Bethesda, Maryland, June 8-12, 2009

    SciTech Connect

    2009-06-08

    Nuclear fusion - the process that powers the sun - offers an environmentally benign, intrinsically safe energy source with an abundant supply of low-cost fuel. It is the focus of an international research program, including the ITE R fusion collaboration, which involves seven parties representing half the world's population. The realization of fusion power would change the economics and ecology of energy production as profoundly as petroleum exploitation did two centuries ago. The 21st century finds fusion research in a transformed landscape. The worldwide fusion community broadly agrees that the science has advanced to the point where an aggressive action plan, aimed at the remaining barriers to practical fusion energy, is warranted. At the same time, and largely because of its scientific advance, the program faces new challenges; above all it is challenged to demonstrate the timeliness of its promised benefits. In response to this changed landscape, the Office of Fusion Energy Sciences (OFES ) in the US Department of Energy commissioned a number of community-based studies of the key scientific and technical foci of magnetic fusion research. The Research Needs Workshop (ReNeW) for Magnetic Fusion Energy Sciences is a capstone to these studies. In the context of magnetic fusion energy, ReNeW surveyed the issues identified in previous studies, and used them as a starting point to define and characterize the research activities that the advance of fusion as a practical energy source will require. Thus, ReNeW's task was to identify (1) the scientific and technological research frontiers of the fusion program, and, especially, (2) a set of activities that will most effectively advance those frontiers. (Note that ReNeW was not charged with developing a strategic plan or timeline for the implementation of fusion power.) This Report presents a portfolio of research activities for US research in magnetic fusion for the next two decades. It is intended to provide a

  11. Fusion of the Dhfr/Mtx and IR/MAR gene amplification methods produces a rapid and efficient method for stable recombinant protein production.

    PubMed

    Noguchi, Chiemi; Araki, Yoshio; Miki, Daisuke; Shimizu, Noriaki

    2012-01-01

    Amplification of the dihydrofolate reductase gene (Dhfr) by methotrexate (Mtx) exposure is commonly used for recombinant protein expression in Chinese hamster ovary (CHO) cells. However, this method is both time- and labor-intensive, and the high-producing cells that are generated are frequently unstable in culture. Another gene amplification method is based on using a plasmid bearing a mammalian replication initiation region (IR) and a matrix attachment region (MAR), which result in the spontaneous initiation of gene amplification in transfected cells. The IR/MAR and Dhfr/Mtx methods of gene amplification are based on entirely different principles. In this study, we combine these two methods to yield a novel method, termed the IR/MAR-Dhfr fusion method, which was used to express three proteins, the Fc receptor, GFP, and recombinant antibody. The fusion method resulted in a dramatic increase in expression of all three proteins in two CHO sub-lines, DXB-11, and DG44. The IR/MAR-Dhfr fusion amplified the genes rapidly and efficiently, and produced larger amounts of antibody than the Dhfr/Mtx or IR/MAR methods alone. While the amplified structure produced by the Dhfr/Mtx method was highly unstable, and the antibody production rate rapidly decreased with the culture time of the cells, the IR/MAR-Dhfr fusion method resulted in stable amplification and generated clonal cells that produced large amounts of antibody protein over a long period of time. In summary, the novel IR/MAR-Dhfr fusion method enables isolation of stable cells that produce larger amounts of a target recombinant protein more rapidly and easily than either the Dhfr/Mtx or IR/MAR methods alone.

  12. Final technical report. 1998 HU CFRT summer fusion high school workshop

    SciTech Connect

    Ali, Halima; Punjabi, Alkesh

    1999-07-01

    The center conducted its third High School Summer Fusion Science Workshop in Summer 1998. The center had only three faculty mentors available only for a part of Summer 1998, The center accepted four scholars in this workshop, Prof. Halima Ali coordinated this workshop. Each student was assigned to a research mentor according to the student's interest in a specific research area and problem. In the workshop in the center, the students received instructions and training in the basics of energy, plasma and fusion sciences. They also received one-on-one instructions and training by their mentors to further their understanding of the subject and to introduce to relevant concepts such as magnetic confinement fusion, tokamaks, diverters and area-preserving maps.

  13. Conference Report on the 2nd International Symposium on Lithium Applications for Fusion Devices

    NASA Astrophysics Data System (ADS)

    Ono, M.; Bell, M. G.; Hirooka, Y.; Kaita, R.; Kugel, H. W.; Mazzitelli, G.; Menard, J. E.; Mirnov, S. V.; Shimada, M.; Skinner, C. H.; Tabares, F. L.

    2012-03-01

    The 2nd International Symposium on Lithium Applications for Fusion Devices (ISLA-2011) was held on 27-29 April 2011 at the Princeton Plasma Physics Laboratory (PPPL) with broad participation from the community working on aspects of lithium research for fusion energy development. This community is expanding rapidly in many areas including experiments in magnetic confinement devices and a variety of lithium test stands, theory and modeling and developing innovative approaches. Overall, 53 presentations were given representing 26 institutions from 10 countries. The latest experimental results from nine magnetic fusion devices were given in 24 presentations, from NSTX (PPPL, USA), LTX (PPPL, USA), FT-U (ENEA, Italy), T-11M (TRINITY, RF), T-10 (Kurchatov Institute, RF), TJ-II (CIEMAT, Spain), EAST (ASIPP, China), HT-7 (ASIPP, China), and RFX (Padova, Italy). Sessions were devoted to: I. Lithium in magnetic confinement experiments (facility overviews), II. Lithium in magnetic confinement experiments (topical issues), III. Special session on liquid lithium technology, IV. Lithium laboratory test stands, V. Lithium theory/modeling/comments, VI. Innovative lithium applications and VII. Panel discussion on lithium PFC viability in magnetic fusion reactors. There was notable participation from the fusion technology communities, including the IFE, IFMIF and TBM communities providing productive exchanges with the physics oriented magnetic confinement lithium research groups. It was agreed to continue future exchanges of ideas and data to help develop attractive liquid lithium solutions for very challenging magnetic fusion issues, such as development of a high heat flux steady-state divertor concept and acceptable plasma disruption mitigation techniques while improving plasma performance with lithium. The next workshop will be held at ENEA, Frascati, Italy in 2013.

  14. Light ion fusion experiment (L. I. F. E. ) concept validation studies. Final report, July 1979-May 1980

    SciTech Connect

    Christensen, T E; Orthel, J L; Thomson, J J

    1980-12-01

    This report reflects the considerable advances made for the objectives of the contractual program, validating by detailed anaytical studies the concept of a new Light Ion Fusion Experiment for Inertial Confinement Fusion. The studies have produced an analytical design of a novel electrostatic accelerator based on separate function and strong channel focusing principles, to launch 3 to 10 MeV, 23 kA, He/sup +/ neutralized beams in 400 ns pulses, delivering on a 5 mm radius target located 10 m downstream, 50 kJ of implosion energy in approx. 20 ns impact times The control, stability and focusing of beams is made by electrostatic quadrupoles, producing overall beam normalized emittance of approx. 3 x 10/sup -5/ m-rad.

  15. API2-MALT1 fusion protein induces transcriptional activation of the API2 gene through NF-{kappa}B binding elements: Evidence for a positive feed-back loop pathway resulting in unremitting NF-{kappa}B activation

    SciTech Connect

    Hosokawa, Yoshitaka . E-mail: yhosokaw@aichi-cc.jp; Suzuki, Hiroko; Nakagawa, Masao; Lee, Tae H.; Seto, Masao

    2005-08-19

    t(11;18)(q21;q21) is a characteristic as well as the most frequent chromosomal translocation in mucosa-associated lymphoid tissue (MALT) type lymphoma, and this translocation results in a fusion transcript, API2-MALT1. Although API2-MALT1 has been shown to enforce activation of NF-{kappa}B signaling, the transcriptional target genes of this fusion protein remains to be identified. Our analyses of the API2-MALT transfectants suggested that one of the target genes may be the apoptotic inhibitor API2 gene. Luciferase reporter assays with deletion and mutational constructs of the API2 promoter and electrophoretic mobility shift assays established that API2-MALT1 induces transcriptional activation of the API2 gene through two NF-{kappa}B binding elements. Moreover, supershift experiments indicated that these elements are recognized by the NF-{kappa}B p50/p65 heterodimer. Taken together, our results strongly indicated that API2-MALT1 possesses a novel mechanism of self-activation by up-regulating its own expression in t(11;18)(q21;q21)-carrying MALT lymphomas, highlighting a positive feedback-loop pathway resulting in unremitting NF-{kappa}B activation.

  16. NAB2-STAT6 gene fusion and STAT6 immunoexpression in extrathoracic solitary fibrous tumors: the association between fusion variants and locations.

    PubMed

    Chuang, I-Chieh; Liao, Kuan-Cho; Huang, Hsuan-Ying; Kao, Yu-Chien; Li, Chien-Feng; Huang, Shih-Chiang; Tsai, Jen-Wei; Chen, Ko-Chin; Lan, Jui; Lin, Po-Chun

    2016-05-01

    Solitary fibrous tumor (SFT) is a rare mesenchymal neoplasm harboring NAB2-STAT6 fusion, which drives STAT6 nuclear relocation. For extrathoracic SFTs, the clinical relevance of this molecular hallmark remains obscure. We assessed STAT6 immunoexpression for 61 extrathoracic SFTs exclusive of the meninges and head and neck, and 25 had analyzable RNAs to distinguish fusion variants by RT-PCR. The immunohistochemical and molecular findings were correlated with clincopathological features and disease-free survival (DFS). Twenty-eight males and 33 females had SFTs in the body cavities (n = 31), extremities (n = 17), and trunk (n = 13), categorized into 53 non-malignant and 8 malignant tumors. The vast majority (n = 57, 93%) exhibited distinctive STAT6 nuclear expression, including malignant ones. The common fusion variants were NAB2ex6-STAT6ex16/17 in 13 SFTs and NAB2ex4-STAT6ex2 in 8, while miscellaneous variants were detected only in 4 SFTs in the limbs and trunk but not in any body cavity-based cases (P = 0.026). The worse DFS was univariately associated with malignant histology (P = 0.04) but unrelated to tumor size, location, or fusion variant. Conclusively, extrathoracic SFTs mostly harbor NAB2ex6-STAT6ex16/17, followed by NAB2ex4-STAT6ex2. Miscellaneous variants are significantly rare in SFTs within the body cavities. The clinical aggressiveness of extrathoraic SFTs is associated with malignant histology but unrelated to the NAB2-STAT6 fusion variants.

  17. Epidermal Growth Factor Receptor Mutation and Anaplastic Lymphoma Kinase Gene Fusion: Detection in Malignant Pleural Effusion by RNA or PNA Analysis

    PubMed Central

    Chen, Yi-Lin; Lee, Chung-Ta; Lu, Cheng-Chan; Yang, Shu-Ching; Chen, Wan-Li; Lee, Yang-Cheng; Yang, Chung-Hsien; Peng, Shu-Ling; Su, Wu-Chou; Chow, Nan-Haw; Ho, Chung-Liang

    2016-01-01

    Analyzing EGFR mutations and detecting ALK gene fusion are indispensable when planning to treat pulmonary adenocarcinoma. Malignant pleural effusion (MPE) is a devastating complication of lung cancer and sometimes the only source for mutation analysis. The percentage of tumor cells in the pleural effusion may be low; therefore, mutant enrichment is required for a successful analysis. The EGFR mutation status in MPE was determined using three methods: (1) PCR sequencing of genomic DNA (direct sequencing), (2) mutant-enriched PCR sequencing of genomic DNA using peptide nucleic acid (PNA-sequencing), and (3) PCR sequencing of cDNA after reverse transcription for cellular RNA (RNA-sequencing). RT-PCR was also used to test cases for ALK gene fusion. PNA-sequencing and RNA-sequencing had similar analytical sensitivities (< 1%), which indicates similar enrichment capabilities. The clinical sensitivity in 133 cases when detecting the common EGFR exon 19 and exon 21 mutations was 56.4% (75/133) for direct sequencing, 63.2% (84/133) for PNA-sequencing, and 65.4% (87/133) for RNA-sequencing. RT-PCR and sequencing showed 5 cases (3.8%) with ALK gene fusion. All had wild-type EGFR. For EGFR analysis of MPE, RNA-sequencing is at least as sensitive as PNA-sequencing but not limited to specific mutations. Detecting ALK fusion can be incorporated in the same RNA workflow. Therefore, RNA is a better source for comprehensive molecular diagnoses in MPE. PMID:27352172

  18. Mutations in the lipoma HMGIC fusion partner-like 5 (LHFPL5) gene cause autosomal recessive nonsyndromic hearing loss.

    PubMed

    Kalay, Ersan; Li, Yun; Uzumcu, Abdullah; Uyguner, Oya; Collin, Rob W; Caylan, Refik; Ulubil-Emiroglu, Melike; Kersten, Ferry F J; Hafiz, Gunter; van Wijk, Erwin; Kayserili, Hulya; Rohmann, Edyta; Wagenstaller, Janine; Hoefsloot, Lies H; Strom, Tim M; Nürnberg, Gudrun; Baserer, Nermin; den Hollander, Anneke I; Cremers, Frans P M; Cremers, Cor W R J; Becker, Christian; Brunner, Han G; Nürnberg, Peter; Karaguzel, Ahmet; Basaran, Seher; Kubisch, Christian; Kremer, Hannie; Wollnik, Bernd

    2006-07-01

    In two large Turkish consanguineous families, a locus for autosomal recessive nonsyndromic hearing loss (ARNSHL) was mapped to chromosome 6p21.3 by genome-wide linkage analysis in an interval overlapping with the loci DFNB53 (COL11A2), DFNB66, and DFNB67. Fine mapping excluded DFNB53 and subsequently homozygous mutations were identified in the lipoma HMGIC fusion partner-like 5 (LHFPL5) gene, also named tetraspan membrane protein of hair cell stereocilia (TMHS) gene, which was recently shown to be mutated in the "hurry scurry" mouse and in two DFNB67-linked families from Pakistan. In one family, we found a homozygous one-base pair deletion, c.649delG (p.Glu216ArgfsX26) and in the other family we identified a homozygous transition c.494C>T (p.Thr165Met). Further screening of index patients from 96 Turkish ARNSHL families and 90 Dutch ARNSHL patients identified one additional Turkish family carrying the c.649delG mutation. Haplotype analysis revealed that the c.649delG mutation was located on a common haplotype in both families. Mutation screening of the LHFPL5 homologs LHFPL3 and LHFPL4 did not reveal any disease causing mutation. Our findings indicate that LHFPL5 is essential for normal function of the human cochlea.

  19. HAWAIIAN SKIRT: An F-Box Gene That Regulates Organ Fusion and Growth in Arabidopsis1[C][W][OA

    PubMed Central

    González-Carranza, Zinnia H.; Rompa, Unchalee; Peters, Janny L.; Bhatt, Anuj M.; Wagstaff, Carol; Stead, Anthony D.; Roberts, Jeremy A.

    2007-01-01

    A fast neutron-mutagenized population of Arabidopsis (Arabidopsis thaliana) Columbia-0 wild-type plants was screened for floral phenotypes and a novel mutant, termed hawaiian skirt (hws), was identified that failed to shed its reproductive organs. The mutation is the consequence of a 28 bp deletion that introduces a premature amber termination codon into the open reading frame of a putative F-box protein (At3g61590). The most striking anatomical characteristic of hws plants is seen in flowers where individual sepals are fused along the lower part of their margins. Crossing of the abscission marker, ProPGAZAT:β-glucuronidase, into the mutant reveals that while floral organs are retained it is not the consequence of a failure of abscission zone cells to differentiate. Anatomical analysis indicates that the fusion of sepal margins precludes shedding even though abscission, albeit delayed, does occur. Spatial and temporal characterization, using ProHWS:β-glucuronidase or ProHWS:green fluorescent protein fusions, has identified HWS expression to be restricted to the stele and lateral root cap, cotyledonary margins, tip of the stigma, pollen, abscission zones, and developing seeds. Comparative phenotypic analyses performed on the hws mutant, Columbia-0 wild type, and Pro35S:HWS ectopically expressing lines has revealed that loss of HWS results in greater growth of both aerial and below-ground organs while overexpressing the gene brings about a converse effect. These observations are consistent with HWS playing an important role in regulating plant growth and development. PMID:17496113

  20. LDRD final report on confinement of cluster fusion plasmas with magnetic fields.

    SciTech Connect

    Argo, Jeffrey W.; Kellogg, Jeffrey W.; Headley, Daniel Ignacio; Stoltzfus, Brian Scott; Waugh, Caleb J.; Lewis, Sean M.; Porter, John Larry, Jr.; Wisher, Matthew; Struve, Kenneth William; Savage, Mark Edward; Quevedo, Hernan J.; Bengtson, Roger

    2011-11-01

    Two versions of a current driver for single-turn, single-use 1-cm diameter magnetic field coils have been built and tested at the Sandia National Laboratories for use with cluster fusion experiments at the University of Texas in Austin. These coils are used to provide axial magnetic fields to slow radial loss of electrons from laser-produced deuterium plasmas. Typical peak field strength achievable for the two-capacitor system is 50 T, and 200 T for the ten-capacitor system. Current rise time for both systems is about 1.7 {mu}s, with peak current of 500 kA and 2 MA, respectively. Because the coil must be brought to the laser, the driver needs to be portable and drive currents in vacuum. The drivers are complete but laser-plasma experiments are still in progress. Therefore, in this report, we focus on system design, initial tests, and performance characteristics of the two-capacitor and ten-capacitors systems. The questions of whether a 200 T magnetic field can retard the breakup of a cluster-fusion plasma, and whether this field can enhance neutron production have not yet been answered. However, tools have been developed that will enable producing the magnetic fields needed to answer these questions. These are a two-capacitor, 400-kA system that was delivered to the University of Texas in 2010, and a 2-MA ten-capacitor system delivered this year. The first system allowed initial testing, and the second system will be able to produce the 200 T magnetic fields needed for cluster fusion experiments with a petawatt laser. The prototype 400-kA magnetic field driver system was designed and built to test the design concept for the system, and to verify that a portable driver system could be built that delivers current to a magnetic field coil in vacuum. This system was built copying a design from a fixed-facility, high-field machine at LANL, but made to be portable and to use a Z-machine-like vacuum insulator and vacuum transmission line. This system was sent to the

  1. Molecular evolution of viral fusion and matrix protein genes and phylogenetic relationships among the Paramyxoviridae.

