Sample records for packaging cell lines

  1. Development and characterization of a packaging cell line for pseudo-infectious yellow fever virus particle generation.

    PubMed

    Queiroz, Sabrina Ribeiro de Almeida; Silva Júnior, José Valter Joaquim; Silva, Andréa Nazaré Monteiro Rangel da; Carvalho, Amanda Gomes de Oliveira; Santos, Jefferson José da Silva; Gil, Laura Helena Vega Gonzales

    2018-01-01

    Pseudo-infectious yellow fever viral particles (YFV-PIVs) have been used to study vaccines and viral packaging. Here, we report the development of a packaging cell line, which expresses the YFV prM/E proteins. HEK293 cells were transfected with YFV prM/E and C (84 nt) genes to generate HEK293-YFV-PrM/E-opt. The cells were evaluated for their ability to express the heterologous proteins and to package the replicon repYFV-17D-LucIRES, generating YFV-PIVs. The expression of prM/E proteins was confirmed, and the cell line trans-packaged the replicon for recovery of a reporter for the YFV-PIVs. HEK293-YFV-prM/E-opt trans-packaging capacity demonstrates its possible biotechnology application.

  2. APOBEC3-mediated hypermutation of retroviral vectors produced from some retrovirus packaging cell lines.

    PubMed

    Miller, A D; Metzger, M J

    2011-05-01

    APOBEC3 proteins are packaged into retrovirus virions and can hypermutate retroviruses during reverse transcription. We found that HT-1080 human fibrosarcoma cells hypermutate retroviruses, and that the HT-1080 cell-derived FLYA13 retrovirus packaging cells also hypermutate a retrovirus vector produced using these cells. We found no hypermutation of the same vector produced by the mouse cell-derived packaging line PT67 or by human 293 cells transfected with the vector and retrovirus packaging plasmids. We expect that avoidance of vector hypermutation will be particularly important for vectors used in gene therapy, wherein mutant proteins might stimulate deleterious immune responses.

  3. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bradbury, Andrew M.

    The invention relates to a novel phagemid display system for packaging phagemid DNA into phagemid particles which completely avoids the use of helper phage. The system of the invention incorporates the use of bacterial packaging cell lines which have been transformed with helper plasmids containing all required phage proteins but not the packaging signals. The absence of packaging signals in these helper plasmids prevents their DNA from being packaged in the bacterial cell, which provides a number of significant advantages over the use of both standard and modified helper phage. Packaged phagemids expressing a protein or peptide of interest, inmore » fusion with a phage coat protein such as g3p, are generated simply by transfecting phagemid into the packaging cell line.« less

  4. West Nile virus infectious replicon particles generated using a packaging-restricted cell line is a safe reporter system.

    PubMed

    Li, Wei; Ma, Le; Guo, Li-Ping; Wang, Xiao-Lei; Zhang, Jing-Wei; Bu, Zhi-Gao; Hua, Rong-Hong

    2017-06-12

    West Nile virus (WNV) is a neurotropic pathogen which causes zoonotic disease in humans. Recently, there have been an increasing number of infected cases and there are no clinically approved vaccines or effective drugs to treat WNV infections in humans. The purpose of this study was to facilitate vaccine and antiviral drug discovery by developing a packaging cell line-restricted WNV infectious replicon particle system. We constructed a DNA-based WNV replicon lacking the C-prM-E coding region and replaced it with a GFP coding sequence. To produce WNV replicon particles, cell lines stably-expressing prM-E and C-prM-E were constructed. When the WNV replicon plasmid was co-transfected with a WNV C-expressing plasmid into the prM-E-expressing cell line or directly transfected the C-prM-E expressing cell line, the replicon particle was able to replicate, form green fluorescence foci, and exhibit cytopathic plaques similar to that induced by the wild type virus. The infectious capacity of the replicon particles was restricted to the packaging cell line as the replicons demonstrated only one round of infection in other permissive cells. Thus, this system provides a safe and convenient reporter WNV manipulating tool which can be used to study WNV viral invasion mechanisms, neutralizing antibodies and antiviral efficacy.

  5. A novel packaging system for the generation of helper-free oncolytic MVM vector stocks.

    PubMed

    Brandenburger, A; Russell, S

    1996-10-01

    MVM-based autonomous parvoviral vectors have been shown to target the expression of heterologous genes in neoplastic cells and are therefore of interest for cancer gene therapy. The traditional method for production of parvoviral vectors requires the cotransfection of vector and helper plasmids into MVM-permissive cell lines, but recombination between the cotransfected plasmids invariably gives rise to vector stocks that are heavily contaminated with wild-type MVM. Therefore, to minimise recombination between the vector and helper genomes we have utilised a cell line in which the MVM helper functions are expressed inducibly from a modified MVM genome that is stably integrated into the host cell chromosome. Using this MVM packaging cell line, we could reproducibly generate MVM vector stocks that contained no detectable helper virus.

  6. Use of the piggyBac transposon to create stable packaging cell lines for the production of clinical-grade self-inactivating γ-retroviral vectors.

    PubMed

    Feldman, Steven A; Xu, Hui; Black, Mary A; Park, Tristen S; Robbins, Paul F; Kochenderfer, James N; Morgan, Richard A; Rosenberg, Steven A

    2014-08-01

    Efforts to improve the biosafety of γ-retroviral-mediated gene therapy have resulted in a shift toward the use of self-inactivating (SIN) γ-retroviral vectors. However, scale-up and manufacturing of such vectors requires significant optimization of transient transfection-based processes or development of novel platforms for the generation of stable producer cell clones. To that end, we describe the use of the piggybac transposon to generate stable producer cell clones for the production of SIN γ-retroviral vectors. The piggybac transposon is a universal tool allowing for the stable integration of SIN γ-retroviral constructs into murine (PG13) and human 293-based Phoenix (GALV and RD114, respectively) packaging cell lines without reverse transcription. Following transposition, a high-titer clone is selected for manufacture of a master cell bank and subsequent γ-retroviral vector supernatant production. Packaging cell clones created using the piggybac transposon have comparable titers to non-SIN vectors generated via conventional methods. We describe herein the use of the piggybac transposon for the production of stable packaging cell clones for the manufacture of clinical-grade SIN γ-retroviral vectors for ex vivo gene therapy clinical trials.

  7. CellTrans: An R Package to Quantify Stochastic Cell State Transitions.

    PubMed

    Buder, Thomas; Deutsch, Andreas; Seifert, Michael; Voss-Böhme, Anja

    2017-01-01

    Many normal and cancerous cell lines exhibit a stable composition of cells in distinct states which can, e.g., be defined on the basis of cell surface markers. There is evidence that such an equilibrium is associated with stochastic transitions between distinct states. Quantifying these transitions has the potential to better understand cell lineage compositions. We introduce CellTrans, an R package to quantify stochastic cell state transitions from cell state proportion data from fluorescence-activated cell sorting and flow cytometry experiments. The R package is based on a mathematical model in which cell state alterations occur due to stochastic transitions between distinct cell states whose rates only depend on the current state of a cell. CellTrans is an automated tool for estimating the underlying transition probabilities from appropriately prepared data. We point out potential analytical challenges in the quantification of these cell transitions and explain how CellTrans handles them. The applicability of CellTrans is demonstrated on publicly available data on the evolution of cell state compositions in cancer cell lines. We show that CellTrans can be used to (1) infer the transition probabilities between different cell states, (2) predict cell line compositions at a certain time, (3) predict equilibrium cell state compositions, and (4) estimate the time needed to reach this equilibrium. We provide an implementation of CellTrans in R, freely available via GitHub (https://github.com/tbuder/CellTrans).