    PubMed

    Westover, K M; Hughes, A L

    2001-10-01

    Phylogenetic relationships among the Paramyxoviridae, a broad family of viruses whose members cause devastating diseases of wildlife, livestock, and humans, were examined with both fusion (F) and matrix (M) protein-coding sequences. Neighbor-joining trees of F and M protein sequences showed that the Paramyxoviridae was divided into the two traditionally recognized subfamilies, the Paramyxovirinae and the Pneumovirinae. Within the Paramyxovirinae, the results also showed groups corresponding to three currently recognized genera: Respirovirus, Morbillivirus, and Rubulavirus. The relationships among the three genera of the Paramyxovirinae were resolved with M protein sequences and there was significant bootstrap support (100%) showing that members of the genus Respirovirus and the genus Morbillivirus were more closely related to each other than to members of the genus Rubulavirus. Both F and M phylogenies showed that Newcastle disease virus (NDV) was more closely related to the genus Rubulavirus than to the other two genera but were consistent with the proposal (B. S. Seal et al., 2000, Virus Res. 66, 1-11) that NDV be classified as a separate genus within the Paramyxovirinae. Both F and M phylogenies were also consistent with the proposal (L. Wang et al., 2000, J. Virol 74, 9972-9979) that Hendra virus be classified as a new genus closely related and basal to the genus Morbillivirus. Rinderpest was most closely related to measles and a more derived virus than to canine distemper virus, phocine distemper virus, or dolphin morbillivirus.

  2. A burning plasma program strategy to advance fusion energy. Report of the Fusion Energy Sciences Advisory Committee, Burning Plasma Strategy Panel

    SciTech Connect

    None, None

    2002-09-01

    Fusion energy shows great promise to contribute to securing the energy future of humanity. The risk of conflicts arising from energy shortages and supply cutoffs, as well as the risk of severe environmental impacts from existing methods of energy production, are strong reasons to pursue fusion energy now. The world effort to develop fusion energy is at the threshold of a new stage in its research: the investigation of burning plasmas. This investigation, at the frontier of the physics of complex systems, would be a huge step in establishing the potential of magnetic fusion energy to contribute to the world’s energy security. The defining feature of a burning plasma is that it is self-heated: the 100 million degree temperature of the plasma is maintained mainly by the heat generated by the fusion reactions themselves, as occurs in burning stars. The fusion-generated alpha particles produce new physical phenomena that are strongly coupled together as a nonlinear complex system. Understanding all elements of this system poses a major challenge to fundamental plasma physics. The technology needed to produce and control a burning plasma presents challenges in engineering science similarly essential to the development of fusion energy.

  3. Fusion reactor materials semiannual progress report for the period ending September 30, 1988

    SciTech Connect

    none,

    1989-04-01

    This paper discusses the following topics on fusion reactor materials: irradiation, facilities, test matrices, and experimental methods; dosimetry, damage parameters, and activation calculations; materials engineering and design requirements; fundamental mechanical behavior; radiation effects; development of structural alloys; solid breeding materials; and ceramics.

  4. Inertial confinement fusion quarterly report, July--September 1994. Volume 4, Number 4

    SciTech Connect

    Honea, E.

    1994-09-01

    The ICF Quarterly continues with six articles in this issue describing recent developments in the Inertial Confinement Fusion (ICF) Program at Lawrence Livermore National Laboratory. The topics include plasma characterization, production of millimeter scale-length plasmas for studying laser-plasma instabilities, hohlraum physics, three-dimensional hydrodynamic modeling, crystal growth, and laser-beam smoothing.

  5. Conference report on the 3rd International Symposium on Lithium Application for Fusion Devices

    DOE PAGES

    Mazzitelli, Guiseppe; Hirooka, Y.; Hu, J. S.; ...

    2015-01-14

    The third International Symposium on Lithium Application for Fusion Device (ISLA-2013) was held on 9-11 October 2013 at ENEA Frascati Centre with growing participation and interest from the community working on more general aspect of liquid metal research for fusion energy development. ISLA-2013 has been confirmed to be the largest and the most important meeting dedicated to liquid metal application for the magnetic fusion research. Overall, 45 presentation plus 5 posters were given, representing 28 institutions from 11 countries. The latest experimental results from nine magnetic fusion devices were presented in 16 presentations from NSTX (PPPL, USA), FTU (ENEA, Italy),more » T-11M (Trinity, RF), T-10 (Kurchatov Institute, RF), TJ-II (CIEMAT, Spain), EAST(ASIPP, China), HT-7 (ASIPP, China), RFX (Padova, Italy), KTM (NNC RK, Kazakhstan). Sessions were devoted to the following: (I) lithium in magnetic confinement experiments (facility overviews), (II) lithium in magnetic confinement experiments (topical issues), (III) special session on liquid lithium technology, (IV) lithium laboratory test stands, (V) Lithium theory/modelling/comments, (VI) innovative lithium applications and (VII) special Session on lithium-safety and lithium handling. There was a wide participation from the fusion technology communities, including IFMIF and TBM communities providing productive exchange with the physics oriented magnetic confinement liquid metal research groups. Furthermore, this international workshop will continue on a biennial basis (alternating with the Plasma-Surface Interactions (PSI) Conference) and the next workshop will be held at CIEMAT, Madrid, Spain, in 2015.« less

  6. Recent Progress on the Construction and Testing of a Fusion Poly(hydroxyalkanoate) Synthase Gene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Poly(hydroxyalkanoates) (PHAs) are biodegradable polyesters produced by some bacteria. Two genes in Allochromatium vinosum, phaE and phaC, respectively code for the two subunits of the enzyme complex, PHA synthase, which catalyzes the polymerization of precursors into PHA. We hypothesized that by ...

  7. Establishment of the first WHO International Standard for etanercept, a TNF receptor II Fc fusion protein: Report of an international collaborative study.

    PubMed

    Wadhwa, Meenu; Bird, Chris; Dilger, Paula; Rigsby, Peter; Jia, Haiyan; Gross, Marie Emmanuelle Behr

    2017-03-10

    Etanercept, a recombinant human tumor necrosis factor (TNF) receptor Fc fusion protein is an effective treatment option in adults with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis or plaque psoriasis and paediatrics with juvenile idiotypic arthritis and plaque psoriasis. Patent expiration in Europe and intense development of various etanercept products worldwide triggered a need for an international reference standard to facilitate determination of biological activity. Therefore, three candidate preparations of etanercept were lyophilized and evaluated in a multi-centre collaborative study comprising twenty eight laboratories from 15 countries for their suitability to serve as an international standard for the bioactivity of TNF receptor II Fc fusion proteins (international nonproprietary name, Etanercept). The preparations were tested for neutralization activity against the third TNF-α international standard (IS) in different in vitro cell-based assays, e.g., cytotoxicity, apoptosis and reporter gene methods. Regardless of the assay and the amount of TNF-α IS used, potency estimates for the different preparations were very similar. An indication of the inhibitory activity of etanercept in terms of the biological activity of the TNF-α IS based on ED50 data derived from a limited number of laboratories using a cytotoxicity assay was also derived. Results indicated that the candidate preparation coded 13/204 was stable and suitable to serve as an international standard for the biological activity of etanercept. Therefore, the preparation coded 13/204 was established by the WHO Expert Committee on Biological Standardization (ECBS) in 2015 as the WHO first International Standard for TNF receptor II Fc fusion protein (INN, etanercept) with an assigned in vitro bioactivity of 10,000IU per ampoule. It should be noted that this first-in-class international standard for a Fc fusion protein, available from the National Institute for Biological

  8. Lateral Lumbar Interbody Fusion for Ossification of the Yellow Ligament in the Lumbar Spine: First Reported Case

    PubMed Central

    Abe, Tetsuya; Funayama, Toru; Noguchi, Hiroshi; Nakayama, Keita; Miura, Kousei; Nagashima, Katsuya; Kumagai, Hiroshi; Yamazaki, Masashi

    2017-01-01

    When ossification of the yellow ligament (OYL) occurs in the lumbar spine and extends to the lateral wall of the spinal canal, facetectomy is required to remove all of the ossified lesion and achieve decompression. Subsequent posterior fixation with interbody fusion will then be necessary to prevent postoperative progression of the ossification and intervertebral instability. The technique of lateral lumbar interbody fusion (LLIF) has recently been introduced. Using this procedure, surgeons can avoid excess blood loss from the extradural venous plexus and detachment of the ossified lesion and the ventral dura mater is avoidable. We present a 55-year-old male patient with OYL at L3/4 and anterior spondylolisthesis of L4 vertebra, with concomitant ossification of the posterior longitudinal ligament, who presented with a severe gait disturbance. He underwent a 2-stage operation without complications: LLIF for L3/4 and L4/5 was performed at the initial surgery, and posterior decompression fixation using pedicle screws from L3 to L5 was performed at the second surgery. His postoperative progress was favorable, and his interbody fusion was deemed successful. Here, we present the first reported case of LLIF for OYL of the lumbar spine. This procedure can be a good option for OYL of the lumbar spine. PMID:28352485

  9. Functional Impact and Evolution of a Novel Human Polymorphic Inversion That Disrupts a Gene and Creates a Fusion Transcript.

    PubMed

    Puig, Marta; Castellano, David; Pantano, Lorena; Giner-Delgado, Carla; Izquierdo, David; Gayà-Vidal, Magdalena; Lucas-Lledó, José Ignacio; Esko, Tõnu; Terao, Chikashi; Matsuda, Fumihiko; Cáceres, Mario

    2015-10-01

    Despite many years of study into inversions, very little is known about their functional consequences, especially in humans. A common hypothesis is that the selective value of inversions stems in part from their effects on nearby genes, although evidence of this in natural populations is almost nonexistent. Here we present a global analysis of a new 415-kb polymorphic inversion that is among the longest ones found in humans and is the first with clear position effects. This inversion is located in chromosome 19 and has been generated by non-homologous end joining between blocks of transposable elements with low identity. PCR genotyping in 541 individuals from eight different human populations allowed the detection of tag SNPs and inversion genotyping in multiple populations worldwide, showing that the inverted allele is mainly found in East Asia with an average frequency of 4.7%. Interestingly, one of the breakpoints disrupts the transcription factor gene ZNF257, causing a significant reduction in the total expression level of this gene in lymphoblastoid cell lines. RNA-Seq analysis of the effects of this expression change in standard homozygotes and inversion heterozygotes revealed distinct expression patterns that were validated by quantitative RT-PCR. Moreover, we have found a new fusion transcript that is generated exclusively from inverted chromosomes around one of the breakpoints. Finally, by the analysis of the associated nucleotide variation, we have estimated that the inversion was generated ~40,000-50,000 years ago and, while a neutral evolution cannot be ruled out, its current frequencies are more consistent with those expected for a deleterious variant, although no significant association with phenotypic traits has been found so far.

  10. Functional Impact and Evolution of a Novel Human Polymorphic Inversion That Disrupts a Gene and Creates a Fusion Transcript

    PubMed Central

    Puig, Marta; Castellano, David; Pantano, Lorena; Giner-Delgado, Carla; Izquierdo, David; Gayà-Vidal, Magdalena; Lucas-Lledó, José Ignacio; Esko, Tõnu; Terao, Chikashi; Matsuda, Fumihiko; Cáceres, Mario

    2015-01-01

    Despite many years of study into inversions, very little is known about their functional consequences, especially in humans. A common hypothesis is that the selective value of inversions stems in part from their effects on nearby genes, although evidence of this in natural populations is almost nonexistent. Here we present a global analysis of a new 415-kb polymorphic inversion that is among the longest ones found in humans and is the first with clear position effects. This inversion is located in chromosome 19 and has been generated by non-homologous end joining between blocks of transposable elements with low identity. PCR genotyping in 541 individuals from eight different human populations allowed the detection of tag SNPs and inversion genotyping in multiple populations worldwide, showing that the inverted allele is mainly found in East Asia with an average frequency of 4.7%. Interestingly, one of the breakpoints disrupts the transcription factor gene ZNF257, causing a significant reduction in the total expression level of this gene in lymphoblastoid cell lines. RNA-Seq analysis of the effects of this expression change in standard homozygotes and inversion heterozygotes revealed distinct expression patterns that were validated by quantitative RT-PCR. Moreover, we have found a new fusion transcript that is generated exclusively from inverted chromosomes around one of the breakpoints. Finally, by the analysis of the associated nucleotide variation, we have estimated that the inversion was generated ~40,000–50,000 years ago and, while a neutral evolution cannot be ruled out, its current frequencies are more consistent with those expected for a deleterious variant, although no significant association with phenotypic traits has been found so far. PMID:26427027

  11. Inertial Conference Fusion Semiannual Report October 1999 - March 2000, Volume 1, Number 1

    SciTech Connect

    Miguel, Al; Carpenter, Jason; Cassady, Cindy

    2000-03-01

    improvements can have important implications for the interpretation of inertial confinement fusion capsule implosions. The fifth article reports on experiments using the OMEGA laser that investigate symmetry control in hohlraums. The experiments explore a control method where different pointings are used for different groups of beams and the beams are staggered in time. This gives a dynamic beam pointing adjustment during the laser pulse. Measurements of the capsule symmetry show agreement with simulations and show the ability to control low-mode drive asymmetries. The sixth article reports on the observation of an intense high-energy proton beam produced by irradiating a thin-foil target with the petawatt laser. This experiment is important for understanding new mechanisms of ion acceleration using high-intensity short-pulse lasers. Proton beams of the type observed here could be of interest for applications ranging from medicine to fast ignition.

  12. Creation of immune 'stealth' genes for gene therapy through fusion with the Gly-Ala repeat of EBNA-1.

    PubMed

    Ossevoort, M; Visser, B M J; van den Wollenberg, D J M; van der Voort, E I H; Offringa, R; Melief, C J M; Toes, R E M; Hoeben, R C

    2003-11-01

    A major obstacle in gene-therapy protocols is T-cell-mediated destruction of transgene-expressing cells. Therefore new approaches are needed to prevent rapid clearance of transduced cells. We exploited the Gly-Ala repeat (GAr) domain of the Epstein-Barr virus nuclear antigen-1, since the GAr prevents cytotoxic T-lymphocyte-epitope generation. Here we show that three different enzymes (viz. the E. coli LacZ gene encoded beta-galactosidase, firefly luciferase, and HSV1 thymidine kinase) fused with the GAr retained their function. Moreover, linking GAr with beta-galactosidase successfully prevented recognition of GAr-LacZ-expressing cells by beta-galactosidase-specific CTL. Nonetheless, vaccination with a GAr-LacZ adenovirus or with an allogeneic cell line expressing GAr-LacZ resulted in the induction of beta-gal-specific CTL. This demonstrates that the GAr domain does not inhibit cross presentation of antigens, but only affects breakdown of endogenously synthesized proteins. These data demonstrate how the GAr domain can be exploited to create immuno'stealth' genes by hiding transgene products from CTL-mediated immune attack.

  13. Energy payback and CO{sub 2} gas emissions from fusion and solar photovoltaic electric power plants. Final report to Department of Energy, Office of Fusion Energy Sciences

    SciTech Connect

    Kulcinski, G.L.

    2002-12-01

    A cradle-to-grave net energy and greenhouse gas emissions analysis of a modern photovoltaic facility that produces electricity has been performed and compared to a similar analysis on fusion. A summary of the work has been included in a Ph.D. thesis titled ''Life-cycle assessment of electricity generation systems and applications for climate change policy analysis'' by Paul J. Meier, and a synopsis of the work was presented at the 15th Topical meeting on Fusion Energy held in Washington, DC in November 2002. In addition, a technical note on the effect of the introduction of fusion energy on the greenhouse gas emissions in the United States was submitted to the Office of Fusion Energy Sciences (OFES).

  14. FINESSE: study of the issues, experiments and facilities for fusion nuclear technology research and development. Interim report. Volume II

    SciTech Connect

    Abdou, M.

    1984-10-01

    The Nuclear Fusion Issues chapter contains a comprehensive list of engineering issues for fusion reactor nuclear components. The list explicitly defines the uncertainties associated with the engineering option of a fusion reactor and addresses the potential consequences resulting from each issue. The next chapter identifies the fusion nuclear technology testing needs up to the engineering demonstration stage. (MOW)

  15. The actin cytoskeleton inhibits pore expansion during PIV5 fusion protein-promoted cell-cell fusion

    SciTech Connect

    Wurth, Mark A.; Schowalter, Rachel M.; Smith, Everett Clinton; Moncman, Carole L.; Ellis Dutch, Rebecca; McCann, Richard O.