  8. [Anti-HBV effects of genetically engineered replication-defective HBV with combined expression of antisense RNA and dominant negative mutants of core protein and construction of first-generation packaging cell line for HBV vector].

    PubMed

    Sun, Dian Xing; Hu, Da Rong; Wu, Guang Hui; Hu, Xue Ling; Li, Juan; Fan, Gong Ren

    2002-08-01

    To explore the possibility of using HBV as a gene delivery vector, and to test the anti-HBV effects by intracellular combined expression of antisense RNA and dominant negative mutants of core protein. Full length of mutant HBV genome, which expresses core-partial P fusion protein and/or antisense RNA, was transfected into HepG2.2.15 cell lines. Positive clones were selected and mixed in respective groups with hygromycin in the culture medium. HBsAg and HBeAg, which exist in the culture medium, were tested by ELISA method. Intracellular HBc related HBV DNA was examined by dot blot hybridization. The existence of recombinant HBV virion in the culture medium was examined by PCR. Free of packaging signal, HBV genome, which express the HBV structural proteins including core, pol and preS/S proteins, was inserted into pCI-neo vector. HepG2 cell lines were employed to transfect with the construct. G418 selection was done at the concentration of 400mug/ml in the culture medium. The G418-resistant clones with the best expression of HBsAg and HBcAg were theoretically considered as packaging cell lines and propagated under the same conditions. It was transfected with plasmid pMEP-CPAS and then selected with G418 and hygromycin in the culture medium. The existence of recombinant HBV virion in the culture medium was examined by PCR. The mean inhibitory rates of HBsAg were 2.74% 3.83%, 40.08 2.05% (t=35.5, P<0.01), 66.54% 4.45% (t=42.3, P<0.01), and 73.68% 5.07% (t=51.9, P<0.01) in group 2.2.15-pMEP4, 2.2.15-CP, 2.2.15-SAS, and 2.2.15-CPAS, respectively. The mean inhibitory rates of HBeAg were 4.46% 4.25%, 52.86% 1.32% (t=36.2, P<0.01), 26.36% 1.69% (t=22.3, P<0.01), and 59.28% 2.10% (t=39.0, P<0.01), respectively. The inhibitory rates of HBc related HBV DNA were 0, 82.0%, 59.9%, and 96.6%, respectively. Recombinant HB virion was detectable in the culture medium of all the three treatment groups. G418-resistant HBV packaging cell line, which harbored an HBV mutant whose packaging signal had been deleted, was generated. Expression of HBsAg and HBcAg was detectable. Transfected with plasmid pMEP-CPAS, it was found to secrete recombinant HB virion and no wild-type HBV was detectable in the culture medium. It has stronger anti-HBV effects by combined expression of antisense RNA and dominant negative mutants than by individual expression of them. With the help of wild-type HBV, the modified HBV genome can form and secret HBV like particles, which provides evidence that the antiviral gene will be hepatotropic expression and the antiviral effects will be amplified. The packaging cell line can provide packaging for replication-defective HBV, but with low efficiency.

  9. Alpharetroviral self-inactivating vectors produced by a superinfection-resistant stable packaging cell line allow genetic modification of primary human T lymphocytes.

    PubMed

    Labenski, Verena; Suerth, Julia D; Barczak, Elke; Heckl, Dirk; Levy, Camille; Bernadin, Ornellie; Charpentier, Emmanuelle; Williams, David A; Fehse, Boris; Verhoeyen, Els; Schambach, Axel

    2016-08-01

    Primary human T lymphocytes represent an important cell population for adoptive immunotherapies, including chimeric-antigen and T-cell receptor applications, as they have the capability to eliminate non-self, virus-infected and tumor cells. Given the increasing numbers of clinical immunotherapy applications, the development of an optimal vector platform for genetic T lymphocyte engineering, which allows cost-effective high-quality vector productions, remains a critical goal. Alpharetroviral self-inactivating vectors (ARV) have several advantages compared to other vector platforms, including a more random genomic integration pattern and reduced likelihood for inducing aberrant splicing of integrated proviruses. We developed an ARV platform for the transduction of primary human T lymphocytes. We demonstrated functional transgene transfer using the clinically relevant herpes-simplex-virus thymidine kinase variant TK.007. Proof-of-concept of alpharetroviral-mediated T-lymphocyte engineering was shown in vitro and in a humanized transplantation model in vivo. Furthermore, we established a stable, human alpharetroviral packaging cell line in which we deleted the entry receptor (SLC1A5) for RD114/TR-pseudotyped ARVs to prevent superinfection and enhance genomic integrity of the packaging cell line and viral particles. We showed that superinfection can be entirely prevented, while maintaining high recombinant virus titers. Taken together, this resulted in an improved production platform representing an economic strategy for translating the promising features of ARVs for therapeutic T-lymphocyte engineering. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Vero/BC-F: an efficient packaging cell line stably expressing F protein to generate single round-infectious human parainfluenza virus type 2 vector.

    PubMed

    Ohtsuka, J; Fukumura, M; Tsurudome, M; Hara, K; Nishio, M; Kawano, M; Nosaka, T

    2014-08-01

    A stable packaging cell line (Vero/BC-F) constitutively expressing fusion (F) protein of the human parainfluenza virus type 2 (hPIV2) was established for production of the F-defective and single round-infectious hPIV2 vector in a strategy for recombinant vaccine development. The F gene expression has not evoked cytostatic or cytotoxic effects on the Vero/BC-F cells and the F protein was physiologically active to induce syncytial formation with giant polykaryocytes when transfected with a plasmid expressing hPIV2 hemagglutinin-neuraminidase (HN). Transduction of the F-defective replicon RNA into the Vero/BC-F cells led to the release of the infectious particles that packaged the replicon RNA (named as hPIV2ΔF) without detectable mutations, limiting the infectivity to a single round. The maximal titer of the hPIV2ΔF was 6.0 × 10(8) median tissue culture infections dose per ml. The influenza A virus M2 gene was inserted into hPIV2ΔF, and the M2 protein was found to be highly expressed in a human lung cancer cell line after transduction. Furthermore, in vivo airway infection experiments revealed that the hPIV2ΔF was capable of delivering transgenes to hamster tracheal cells. Thus, non-transmissible or single round-infectious hPIV2 vector will be potentially applicable to human gene therapy or recombinant vaccine development.

  11. RD2-MolPack-Chim3, a packaging cell line for stable production of lentiviral vectors for anti-HIV gene therapy.

    PubMed

    Stornaiuolo, Anna; Piovani, Bianca Maria; Bossi, Sergio; Zucchelli, Eleonora; Corna, Stefano; Salvatori, Francesca; Mavilio, Fulvio; Bordignon, Claudio; Rizzardi, Gian Paolo; Bovolenta, Chiara

    2013-08-01

    Over the last two decades, several attempts to generate packaging cells for lentiviral vectors (LV) have been made. Despite different technologies, no packaging clone is currently employed in clinical trials. We developed a new strategy for LV stable production based on the HEK-293T progenitor cells; the sequential insertion of the viral genes by integrating vectors; the constitutive expression of the viral components; and the RD114-TR envelope pseudotyping. We generated the intermediate clone PK-7 expressing constitutively gag/pol and rev genes and, by adding tat and rd114-tr genes, the stable packaging cell line RD2-MolPack, which can produce LV carrying any transfer vector (TV). Finally, we obtained the RD2-MolPack-Chim3 producer clone by transducing RD2-MolPack cells with the TV expressing the anti-HIV transgene Chim3. Remarkably, RD114-TR pseudovirions have much higher potency when produced by stable compared with transient technology. Most importantly, comparable transduction efficiency in hematopoietic stem cells (HSC) is obtained with 2-logs less physical particles respect to VSV-G pseudovirions produced by transient transfection. Altogether, RD2-MolPack technology should be considered a valid option for large-scale production of LV to be used in gene therapy protocols employing HSC, resulting in the possibility of downsizing the manufacturing scale by about 10-fold in respect to transient technology.