    2010-08-15

    Paramyxovirus fusion (F) proteins promote both virus-cell fusion, required for viral entry, and cell-cell fusion, resulting in syncytia formation. We used the F-actin stabilizing drug, jasplakinolide, and the G-actin sequestrant, latrunculin A, to examine the role of actin dynamics in cell-cell fusion mediated by the parainfluenza virus 5 (PIV5) F protein. Jasplakinolide treatment caused a dose-dependent increase in cell-cell fusion as measured by both syncytia and reporter gene assays, and latrunculin A treatment also resulted in fusion stimulation. Treatment with jasplakinolide or latrunculin A partially rescued a fusion pore opening defect caused by deletion of the PIV5 F protein cytoplasmic tail, but these drugs had no effect on fusion inhibited at earlier stages by either temperature arrest or by a PIV5 heptad repeat peptide. These data suggest that the cortical actin cytoskeleton is an important regulator of fusion pore enlargement, an energetically costly stage of viral fusion protein-mediated membrane merger.

  16. A lacZ reporter fusion method for the genetic analysis of regulatory mutations in pathways of fungal secondary metabolism and its application to the Aspergillus nidulans penicillin pathway.

    PubMed Central

    Pérez-Esteban, B; Gómez-Pardo, E; Peñalva, M A

    1995-01-01

    Secondary metabolism, usually superfluous under laboratory conditions, is intrinsically elusive to genetic analysis of its regulation. We describe here a method of analyzing regulatory mutations affecting expression of secondary metabolic genes, with an Aspergillus nidulans penicillin structural gene (ipnA [encoding isopenicillin N-synthase]) as a model. The method is based on a targeted double integration of a lacZ fusion reporter gene in a chromosome different from that containing the penicillin gene cluster. The trans-acting regulatory mutations simultaneously affect lacZ expression and penicillin biosynthesis. One of these mutations (npeE1) has been analyzed in detail. This mutation is recessive, prevents penicillin production and ipnA'::'lacZ expression, and results in very low levels of the ipnA message at certain times of growth. This indicates that npeE positively controls ipnA transcription. We also show that this tandem reporter fusion allows genetic analysis of npeE1 by using the sexual and parasexual cycles and that lacZ expression is an easily scorable phenotype. Haploidization analysis established that npeE is located in chromosome IV, but npeE1 does not show meiotic linkage to a number of known chromosome IV markers. This method might be of general applicability to genetic analysis of regulation of other fungal secondary metabolic pathways. PMID:7592369

  17. Heavy Ion Fusion Accelerator Research (HIFAR) year-end report, October 1, 1987--March 31, 1988

    SciTech Connect

    Not Available

    1988-06-01

    The basic objective of the Heavy Ion Fusion Accelerator Research (HIFAR) program is to assess the suitability of heavy ion accelerators as igniters for Inertial Confinement Fusion (ICF). A specific accelerator technology, the induction linac, has been studied at Lawrence Berkeley Laboratory and has reached the point at which its viability for ICF applications can be assessed over the next few years. The HIFAR program addresses the generation of high-power, high-brightness beams of heavy ions, the understanding of the scaling laws in this novel physics regime, and the validation of new accelerator strategies, to cut costs. Key elements to be addressed include: beam quality limits set by transverse and longitudinal beam physics; development of induction accelerating modules, and multiple-beam hardware, at affordable costs; acceleration of multiple beams with current amplification -- both new features in a linac -- without significant dilution of the optical quality of beams; and final bunching, transport, and accurate focusing on a small target.

  18. Heavy Ion Fusion Accelerator Research (HIFAR) year-end report, April 1--September 30, 1988

    SciTech Connect

    Not Available

    1988-12-01

    The basic objective of the Heavy Ion Fusion Accelerator Research (HIFAR) program is to assess the suitability of heavy ion accelerators as igniters for Inertial Confinement Fusion (ICF). A specific accelerator technology, the induction linac, has been studied at the Lawrence Berkeley Laboratory and has reached the point at which its viability for ICF applications can be assessed over the next few years. The HIFAR program addresses the generation of high power, high-brightness beams of heavy ions, the understanding of the scaling laws in this novel physics regime, and the validation of new accelerator strategies, to cut costs. Key elements to be addressed include: beam quality limits set by transverse and longitudinal beam physics; development of induction accelerating modules, and multiple-beam hardware, at affordable costs; acceleration of multiple beams with current amplification --both new features in a linac -- without significant dilution of the optical quality of the beams; final bunching, transport, and accurate focusing on a small target.

  19. Fluid mechanics of fusion lasers. Final report, September 11, 1978-June 5, 1980

    SciTech Connect

    Shwartz, J; Kulkarny, V A; Ausherman, D A; Legner, H H; Sturtevant, B

    1980-01-01

    Flow loop components required to operate continuous-flow, repetitively-pulsed CO/sub 2/ and KrF laser drivers for ICF were identified and their performance requirements were specified. It was found that the laser flow loops can have a major effect on the laser beam quality and overall efficiency. The pressure wave suppressor was identified as the most critical flow loop component. The performance of vented side-wall suppressors was evaluated both analytically and experimentally and found capable of meeting the performance requirements of the CO/sub 2/ and KrF fusion lasers. All other laser flow loop components are essentially similar to those used in conventional, low speed wind tunnels and are therefore well characterized and can be readily incorporated into fusion laser flow systems designs.

  20. Inertial confinement fusion quarterly report, April--June 1994. Volume 4, Number 3

    SciTech Connect

    Shaw, M.J.

    1994-06-01

    This issue of the ICF Quarterly contains six articles covering a wide range of activities within the Inertial Confinement Fusion (ICF) Program. It concentrates on target design; theoretical spectral analysis of ICF capsule surfaces; laser fusion experimental methods; and an alternative ICF design, based on ultrafast, ultrapowerful lasers. A key issue for the success of the ICF process is the hydrodynamic stability of the imploding capsule. There are two primary sources of instability growth in the ICF process: (1) asymmetries in the x-ray flux that drive the compression lead to asymmetric in the imploding surface; (2) imperfections on the capsule surface can grow into large perturbations, degrading the capsule performance. In recent years, a great deal of effort, both experimentally and theoretically, has been spent to enhance the Program`s ability to measure, model, and minimize instability growth during an implosion. Four the articles in this issue discuss this subject.

  1. Q289P mutation in the FGFR2 gene: first report in a patient with type 1 Pfeiffer syndrome.

    PubMed

    Piccione, Maria; Antona, Vincenzo; Niceta, Marcello; Fabiano, Carmelo; Martines, Manuela; Bianchi, Alberto; Corsello, Giovanni

    2009-09-01

    When normal development and growth of the calvarial sutures is disrupted, craniosynostosis (premature calvarial suture fusion) may result. Classical craniosynostosis syndromes are autosomal dominant traits and include Apert, Pfeiffer, Crouzon, Jackson-Weiss, and Saethre-Chotzen syndromes. In these conditions, there is premature fusion of skull bones leading to an abnormal head shape, ocular hypertelorism with proptosis, and midface hypoplasia. It is known that mutations in the fibroblast growth factor receptors 1, 2, and 3 cause craniosynostosis. We report on a child with a clinically diagnosed Pfeiffer syndrome that shows the missense point mutation Q289P in exon 8 of the FGFR2 gene. This is a mutation not previously described in the Pfeiffer syndrome but reported in the Crouzon, Jackson-Weiss, and Saethre-Chotzen syndromes. In this paper, we propose the concept that these disorders may represent one genetic condition with phenotypic variability.

  2. IFMIF - International Fusion Materials Irradiation Facility Conceptual Design Activity/Interim Report

    SciTech Connect

    Rennich, M.J.

    1995-12-01

    Environmental acceptability, safety, and economic viability win ultimately be the keys to the widespread introduction of fusion power. This will entail the development of radiation- resistant and low- activation materials. These low-activation materials must also survive exposure to damage from neutrons having an energy spectrum peaked near 14 MeV with annual radiation doses in the range of 20 displacements per atom (dpa). Testing of candidate materials, therefore, requires a high-flux source of high energy neutrons. The problem is that there is currently no high-flux source of neutrons in the energy range above a few MeV. The goal, is therefore, to provide an irradiation facility for use by fusion material scientists in the search for low-activation and damage-resistant materials. An accellerator-based neutron source has been established through a number of international studies and workshops` as an essential step for materials development and testing. The mission of the International Fusion Materials Irradiation Facility (IFMIF) is to provide an accelerator-based, deuterium-lithium (D-Li) neutron source to produce high energy neutrons at sufficient intensity and irradiation volume to test samples of candidate materials up to about a full lifetime of anticipated use in fusion energy reactors. would also provide calibration and validation of data from fission reactor and other accelerator-based irradiation tests. It would generate material- specific activation and radiological properties data, and support the analysis of materials for use in safety, maintenance, recycling, decommissioning, and waste disposal systems.

  3. Maryland controlled fusion research program. Progress report, November 1, 1992--October 31, 1993

    SciTech Connect

    Antonsen, T.M. Jr.; Drake, J.F.; Finn, J.M.; Guzdar, P.; Hassam, A.; Liu, C.S.; Ott, E.

    1993-05-01

    In recent years, members of the Maryland Plasma Theory Group have made significant contributions to the national fusion theory program, and, in many cases, these theoretical developments helped to interpret experimental results and to design new experimental programs. In this paper, we summarize the technical progress in four major areas of tokamak research: (a) L/H transition and edge turbulence and transport; (b) active control of micro-turbulence and transport; (c) major disruptions; and (d) the sawtooth crash.

  4. Muon-catalyzed fusion experiment target and detector system. Preliminary design report

    SciTech Connect

    Jones, S.E.; Watts, K.D.; Caffrey, A.J.; Walter, J.B.

    1982-03-01

    We present detailed plans for the target and particle detector systems for the muon-catalyzed fusion experiment. Requirements imposed on the target vessel by experimental conditions and safety considerations are delineated. Preliminary designs for the target vessel capsule and secondary containment vessel have been developed which meet these requirements. In addition, the particle detection system is outlined, including associated fast electronics and on-line data acquisition. Computer programs developed to study the target and detector system designs are described.

  5. Low-density hydrocarbon foams for laser fusion targets: Progress report, 1986

    SciTech Connect

    Chen, C.; Cook, R.C.; Haendler, B.L.; Hair, L.M.; Kong, F.M.; Letts, S.A.

    1987-06-01

    We describe progress made during 1986 in the development of direct-drive hydrocarbon foam targets for laser fusion. The foam materials are polystyrene and resorcinolformaldehyde. The processes for making the foams, their properties, characterization techniques, and the relationships of their properties to target specifications are presented. In the final section, we discuss statistical experimental design techniques that are being used to optimize the foams. 12 refs., 14 figs., 2 tabs.

  6. Helium Find Thaws the Cold Fusion Trail.

    ERIC Educational Resources Information Center

    Pennisi, E.

    1991-01-01

    Reported is a study of cold fusion in which trace amounts of helium, possible evidence of an actual fusion reaction, were found. Research methodology is detailed. The controversy over the validity of experimental results with cold fusion are reviewed. (CW)

  7. Inertial confinement fusion. 1995 ICF annual report, October 1994--September 1995

    SciTech Connect

    1996-06-01

    Lawrence Livermore National Laboratory`s (LLNL`s) Inertial Confinement Fusion (ICF) Program is a Department of Energy (DOE) Defense Program research and advanced technology development program focused on the goal of demonstrating thermonuclear fusion ignition and energy gain in the laboratory. During FY 1995, the ICF Program continued to conduct ignition target physics optimization studies and weapons physics experiments in support of the Defense Program`s stockpile stewardship goals. It also continued to develop technologies in support of the performance, cost, and schedule goals of the National Ignition Facility (NIF) Project. The NIF is a key element of the DOE`s Stockpile Stewardship and Management Program. In addition to its primary Defense Program goals, the ICF Program provides research and development opportunities in fundamental high-energy-density physics and supports the necessary research base for the possible long-term application to inertial fusion energy (IFE). Also, ICF technologies have had spin-off applications for industrial and governmental use. Selected papers are indexed separately for inclusion in the Energy Science and Technology Database.

  8. [Aplication of demineralized human bone matrix in the surgical dental fusion treatment. Report of a case].

    PubMed

    Mora-Rincones, Oscar A; Corona-Rodríguez, Julio C; Díaz-Carvajal, Alvaro L; Franco-Carrero, Isabel C

    2008-06-01

    The purpose of this work is to present a surgical alternative in the treatment of the dental fusions through the placement of demineralized human bone matrix (DHBM) (Grafton Putty)*, immediately after the separation and extraction of the fused tooth to the permanent one. The dental fusion is a dental anomaly of union, that consists in the union of two dental germs during development. It could happen at any of the dental germ evolution stages from the dental sheet or from more advanced processes of differentiation. For the clinical treatment, an allograft of DHBM with osteoinductive and osteoconductive properties was used. This had several factors of bone growth, it allowed the gradual growth of a new bone that helped to correct the bone defects post-extraction and to cover the exposed distal wall of the remaining permanent tooth. The clinic evaluation and the periapical and panoramic radiographies images were used for the clinical control. It can be concluded that the surgical separation and the extraction of the tooth with less anatomical likeness to the contralateral and the placement of the DHBM, represent a surgical treatment alternative of the dental fusion.

  9. Differential roles of SS18-SSX fusion gene and insulin-like growth factor-1 receptor in synovial sarcoma cell growth

    SciTech Connect

    Toernkvist, Maria; Natalishvili, Natalia; Xie Yuntao; Girnita, Ada; D'Arcy, Padraig; Brodin, Bertha; Axelson, Magnus; Girnita, Leonard

    2008-04-11

    Recently we demonstrated that the synovial sarcoma specific fusion gene SS18-SSX is crucial for cyclin D1 expression and is linked to cell proliferation. In this report we explore the role of SS18-SSX and IGF-1R for their potential functions in cellular proliferation and survival in cultured synovial sarcoma cells. We found that targeting of SS18-SSX mRNA by antisense oligonucleotide treatment drastically and rapidly decreased cell proliferation but caused only a slight increase of apoptosis. The synovial sarcoma cells were confirmed to express IGF-1R, and treatment with an IGF-1R inhibitor resulted in substantially reduced cell viability by inducing apoptosis in these cells. Conversely, inhibition of the IGF-1R resulted only in a slight to moderate decrease in DNA synthesis. In conclusion, SS18-SSX and IGF-1R seem to play important but different roles in maintaining malignant growth of synovial sarcoma cells. Whereas SS18-SSX maintains cyclin D1 and cell proliferation, IGF-1R protects from apoptosis.

  10. Transformation of tobacco plants by Yali PPO-GFP fusion gene and observation of subcellular localization.

    PubMed

    Qi, Jing; Li, Gui-Qin; Dong, Zhen; Zhou, Wei

    2016-01-01

    To explore the subcellular localization of Polyphenol oxidase (PPO) from Pyrus bretschneideri, the 1779 bp cDNA of PPO gene excluding the termination codon TAA was cloned and fused with GFP to construct a binary vector pBI121-PPO-GFP. Then, the binary vector was transformed into Nicotiana tabacum by the tumefanciens-mediated method. Using confocal laser scanning microscopy, green fluorescent signals were localized in chloroplasts of the transformed Nicotiana tabacum cell, suggesting that the Polyphenol oxidase from Pyrus bretschneideri was a chloroplast protein.

  11. Graphite for fusion energy applications

    SciTech Connect

    Eatherly, W.P.; Clausing, R.E.; Strehlow, R.A.; Kennedy, C.R.; Mioduszewski, P.K.

    1987-03-01

    Graphite is in widespread and beneficial use in present fusion energy devices. This report reflects the view of graphite materials scientists on using graphite in fusion devices. Graphite properties are discussed with emphasis on application to fusion reactors. This report is intended to be introductory and descriptive and is not intended to serve as a definitive information source. (JDH)

  12. Genes encoding norcoclaurine synthase occur as tandem fusions in the Papaveraceae

    PubMed Central

    Li, Jing; Lee, Eun-Jeong; Chang, Limei; Facchini, Peter J.

    2016-01-01

    Norcoclaurine synthase (NCS) catalyzes the enantioselective Pictet-Spengler condensation of dopamine and 4-hydroxyphenylacetaldehyde as the first step in benzylisoquinoline alkaloid (BIA) biosynthesis. NCS orthologs in available transcriptome databases were screened for variants that might improve the low yield of BIAs in engineered microorganisms. Databases for 21 BIA-producing species from four plant families yielded 33 assembled contigs with homology to characterized NCS genes. Predicted translation products generated from nine contigs consisted of two to five sequential repeats, each containing most of the sequence found in single-domain enzymes. Assembled contigs containing tandem domain repeats were detected only in members of the Papaveraceae family, including opium poppy (Papaver somniferum). Fourteen cDNAs were generated from 10 species, five of which encoded NCS orthologs with repeated domains. Functional analysis of corresponding recombinant proteins yielded six active NCS enzymes, including four containing either two, three or four repeated catalytic domains. Truncation of the first 25 N-terminal amino acids from the remaining polypeptides revealed two additional enzymes. Multiple catalytic domains correlated with a proportional increase in catalytic efficiency. Expression of NCS genes in Saccharomyces cereviseae also produced active enzymes. The metabolic conversion capacity of engineered yeast positively correlated with the number of repeated domains. PMID:27991536

  13. Genes encoding norcoclaurine synthase occur as tandem fusions in the Papaveraceae.

    PubMed

    Li, Jing; Lee, Eun-Jeong; Chang, Limei; Facchini, Peter J

    2016-12-19

    Norcoclaurine synthase (NCS) catalyzes the enantioselective Pictet-Spengler condensation of dopamine and 4-hydroxyphenylacetaldehyde as the first step in benzylisoquinoline alkaloid (BIA) biosynthesis. NCS orthologs in available transcriptome databases were screened for variants that might improve the low yield of BIAs in engineered microorganisms. Databases for 21 BIA-producing species from four plant families yielded 33 assembled contigs with homology to characterized NCS genes. Predicted translation products generated from nine contigs consisted of two to five sequential repeats, each containing most of the sequence found in single-domain enzymes. Assembled contigs containing tandem domain repeats were detected only in members of the Papaveraceae family, including opium poppy (Papaver somniferum). Fourteen cDNAs were generated from 10 species, five of which encoded NCS orthologs with repeated domains. Functional analysis of corresponding recombinant proteins yielded six active NCS enzymes, including four containing either two, three or four repeated catalytic domains. Truncation of the first 25 N-terminal amino acids from the remaining polypeptides revealed two additional enzymes. Multiple catalytic domains correlated with a proportional increase in catalytic efficiency. Expression of NCS genes in Saccharomyces cereviseae also produced active enzymes. The metabolic conversion capacity of engineered yeast positively correlated with the number of repeated domains.