  12. Alternative cell lines to improve the rescue of infectious human astrovirus from a cDNA clone.

    PubMed

    Velázquez-Moctezuma, Rodrigo; Baños-Lara, Ma del Rocío; Acevedo, Yunuén; Méndez, Ernesto

    2012-02-01

    A reverse genetics system for human astrovirus (HAstV) was established previously; however, it has not been exploited mainly because cells used for virus packaging are not permissive, requiring several rounds of replication to obtain acceptable infectious virus. In this work, in the search for alternative permissive cell lines to be used as packaging cells, Hek-293 and Huh7.5.1 were tested. Given that HAstV infection in Hek-293 showed differences with that in Caco-2, the gold standard for HAstV growth but scarcely transfectable, and it was more similar to that observed in the hepatoma Huh7.5.1 cell line, these last cells were further used to transfect viral RNA. Virus titers near to 10(8) infectious particles per ml (ffu/ml) were obtained at 16-20 h after transfection with RNA from infected cells. However, virus titers close to 10(6) ffu/ml were obtained by using in vitro transcribed RNA from a cDNA HAstV-1 clone. In contrast, virus recovery in BHK-21, reported previously as the packaging cells, from this RNA was of about 10(4) ffu/ml, two logarithms less than in Huh7.5.1. Apparently, the 5'-end modification of the viral RNA determined its specific infectivity, since virus recovery was abolished when the total RNA was treated with proteinase-K, probably by removing a protein-linked genome protein, but it increased when capping of the in vitro transcribed RNA was more efficient. Thus, an alternative and more efficient reverse genetics system for HAstV was established by using Huh7.5.1 cells. Copyright © 2011. Published by Elsevier B.V.

  13. Reference Gene Validation for RT-qPCR, a Note on Different Available Software Packages

    PubMed Central

    De Spiegelaere, Ward; Dern-Wieloch, Jutta; Weigel, Roswitha; Schumacher, Valérie; Schorle, Hubert; Nettersheim, Daniel; Bergmann, Martin; Brehm, Ralph; Kliesch, Sabine; Vandekerckhove, Linos; Fink, Cornelia

    2015-01-01

    Background An appropriate normalization strategy is crucial for data analysis from real time reverse transcription polymerase chain reactions (RT-qPCR). It is widely supported to identify and validate stable reference genes, since no single biological gene is stably expressed between cell types or within cells under different conditions. Different algorithms exist to validate optimal reference genes for normalization. Applying human cells, we here compare the three main methods to the online available RefFinder tool that integrates these algorithms along with R-based software packages which include the NormFinder and GeNorm algorithms. Results 14 candidate reference genes were assessed by RT-qPCR in two sample sets, i.e. a set of samples of human testicular tissue containing carcinoma in situ (CIS), and a set of samples from the human adult Sertoli cell line (FS1) either cultured alone or in co-culture with the seminoma like cell line (TCam-2) or with equine bone marrow derived mesenchymal stem cells (eBM-MSC). Expression stabilities of the reference genes were evaluated using geNorm, NormFinder, and BestKeeper. Similar results were obtained by the three approaches for the most and least stably expressed genes. The R-based packages NormqPCR, SLqPCR and the NormFinder for R script gave identical gene rankings. Interestingly, different outputs were obtained between the original software packages and the RefFinder tool, which is based on raw Cq values for input. When the raw data were reanalysed assuming 100% efficiency for all genes, then the outputs of the original software packages were similar to the RefFinder software, indicating that RefFinder outputs may be biased because PCR efficiencies are not taken into account. Conclusions This report shows that assay efficiency is an important parameter for reference gene validation. New software tools that incorporate these algorithms should be carefully validated prior to use. PMID:25825906

  14. Reference gene validation for RT-qPCR, a note on different available software packages.

    PubMed

    De Spiegelaere, Ward; Dern-Wieloch, Jutta; Weigel, Roswitha; Schumacher, Valérie; Schorle, Hubert; Nettersheim, Daniel; Bergmann, Martin; Brehm, Ralph; Kliesch, Sabine; Vandekerckhove, Linos; Fink, Cornelia

    2015-01-01

    An appropriate normalization strategy is crucial for data analysis from real time reverse transcription polymerase chain reactions (RT-qPCR). It is widely supported to identify and validate stable reference genes, since no single biological gene is stably expressed between cell types or within cells under different conditions. Different algorithms exist to validate optimal reference genes for normalization. Applying human cells, we here compare the three main methods to the online available RefFinder tool that integrates these algorithms along with R-based software packages which include the NormFinder and GeNorm algorithms. 14 candidate reference genes were assessed by RT-qPCR in two sample sets, i.e. a set of samples of human testicular tissue containing carcinoma in situ (CIS), and a set of samples from the human adult Sertoli cell line (FS1) either cultured alone or in co-culture with the seminoma like cell line (TCam-2) or with equine bone marrow derived mesenchymal stem cells (eBM-MSC). Expression stabilities of the reference genes were evaluated using geNorm, NormFinder, and BestKeeper. Similar results were obtained by the three approaches for the most and least stably expressed genes. The R-based packages NormqPCR, SLqPCR and the NormFinder for R script gave identical gene rankings. Interestingly, different outputs were obtained between the original software packages and the RefFinder tool, which is based on raw Cq values for input. When the raw data were reanalysed assuming 100% efficiency for all genes, then the outputs of the original software packages were similar to the RefFinder software, indicating that RefFinder outputs may be biased because PCR efficiencies are not taken into account. This report shows that assay efficiency is an important parameter for reference gene validation. New software tools that incorporate these algorithms should be carefully validated prior to use.

  15. A defective retroviral vector encoding human interferon-alpha2 can transduce human leukemic cell lines.

    PubMed

    Austruy, E; Bagnis, C; Carbuccia, N; Maroc, C; Birg, F; Dubreuil, P; Mannoni, P; Chabannon, C

    1998-01-01

    Using the LXSN backbone, a defective retroviral vector (LISN) was constructed that encodes the human interferon (IFN)-alpha2 (hIFN-alpha2) gene and the neomycin resistance gene; the hIFN-alpha2 gene was cloned from human placental genomic DNA. High titers of the LISN retrovirus were produced by the amphotropic packaging cell line GP+envAM12. LISN is able to infect three human hematopoietic and leukemic cell lines: K562, LAMA-84, and TF-1. G418-resistant cells were detected in a similar proportion after infection with either the LISN retroviral vector or the LnLSN retroviral vector (encoding the nlsLacZ gene instead of hIFN-alpha2), suggesting that hIFN-alpha2 does not inhibit (or only partially inhibits) the production of retroviral particles by the packaging cell line and the infection of human cells. LISN-infected cells express and secrete hIFN-alpha2 as demonstrated by Northern blot analysis of poly(A)+ RNA, detection of the intracellular protein by fluorescence-activated cell sorter analysis, and detection of secreted hIFN-alpha in cell supernatants using an enzyme-linked immunosorbent assay. Retrovirally produced hIFN-alpha2 is biologically active, as demonstrated by the partial inhibition of the growth of K562 and TF-1, the modulation of the expression of cell surface antigens, the induction of the (2'-5') oligoadenylate synthetase, and, for LAMA-84, the down-modulation of the BCR-ABL protein. We conclude that the infection of human leukemic cell lines with a retroviral vector encoding hIFN-alpha2 is feasible and induces the expected biological effects. This experimental model will be useful in investigating the possibility of transducing normal and leukemic cells and hematopoietic progenitors and in determining the consequences of the autocrine production of hIFN-alpha2 on the behavior of these cells.