  14. Mutations in the Drosophila Pushover Gene Confer Increased Neuronal Excitability and Spontaneous Synaptic Vesicle Fusion

    PubMed Central

    Richards, S.; Hillman, T.; Stern, M.

    1996-01-01

    We describe the identification of a gene called pushover (push), which affects both behavior and synaptic transmission at the neuromuscular junction. Adults carrying either of two mutations in push exhibit sluggishness, uncoordination, a defective escape response, and male sterility. Larvae defective in push exhibit increased release of transmitter at the neuromuscular junction. In particular, the frequency of spontaneous transmitter release and the amount of transmitter release evoked by nerve stimulation are each increased two- to threefold in push mutants at the lowest external [Ca(2+)] tested (0.15 mM). Furthermore, these mutants are more sensitive than wild type to application of the potassium channel-blocking drug quinidine: following qunidine application, push mutants, but not wild-type, display repetitive firing of the motor axon, leading to repetitive muscle postsynaptic potentials. The push gene thus might affect both neuronal excitability and the transmitter release process. Complementation tests and recombinational mapping suggest that the push mutations are allelic to a previously identified P-element-induced mutation, which also causes behavioral abnormalities and male sterility. PMID:8846899

  15. Electrochemical determination of BCR/ABL fusion gene based on in situ synthesized gold nanoparticles and cerium dioxide nanoparticles.

    PubMed

    Li, Shenfeng; Wang, Li; Li, Yajuan; Zhu, Xiaoying; Zhong, Liang; Lu, Lingsong; Zhang, Wei; Liu, Bei; Xie, Guoming; Feng, Wenli

    2013-12-01

    An efficient DNA electrochemical biosensor, based on the gold nanoparticles (GNPs) in situ synthesized at the surface of multiwalled carbon nanotubes (MWCNTs), cerium dioxide (CeO2) and chitosan (Chits) composite membrane, was developed for the detection of BCR/ABL fusion gene in chronic myelogenous leukemia (CML). The capture probe was attached onto the nanocomposite membrane modified glassy carbon electrode (GCE) through the conjugated structure. Owing to the synergistic effects of CeO2 nanoparticles with a strong adsorption ability and MWCNTs with a large surface area and excellent electron transfer ability, the prepared composite membrane was demonstrated an efficient electron transfer ability. The biosensor was electrochemically characterized by cyclic voltammogram (CV) and differential pulse voltammetry (DPV), and the decrease of the peak currents upon hybridization was observed using methylene blue (MB) as the electroactive indicator. Under the optimized conditions, peak currents were linear over the range from 1 × 10(-9) M to 1 × 10(-)(12) M, with a detection limit of 5 × 10(-)(13) M (based on the 3σ). And the proposed method was successfully applied for the detection of PCR real samples with satisfactory results. Furthermore, the developed DNA biosensor was demonstrated a good selectivity, a reasonable stability and a favorable reproducibility, which could be regenerated easily.

  16. Novel real-time polymerase chain reaction assay for simultaneous detection of recurrent fusion genes in acute myeloid leukemia.

    PubMed

    Dolz, Sandra; Barragán, Eva; Fuster, Óscar; Llop, Marta; Cervera, José; Such, Esperanza; De Juan, Inmaculada; Palanca, Sarai; Murria, Rosa; Bolufer, Pascual; Luna, Irene; Gómez, Inés; López, María; Ibáñez, Mariam; Sanz, Miguel A

    2013-09-01

    The recent World Health Organization classification recognizes different subtypes of acute myeloid leukemia (AML) according to the presence of several recurrent genetic abnormalities. Detection of these abnormalities and other molecular changes is of increasing interest because it contributes to a refined diagnosis and prognostic assessment in AML and enables monitoring of minimal residual disease. These genetic abnormalities can be detected using single RT-PCR, although the screening is still labor intensive and costly. We have developed a novel real-time RT-PCR assay to simultaneously detect 15 AML-associated rearrangements that is a simple and easily applicable method for use in clinical diagnostic laboratories. This method showed 100% specificity and sensitivity (95% confidence interval, 91% to 100% and 92% to 100%, respectively). The procedure was validated in a series of 105 patients with AML. The method confirmed all translocations detected using standard cytogenetics and fluorescence in situ hybridization and some additional undetected rearrangements. Two patients demonstrated two molecular rearrangements simultaneously, with BCR-ABL1 implicated in both, in addition to RUNX1-MECOM in one patient and PML-RARA in another. In conclusion, this novel real-time RT-PCR assay for simultaneous detection of multiple AML-associated fusion genes is a versatile and sensitive method for reliable screening of recurrent rearrangements in AML.

  17. Targeting of the CNS in MPS-IH Using a Nonviral Transferrin-α-l-iduronidase Fusion Gene Product

    PubMed Central

    Osborn, Mark J; McElmurry, Ron T; Peacock, Brandon; Tolar, Jakub; Blazar, Bruce R

    2008-01-01

    Mucopolysaccharidosis type I (Hurler syndrome) is caused by a deficiency of the enzyme α-l-iduronidase (IDUA), and is characterized by widespread lysosomal glycosaminoglycan (GAG) accumulation. Successful treatment of central nervous system (CNS) diseases is limited by the presence of the blood–brain barrier, which prevents penetration of the therapeutic enzyme. Given that the brain capillary endothelial cells that form this barrier express high levels of the transferrin receptor (TfR), we hypothesized that the coupling of IDUA to transferrin (Tf) would facilitate IDUA delivery to the CNS. A plasmid bearing a fusion gene consisting of Tf and IDUA was constructed which, when delivered in vivo, resulted in the production of high levels of an enzymatically active protein that was transported into the CNS by TfR-mediated endocytosis. Short-term treatment resulted in a decrease in GAGs in the cerebellum of mucopolysaccharidosis type I (MPS I) mice. This approach, therefore, represents a potential strategy for the delivery of therapeutic enzyme to the CNS. PMID:18523448

  18. CONFERENCE REPORT: Summary of the 5th IAEA Technical Meeting on Steady State Operation of Magnetic Fusion Devices (Daejeon, Republic of Korea, 14 17 May 2007)

    NASA Astrophysics Data System (ADS)

    Lee, G. S.; Na, Yong-Su; Becoulet, A.; Ide, S.; Kessel, C. E.; Komori, A.; Kuteev, B. V.; Mank, G.; Olstad, R. A.; Sarkar, B.; Sips, A. C. C.; van Houtte, D.; Vdovin, V. L.

    2008-08-01

    This report summarizes the contributions presented at the 5th IAEA Technical Meeting on Steady State Operation of Magnetic Fusion Devices, held in Daejeon, Republic of Korea, 14-17 May 2007. The main topics of the meeting were overview and superconducting devices, long pulse operation and advanced tokamak, steady state fusion technology, heating and current drive, particle control and power exhaust and ITER-related issues.

  19. A simple, rapid, low-cost technique for naked-eye detection of urine-isolated TMPRSS2:ERG gene fusion RNA

    PubMed Central

    Koo, Kevin M.; Wee, Eugene J. H.; Mainwaring, Paul N.; Trau, Matt

    2016-01-01

    The TMPRSS2:ERG gene fusion is one of a series of highly promising prostate cancer (PCa) biomarker alternatives to the controversial serum PSA. Current methods for detecting TMPRSS2:ERG are limited in terms of long processing time, high cost and the need for specialized equipment. Thus, there is an unmet need for less complex, faster, and cheaper methods to enable gene fusion detection in the clinic. We describe herein a simple, rapid and inexpensive assay which combines robust isothermal amplification technique with a novel visualization method for evaluating urinary TMPRSS2:ERG status at less than USD 5 and with minimal equipment. The assay is sensitive, and rapidly detects as low as 105 copies of TMPRSS2:ERG transcripts while maintaining high levels of specificity. PMID:27470540

  20. CONFERENCE REPORT: Summary of the 17th International Atomic Energy Agency Technical Meeting on 'Research Using Small Fusion Devices'

    NASA Astrophysics Data System (ADS)

    Varandas, C. A. F.; Silva, C.; Gribkov, V. A.; Malaquias, A.; Van Oost, G.

    2008-07-01

    This paper presents a summary of the recent results reported on several topics on magnetic confinement, dense magnetized plasmas and materials studies. The main topics covered on magnetic confinement devices are configuration studies, diagnostics developments, alternative fuelling techniques, turbulence and transport studies, confinement experiments, plasma facing materials, acquisition and control systems and integrated modelling. For the dense magnetized plasma devices results on development and commissioning of several devices (plasma focus, Z-pinch and plasma discharge type), material tests, scaling laws for plasma focus energy density from a few millijoule to megajoule, modelling of neutron and x-ray production mechanisms and fast diagnostic and signal formatting techniques were reported. Auxiliary heating systems for ITER, dedicated materials modelling and experimental studies relevant for several fusion applications and control and data acquisition systems have also been reported on in dedicated papers.

  1. ETS Gene Fusions as Predictive Biomarkers of Resistance to Radiation Therapy for Prostate Cancer

    DTIC Science & Technology

    2014-08-01

    resistance  ERG is diffusely localized through the prostate cancer cell and does not redistribute upon genotoxic stress Reportable Outcomes: The past...JL, Schrecengost RS, Han S, Den RB, Dicker AP, Feng FY, and Knudsen KE. A hormone-DNA repair circuit governs the response to genotoxic insult

  2. Secretory Carcinoma of the Skin Harboring ETV6 Gene Fusions: A Cutaneous Analogue to Secretory Carcinomas of the Breast and Salivary Glands.

    PubMed

    Bishop, Justin A; Taube, Janis M; Su, Albert; Binder, Scott W; Kazakov, Dmitry V; Michal, Michal; Westra, William H

    2017-01-01

    Mammary analogue secretory carcinoma is a low-grade salivary gland carcinoma that exhibits analogous features to secretory carcinoma of the breast including the presence of a t(12;15) translocation resulting in the ETV6-NTRK3 gene fusion. Rare cases of purported secretory carcinoma of the skin adnexa have been reported, but their relationship to true secretory carcinoma of the breast and salivary glands is unclear, as they generally do not harbor ETV6 rearrangements. Cases of cutaneous neoplasms with histologic features identical to secretory carcinoma of the breast and salivary glands were identified from the consultation files of 3 academic medical institutions. Immunohistochemistry was performed for S100 protein, mammaglobin and STAT5a. Break-apart fluorescence in situ hybridization was used evaluate for disruption of the ETV6 gene. Six cases of cutaneous secretory carcinoma were identified. The tumors arose in 4 women and 2 men, ranging from 24 to 71 years in age (mean, 47 y). The carcinomas presented in the skin of the axilla (n=4), ventral neck (n=1), and cheek (n=1). The tumors arose in the superficial dermis in association with adnexal structures. None of the patients had a prior or concurrent breast or salivary gland tumor. They were histologically characterized by well-circumscribed but unencapsulated proliferations of bland, eosinophilic cells arranged in microcysts and follicles with intraluminal secretions. Ectopic breast or salivary gland tissue was not identified. The cases were diffusely positive for S100 protein (6 of 6), mammaglobin (6 of 6), and STAT5a (5 of 5). All 6 cases harbored rearrangements of ETV6. All tumors were treated by simple excision alone. No recurrences or metastases developed in the 2 cases with follow-up. Secretory carcinoma of the skin represents a phenotypic, immunohistochemical, and genetic counterpart to secretory carcinoma of the breast and salivary glands. This tumor entity is less anatomically restricted than previously

  3. Multiplex PCR assay for detection of recombinant genes encoding fatty acid desaturases fused with lichenase reporter protein in GM plants.

    PubMed

    Berdichevets, Iryna N; Shimshilashvili, Hristina R; Gerasymenko, Iryna M; Sindarovska, Yana R; Sheludko, Yuriy V; Goldenkova-Pavlova, Irina V

    2010-07-01

    Thermostable lichenase encoded by licB gene of Clostridium thermocellum can be used as a reporter protein in plant, bacterial, yeast, and mammalian cells. It has important advantages of high sensitivity and specificity in qualitative and quantitative assays. Deletion variants of LicB (e.g., LicBM3) retain its enzymatic activity and thermostability and can be expressed in translational fusion with target proteins without compromising with their properties. Fusion with the lichenase reporter is especially convenient for the heterologous expression of proteins whose analysis is difficult or compromised by host enzyme activities, as it is in case of fatty acid desaturases occurring in all groups of organisms. Recombinant desaturase-lichenase genes can be used for creating genetically modified (GM) plants with improved chill tolerance. Development of an analytical method for detection of fused desaturase-lichenase transgenes is necessary both for production of GM plants and for their certification. Here, we report a multiplex polymerase chain reaction method for detection of desA and desC desaturase genes of cyanobacteria Synechocystis sp. PCC6803 and Synechococcus vulcanus, respectively, fused to licBM3 reporter in GM plants.

  4. The Neurospora Transcription Factor ADV-1 Transduces Light Signals and Temporal Information to Control Rhythmic Expression of Genes Involved in Cell Fusion

    PubMed Central

    Dekhang, Rigzin; Wu, Cheng; Smith, Kristina M.; Lamb, Teresa M.; Peterson, Matthew; Bredeweg, Erin L.; Ibarra, Oneida; Emerson, Jillian M.; Karunarathna, Nirmala; Lyubetskaya, Anna; Azizi, Elham; Hurley, Jennifer M.; Dunlap, Jay C.; Galagan, James E.; Freitag, Michael; Sachs, Matthew S.; Bell-Pedersen, Deborah

    2016-01-01

    Light and the circadian clock have a profound effect on the biology of organisms through the regulation of large sets of genes. Toward understanding how light and the circadian clock regulate gene expression, we used genome-wide approaches to identify the direct and indirect targets of the light-responsive and clock-controlled transcription factor ADV-1 in Neurospora crassa. A large proportion of ADV-1 targets were found to be light- and/or clock-controlled, and enriched for genes involved in development, metabolism, cell growth, and cell fusion. We show that ADV-1 is necessary for transducing light and/or temporal information to its immediate downstream targets, including controlling rhythms in genes critical to somatic cell fusion. However, while ADV-1 targets are altered in predictable ways in Δadv-1 cells in response to light, this is not always the case for rhythmic target gene expression. These data suggest that a complex regulatory network downstream of ADV-1 functions to generate distinct temporal dynamics of target gene expression relative to the central clock mechanism. PMID:27856696

  5. Chromatin-prebound Crm1 recruits Nup98-HoxA9 fusion to induce aberrant expression of Hox cluster genes.

    PubMed

    Oka, Masahiro; Mura, Sonoko; Yamada, Kohji; Sangel, Percival; Hirata, Saki; Maehara, Kazumitsu; Kawakami, Koichi; Tachibana, Taro; Ohkawa, Yasuyuki; Kimura, Hiroshi; Yoneda, Yoshihiro

    2016-01-07

    The nucleoporin Nup98 is frequently rearranged to form leukemogenic Nup98-fusion proteins with various partners. However, their function remains largely elusive. Here, we show that Nup98-HoxA9, a fusion between Nup98 and the homeobox transcription factor HoxA9, forms nuclear aggregates that frequently associate with facultative heterochromatin. We demonstrate that stable expression of Nup98-HoxA9 in mouse embryonic stem cells selectively induces the expression of Hox cluster genes. Genome-wide binding site analysis revealed that Nup98-HoxA9 is preferentially targeted and accumulated at Hox cluster regions where the export factor Crm1 is originally prebound. In addition, leptomycin B, an inhibitor of Crm1, disassembled nuclear Nup98-HoxA9 dots, resulting in the loss of chromatin binding of Nup98-HoxA9 and Nup98-HoxA9-mediated activation of Hox genes. Collectively, our results indicate that highly selective targeting of Nup98-fusion proteins to Hox cluster regions via prebound Crm1 induces the formation of higher order chromatin structures that causes aberrant Hox gene regulation.