  16. Efficient Transduction of Human and Rhesus Macaque Primary T Cells by a Modified Human Immunodeficiency Virus Type 1-Based Lentiviral Vector.

    PubMed

    He, Huan; Xue, Jing; Wang, Weiming; Liu, Lihong; Ye, Chaobaihui; Cong, Zhe; Kimata, Jason T; Qin, Chuan; Zhou, Paul

    2017-03-01

    Human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors efficiently transduce genes to human, but not rhesus, primary T cells and hematopoietic stem cells (HSCs). The poor transduction of HIV-1 vectors to rhesus cells is mainly due to species-specific restriction factors such as rhesus TRIM5α. Previously, several strategies to modify HIV-1 vectors were developed to overcome rhesus TRIM5α restriction. While the modified HIV-1 vectors efficiently transduce rhesus HSCs, they remain suboptimal for rhesus primary T cells. Recently, HIV-1 variants that encode combinations of LNEIE mutations in capsid (CA) protein and SIVmac239 Vif were found to replicate efficiently in rhesus primary T cells. Thus, the present study tested whether HIV-1 vectors packaged by a packaging construct containing these CA substitutions could efficiently transduce both human and rhesus primary CD4 T cells. To accomplish this, LNEIE mutations were made in the packaging construct CEMΔ8.9, and recombinant HIV-1 vectors packaged by Δ8.9 WT or Δ8.9 LNEIE were generated. Transduction rates, CA stability, and vector integration in CEMss-CCR5 and CEMss-CCR5-rhTRIM5α/green fluorescent protein cells, as well as transduction rates in human and rhesus primary CD4 T cells by Δ8.9 WT or Δ8.9 LNEIE-packaged HIV-1 vectors, were compared. Finally, the influence of rhesus TRIM5α variations in transduction rates to primary CD4 T cells from a cohort of 37 Chinese rhesus macaques was studied. While it maintains efficient transduction for human T-cell line and primary CD4 T cells, Δ8.9 LNEIE-packaged HIV-1 vector overcomes rhesus TRIM5α-mediated CA degradation, resulting in significantly higher transduction efficiency of rhesus primary CD4 T cells than Δ8.9 WT-packaged HIV-1 vector. Rhesus TRIM5α variations strongly influence transduction efficiency of rhesus primary CD4 T cells by both Δ8.9 WT or Δ8.9 LNEIE-packaged HIV-1 vectors. Thus, it is concluded that Δ8.9 LNEIE-packaged HIV-1 vector overcomes rhesus TRIM5α restriction and efficiently transduces both human and rhesus primary T cells.

  17. Efficient Transduction of Human and Rhesus Macaque Primary T Cells by a Modified Human Immunodeficiency Virus Type 1–Based Lentiviral Vector

    PubMed Central

    He, Huan; Xue, Jing; Wang, Weiming; Liu, Lihong; Ye, Chaobaihui; Cong, Zhe; Kimata, Jason T.; Qin, Chuan; Zhou, Paul

    2017-01-01

    Human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors efficiently transduce genes to human, but not rhesus, primary T cells and hematopoietic stem cells (HSCs). The poor transduction of HIV-1 vectors to rhesus cells is mainly due to species-specific restriction factors such as rhesus TRIM5α. Previously, several strategies to modify HIV-1 vectors were developed to overcome rhesus TRIM5α restriction. While the modified HIV-1 vectors efficiently transduce rhesus HSCs, they remain suboptimal for rhesus primary T cells. Recently, HIV-1 variants that encode combinations of LNEIE mutations in capsid (CA) protein and SIVmac239 Vif were found to replicate efficiently in rhesus primary T cells. Thus, the present study tested whether HIV-1 vectors packaged by a packaging construct containing these CA substitutions could efficiently transduce both human and rhesus primary CD4 T cells. To accomplish this, LNEIE mutations were made in the packaging construct CEMΔ8.9, and recombinant HIV-1 vectors packaged by Δ8.9 WT or Δ8.9 LNEIE were generated. Transduction rates, CA stability, and vector integration in CEMss-CCR5 and CEMss-CCR5-rhTRIM5α/green fluorescent protein cells, as well as transduction rates in human and rhesus primary CD4 T cells by Δ8.9 WT or Δ8.9 LNEIE-packaged HIV-1 vectors, were compared. Finally, the influence of rhesus TRIM5α variations in transduction rates to primary CD4 T cells from a cohort of 37 Chinese rhesus macaques was studied. While it maintains efficient transduction for human T-cell line and primary CD4 T cells, Δ8.9 LNEIE-packaged HIV-1 vector overcomes rhesus TRIM5α-mediated CA degradation, resulting in significantly higher transduction efficiency of rhesus primary CD4 T cells than Δ8.9 WT-packaged HIV-1 vector. Rhesus TRIM5α variations strongly influence transduction efficiency of rhesus primary CD4 T cells by both Δ8.9 WT or Δ8.9 LNEIE-packaged HIV-1 vectors. Thus, it is concluded that Δ8.9 LNEIE-packaged HIV-1 vector overcomes rhesus TRIM5α restriction and efficiently transduces both human and rhesus primary T cells. PMID:28042947

  18. Large-scale Clinical-grade Retroviral Vector Production in a Fixed-Bed Bioreactor

    PubMed Central

    Wang, Xiuyan; Olszewska, Malgorzata; Qu, Jinrong; Wasielewska, Teresa; Bartido, Shirley; Hermetet, Gregory; Sadelain, Michel

    2015-01-01

    The successful genetic engineering of patient T cells with γ-retroviral vectors expressing chimeric antigen receptors or T-cell receptors for phase II clinical trials and beyond requires the large-scale manufacture of high-titer vector stocks. The production of retroviral vectors from stable packaging cell lines using roller bottles or 10- to 40-layer cell factories is limited by a narrow harvest window, labor intensity, open-system operations, and the requirement for significant incubator space. To circumvent these shortcomings, we optimized the production of vector stocks in a disposable fixed-bed bioreactor using good manufacturing practice–grade packaging cell lines. High-titer vector stocks were harvested over 10 days, representing a much broader harvest window than the 3-day harvest afforded by cell factories. For PG13 and 293Vec packaging cells, the average vector titer and the vector stocks’ yield in the bioreactor were higher by 3.2- to 7.3-fold, and 5.6- to 13.1-fold, respectively, than those obtained in cell factories. The vector production was 10.4 and 18.6 times more efficient than in cell factories for PG13 and 293Vec cells, respectively. Furthermore, the vectors produced from the fixed-bed bioreactors passed the release test assays for clinical applications. Therefore, a single vector lot derived from 293Vec is suitable to transduce up to 500 patients cell doses in the context of large clinical trials using chimeric antigen receptors or T-cell receptors. These findings demonstrate for the first time that a robust fixed-bed bioreactor process can be used to produce γ-retroviral vector stocks scalable up to the commercialization phase. PMID:25751502

  19. Mixomics analysis of breast cancer: Long non-coding RNA linc01561 acts as ceRNA involved in the progression of breast cancer.