  6. Chromatin-prebound Crm1 recruits Nup98-HoxA9 fusion to induce aberrant expression of Hox cluster genes

    PubMed Central

    Oka, Masahiro; Mura, Sonoko; Yamada, Kohji; Sangel, Percival; Hirata, Saki; Maehara, Kazumitsu; Kawakami, Koichi; Tachibana, Taro; Ohkawa, Yasuyuki; Kimura, Hiroshi; Yoneda, Yoshihiro

    2016-01-01

    The nucleoporin Nup98 is frequently rearranged to form leukemogenic Nup98-fusion proteins with various partners. However, their function remains largely elusive. Here, we show that Nup98-HoxA9, a fusion between Nup98 and the homeobox transcription factor HoxA9, forms nuclear aggregates that frequently associate with facultative heterochromatin. We demonstrate that stable expression of Nup98-HoxA9 in mouse embryonic stem cells selectively induces the expression of Hox cluster genes. Genome-wide binding site analysis revealed that Nup98-HoxA9 is preferentially targeted and accumulated at Hox cluster regions where the export factor Crm1 is originally prebound. In addition, leptomycin B, an inhibitor of Crm1, disassembled nuclear Nup98-HoxA9 dots, resulting in the loss of chromatin binding of Nup98-HoxA9 and Nup98-HoxA9-mediated activation of Hox genes. Collectively, our results indicate that highly selective targeting of Nup98-fusion proteins to Hox cluster regions via prebound Crm1 induces the formation of higher order chromatin structures that causes aberrant Hox gene regulation. DOI: http://dx.doi.org/10.7554/eLife.09540.001 PMID:26740045

  7. Bis-three-way junction nanostructure and DNA machineries for ultrasensitive and specific detection of BCR/ABL fusion gene by chemiluminescence imaging

    PubMed Central

    Xu, Yongjie; Bian, Xintong; Sang, Ye; Li, Yujian; Li, Dandan; Cheng, Wei; Yin, Yibing; Ju, Huangxian; Ding, Shijia

    2016-01-01

    A novel G-quadruplex DNAzyme-driven chemiluminescence (CL) imaging method has been developed for ultrasensitive and specific detection of BCR/ABL fusion gene based on bis-three-way junction (bis-3WJ) nanostructure and cascade DNA machineries. Bis-3WJ probes are designed logically to recognize BCR/ABL fusion gene, which forms the stable bis-3WJ nanostructure for the activation of polymerase/nicking enzyme machineries in cascade, resulting in synthesis of DNAzyme subunits. These DNAzyme subunits can form integrated DNAzyme by self-assembly to catalyze CL substrate, thus providing an amplified signal for the sensing events or outputs for AND logic operation. The imaging method achieved ultrasensitive detection of BCR/ABL fusion gene with a low detection limit down to 23 fM. And this method exhibited wide linear ranges over seven orders of magnitude and excellent discrimination ability toward target. In addition, an acceptable recovery was obtained in complex matrix. It is notable that this biosensing strategy possesses merits of homogenous, isothermal and label-free assay system. Therefore, these merits endow the developed imaging method with a potential tool for CML diagnosis. PMID:27577607

  8. ETS Gene Fusions as Predictive Biomarkers of Resistance to Radiation Therapy for Prostate Cancer

    DTIC Science & Technology

    2011-08-01

    radiation resistance can be reversed with DNAPK inhibition. These findings suggest that DNA-PK inhibition should be explored as a clinical strategy for...PK inhibition should be explored as a clinical strategy for radiosensitizing prostate cancers. In addition, we have discovered that ERG interacts...DOD Annual Report review committee to consider allowing us to expand the aims of this grant to assess PARP1 inhibition as a therapeutic strategy for

  9. Identification and Analysis of Bacterial Protein Secretion Inhibitors Utilizing a SecA-LacZ Reporter Fusion System

    PubMed Central

    Alksne, L. E.; Burgio, P.; Hu, W.; Feld, B.; Singh, M. P.; Tuckman, M.; Petersen, P. J.; Labthavikul, P.; McGlynn, M.; Barbieri, L.; McDonald, L.; Bradford, P.; Dushin, R. G.; Rothstein, D.; Projan, S. J.

    2000-01-01

    Protein secretion is an essential process for bacterial growth, yet there are few if any antimicrobial agents which inhibit secretion. An in vivo, high-throughput screen to detect secretion inhibitors was developed based on the translational autoregulation of one of the central protein components, SecA. The assay makes use of a SecA-LacZ fusion reporter construct in Escherichia coli which is induced when secretion is perturbed. Several compounds, including two natural product extracts, which had the ability to induce the reporter fusion were identified and the MICs of these compounds for Staphylococcus aureus strain MN8 were found to be ≤128 μg/ml. Enzyme-linked immunosorbent assay, Western blotting, and immunoprecipitation techniques were used to analyze the affects of these compounds on protein secretion. Six representative compounds presented here appear to be bona fide secretion inhibitors but were found to have deleterious effects on membranes. It was concluded that, while the method described here for identifying inhibitors of secretion is valid, screens such as this, which are directed against the membrane-bound portion of a pathway, may preferentially identify compounds which affect membrane integrity. PMID:10817687

  10. Inertial confinement fusion target component fabrication and technology development support. Annual report, October 1, 1994--September 30, 1995

    SciTech Connect

    Hoppe, M.

    1996-05-01

    On December 30, 1990, the US Department of Energy entered into a contract with General Atomics (GA) to be the Inertial Confinement Fusion (ICF) Target Component Fabrication and Technology Development Support contractor. This report documents the technical activities of the period October 1, 1994 through September 30, 1995. During this period, GA was assigned 15 tasks in support of the Inertial Confinement Fusion program and its laboratories. A portion of the effort on these tasks included providing direct ``Onsite Support`` at Lawrence Livermore National Laboratory (LLNL), Los Alamos National Laboratory (LANL), and Sandia National Laboratory Albuquerque (SNLA). The ICF program is anticipating experiments at the National Ignition Facility (NIF) and the OMEGA Upgrade. Both facilities will require capsules containing layered D{sub 2} or deuterium-tritium (D-T) fuel. The authors are part of the National Cryogenic Target Program to create and demonstrate viable ways to generate and characterize cryogenic layers. Progress has been made on ways to both create viable layers and to characterize them. They continued engineering, assembly and testing of equipment for a cryogenic target handling system for University of Rochester`s Laboratory for Laser Energetics (UR/LLE) that will fill, transport, layer, and characterize targets filled with cryogenic fuel, and insert these cryogenic targets into the OMEGA Upgrade target chamber for laser implosion experiments. This report summarizes and documents the technical progress made on these tasks.

  11. Recombinant Carcinoembryonic Antigen as a Reporter Gene for Molecular Imaging

    PubMed Central

    Kenanova, Vania; Barat, Bhaswati; Olafsen, Tove; Chatziioannou, Arion; Herschman, Harvey R.; Braun, Jonathan; Wu, Anna M.

    2009-01-01

    Purpose Reporter genes can provide a way of non-invasively assessing gene activity in vivo. However, current reporter gene strategies may be limited by the immunogenicity of foreign reporter proteins, endogenous expression or unwanted biological activity. We have developed a reporter gene based on carcinoembryonic antigen (CEA), a human protein with limited normal tissue expression. Methods To construct a CEA reporter gene for PET, a CEA minigene (N-A3) was fused to the extracellular and transmembrane domains of the human FcγRIIb receptor. The NA3-FcγRIIb recombinant gene, driven by a CMV promoter, was transfected in Jurkat (human T cell leukemia) cells. Expression was analyzed by flow cytometry, immunohistochemistry (IHC), and microPET imaging. Results Flow cytometry identified Jurkat clones stably expressing NA3-FcγRIIb at low, medium, and high levels. High and medium NA3-FcγRIIb expression could also be detected by Western blot. Reporter gene positive and negative Jurkat cells were used to establish xenografts in athymic mice. IHC showed staining of the tumor with high reporter gene expression; medium and low N-A3 expression was not detected. MicroPET imaging, using an anti-CEA 124I-labeled scFv-Fc antibody fragment, demonstrated that only high N-A3 expression could be detected. Specific accumulation of activity was visualized at the N-A3 positive tumor as early as 4h. MicroPET image quantitation showed tumor activity of 1.8(±0.2), 15.2(±1.3) and 4.6(±1.2) %ID/g at 4h, 20h and 48h, respectively. Biodistribution at 48h, demonstrated tumor uptake of 4.8(±0.8) %ID/g. Conclusion The CEA N-A3 minigene has the potential to be used as a reporter gene for imaging cells in vivo. PMID:18719907

  12. Enhanced immune response with foot and mouth disease virus VP1 and interleukin-1 fusion genes.

    PubMed

    Park, Jong Hyeon; Kim, Sun Jin; Oem, Jae Ku; Lee, Kwang Nyeong; Kim, Yong Joo; Kye, Soo Jeong; Park, Jee Yong; Joo, Yi Seok

    2006-09-01

    The capsid of the foot and mouth disease (FMD) virus carries the epitopes that are critical for inducing the immune response. In an attempt to enhance the specific immune response, plasmid DNA was constructed to express VP1/interleukin-1alpha (IL-1alpha) and precursor capsid (P1) in combination with 2A (P1-2A)/IL-1alpha under the control of the human cytomegalovirus (HCMV) immediateearly promoter and intron. After DNA transfection into MA104 (monkey kidney) cells, Western blotting and an immunofluorescence assay were used to confirm the expression of VP1 or P1-2A and IL-1alpha. Mice were inoculated with the encoding plasmids via the intradermal route, and the IgG1 and IgG2a levels were used to determine the immune responses. These results show that although the immunized groups did not carry a high level of neutralizing antibodies, the plasmids encoding the VP1/ IL-1alpha, and P1-2A /IL-1alpha fused genes were effective in inducing an enhanced immune response.

  13. Heavy Ion Fusion Accelerator Research (HIFAR) half-year report, October 1, 1988--March 31, 1989

    SciTech Connect

    Not Available

    1989-06-01

    The basic objective of the Heavy Ion Fusion Accelerator Research (HIFAR) program is to assess the suitability of heavy ion accelerators as igniters for Inertial Confinement Fusion (ICF). A specific accelerator technology, the induction linac, has been studied at the Lawrence Berkeley Laboratory and has reached the point at which its viability for ICF applications can be assessed over the next few years. The HIFAR program addresses the generation of high-power, high-brightness beams of heavy ions, the understanding of the scaling laws in this novel physics regime, and the validation of new accelerator strategies, to cut costs. Key elements to be addressed include: beam quality limits set by transverse and longitudinal beam physics; development of induction accelerating modules, and multiple-beam hardware, at affordable costs; acceleration of multiple beams with current amplification --both new features in a linac -- without significant dilution of the optical quality of the beams; and final bunching, transport, and accurate focusing on a small target.

  14. Materials recycle and waste management in fusion power reactors. Progress report for 1982

    SciTech Connect

    Vogler, S.; Jung, J.; Steindler, M.J.; Maya, I.; Levine, H.E.; Peterman, D.D.; Strausburg, S.; Schultz, K.R.

    1983-01-01

    Several components of a STARFIRE fusion reactor have been studied. The breeding ratios were calculated as a function of lithium enrichment and neutron multiplier for systems containing either Li/sub 2/O or LiAlO/sub 2/. The lithium requirements for a fusion economy were also estimated for those cases and the current US resources were found to be adequate. However, competition with other lithium demands in the future emphasizes the need for recovering and reusing lithium. The radioactivities induced in the breeder and the impurities responsible for their formation were determined. The residual radioactivities of several low-activation structural materials were compared with the radioactivity from the prime candidate alloy (PCA) a titanium modified Type 316 stainless steel used in STARFIRE. The impurities responsible for the radioactivity levels were identified. From these radioactive impurity levels it was determined that V15Cr5Ti could meet the requirements for shallow land burial as specified by the Nuclear Regulatory Commission (10CFR61), whereas PCA would require a more restrictive disposal mode, i.e. in a geologic medium. The costs for each of these disposal modes were then estimated.

  15. A novel hNIS/tdTomato fusion reporter for visualizing the relationship between the cellular localization of sodium iodide symporter and its iodine uptake function under heat shock treatment.

    PubMed

    Yeom, Chan Joo; Chung, Taemoon; Youn, Hyewon; Kang, Keon Wook; Lee, Dong Soo; Chung, June-Key

    2015-01-01

    The function of membrane-localized sodium iodide symporter (NIS) determines the efficacy of radioiodine therapy in thyroid cancer. Here, we describe a dual mode reporter fused with human NIS (hNIS) and a red fluorescent protein named tandem dimeric Tomato (tdTomato) for the in vitro and in vivo imaging of hNIS protein expression, localization, and iodide uptake function. Human cervical epithelial adenocarcinoma cell line (HeLa)-hNIS/tdTomato cells were established by transducing a fusion gene expressing hNIS/tdTomato under the control of a cytomegalovirus promoter. Fluorescence imaging, confocal microscopy, and an 125I uptake assay were performed to validate the integrity of the fusion protein. Actinomycin D and cycloheximide were used to block newly synthesized hNIS proteins. In vivo images were acquired using a gamma camera and a Maestro fluorescence imaging device. The fluorescence intensity of membrane-localized hNIS and 125I uptake both were increased after heat shock. Scintigraphy and fluorescence imaging indicated specific accumulation of the hNIS/tdTomato fusion protein in xenografted tumors, supporting the utility of this system for in vivo monitoring of hNIS expression and activity. We developed a novel hNIS/tdTomato dual mode reporter that enables visualization of the expression, localization, and iodine uptake function of hNIS in vitro and in vivo.

  16. The glpD gene is a novel reporter gene for E. coli that is superior to established reporter genes like lacZ and gusA.

    PubMed

    Wegener, Marius; Vogtmann, Kristina; Huber, Madeleine; Laass, Sebastian; Soppa, Jörg

    2016-12-01

    Reporter genes facilitate the characterization of promoter activities, transcript stabilities, translational efficiencies, or intracellular localization. Various reporter genes for Escherichia coli have been established, however, most of them have drawbacks like transcript instability or the inability to be used in genetic selections. Therefore, the glpD gene encoding glycerol-3-phosphate dehydrogenase was introduced as a novel reporter gene for E. coli. The enzymatic assay was optimized, and it was verified that growth on glycerol strictly depends on the presence of GlpD. The 5'-UTRs of three E. coli genes were chosen and cloned upstream of the new reporter gene glpD as well as the established reporter genes lacZ and gusA. Protein and transcript levels were quantified and translational efficiencies were calculated. The lacZ transcript was very unstable and its level highly depended on its translation, compromising its use as a reporter. The results obtained with gusA and glpD were similar, however, only glpD can be used for genetic selections. Therefore, glpD was found to be a superior novel reporter gene compared to the established reporter genes lacZ and gusA.

  17. Fusion reactor systems studies. Progress report for the period November 1, 1996--October 31, 1997, and final report

    SciTech Connect

    El-Guebaly, L.A.; Blanchard, J.P.; Kulcinski, G.L.

    1997-08-01

    During FY97, the University of Wisconsin Fusion Technology Institute personnel have participated in the ARIES-RS and the ARIES-ST projects. The main areas of effort are: (1) neutronics analysis; (2) shielding of components and personnel; (3) neutron wall loading distribution; (4) radiation damage to in-vessel components; (5) components lifetimes; (6) embrittled materials designs issues; (7) stress and structural analysis; (8) activation, LOCA, and safety analysis; (9) support and fabrication of components; (10) vacuum system; and (11) maintenance. Progress made in these areas are summarized.

  18. Two novel alkane hydroxylase-rubredoxin fusion genes isolated from a Dietzia bacterium and the functions of fused rubredoxin domains in long-chain n-alkane degradation.

    PubMed

    Nie, Yong; Liang, Jieliang; Fang, Hui; Tang, Yue-Qin; Wu, Xiao-Lei

    2011-10-01

    Two alkane hydroxylase-rubredoxin fusion gene homologs (alkW1 and alkW2) were cloned from a Dietzia strain, designated DQ12-45-1b, which can grow on crude oil and n-alkanes ranging in length from 6 to 40 carbon atoms as sole carbon sources. Both AlkW1 and AlkW2 have an integral-membrane alkane monooxygenase (AlkB) conserved domain and a rubredoxin (Rd) conserved domain which are fused together. Phylogenetic analysis showed that these two AlkB-fused Rd domains formed a novel third cluster with all the Rds from the alkane hydroxylase-rubredoxin fusion gene clusters in Gram-positive bacteria and that this third cluster was distant from the known AlkG1- and AlkG2-type Rds. Expression of the alkW1 gene in DQ12-45-1b was induced when cells were grown on C(8) to C(32) n-alkanes as sole carbon sources, but expression of the alkW2 gene was not detected. Functional heterologous expression in an alkB deletion mutant of Pseudomonas fluorescens KOB2Δ1 suggested the alkW1 could restore the growth of KOB2Δ1 on C(14) and C(16) n-alkanes and induce faster growth on C(18) to C(32) n-alkanes than alkW1ΔRd, the Rd domain deletion mutant gene of alkW1, which also caused faster growth than KOB2Δ1 itself. In addition, the artificial fusion of AlkB from the Gram-negative P. fluorescens CHA0 and the Rds from both Gram-negative P. fluorescens CHA0 and Gram-positive Dietzia sp. DQ12-45-1b significantly increased the degradation of C(32) alkane compared to that seen with AlkB itself. In conclusion, the alkW1 gene cloned from Dietzia species encoded an alkane hydroxylase which increased growth on and degradation of n-alkanes up to C(32) in length, with its fused rubredoxin domain being necessary to maintain the functions. In addition, the fusion of alkane hydroxylase and rubredoxin genes from both Gram-positive and -negative bacteria can increase the degradation of long-chain n-alkanes (such as C(32)) in the Gram-negative bacterium.