    PubMed

    Jiang, Rui; Zhao, Chunming; Gao, Binbin; Xu, Jiawen; Song, Wei; Shi, Peng

    2018-06-08

    This study aimed at finding the long non-coding RNA (lncRNA), miRNA and mRNA which played critical roles in breast cancer (BrCa) by using mixOmics R package. The BrCa dataset were obtained from TCGA and then analyzed using "DESeq2" R package. Multivariate analyses were performed with the "mixOmics" R package and the first component of the stacked partial least-Squares discriminant analysis results were used for searching the interested lncRNA, miRNA and mRNA. qRT-PCR was applied to identify the bioinformatics results in four BrCa cell lines (MCF7, BT-20, ZR-75-1, and MX-1) and the breast epithelial cell line MCF-10 A. Then cells (MCF-1 and MX-1) were transfected with si-linc01561, miR-145-5p mimics and si-MMP11 to further investigate the effects of linc01561, miR-145-5p and MMP11 on the BrCa cells proliferation and apoptosis. MixOmics results showed that linc01561, miR-145-5p and MMP11 might play important roles in BrCa. qRT-PCR results identified that in BrCa cell lines, linc01561 and MMP11 were higher expressed while miR-145-5p was lower expressed compared with those in epithelial cell line. The linc01561 inhibition elevated miR-145-5p expression and then suppressed MMP11 expression. Moreover, linc01561 inhibition suppressed the BrCa cells proliferation and promoted the apoptosis, which was realized by up-regulating expression of miR-145-5p and down-regulating expression of MMP11. In summary, the findings of this study, based on ceRNA theory, combining the research foundation of miR-145-5p and MMP11, and taking linc01561 as a new study point, provide new insight into molecular-level reversing proliferation and apoptosis of BrCa. Copyright © 2018 Elsevier Ltd. All rights reserved.

  20. An avian cell line designed for production of highly attenuated viruses.

    PubMed

    Jordan, Ingo; Vos, Ad; Beilfuss, Stefanie; Neubert, Andreas; Breul, Sabine; Sandig, Volker

    2009-01-29

    Several viral vaccines, including highly promising vectors such as modified vaccinia Ankara (MVA), are produced on chicken embryo fibroblasts. Dependence on primary cells complicates production especially in large vaccination programs. With primary cells it is also not possible to create packaging lines for replication-deficient vectors that are adapted to proliferation in an avian host. To obviate requirement for primary cells permanent lines from specific tissues of muscovy duck were derived (AGE1.CR, CS, and CA) and further modified: we demonstrate that stable expression of the structural gene pIX from human adenovirus increases titers for unrelated poxvirus in the avian cells. This augmentation appears to be mediated via induction of heat shock and thus provides a novel cellular substrate that may allow further attenuation of vaccine strains.

  1. Functional characterization of adenoviral/retroviral chimeric vectors and their use for efficient screening of retroviral producer cell lines.

    PubMed

    Duisit, G; Salvetti, A; Moullier, P; Cosset, F L

    1999-01-20

    We have generated three different E1-deleted replication-defective adenoviral vectors expressing either Moloney murine leukemia virus (Mo-MuLV) Gag-Pol core particle proteins, gibbon ape leukemia virus (GALV) envelope glycoproteins, or an MuLV-derived retroviral vector genome encoding mCD2 antigen, a murine cell surface marker easily detectable by flow cytometry. Each of the three vectors was first characterized individually by infection of cells providing the complementary retroviral function(s) and able to induce the production of retroviral vectors with an efficiency similar to or higher than that of FLY stable retroviral packaging cells [Cosset, F.-L., Takeuchi, Y., Battini, J.-L., Weiss, R.A., and Collins, M.K.L., (1995). J. Virol. 69, 7430-7436]. In small-scale pilot experiments, TE671 cells simultaneously coinfected with the three adenoviral vectors efficiently released helper-free retroviral vectors in their supernatant, with titers greater than 10(6) infectious particles per milliliter by end-point titrations. Our results also indicated that in contrast to retroviral vector-packageable RNAs, the adenovirus-mediated overexpression of both Gag-Pol and Env packaging functions had limited impact on retroviral titers. The primary mechanism suspected is the premature intracellular cleavage of the Pr65gag precursor that we found in gag-pol-expressing cells, which in turn may impair the normal incorporation of high loads of functional Env. Last, the characterization of the adenoviral/retroviral chimeric vectors allowed the screening of various primate cells for retroviral production and we found that three hepatocyte-derived cell lines were highly efficient in the assembly and release of infectious retroviral particles.

  2. Extensive characterization of a lentiviral-derived stable cell line expressing rabbit hemorrhagic disease virus VPg protein.

    PubMed

    Zhu, Jie; Miao, Qiuhong; Tan, Yonggui; Guo, Huimin; Li, Chuanfeng; Chen, Zongyan; Liu, Guangqing

    2016-11-01

    Rabbit hemorrhagic disease virus (RHDV) is an important member of the caliciviridae family. Currently, no suitable tissue culture system is available for proliferating RHDV, which limits the study of its pathogenesis. To bypass this obstacle, we established a cell line, RK13-VPg, stably expressing the VPg gene with a lentivirus packaging system in this study. In addition, the recently constructed RHDV replicon in our laboratory provided an appropriate model for studying the pathogenesis of RHDV without in vitro RHDV propagation and culture. Using this RHDV replicon and RK13-VPg cell line, we further demonstrated that the presence of VPg protein is essential for efficient translation of an RHDV replicon. Therefore, the RK13-VPg cell line is a powerful tool for studying the replication and translation mechanisms of RHDV. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. A Promising IFN-Deficient System to Manufacture IFN-Sensitive Influenza Vaccine Virus.

    PubMed

    Chen, Can; Fan, Wenhui; Li, Jing; Zheng, Weinan; Zhang, Shuang; Yang, Limin; Liu, Di; Liu, Wenjun; Sun, Lei

    2018-01-01

    Interferon (IFN)-sensitive and replication-incompetent influenza viruses are likely to be the alternatives to inactivated and attenuated virus vaccines. Some IFN-sensitive influenza vaccine candidates with modified non-structural protein 1 (NS1) are highly attenuated in IFN-competent hosts but induce robust antiviral immune responses. However, little research has been done on the manufacturability of these IFN-sensitive vaccine viruses. Here, RIG-I-knockout 293T cells were used to package the IFN-sensitive influenza A/WSN/33 (H1N1) virus expressing the mutant NS1 R38A/K41A. We found that the packaging efficiency of the NS1 R38A/K41A virus in RIG-I-knockout 293T cells was much higher than that in 293T cells. Moreover, the NS1 R38A/K41A virus almost lost its IFN antagonist activity and could no longer replicate in A549, MDCK, and Vero cells after 3-6 passages. This indicated that the replication of NS1 R38A/K41A virus is limited in conventional cells. Therefore, we further established a stable Vero cell line expressing the wild-type (WT) NS1 of the WSN virus, based on the Tet-On 3G system. The NS1 R38A/K41A virus was able to steadily propagate in this IFN-deficient cell line for at least 20 passages. In a mouse model, the NS1 R38A/K41A virus showed more than a 4-log reduction in lung virus titers compared to the WT virus at 3 and 5 days post infection. Furthermore, we observed that the NS1 R38A/K41A virus triggered high-level of IFN-α/β production in lung tissues and was eliminated from the host in a relatively short period of time. Additionally, this virus induced high-titer neutralizing antibodies against the WT WSN, A/Puerto Rico/8/1934 (PR8), or A/California/04/2009 (CA04) viruses and provided 100% protection against the WT WSN virus. Thus, we found that the replication of the NS1 R38A/K41A virus was limited in IFN-competent cells and mice. We also presented a promising IFN-deficient system, involving a RIG-I-knockout 293T cell line to package the IFN-sensitive vaccine virus and a stable Vero cell line expressing NS1 to propagate the IFN-sensitive vaccine virus. The IFN-deficient system is applicable for the manufacture of IFN-sensitive vaccine virus.

  4. Differential Expression of HERV-K (HML-2) Proviruses in Cells and Virions of the Teratocarcinoma Cell Line Tera-1

    PubMed Central

    Bhardwaj, Neeru; Montesion, Meagan; Roy, Farrah; Coffin, John M.

    2015-01-01

    Human endogenous retrovirus (HERV-K (HML-2)) proviruses are among the few endogenous retroviral elements in the human genome that retain coding sequence. HML-2 expression has been widely associated with human disease states, including different types of cancers as well as with HIV-1 infection. Understanding of the potential impact of this expression requires that it be annotated at the proviral level. Here, we utilized the high throughput capabilities of next-generation sequencing to profile HML-2 expression at the level of individual proviruses and secreted virions in the teratocarcinoma cell line Tera-1. We identified well-defined expression patterns, with transcripts emanating primarily from two proviruses located on chromosome 22, only one of which was efficiently packaged. Interestingly, there was a preference for transcripts of recently integrated proviruses, over those from other highly expressed but older elements, to be packaged into virions. We also assessed the promoter competence of the 5’ long terminal repeats (LTRs) of expressed proviruses via a luciferase assay following transfection of Tera-1 cells. Consistent with the RNASeq results, we found that the activity of most LTRs corresponded to their transcript levels. PMID:25746218

  5. An efficient deletion mutant packaging system for defective herpes simplex virus vectors: Potential applications to human gene therapy and neuronal physiology

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Geller, A.I.; Keyomarsi, K.; Bryan, J.

    1990-11-01

    The authors have previously described a defective herpes simplex virus (HSV-1) vector system that permits that introduction of virtually any gene into nonmitotic cells. pHSVlac, the prototype vector, stably expresses Escherichia coli {beta}-galactosidase from a constitutive promoter in many human cell lines, in cultured rat neurons from throughout the nervous system, and in cells in the adult rat brain. HSV-1 vectors expressing other genes may prove useful for studying neuronal physiology or performing human gene therapy for neurological diseases, such as Parkinson disease or brain tumors. A HSV-1 temperature-sensitive (ts) mutant, ts K, has been used as helper virus; tsmore » mutants revert to wild type. In contrast, HSV-1 deletion mutants essentially cannot revert to wild type; therefore, use of a deletion mutant as helper virus might permit human gene therapy with HSV-1 vectors. They now report an efficient packaging system for HSV-1 VECTORS USING A DELETION MUTANT, d30EBA, as helper virus; virus is grown on the complementing cell line M64A. pHSVlac virus prepared using the deletion mutant packaging system stably expresses {beta}-galactosidase in cultured rat sympathetic neurons and glia. Both D30EBA and ts K contain a mutation in the IE3 gene of HSV-1 strain 17 and have the same phenotype; therefore, changing the helper virus from ts K to D30EBA does not alter the host range or other properties of the HSV-1 vector system.« less

  6. Noninfectious virus-like particles produced by Moloney murine leukemia virus-based retrovirus packaging cells deficient in viral envelope become infectious in the presence of lipofection reagents

    PubMed Central

    Sharma, Sanjai; Murai, Fukashi; Miyanohara, Atsushi; Friedmann, Theodore

    1997-01-01

    Retrovirus packaging cell lines expressing the Moloney murine leukemia virus gag and pol genes but lacking virus envelope genes produce virus-like particles constitutively, whether or not they express a transcript from an integrated retroviral provirus. In the absence of a proviral transcript, the assembled particles contain processed gag and reverse transcriptase, and particles made by cells expressing an integrated lacZ provirus also contain viral RNA. The virus-like particles from both cell types are enveloped and are secreted/budded into the extracellular space but are noninfectious. Their physicochemical properties are similar to those of mature retroviral particles. The noninfectious gag pol RNA particles can readily be made infectious by the addition of lipofection reagents to produce preparations with titers of up to 105 colony-forming units per ml. PMID:9380714

  7. Efficient construction of producer cell lines for a SIN lentiviral vector for SCID-X1 gene therapy by concatemeric array transfection

    PubMed Central

    Throm, Robert E.; Ouma, Annastasia A.; Zhou, Sheng; Chandrasekaran, Anantharaman; Lockey, Timothy; Greene, Michael; De Ravin, Suk See; Moayeri, Morvarid; Malech, Harry L.; Sorrentino, Brian P.

    2009-01-01

    Retroviral vectors containing internal promoters, chromatin insulators, and self-inactivating (SIN) long terminal repeats (LTRs) may have significantly reduced genotoxicity relative to the conventional retroviral vectors used in recent, otherwise successful clinical trials. Large-scale production of such vectors is problematic, however, as the introduction of SIN vectors into packaging cells cannot be accomplished with the traditional method of viral transduction. We have derived a set of packaging cell lines for HIV-based lentiviral vectors and developed a novel concatemeric array transfection technique for the introduction of SIN vector genomes devoid of enhancer and promoter sequences in the LTR. We used this method to derive a producer cell clone for a SIN lentiviral vector expressing green fluorescent protein, which when grown in a bioreactor generated more than 20 L of supernatant with titers above 107 transducing units (TU) per milliliter. Further refinement of our technique enabled the rapid generation of whole populations of stably transformed cells that produced similar titers. Finally, we describe the construction of an insulated, SIN lentiviral vector encoding the human interleukin 2 receptor common γ chain (IL2RG) gene and the efficient derivation of cloned producer cells that generate supernatants with titers greater than 5 × 107 TU/mL and that are suitable for use in a clinical trial for X-linked severe combined immunodeficiency (SCID-X1). PMID:19286997

  8. fgui: A Method for Automatically Creating Graphical User Interfaces for Command-Line R Packages

    PubMed Central

    Hoffmann, Thomas J.; Laird, Nan M.

    2009-01-01

    The fgui R package is designed for developers of R packages, to help rapidly, and sometimes fully automatically, create a graphical user interface for a command line R package. The interface is built upon the Tcl/Tk graphical interface included in R. The package further facilitates the developer by loading in the help files from the command line functions to provide context sensitive help to the user with no additional effort from the developer. Passing a function as the argument to the routines in the fgui package creates a graphical interface for the function, and further options are available to tweak this interface for those who want more flexibility. PMID:21625291

  9. Design of Control System for Flexible Packaging Bags Palletizing Production Line Based on PLC

    NASA Astrophysics Data System (ADS)

    Zheng, Huiping; Chen, Lin; Zhao, Xiaoming; Liu, Zhanyang

    Flexible packaging bags palletizing production line is to put the bags in the required area according to particular order and size, in order to finish handling, storage, loading and unloading, transportation and other logistics work of goods. Flexible packaging bags palletizing line is composed of turning bags mechanism, shaping mechanism, indexing mechanism, marshalling mechanism, pushing bags mechanism, pressing bags mechanism, laminating mechanism, elevator, tray warehouse, tray conveyor and loaded tray conveyor. Whether the whole production line can smoothly run depends on each of the above equipment and precision control among them. In this paper the technological process and the control logic of flexible packaging bags palletizing production line is introduced. Palletizing process of the production line realized automation by means of a control system based on programmable logic controller (PLC). It has the advantages of simple structure, reliable and easy maintenance etc.