  19. FES Science Network Requirements - Report of the Fusion Energy Sciences Network Requirements Workshop Conducted March 13 and 14, 2008

    SciTech Connect

    Tierney, Brian; Dart, Eli; Tierney, Brian

    2008-07-10

    The Energy Sciences Network (ESnet) is the primary provider of network connectivity for the U.S. Department of Energy Office of Science, the single largest supporter of basic research in the physical sciences in the United States of America. In support of the Office of Science programs, ESnet regularly updates and refreshes its understanding of the networking requirements of the instruments, facilities, scientists, and science programs that it serves. This focus has helped ESnet to be a highly successful enabler of scientific discovery for over 20 years. In March 2008, ESnet and the Fusion Energy Sciences (FES) Program Office of the DOE Office of Science organized a workshop to characterize the networking requirements of the science programs funded by the FES Program Office. Most sites that conduct data-intensive activities (the Tokamaks at GA and MIT, the supercomputer centers at NERSC and ORNL) show a need for on the order of 10 Gbps of network bandwidth for FES-related work within 5 years. PPPL reported a need for 8 times that (80 Gbps) in that time frame. Estimates for the 5-10 year time period are up to 160 Mbps for large simulations. Bandwidth requirements for ITER range from 10 to 80 Gbps. In terms of science process and collaboration structure, it is clear that the proposed Fusion Simulation Project (FSP) has the potential to significantly impact the data movement patterns and therefore the network requirements for U.S. fusion science. As the FSP is defined over the next two years, these changes will become clearer. Also, there is a clear and present unmet need for better network connectivity between U.S. FES sites and two Asian fusion experiments--the EAST Tokamak in China and the KSTAR Tokamak in South Korea. In addition to achieving its goal of collecting and characterizing the network requirements of the science endeavors funded by the FES Program Office, the workshop emphasized that there is a need for research into better ways of conducting remote

  20. Static and dynamic analyses on the MFTF (Mirror Fusion Test Facility)-B Axicell Vacuum Vessel System: Final report

    SciTech Connect

    Ng, D.S.

    1986-09-01

    The Mirror Fusion Test Facility (MFTF-B) at Lawrence Livermore National Laboratory (LLNL) is a large-scale, tandem-mirror-fusion experiment. MFTF-B comprises many highly interconnected systems, including a magnet array and a vacuum vessel. The vessel, which houses the magnet array, is supported by reinforced concrete piers and steel frames resting on an array of foundations and surrounded by a 7-ft-thick concrete shielding vault. The Pittsburgh-Des Moines (PDM) Corporation, which was awarded the contract to design and construct the vessel, carried out fixed-base static and dynamic analyses of a finite-element model of the axicell vessel and magnet systems, including the simulation of various loading conditions and three postulated earthquake excitations. Meanwhile, LLNL monitored PDM's analyses with modeling studies of its own, and independently evaluated the structural responses of the vessel in order to define design criteria for the interface members and other project equipment. The assumptions underlying the finite-element model and the behavior of the axicell vessel are described in detail in this report, with particular emphasis placed on comparing the LLNL and PDM studies and on analyzing the fixed-base behavior with the soil-structure interaction, which occurs between the vessel and the massive concrete vault wall during a postulated seismic event. The structural members that proved sensitive to the soil effect are also reevaluated.

  1. Fusion of a supernumerary tooth to right mandibular second molar: a case report and literature review

    PubMed Central

    Zhu, Min; Liu, Chao; Ren, Shuangshuang; Lin, Zintong; Miao, Leiying; Sun, Weibin

    2015-01-01

    Gemination or fusion is a rare occurrence in the mandibular posterior teeth. The aim of this article is to describe the problems encountered and the strategy employed in treating such cases. A 34 years old patient came with the complaint of spontaneous and radiating pain in the right mandibular posterior region. The tooth in concern was an anomalous ‘double’ second mandibular molar diagnosed as having necrotic pulp with chronic apical abscess of endodontic origin. The present case emphasizes the importance of identifying anatomical anomalies during treatment of fused teeth with supernumerary tooth, and the need for the use of advanced imaging modalities like CBCT which is a critical aid in the diagnosis of such cases. Fused teeth can be managed quite efficiently by an overall combined treatment including both endodontic and periodontal therapy. PMID:26550101

  2. HYFIRE II: fusion/high-temperature electrolysis conceptual-design study. Annual report

    SciTech Connect

    Fillo, J.A.

    1983-08-01

    As in the previous HYFIRE design study, the current study focuses on coupling a Tokamak fusion reactor with a high-temperature blanket to a High-Temperature Electrolyzer (HTE) process to produce hydrogen and oxygen. Scaling of the STARFIRE reactor to allow a blanket power to 6000 MW(th) is also assumed. The primary difference between the two studies is the maximum inlet steam temperature to the electrolyzer. This temperature is decreased from approx. 1300/sup 0/ to approx. 1150/sup 0/C, which is closer to the maximum projected temperature of the Westinghouse fuel cell design. The process flow conditions change but the basic design philosophy and approaches to process design remain the same as before. Westinghouse assisted in the study in the areas of systems design integration, plasma engineering, balance-of-plant design, and electrolyzer technology.

  3. A Concept Exploration Program in Fast Ignition Inertial Fusion — Final Report

    SciTech Connect

    Stephens, Richarad Burnite; Freeman, Richard R.; Van Woekom, L. D.; Key, M.; MacKinnon, Andrew J.; Wei, Mingsheng

    2014-02-27

    The Fast Ignition (FI) approach to Inertial Confinement Fusion (ICF) holds particular promise for fusion energy because the independently generated compression and ignition pulses allow ignition with less compression, resulting in (potentially) higher gain. Exploiting this concept effectively requires an understanding of the transport of electrons in prototypical geometries and at relevant densities and temperatures. Our consortium, which included General Atomics (GA), The Ohio State University (OSU), the University of California, San Diego (UCSD), University of California, Davis (UC-Davis), and Princeton University under this grant (~$850K/yr) and Lawrence Livermore National Laboratory (LLNL) under a companion grant, won awards in 2000, renewed in 2005, to investigate the physics of electron injection and transport relevant to the FI concept, which is crucial to understand electron transport in integral FI targets. In the last two years we have also been preparing diagnostics and starting to extend the work to electron transport into hot targets. A complementary effort, the Advanced Concept Exploration (ACE) program for Fast Ignition, was funded starting in 2006 to integrate this understanding into ignition schemes specifically suitable for the initial fast ignition attempts on OMEGA and National Ignition Facility (NIF), and during that time these two programs have been managed as a coordinated effort. This result of our 7+ years of effort has been substantial. Utilizing collaborations to access the most capable laser facilities around the world, we have developed an understanding that was summarized in a Fusion Science & Technology 2006, Special Issue on Fast Ignition. The author lists in the 20 articles in that issue are dominated by our group (we are first authors in four of them). Our group has published, or submitted 67 articles, including 1 in Nature, 2 Nature Physics, 10 Physical Review Letters, 8 Review of Scientific Instruments, and has been invited to

  4. Feasibility study of a railgun as a driver for impact fusion: Final report

    SciTech Connect

    Thio. Y.C.

    1986-06-01

    The feasibility of a railgun as a driver for impact fusion is studied through a series of theoretical and experimental investigations. The results of both the theoretical and experimental investigations presented here have helped to identify the potential problems of the railgun launcher to attain velocity in excess of 100 km/s. These include ablation, viscous drag, and secondary arc formation due to either armature dispersion (instability) or restrike. These problems are analyzed and examined experimentally. The behavior of the conventional open-plasma-armature driven railguns have been shown to be quite complex and not easily controllable in the domain of ultrahigh velocity (>6 km/s). Methods to overcome these problems are proposed, analyzed in regards to their technological feasibility, and tested experimentally wherever possible. Techniques for reducing radiative ablation, the concept of a mechanically controlled plasma armature, and the concept of achieving super high augmentation by the technique of trans-augmentation are presented.

  5. Development of high power radio frequency components for fusion plasma heating. Final report, Revision 3

    SciTech Connect

    1997-09-11

    The purpose of this CRADA was to develop advanced microwave heating systems for both ion cyclotron heating and electron cyclotron heating for magnetic fusion reactors. This involved low-frequency (UHF), high-power (millimeter-wave) microwave components, such as antennas, windows, and matching elements. This CRADA also involved developing conceptual designs for new microwave sources. General Atomics built and tested the distributed cooled window and provided LLNL with transmission and reflection test data in order to then benchmark the EM computer codes. The combline antenna built and analyzed by LLNL was based on a GA design. GA provided LLNL with a number of niobium plates for hot pressing and provided the necessary guidance to allow successful bonding. GA representatives were on site at LLNL on numerous occasions to consult and give guidance on the ferroelectric tuner, combline antenna and distributed window analysis.

  6. Inertial Confinement Fusion Quarterly Report: April--June 1993. Volume 3, Number 3

    SciTech Connect

    MacGowan, B.J.; Kotowski, M.; Schleich, D.

    1993-11-01

    This issue of the ICF Quarterly contains six articles describing recent advances in Lawrence Livermore National Laboratory`s inertial confinement fusion (ICF) program. The current emphasis of the ICF program is in support of DOE`s National Ignition Facility (NIF) initiative for demonstrating ignition and gain with a 1-2 MJ glass laser. The articles describe recent Nova experiments and investigations tailored towards enhancing understanding of the key physics and technological issues for the NIF. Titles of the articles are: development of large-aperture KDP crystals; inner-shell photo-ionized X-ray lasers; X-ray radiographic measurements of radiation-driven shock and interface motion in solid density materials; the role of nodule defects in laser-induced damage of multilayer optical coatings; techniques for Mbar to near-Gbar equation-of-state measurements with the Nova laser; parametric instabilities and laser-beam smoothing.

  7. Epigenome Mapping Reveals Distinct Modes of Gene Regulation and Widespread Enhancer Reprogramming by the Oncogenic Fusion Protein EWS-FLI1

    PubMed Central

    Tomazou, Eleni M.; Sheffield, Nathan C.; Schmidl, Christian; Schuster, Michael; Schönegger, Andreas; Datlinger, Paul; Kubicek, Stefan; Bock, Christoph; Kovar, Heinrich

    2015-01-01

    Summary Transcription factor fusion proteins can transform cells by inducing global changes of the transcriptome, often creating a state of oncogene addiction. Here, we investigate the role of epigenetic mechanisms in this process, focusing on Ewing sarcoma cells that are dependent on the EWS-FLI1 fusion protein. We established reference epigenome maps comprising DNA methylation, seven histone marks, open chromatin states, and RNA levels, and we analyzed the epigenome dynamics upon downregulation of the driving oncogene. Reduced EWS-FLI1 expression led to widespread epigenetic changes in promoters, enhancers, and super-enhancers, and we identified histone H3K27 acetylation as the most strongly affected mark. Clustering of epigenetic promoter signatures defined classes of EWS-FLI1-regulated genes that responded differently to low-dose treatment with histone deacetylase inhibitors. Furthermore, we observed strong and opposing enrichment patterns for E2F and AP-1 among EWS-FLI1-correlated and anticorrelated genes. Our data describe extensive genome-wide rewiring of epigenetic cell states driven by an oncogenic fusion protein. PMID:25704812

  8. Multisensor data fusion algorithm development

    SciTech Connect

    Yocky, D.A.; Chadwick, M.D.; Goudy, S.P.; Johnson, D.K.

    1995-12-01

    This report presents a two-year LDRD research effort into multisensor data fusion. We approached the problem by addressing the available types of data, preprocessing that data, and developing fusion algorithms using that data. The report reflects these three distinct areas. First, the possible data sets for fusion are identified. Second, automated registration techniques for imagery data are analyzed. Third, two fusion techniques are presented. The first fusion algorithm is based on the two-dimensional discrete wavelet transform. Using test images, the wavelet algorithm is compared against intensity modulation and intensity-hue-saturation image fusion algorithms that are available in commercial software. The wavelet approach outperforms the other two fusion techniques by preserving spectral/spatial information more precisely. The wavelet fusion algorithm was also applied to Landsat Thematic Mapper and SPOT panchromatic imagery data. The second algorithm is based on a linear-regression technique. We analyzed the technique using the same Landsat and SPOT data.

  9. Characterization of a Cellulomonas fimi exoglucanase/xylanase-endoglucanase gene fusion which improves microbial degradation of cellulosic biomass.

    PubMed

    Duedu, Kwabena O; French, Christopher E

    2016-11-01

    Effective degradation of cellulose requires multiple classes of enzyme working together. However, naturally occurring cellulases with multiple catalytic domains seem to be rather rare in known cellulose-degrading organisms. A fusion protein made from Cellulomonas fimi exo- and endo- glucanases, Cex and CenA which improves breakdown of cellulose is described. A homologous carbohydrate binding module (CBM-2) present in both glucanases was fused to give a fusion protein CxnA. CxnA or unfused constructs (Cex+CenA, Cex, or CenA) were expressed in Escherichia coli and Citrobacter freundii. The latter recombinant strains were cultured at the expense of cellulose filter paper. The expressed CxnA had both exo- and endo- glucanase activities. It was also exported to the supernatant as were the non-fused proteins. In addition, the hybrid CBM from the fusion could bind to microcrystalline cellulose. Growth of C. freundii expressing CxnA was superior to that of cells expressing the unfused proteins. Physical degradation of filter paper was also faster with the cells expressing fusion protein than the other constructs. Our results show that fusion proteins with multiple catalytic domains can improve the efficiency of cellulose degradation. Such fusion proteins could potentially substitute cloning of multiple enzymes as well as improving product yields.

  10. The 3; 21 translocation in myelodysplasia results in a fusion transcript between the AML1 gene and the gene for EAP, a highly conserved protein associated with the Epstein-Barr virus small RNA EBER 1

    SciTech Connect

    Nucifora, G.; Begy, C.R.; Rowley, J.D. ); Erickson, P.; Drabkin, H.A. )

    1993-08-15

    In the 8;21 translocation, the AML1 gene, located at chromosome band 21q22, is translocated to chromosome 8 (q22), where it is fused to the ETO gene and transcribed as a chimeric gene. AML1 is the human homolog of the recently cloned mouse gene pebp2[alpha]B, homologous to the DNA binding [alpha] subunit of the polyoma enhancer factor pebp2. AML1 is also involved in a translocation with chromosome 3 that is seen in patients with therapy-related acute myeloid leukemia and myelodysplastic syndrome and in chronic myelogenous leukemia in blast crisis. The authors have isolated a fusion cDNA clone from a t(3;21) library derived from a patient with therapy-related myelodysplastic syndrome; this clone contains sequences from AML1 and from EAP, which have now been localized to ban 3q26. EAP has previously been characterized as a highly expressed small nuclear protein of 128 residues (EBER 1) associated with Epstein-Barr virus small RNA. The fusion clone contains the DNA binding 5[prime] part of AML1 that is fused to ETO in the t(8;21) and, in addition, at least one other exon. The translocation replaces the last nine codons of AML1 with the last 96 codons of EAP. The fusion does not maintain the correct reading frame of EAP and may not lead to a functional chimeric protein. 23 refs., 6 figs.

  11. Trans-activation function of a 3 prime truncated X gene-cell fusion product from integrated hepatitis B virus DNA in chronic hepatitis tissues

    SciTech Connect

    Takada, Shinako; Koike, Katsuro )

    1990-08-01

    To investigate the expression and transactivation function of the X gene in integrated hepatitis B virus (HBV) DNA from chronic hepatitis tissues, a series of transfectants containing cloned integrated HBV DNAs was made and analyzed for X mRNA expression and trans-activation activity by using a chloramphenicol acetyltransferase assay. Most of the integrated HBV DNAs expressed X mRNA and encoded a product with trans-activation activity in spite of the loss of the 3{prime} end region of the X gene due to integration. From cDNA cloning and sequence analysis of X mRNA transcribed from native or integrated HBV DNA, the X protein was found to be translated from the X open reading frame without splicing. For integrated HBV DNA, transcription was extended to a cellular flanking DNA and an X gene-cell fusion transcript was terminated by using a cellular poly(A) signal. The amino acid sequence deduced from an X-cell fusion transcript indicated truncation of the carboxyl-terminal five amino acids, but the upstream region of seven amino acids conserved among hepadnaviruses was retained in the integrated HBV DNA, suggesting that this conserved region is essential for the transactivation function of the X protein. These findings support the following explanation for hepatocarcinogenesis by HBV DNA integration: the expression of a cellular oncogene(s) is transactivated at the time of chronic infection by the increasing amounts of the integrated HBV gene product(s), such as the X-cell fusion product.

  12. Detection of Echinoderm Microtubule Associated Protein Like 4-Anaplastic Lymphoma Kinase Fusion Genes in Non-small Cell Lung Cancer Clinical Samples by a Real-time Quantitative Reverse Transcription Polymerase Chain Reaction Method.

    PubMed

    Zhao, Jing; Zhao, Jin-Yin; Chen, Zhi-Xia; Zhong, Wei; Li, Long-Yun; Liu, Li-Cheng; Hu, Xiao-Xu; Chen, Wei-Jun; Wang, Meng-Zhao

    2016-12-20

    Objective To establish a real-time quantitative reverse transcription polymerase chain reaction assay (qRT-PCR) for the rapid, sensitive, and specific detection of echinoderm microtubule associated protein like 4-anaplastic lymphoma kinase (EML4-ALK) fusion genes in non-small cell lung cancer. Methods The specific primers for the four variants of EML4-ALK fusion genes (V1, V2, V3a, and V3b) and Taqman fluorescence probes for the detection of the target sequences were carefully designed by the Primer Premier 5.0 software. Then, using pseudovirus containing EML4-ALK fusion genes variants (V1, V2, V3a, and V3b) as the study objects, we further analyzed the lower limit, sensitivity, and specificity of this method. Finally, 50 clinical samples, including 3 ALK-fluorescence in situ hybridization (FISH) positive specimens, were collected and used to detect EML4-ALK fusion genes using this method. Results The lower limit of this method for the detection of EML4-ALK fusion genes was 10 copies/μl if no interference of background RNA existed. Regarding the method's sensitivity, the detection resolution was as high as 1% and 0.5% in the background of 500 and 5000 copies/μl wild-type ALK gene, respectively. Regarding the method's specificity, no non-specific amplification was found when it was used to detect EML4-ALK fusion genes in leukocyte and plasma RNA samples from healthy volunteers. Among the 50 clinical samples, 47 ALK-FISH negative samples were also negative. Among 3 ALK-FISH positive samples, 2 cases were detected positive using this method, but another was not detected because of the failure of RNA extraction. Conclusion The proposed qRT-PCR assay for the detection of EML4-ALK fusion genes is rapid, simple, sensitive, and specific, which is deserved to be validated and widely used in clinical settings.