  10. Experimental Verification of the Use of Metal Filled Via Hole Fences for Crosstalk Control of Microstrip Lines in LTCC Packages

    NASA Technical Reports Server (NTRS)

    Ponchak, George E.; Chun, Donghoon; Katehi, Linda P. B.; Yook, Jong-Gwan

    1999-01-01

    Coupling between microstrip lines in dense RF packages is a common problem that degrades circuit performance. Prior 3D-FEM electromagnetic simulations have shown that metal filled via hole fences between two adjacent microstrip lines actually increases coupling between the lines; however, if the top of the via posts are connected by a metal Strip, coupling is reduced. In this paper, experimental verification of the 3D-FEM simulations Is demonstrated for commercially fabricated LTCC packages.

  11. The GRIDView Visualization Package

    NASA Astrophysics Data System (ADS)

    Kent, B. R.

    2011-07-01

    Large three-dimensional data cubes, catalogs, and spectral line archives are increasingly important elements of the data discovery process in astronomy. Visualization of large data volumes is of vital importance for the success of large spectral line surveys. Examples of data reduction utilizing the GRIDView software package are shown. The package allows users to manipulate data cubes, extract spectral profiles, and measure line properties. The package and included graphical user interfaces (GUIs) are designed with pipeline infrastructure in mind. The software has been used with great success analyzing spectral line and continuum data sets obtained from large radio survey collaborations. The tools are also important for multi-wavelength cross-correlation studies and incorporate Virtual Observatory client applications for overlaying database information in real time as cubes are examined by users.

  12. High power VCSEL devices for atomic clock applications

    NASA Astrophysics Data System (ADS)

    Watkins, L. S.; Ghosh, C.; Seurin, J.-F.; Zhou, D.; Xu, G.; Xu, B.; Miglo, A.

    2015-09-01

    We are developing VCSEL technology producing >100mW in single frequency at wavelengths 780nm, 795nm and 850nm. Small aperture VCSELs with few mW output have found major applications in atomic clock experiments. Using an external cavity three-mirror configuration we have been able to operate larger aperture VCSELs and obtain >70mW power in single frequency operation. The VCSEL has been mounted in a fiber pigtailed package with the external mirror mounted on a shear piezo. The package incorporates a miniature Rb cell locker to lock the VCSEL wavelength. This VCSEL operates in single frequency and is tuned by a combination of piezo actuator, temperature and current. Mode-hop free tuning over >30GHz frequency span is obtained. The VCSEL has been locked to the Rb D2 line and feedback control used to obtain line-widths of <100kHz.

  13. Establishing a store baseline during interim storage of waste packages and a review of potential technologies for base-lining

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    McTeer, Jennifer; Morris, Jenny; Wickham, Stephen

    Interim storage is an essential component of the waste management lifecycle, providing a safe, secure environment for waste packages awaiting final disposal. In order to be able to monitor and detect change or degradation of the waste packages, storage building or equipment, it is necessary to know the original condition of these components (the 'waste storage system'). This paper presents an approach to establishing the baseline for a waste-storage system, and provides guidance on the selection and implementation of potential base-lining technologies. The approach is made up of two sections; assessment of base-lining needs and definition of base-lining approach. Duringmore » the assessment of base-lining needs a review of available monitoring data and store/package records should be undertaken (if the store is operational). Evolutionary processes (affecting safety functions), and their corresponding indicators, that can be measured to provide a baseline for the waste-storage system should then be identified in order for the most suitable indicators to be selected for base-lining. In defining the approach, identification of opportunities to collect data and constraints is undertaken before selecting the techniques for base-lining and developing a base-lining plan. Base-lining data may be used to establish that the state of the packages is consistent with the waste acceptance criteria for the storage facility and to support the interpretation of monitoring and inspection data collected during store operations. Opportunities and constraints are identified for different store and package types. Technologies that could potentially be used to measure baseline indicators are also reviewed. (authors)« less

  14. Experimental Verification of the Use of Metal Filled Via Hole Fences for Crosstalk Control of Microstrip Lines in LTCC Packages

    NASA Technical Reports Server (NTRS)

    Ponchak, George E.; Chun, Donghoon; Yook, Jong-Gwan; Katehi, Linda P. B.

    2001-01-01

    Coupling between microstrip lines in dense RF packages is a common problem that degrades circuit performance. Prior three-dimensional-finite element method (3-D-FEM) electromagnetic simulations have shown that metal filled via hole fences between two adjacent microstrip lines actually Increases coupling between the lines: however, if the top of the via posts are connected by a metal strip, coupling is reduced. In this paper, experimental verification of the 3-D-FEM simulations is demonstrated for commercially fabricated low temperature cofired ceramic (LTCC) packages. In addition, measured attenuation of microstrip lines surrounded by the shielding structures is presented and shows that shielding structures do not change the attenuation characteristics of the line.

  15. PharmacoGx: an R package for analysis of large pharmacogenomic datasets.

    PubMed

    Smirnov, Petr; Safikhani, Zhaleh; El-Hachem, Nehme; Wang, Dong; She, Adrian; Olsen, Catharina; Freeman, Mark; Selby, Heather; Gendoo, Deena M A; Grossmann, Patrick; Beck, Andrew H; Aerts, Hugo J W L; Lupien, Mathieu; Goldenberg, Anna; Haibe-Kains, Benjamin

    2016-04-15

    Pharmacogenomics holds great promise for the development of biomarkers of drug response and the design of new therapeutic options, which are key challenges in precision medicine. However, such data are scattered and lack standards for efficient access and analysis, consequently preventing the realization of the full potential of pharmacogenomics. To address these issues, we implemented PharmacoGx, an easy-to-use, open source package for integrative analysis of multiple pharmacogenomic datasets. We demonstrate the utility of our package in comparing large drug sensitivity datasets, such as the Genomics of Drug Sensitivity in Cancer and the Cancer Cell Line Encyclopedia. Moreover, we show how to use our package to easily perform Connectivity Map analysis. With increasing availability of drug-related data, our package will open new avenues of research for meta-analysis of pharmacogenomic data. PharmacoGx is implemented in R and can be easily installed on any system. The package is available from CRAN and its source code is available from GitHub. bhaibeka@uhnresearch.ca or benjamin.haibe.kains@utoronto.ca Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  16. The expression of full length Gp91-phox protein is associated with reduced amphotropic retroviral production.