  13. Osteopathia striata with cranial sclerosis (OSCS): review of the literature and case report demonstrating challenges of spinal fusion after trauma.

    PubMed

    Katsevman, Gennadiy A; Turner, Ryan C; Lucke-Wold, Brandon P; Sedney, Cara L; Bhatia, Sanjay

    2016-06-01

    Osteopathia striata with cranial sclerosis (OSCS) is a rare but well-described pathology characterized by abnormalities in bone deposition in the axial and cranial skeleton as well as other abnormalities and associated deficits. These skeletal abnormalities can lead to significant intra-operative challenges for the surgeon and influence outcomes for the patient. In this report, we present a case of a patient with OSCS who was involved in a traumatic motor vehicle crash and underwent posterior cervico-thoracic fusion for a T4 chance fracture. Bony abnormalities in the cervico-thoracic spine presented a significant operative challenge due to alterations in bony anatomy and bone architecture. This case serves as an example of the challenges that the spine surgeon faces when dealing with OSCS, and highlights the differences between OSCS and commoner skeletal hyperplasias such as osteopetrosis.

  14. Fusion Plasma Theory Grant: Task 3, Auxiliary Radiofrequency Heating of Tokamaks. Annual report, November 16, 1992--November 15, 1993

    SciTech Connect

    Scharer, J.E.

    1993-06-01

    The research performed under this grant during the past year has been concentrated on the following several key tokamak ICRF (Ion Cyclotron Range of Frequencies) coupling, heating and current drive issues. We have made progress in developing a ``3-D`` cavity backed antenna array code to examine ICRF coupling to general plasma edge profiles. The effects of the finite antenna length and feeders as well as Faraday shield blade angle are being examined. We are also developing an analysis to examine large k{perpendicular}{rho} gyroradius interaction between alpha or beam particles and ICRF waves. This topic has important applications in the areas of ICRF heating for deuterium-tritium fusion plasmas, TAE modes, ash removal and minority ion current drive. Research progress, publications, and conference and workshop presentations are summarized in this report.

  15. Fusion Plasma Theory: Task 3, Auxiliary radiofrequency heating of tokamaks. Annual report, November 16, 1991--November 15, 1992

    SciTech Connect

    Scharer, J.E.

    1992-12-31

    The research performed under this grant during the past year has been concentrated on the following several key tokamak ICRF (Ion Cyclotron Range of Frequencies) coupling, heating and current drive issues: Efficient coupling during the L- to H- mode transition by analysis and computer simulation of ICRF antennas; analysis of ICRF cavity-backed coil antenna coupling to plasma edge profiles including fast and ion Bernstein wave coupling for heating and current drive; benchmarking the codes to compare with current JET, D-IIID and ASDEX experimental results and predictions for advanced tokamaks such as BPX and SSAT (Steady-State Advanced Tokamak); ICRF full-wave field solutions, power conservation, heating analyses and minority ion current drive; and the effects of fusion alpha particle or ion tail populations on the ICRF absorption. Research progress, publications, and conference and workshop presentations are summarized in this report.

  16. The TMPRSS2-ERG gene fusion blocks XRCC4-mediated non-homologous end-joining repair and radiosensitizes prostate cancer cells to PARP inhibition

    PubMed Central

    Chatterjee, Payel; Choudhary, Gaurav S.; Alswillah, Turkeyah; Xiong, Xiahui; Heston, Warren D.; Magi-Galuzzi, Cristina; Zhang, Junran; Klein, Eric A.; Almasan, Alexandru

    2015-01-01

    Exposure to genotoxic agents, such as ionizing radiation (IR) produces DNA damage leading to DNA double-strand breaks (DSBs); IR toxicity is augmented when the DNA repair is impaired. We reported that radiosensitization by a PARP inhibitor (PARPi) was highly prominent in prostate cancer (PCa) cells expressing the TMPRSS2-ERG gene fusion protein. Here, we show that TMPRSS2-ERG blocks non-homologous end-joining (NHEJ) DNA repair by inhibiting DNA-PKcs. VCaP cells, which harbor TMPRSS2-ERG and PC3 cells that stably express it displayed γH2AX and 53BP1 foci constitutively, indicating persistent DNA damage that was absent if TMPRSS2-ERG was depleted by siRNA in VCaP cells. The extent of DNA damage was enhanced and associated with TMPRSS2-ERG’s ability to inhibit DNA-PKcs function, as indicated by its own phosphorylation (Thr2609, Ser2056) and that of its substrate, Ser1778-53BP1. DNA-PKcs deficiency caused by TMPRSS2-ERG destabilized critical NHEJ components on chromatin. Thus, XRCC4 was not recruited to chromatin, with retention of other NHEJ core factors being reduced. DNA-PKcs autophosphorylation was restored to the level of parental cells when TMPRSS2-ERG was depleted by siRNA. Following IR, TMPRSS2-ERG-expressing PC3 cells had elevated Rad51 foci and homologous recombination (HR) activity, indicating that HR compensated for defective NHEJ in these cells, hence addressing why TMPRSS2-ERG alone did not lead to radiosensitization. However, the presence of TMPRSS2-ERG, by inhibiting NHEJ DNA repair, enhanced PARPi-mediated radiosensitization. IR in combination with PARPi resulted in enhanced DNA damage in TMPRSS2-ERG-expressing cells. Thus, by inhibiting NHEJ, TMPRSS2-ERG provides a synthetic lethal interaction with PARPi in PCa patients expressing TMPRSS2-ERG. PMID:26026052

  17. Spinal Fusion

    MedlinePlus

    ... concept of fusion is similar to that of welding in industry. Spinal fusion surgery, however, does not ... bone taken from the patient has a long history of use and results in predictable healing. Autograft ...

  18. Accelerators for heavy ion fusion

    SciTech Connect

    Bangerter, R.O.

    1985-10-01

    Large fusion devices will almost certainly produce net energy. However, a successful commercial fusion energy system must also satisfy important engineering and economic constraints. Inertial confinement fusion power plants driven by multi-stage, heavy-ion accelerators appear capable of meeting these constraints. The reasons behind this promising outlook for heavy-ion fusion are given in this report. This report is based on the transcript of a talk presented at the Symposium on Lasers and Particle Beams for Fusion and Strategic Defense at the University of Rochester on April 17-19, 1985.

  19. Inertial Confinement Fusion Target Component Fabrication and Technology Development Support. Annual report, January 1, 1991--September 30, 1992

    SciTech Connect

    Steinman, D.

    1993-03-01

    On December 31, 1990, the US Department of Energy entered into a contract with General Atomics (GA) to be the Inertial Confinement Fusion (ICF) Target Component Fabrication and Technology Development Support contractor. This report documents the technical activities of the period January 1, 1991 through September 30, 1992. During this period, GA was assigned 15 tasks in support of the Inertial Confinement Fusion program and its laboratories. These tasks included Facilities Activation, Staff Development, and Capabilities Validation to establish facilities and equipment, and demonstrate capability to perform ICF target fabrication research, development and production activities. The capabilities developed and demonstrated are those needed for fabrication and precise characterization of polymer shells and polymer coatings. We made progress toward production capability for glass shells, barrier layer coatings, and gas idling of shells. We fabricated over 1000 beam diagnostic foil targets for Sandia National Laboratory Albuquerque and provided full-time on-site engineering support for target fabrication and characterization. We initiated development of methods to fabricate polymer shells by a controlled mass microencapsulation technique, and performed chemical syntheses of several chlorine- and silicon-doped polymer materials for the University of Rochester`s Laboratory for Laser Energetics (UR/LLE). We performed the conceptual design of a cryogenic target handling system for UR/LLE that will fill, transport, layer, and characterize targets filled with cryogenic deuterium or deuterium-tritium fuel, and insert these cryogenic targets into the OMEGA-Upgrade target chamber for laser implosion experiments. This report summarizes and documents the technical progress made on these tasks.

  20. Inertial confinement fusion target component fabrication and technology development support: Annual report, October 1, 1995--September 30, 1996

    SciTech Connect

    Hoppe, M.

    1997-02-01

    On December 30, 1990, the U.S. Department of Energy entered into a contract with General Atomics (GA) to be the Inertial Confinement Fusion (ICF) Target Component Fabrication and Technology Development Support contractor. In September 1995 this contract ended and a second contract was issued for us to continue this ICF target support work. This report documents the technical activities of the period October 1, 1995 through September 30, 1996. During this period, GA and our partners WJ Schafer Associates (WJSA) and Soane Technologies, Inc. (STI) were assigned 14 formal tasks in support of the Inertial Confinement Fusion program and its five laboratories. A portion of the effort on these tasks included providing direct {open_quotes}Onsite Support{close_quotes} at Lawrence Livermore National Laboratory (LLNL), Los Alamos National Laboratory (LANL), and Sandia National Laboratory Albuquerque (SNLA). We fabricated and delivered over 800 gold-plated hohlraum mandrels to LLNL, LANL and SNLA. We produced nearly 1,200 glass and plastic target capsules for LLNL, LANL, SNLA and University of Rochester/Laboratory for Laser Energetics (UR/LLE). We also delivered over 100 flat foil targets for Naval Research Lab (NRL) and SNLA in FY96. This report describes these target fabrication activities and the target fabrication and characterization development activities that made the deliveries possible. The ICF program is anticipating experiments at the OMEGA laser and the National Ignition Facility (NIF) which will require capsules containing cryogenic layered D{sub 2} or deuterium-tritium (DT) fuel. We are part of the National Cryogenic Target Program to create and demonstrate viable ways to generate and characterize cryogenic layers. Substantial progress has been made on ways to both create and characterize viable layers. During FY96, significant progress was made in the design of the OMEGA Cryogenic Target System that will field cryogenic targets on OMEGA.

  1. Fusion programs in applied plasma physics. Final report, fiscal years 1989--1991

    SciTech Connect

    Not Available

    1992-02-01

    The objectives of the theoretical science program are: To support the interpretation of present experiments and predict the outcome of future planned experiments; to improve on existing models and codes and validate against experimental results; and to conduct theoretical physics development of advanced concepts with applications for DIII-D and future devices. Major accomplishments in FY91 include the corroboration between theory and experiment on MHD behavior in the second stable regime of operation on DIII-D, and the frequency and mode structure of toroidal Alfven eigenmodes in high beta, shaped plasmas. We have made significant advances in the development of the gyro-Landau fluid approach to turbulence simulation which more accurately models kinetic drive and damping mechanisms. Several theoretical models to explain the bifurcation phenomenon in L- to H-mode transition were proposed providing the theoretical basis for future experimental verification. The capabilities of new rf codes have been upgraded in response to the expanding needs of the rf experiments. Codes are being employed to plan for a fully non-inductive current drive experiment in a high beta, enhanced confinement regime. GA`s experimental effort in Applied Physics encompasses two advanced diagnostics essential for the operation of future fusion experiments: Alpha particle diagnostic, and current and density profile diagnostics. This paper discusses research in all these topics.

  2. The Fusion Science Research Plan for the Major U.S. Tokamaks. Advisory report

    SciTech Connect

    none,

    1996-05-31

    In summary, the community has developed a research plan for the major tokamak facilities that will produce impressive scientific benefits over the next two years. The plan is well aligned with the new mission and goals of the restructured fusion energy sciences program recommended by FEAC. Budget increases for all three facilities will allow their programs to move forward in FY 1997, increasing their rate of scientific progress. With a shutdown deadline now established, the TFTR will forego all but a few critical upgrades and maximize operation to achieve a set of high-priority scientific objectives with deuterium-tritium plasmas. The DIII-D and Alcator C-Mod facilities will still fall well short of full utilization. Increasing the run time in – vii – DIII-D is recommended to increase the scientific output using its existing capabilities, even if scheduled upgrades must be further delayed. An increase in the Alcator C-Mod budget is recommended, at the expense of equal and modest reductions (~1%) in the other two facilities if necessary, to develop its capabilities for the long-term and increase its near-term scientific output.

  3. Fusion welding of advanced borated stainless steels. Final report: CRADA No. CR1042

    SciTech Connect

    Robino, C.V.; Cieslak, M.J.

    1994-02-01

    This work addressed two major areas concerning joining of advanced borated stainless steels. These areas included the development of a understanding of the physical metallurgy of borated stainless steels and the development of welding processes and post-weld heat treatments for these alloys. Differential thermal analysis experiments were conducted on ten heats of borated stainless steel to determine the transformation temperatures and melting behavior of the alloys. On-heating solidus temperatures were measured for all of the alloys and were used to define the temperatures associated with the fusion line during welding. Isothermal heat treatments designed to evaluate the effects of elevated temperature exposures on the toughness of the borated grades were conducted. These tests were used to determine if significant changes in the microstructure or mechanical properties of weld heat-affected zones (HAZ) occur. Specifically, the tests addressed the solid-state region of the HAZ. The test matrix included a variety of alloy compositions and thermal exposures at temperatures near the on-heating solidus (as determined by the DTA experiments). Welding experiments designed to assess the mechanical properties and microstructure of gas-tungsten arc and electron beam welds were conducted.

  4. Multimodality Imaging of Gene Transfer with a Receptor-Based Reporter Gene

    PubMed Central

    Chen, Ron; Parry, Jesse J.; Akers, Walter J.; Berezin, Mikhail Y.; El Naqa, Issam M.; Achilefu, Samuel; Edwards, W. Barry; Rogers, Buck E.

    2010-01-01

    Gene therapy trials have traditionally used tumor and tissue biopsies for assessing the efficacy of gene transfer. Non-invasive imaging techniques offer a distinct advantage over tissue biopsies in that the magnitude and duration of gene transfer can be monitored repeatedly. Human somatostatin receptor subtype 2 (SSTR2) has been used for the nuclear imaging of gene transfer. To extend this concept, we have developed a somatostatin receptor–enhanced green fluorescent protein fusion construct (SSTR2-EGFP) for nuclear and fluorescent multimodality imaging. Methods An adenovirus containing SSTR2-EGFP (AdSSTR2-EGFP) was constructed and evaluated in vitro and in vivo. SCC-9 human squamous cell carcinoma cells were infected with AdEGFP, AdSSTR2, or AdSSTR2-EGFP for in vitro evaluation by saturation binding, internalization, and fluorescence spectroscopy assays. In vivo biodistribution and nano-SPECT imaging studies were conducted with mice bearing SCC-9 tumor xenografts directly injected with AdSSTR2-EGFP or AdSSTR2 to determine the tumor localization of 111In-diethylenetriaminepentaacetic acid (DTPA)-Tyr3-octreotate. Fluorescence imaging was conducted in vivo with mice receiving intratumoral injections of AdSSTR2, AdSSTR2-EGFP, or AdEGFP as well as ex vivo with tissues extracted from mice. Results The similarity between AdSSTR2-EGFP and wild-type AdSSTR2 was demonstrated in vitro by the saturation binding and internalization assays, and the fluorescence emission spectra of cells infected with AdSSTR2-EGFP was almost identical to the spectra of cells infected with wild-type AdEGFP. Biodistribution studies demonstrated that the tumor uptake of 111In-DTPA-Tyr3-octreotate was not significantly different (P > 0.05) when tumors (n = 5) were injected with AdSSTR2 or AdSSTR2-EGFP but was significantly greater than the uptake in control tumors. Fluorescence was observed in tumors injected with AdSSTR2-EGFP and AdEGFP in vivo and ex vivo but not in tumors injected with AdSSTR2

  5. The β-actin gene promoter of rohu carp (Labeo rohita) drives reporter gene expressions in transgenic rohu and various cell lines, including spermatogonial stem cells.

    PubMed

    Barman, Hirak Kumar; Mohanta, Ramya; Patra, Swagat Kumar; Chakrapani, Vemulawada; Panda, Rudra Prasanna; Nayak, Swapnarani; Jena, Sasmita; Jayasankar, Pallipuram; Nandanpawar, Priyanka

    2015-06-01

    We previously characterized the β-actin gene promoter of Indian domesticated rohu carp (Labeo rohita) and made a reporter construct via fusion to green fluorescence protein (GFP) cDNA. In this study, the same construct was used to breed transgenic rohu fish. About 20% of the transgenic offspring showed ubiquitous expression of the reporter GFP gene. In a few of the transgenic fish, we documented massive epithelial and/or muscular expression with visible green color under normal light. The expression of GFP mRNA was higher in the muscle tissue of transgenic fish than in that of non-transgenic fish. A highly efficient nucleofection protocol was optimized to transfect proliferating spermatogonial cells of rohu using this reporter construct. The β-actin promoter also drove expressions in HEK293 (derived from human embryonic kidney cells), K562 (human leukemic cells) and SF21 (insect ovarian cells) lines. These findings imply conserved regulatory mechanisms of β-actin gene expression across eukaryotes. Furthermore, the isolated β-actin promoter with consensus regulatory elements has the potential to be used in generating transgenic carp with genes of interest and in basic biology research.

  6. Development of Plant Gene Vectors for Tissue-Specific Expression Using GFP as a Reporter Gene

    NASA Technical Reports Server (NTRS)

    Jackson, Jacquelyn; Egnin, Marceline; Xue, Qi-Han; Prakash, C. S.

    1997-01-01

    Reporter genes are widely employed in plant molecular biology research to analyze gene expression and to identify promoters. Gus (UidA) is currently the most popular reporter gene but its detection requires a destructive assay. The use of jellyfish green fluorescent protein (GFP) gene from Aequorea Victoria holds promise for noninvasive detection of in vivo gene expression. To study how various plant promoters are expressed in sweet potato (Ipomoea batatas), we are transcriptionally fusing the intron-modified (mGFP) or synthetic (modified for codon-usage) GFP coding regions to these promoters: double cauliflower mosaic virus 35S (CaMV 35S) with AMV translational enhancer, ubiquitin7-intron-ubiquitin coding region (ubi7-intron-UQ) and sporaminA. A few of these vectors have been constructed and introduced into E. coli DH5a and Agrobacterium tumefaciens EHA105. Transient expression studies are underway using protoplast-electroporation and particle bombardment of leaf tissues.

  7. Renal Cell Carcinoma Associated with Xp11.2 Translocation/TFE3 Gene Fusions: Clinical Features, Treatments and Prognosis

    PubMed Central

    Gan, Weidong; Xiong, Lei; Miao, Baolei; Chen, Xiancheng; Guo, Hongqian; Li, Dongmei

    2016-01-01

    To investigate the clinical characteristics, treatments and prognosis of renal cell carcinoma associated with Xp11.2 translocation/TFE3 gene fusions (Xp11.2 tRCC), the epidemiological features and treatment results of 34 cases of Xp11.2 tRCC, which were diagnosed by immunohistochemistry staining of TFE3 and fluorescence in situ hybridization at our center, were retrospectively reviewed. The 34 patients included 21 females and 13 males aged 3 to 64 years (median age: 27 years). Four patients were children or adolescents (<18 years of age), and 26 patients were young or middle-aged adults (18–45 years). Radical nephrectomy was performed on 25 patients. Laparoscopic nephron-sparing surgery was performed on 9 patients who presented with an isolated mass with a small diameter (<7 cm) and well-defined boundary on computed tomography imaging. Postoperative staging showed that 25 cases (73.53%) were at stage I/II, while 9 cases (26.47%) were at stage III/IV. All stage I/II patients received a favorable prognosis with a three-year overall survival rate of 100%, including the patients who underwent laparoscopic nephron-sparing surgery. With the exception of 2 children, the other 7 stage III/IV patients died or developed recurrence with a median follow-up of 29 months. On univariate analysis, maximum diameter, adjuvant treatment, TNM stage, lymph node metastasis, inferior vena cava tumor thrombosis and tumor boundary were identified as statistically significant factors impacting survival (P<0.05). Multivariate analysis indicated that TNM stage and inferior vena cava tumor thrombosis were independent prognostic factors (P<0.05). In conclusion, Xp11.2 tRCC is a rare subtype of renal cell carcinoma that mainly occurs in young females. Nephron-sparing surgery was confirmed effective preliminarily in the treatment of small Xp11.2 tRCCs with clear rims. Advanced TNM stage and inferior vena cava tumor thrombosis were associated with poor prognosis. PMID:27893792

  8. Developmental and environmental regulation of a phenylalanine ammonia-lyase-beta-glucuronidase gene fusion in transgenic tobacco plants.

    PubMed Central

    Liang, X W; Dron, M; Schmid, J; Dixon, R A; Lamb, C J

    1989-01-01

    A 1.1-kilobase promoter fragment of the bean (Phaseolus vulgaris L.) phenylalanine ammonia-lyase (EC 4.3.1.5) gene PAL2 was translationally fused to the beta-glucuronidase reporter gene and transferred to tobacco by Agrobacterium tumefaciens-mediated leaf disk transformation. The distribution of beta-glucuronidase activity in these transgenic plants is very similar to that of endogenous PAL2 transcripts in bean, with very high levels in petals; marked accumulation in anthers, stigmas, roots, and shoots; and low levels in sepals, ovaries, and leaves. Histochemical analysis of the spatial pattern of beta-glucuronidase activity showed that the PAL2 promoter is highly active in the shoot apical meristem, the zone of cell proliferation immediately adjacent to the root apical meristem, and in the early stages of vascular development at the inception of xylem differentiation. Wounding and light evoke specific changes in the spatial pattern of beta-glucuronidase activity in stems, including induction in the epidermis. These data indicate that the PAL2 promoter transduces a complex set of developmental and environmental cues into an integrated spatial and temporal program of gene expression to regulate the synthesis of a diverse array of phenylpropanoid natural products. Images PMID:2594769

  9. Developing a Novel Gene-Delivery Vector System Using the Recombinant Fusion Protein of Pseudomonas Exotoxin A and Hyperthermophilic Archaeal Histone HPhA.

    PubMed

    Deng, Xin; Zhang, Guoli; Zhang, Ling; Feng, Yan; Li, Zehong; Wu, GuangMou; Yue, Yuhuan; Li, Gensong; Cao, Yu; Zhu, Ping

    2015-01-01

    Non-viral gene delivery system with many advantages has a great potential for the future of gene therapy. One inherent obstacle of such approach is the uptake by endocytosis into vesicular compartments. Receptor-mediated gene delivery method holds promise to overcome this obstacle. In this study, we developed a receptor-mediated gene delivery system based on a combination of the Pseudomonas exotoxin A (PE), which has a receptor binding and membrane translocation domain, and the hyperthermophilic archaeal histone (HPhA), which has the DNA binding ability. First, we constructed and expressed the rPE-HPhA fusion protein. We then examined the cytotoxicity and the DNA binding ability of rPE-HPhA. We further assessed the efficiency of transfection of the pEGF-C1 plasmid DNA to CHO cells by the rPE-HPhA system, in comparison to the cationic liposome method. The results showed that the transfection efficiency of rPE-HPhA was higher than that of cationic liposomes. In addition, the rPE-HPhA gene delivery system is non-specific to DNA sequence, topology or targeted cell type. Thus, the rPE-HPhA system can be used for delivering genes of interest into mammalian cells and has great potential to be applied for gene therapy.

  10. Checkpoint genes and Exo1 regulate nearby inverted repeat fusions that form dicentric chromosomes in Saccharomyces cerevisiae.

    PubMed

    Kaochar, Salma; Shanks, Lisa; Weinert, Ted

    2010-12-14

    Genomic rearrangements are common, occur by largely unknown mechanisms, and can lead to human diseases. We previously demonstrated that some genome rearrangements occur in budding yeast through the fusion of two DNA sequences that contain limited sequence homology, lie in inverted orientation, and are within 5 kb of one another. This inverted repeat fusion reaction forms dicentric chromosomes, which are well-known intermediates to additional rearrangements. We have previously provided evidence indicating that an error of stalled or disrupted DNA replication forks can cause inverted repeat fusion. Here we analyze how checkpoint protein regulatory pathways known to stabilize stalled forks affect this form of instability. We find that two checkpoint pathways suppress inverted repeat fusion, and that their activities are distinguishable by their interactions with exonuclease 1 (Exo1). The checkpoint kinase Rad53 (Chk2) and recombination protein complex MRX(MRN) inhibit Exo1 in one pathway, whereas in a second pathway the ATR-like kinases Mec1 and Tel1, adaptor protein Rad9, and effector kinases Chk1 and Dun1 act independently of Exo1 to prevent inverted repeat fusion. We provide a model that indicates how in Rad53 or MRX mutants, an inappropriately active Exo1 may facilitate faulty template switching between nearby inverted repeats to form dicentric chromosomes. We further investigate the role of Rad53, using hypomorphic alleles of Rad53 and null mutations in Rad9 and Mrc1, and provide evidence that only local, as opposed to global, activity of Rad53 is sufficient to prevent inverted repeat fusion.

  11. Accelerator and Fusion Research Division annual report, October 1980-September 1981. Fiscal year, 1981

    SciTech Connect

    Johnson, R.K.; Thomson, H.A.

    1982-04-01

    Major accomplishments during fiscal year 1981 are presented. During the Laboratory's 50th anniversary celebrations, AFRD and the Nuclear Science Division formally dedicated the new (third) SuperHILAC injector that adds ions as heavy as uranium to the ion repertoire at LBL's national accelerator facilities. The Bevalac's new multiparticle detectors (the Heavy Ion Spectrometer System and the GSI-LBL Plastic Ball/Plastic Wall) were completed in time to take data before the mid-year shutdown to install the new vacuum liner, which passed a milestone in-place test with flying colors in September. The Bevalac biomedical program continued patient treatment with neon beams aimed at establishing a complete data base for a dedicated biomedical accelerator, the design of which NCI funded during the year. Our program to develop alternative Isabelle superconducting dipole magnets, which DOE initiated in FY80, proved the worth of a new magnet construction technique and set a world record - 7.6 Tesla at 1.8 K - with a model magnet in our upgraded test facility. Final test results at LBL were obtained by the Magnetic Fusion Energy Group on the powerful neutral beam injectors developed for Princeton's TFTR. The devices exceeded the original design requirements, thereby completing the six-year, multi-million-dollar NBSTF effort. The group also demonstrated the feasibility of efficient negative-ion-based neutral beam plasma heating for the future by generating 1 A of negative ions at 34 kV for 7 seconds using a newly developed source. Collaborations with other research centers continued, including: (1) the design of LBL/Exxon-dedicated beam lines for the Stanford Synchrotron Radiation Laboratory; (2) beam cooling tests at Fermilab and the design of a beam cooling system for a proton-antiproton facility there; and (3) the development of a high-current betatron for possible application to a free electron laser.

  12. A quasi-optical electron cyclotron maser for fusion reactor heating. Final report

    SciTech Connect

    Morse, E.C.

    1990-12-31

    High power microwave and millimeter sources, such as the quasi-optical electron cyclotron maser (QOECM) are important in fusion research as well as in high-energy physics and in other applications. The interaction between the electromagnetic modes of a Fabry-Perot resonator and an electron beam gyrating through a magnetic field has been studied for both the cases of beams parallel and perpendicular to the resonator. The parallel case was theoretically first studied by Kurin for forward and backward wave interaction, and experimentally by Komlev and Kurin. Kreischer and Temkin reviewed the general case of the linear small signal interaction parallel and perpendicular to the resonator. Sprangle, et al discussed the perpendicular case in a self-consistent linear and nonlinear theoretical study using the Gaussian transverse profile of an open resonator with a single longitudinal mode. Experimental verification of the devices operation was first mentioned in work at the Naval Research Laboratory. Theoretical studies using a time-dependent analysis of a large number of longitudinal modes with similar transverse mode profiles have demonstrated that single longitudinal-mode operation can be achieved at equilibrium and that performance can be enhanced by prebunching the electron beam and tapering the magnetic field. The use of output coupling apertures in the mirrors has been studied theoretically in relation to the structure of the modes for both confocal and nonconfocal resonators by Permnoud; use of an open resonator with stepped mirrors has been studied in order to choose a particular longitudinal mode. Studies at the Naval Research Laboratory mirror used configurations that diffraction couple the energy from around the mirror edges, so that the transverse profile inside the resonator can be selective to the fundamental mode.

  13. Ion Fast Ignition-Establishing a Scientific Basis for Inertial Fusion Energy --- Final Report

    SciTech Connect

    Stephens, Richard Burnite; Foord, Mark N.; Wei, Mingsheng; Beg, Farhat N.; Schumacher, Douglass W.

    2013-10-31

    The Fast Ignition (FI) Concept for Inertial Confinement Fusion (ICF) has the potential to provide a significant advance in the technical attractiveness of Inertial Fusion Energy reactors. FI differs from conventional ?central hot spot? (CHS) target ignition by decoupling compression from heating: using a laser (or heavy ion beam or Z pinch) drive pulse (10?s of nanoseconds) to create a dense fuel and a second, much shorter (~10 picoseconds) high intensity pulse to ignite a small volume within the dense fuel. The compressed fuel is opaque to laser light. The ignition laser energy must be converted to a jet of energetic charged particles to deposit energy in the dense fuel. The original concept called for a spray of laser-generated hot electrons to deliver the energy; lack of ability to focus the electrons put great weight on minimizing the electron path. An alternative concept, proton-ignited FI, used those electrons as intermediaries to create a jet of protons that could be focused to the ignition spot from a more convenient distance. Our program focused on the generation and directing of the proton jet, and its transport toward the fuel, none of which were well understood at the onset of our program. We have developed new experimental platforms, diagnostic packages, computer modeling analyses, and taken advantage of the increasing energy available at laser facilities to create a self-consistent understanding of the fundamental physics underlying these issues. Our strategy was to examine the new physics emerging as we added the complexity necessary to use proton beams in an inertial fusion energy (IFE) application. From the starting point of a proton beam accelerated from a flat, isolated foil, we 1) curved it to focus the beam, 2) attached the foil to a superstructure, 3) added a side sheath to protect it from the surrounding plasma, and finally 4) studied the proton beam behavior as it passed through a protective end cap into plasma. We built up, as we proceeded

  14. Fusion of platelet-derived growth receptor {beta} to a novel ets-like gene, tel, in chronic myelomonocytic leukemia with t(5;12) chromosomal translocation

    SciTech Connect

    Golub, T.; Barker, G.; Gilliland, D.G.

    1994-09-01

    Chronic myelomonocytic leukemia (CMML) is a myelodysplastic syndrome characterized by abnormal clonal myeloid proliferation, and by progression to acute myelogenous leukemia (AML). A recently recognized subgroup of CMML has a t(5;12) (q33;p13) balanced translocation. Fluorescence in situ hybridization (FISH) localized the translocation breakpoint near the CSF1 receptor (CSF1R) locus on chromosome 5q. Pulsed-field gel electrophoresis confirmed rearrangements near CSF1R, but involvement of CSF1R itself was excluded. Southern blotting showed a rearrangement within the closely linked PDGF receptor {beta} (PDGFR{beta}) gene. Ribonuclease protection assays localized the translocation breakpoint to nucleotide 1766 in PDGFR{beta} RNA. Anchored PCR was used to identify the chromosome 12 fusion partner, a novel ets-like protein, tel. Tel contains a highly conserved carboxy terminal ets-like DNA-binding domain, and an amino terminal domain with a predicted helix-loop-helix (HLH) secondary structure. The consequence of the t(5;12) translocation is fusion of the tel HLH domain to the PDGFR{beta} transmembrane and tyrosine kinase domains. The tel HLH domain may contribute a dimerization motif which serves to constitutively activate PDGFR{beta} tyrosine kinase activity. The tel-PDGFR{beta} fusion demonstrates the oncogenic potential of PDGFR{beta}, and may provide a paradigm for early events in the pathogenesis of AML.

  15. Establishment of an Institute for Fusion Studies. Technical progress report, 1 November 1993--31 October 1994

    SciTech Connect

    Hazeltine, R.D.

    1994-07-01

    The Institute for Fusion Studies is a national center for theoretical fusion plasma physics research. Its purposes are: (1) to conduct research on theoretical questions concerning the achievement of controlled fusion energy by means of magnetic confinement--including both fundamental problems of long-range significance, as well as shorter-term issues; (2) to serve as a national and international center for information exchange by hosting exchange visits, conferences, and workshops; (3) and to train students and postdoctoral research personnel for the fusion energy program and plasma physics research areas. The theoretical research results obtained by the Institute contribute to the progress of nuclear fusion research, whose goal is the development of fusion power as a basic energy source. Close collaborative relationships have been developed with other university and national laboratory fusion groups, both in the US and abroad. In addition to its primary focus on mainstream fusion physics, the Institute is also involved with research in fusion-sidestream fields, such as advanced computing techniques, nonlinear dynamics, space plasmas and astrophysics, statistical mechanics, fluid dynamics, and accelerator physics. Important research discoveries are briefly described.

  16. Reporter gene assays for algal-derived toxins.

    PubMed

    Fairey, E R; Ramsdell, J S

    1999-01-01

    We have modified the cell-based directed cytotoxicity assay for sodium channel and calcium channel active phycotoxins using a c-fos-luciferase reporter gene construct. In this report we describe the conceptual basis to the development of reporter gene assays for algal-derived toxins and summarize both published and unpublished data using this method. N2A mouse neuroblastoma cells, which express voltage-dependent sodium channels, were stably transfected with the reporter gene c-fos-luc, which contains the firefly luciferase gene under the transcriptional regulation of the human c-fos response element. The characteristics of the N2A reporter gene assay were determined by dose response with brevetoxin and ciguatoxin. Brevetoxin-1 and ciguatoxin-1 induced c-fos-luc with an EC50 of 4.6 and 3.0 ng ml(-1), respectively. Saxitoxin caused a concentration-dependent inhibition of brevetoxin-1 induction of c-fos-luc with an EC50 of 3.5 ng ml(-1). GH4C1 rat pituitary cells, which lack voltage-dependent sodium channels but express voltage-dependent calcium channels, were also stably transfected with the c-fos-luc. GH4C1 cells expressing c-fos-luciferase were responsive to maitotoxin (1 ng ml(-1)) and a putative toxin produced by Pfiesteria piscicida. Although reporter gene assays are not designed to replace existing detection methods used to measure toxin activity in seafood, they do provide a valuable means to screen algal cultures for toxin activity, to conduct assay-guided fractionation and to characterize pharmacologic properties of algal toxins.

  17. Radioscapholunate Fusions

    PubMed Central

    McGuire, Duncan Thomas; Bain, Gregory Ian

    2012-01-01

    Radiocarpal fusions are performed for a variety of indications, most commonly for debilitating painful arthritis. The goal of a wrist fusion is to fuse the painful, diseased joints and to preserve motion through the healthy joints. Depending on the extent of the disease process, radiocarpal fusions may take the form of radiolunate, radioscapholunate, or total wrist fusions. Surgical techniques and instrumentation have advanced over the last few decades, and consequently the functional outcomes have improved and complications decreased. Techniques for partial carpal fusions have improved and now include distal scaphoid and triquetrum excision, which improves range of motion and fusion rates. In this article we discuss the various surgical techniques and fixation methods available and review the corresponding evidence in the literature. The authors' preferred surgical technique of radioscapholunate fusion with distal scaphoid and triquetrum excision is outlined. New implants and new concepts are also discussed. PMID:24179717

  18. [Fusion of the labia minora as a cau