    PubMed

    Bellantuono, I; Lashford, L S; Rafferty, J A; Fairbairn, L J

    2000-05-01

    As a single gene defect in mature bone marrow cells, chronic granulomatous disease (X-CGD) represents a disorder which may be amenable to gene therapy by the transfer of the missing subunit into hemopoietic stem cells. In the majority of cases lack of Gp91-phox causes the disease. So far, studies involving transfer of Gp91-phox cDNA, including a phase I clinical trial, have yielded disappointing results. Most often, low titers of virus have been reported. In the present study we investigated the possible reasons for low titer amphotropic viral production. To investigate the effect of Gp91 cDNA on the efficiency of retroviral production from the packaging cell line, GP+envAm12, we constructed vectors containing either the native cDNA, truncated versions of the cDNA or a mutated form (LATG) in which the natural translational start codon was changed to a stop codon. Following derivation of clonal packaging cell lines, these were assessed for viral titer by RNA slot blot and analyzed by non-parametrical statistical analysis (Whitney-Mann U-test). An improvement in viral titer of just over two-fold was found in packaging cells containing the start-codon mutant of Gp91 and no evidence of truncated viral RNA was seen in these cells. Further analysis revealed the presence of rearranged forms of the provirus in Gp91-expressing cells, and the production of truncated, unpackaged viral RNA. Protein analysis revealed that LATG-transduced cells did not express full-length Gp91-phox, whereas those containing the wild-type cDNA did. However, a truncated protein was seen in ATG-transduced cells which was also present in wild type cells. No evidence for the presence of a negative transcriptional regulatory element was found from studies with the deletion mutants. A statistically significant effect of protein production on the production of virus from Gp91-expressing cells was found. Our data point to a need to restrict expression of the Gp91-phox protein and its derivatives in order to enhance retroviral production and suggest that improvements in current vectors for CGD gene therapy may need to include controlled, directed expression only in mature neutrophils.

  17. LentiPro26: novel stable cell lines for constitutive lentiviral vector production.

    PubMed

    Tomás, H A; Rodrigues, A F; Carrondo, M J T; Coroadinha, A S

    2018-03-27

    Lentiviral vectors (LVs) are excellent tools to promote gene transfer and stable gene expression. Their potential has been already demonstrated in gene therapy clinical trials for the treatment of diverse disorders. For large scale LV production, a stable producer system is desirable since it allows scalable and cost-effective viral productions, with increased reproducibility and safety. However, the development of stable systems has been challenging and time-consuming, being the selection of cells presenting high expression levels of Gag-Pro-Pol polyprotein and the cytotoxicity associated with some viral components, the main limitations. Hereby is described the establishment of a new LV producer cell line using a mutated less active viral protease to overcome potential cytotoxic limitations. The stable transfection of bicistronic expression cassettes with re-initiation of the translation mechanism enabled the generation of LentiPro26 packaging populations supporting high titers. Additionally, by skipping intermediate clone screening steps and performing only one final clone screening, it was possible to save time and generate LentiPro26-A59 cell line, that constitutively produces titers above 10 6 TU.mL -1 .day -1 , in less than six months. This work constitutes a step forward towards the development of improved LV producer cell lines, aiming to efficiently supply the clinical expanding gene therapy applications.

  18. [Construction of BAD Lentivirus Vector and Its Effect on Proliferation in A549 Cell Lines].

    PubMed

    Huang, Na; He, Yan-qi; Zhu, Jing; Li, Wei-min

    2015-05-01

    To construct the recombinant lentivirus expressing vector BAD (Bcl-2-associated death protein) gene and to study its effect on A549 cell proliferation. The BAD gene was amplified from plasmid pAV-MCMV-BAD-GFP by PCR. The purified BAD gene fragment was inserted into a lentivirus vector (pLVX-IRES-ZsGreen 1), and the insertion was identified by PCR, restriction endonuclease analysis and DNA sequencing. A549 cells were then transfected with the packaged recombinant lentivirus, and resistant cell clones were selected with flow cytometry. The expression of BAD in A549 cell lines stably transduction with a lentivirus was examined using Western blot. The effect of BAD overexpression on proliferation of A549 cells was evaluated by using CCK-8 kit. Restriction enzyme digestion and DNA sequencing showed that the full-length BAD gene (507 bp) had been successfully subcloned into the lentiviral vector to result in the recombinant vector pLVX-IRES-ZsGreen 1. Monoclonal cell lines BAD-A549 was produced after transfection with the recombinant lentivirus and selected with flow cytometry. Stable expression of BAD protein was verified by Western blot. In vitro, the OD value in BAD group was significantly lower than that of control groups from 120-144 h (P<0. 05). A549 cell lines stably transduced with a lentivirus expressing the BAD gene had been successfully generated. In vitro, BAD overexpression significantly inhibited A549 cells proliferation.

  19. Nonaqueous Electrical Storage Device

    DOEpatents

    McEwen, Alan B.; Evans, David A.; Blakley, Thomas J.; Goldman, Jay L.

    1999-10-26

    An electrochemical capacitor is disclosed that features two, separated, high surface area carbon cloth electrodes sandwiched between two current collectors fabricated of a conductive polymer having a flow temperature greater than 130.degree. C., the perimeter of the electrochemical capacitor being sealed with a high temperature gasket to form a single cell device. The gasket material is a thermoplastic stable at temperatures greater than 100.degree. C., preferably a polyester or a polyurethane, and having a reflow temperature above 130.degree. C. but below the softening temperature of the current collector material. The capacitor packaging has good mechanical integrity over a wide temperature range, contributes little to the device equivalent series resistance (ESR), and is stable at high potentials. In addition, the packaging is designed to be easily manufacturable by assembly line methods. The individual cells can be stacked in parallel or series configuration to reach the desired device voltage and capacitance.

  20. WCPP-THE WOLF PLOTTING AND CONTOURING PACKAGE

    NASA Technical Reports Server (NTRS)

    Masaki, G. T.

    1994-01-01

    The WOLF Contouring and Plotting Package provides the user with a complete general purpose plotting and contouring capability. This package is a complete system for producing line printer, SC4020, Gerber, Calcomp, and SD4060 plots. The package has been designed to be highly flexible and easy to use. Any plot from a quick simple plot (which requires only one call to the package) to highly sophisticated plots (including motion picture plots) can be easily generated with only a basic knowledge of FORTRAN and the plot commands. Anyone designing a software system that requires plotted output will find that this package offers many advantages over the standard hardware support packages available. The WCPP package is divided into a plot segment and a contour segment. The plot segment can produce output for any combination of line printer, SC4020, Gerber, Calcomp, and SD4060 plots. The line printer plots allow the user to have plots available immediately after a job is run at a low cost. Although the resolution of line printer plots is low, the quick results allows the user to judge if a high resolution plot of a particular run is desirable. The SC4020 and SD4060 provide high speed high resolution cathode ray plots with film and hard copy output available. The Gerber and Calcomp plotters provide very high quality (of publishable quality) plots of good resolution. Being bed or drum type plotters, the Gerber and Calcomp plotters are usually slow and not suited for large volume plotting. All output for any or all of the plotters can be produced simultaneously. The types of plots supported are: linear, semi-log, log-log, polar, tabular data using the FORTRAN WRITE statement, 3-D perspective linear, and affine transformations. The labeling facility provides for horizontal labels, vertical labels, diagonal labels, vector characters of a requested size (special character fonts are easily implemented), and rotated letters. The gridding routines label the grid lines according to user specification. Special line features include multiple lines, dashed lines, and tic marks. The contour segment of this package is a collection of subroutines which can be used to produce contour plots and perform related functions. The package can contour any data which can be placed on a grid or data which is regularly spaced, including any general affine or polar grid data. The package includes routines which will grid random data. Contour levels can be specified at any values desired. Input data can be smoothed with undefined points being acceptable where data is unreliable or unknown. Plots which are extremely large or detailed can be automatically output in parts to improve resolution or overcome plotter size limitations. The contouring segment uses the plot segment for actual plotting, thus all the features described for the plotting segment are available to the user of the contouring segment. Included with this package are two data bases for producing world map plots in Mercator projection. One data base provides just continent outlines and another provides continent outlines and national borders in great detail. This package is written in FORTRAN IV and IBM OS ASSEMBLER and has been implemented on an IBM 360 with a central memory requirement of approximately 140K of 8 bit bytes. The ASSEMBLER routines are basic plotter interface routines. The WCPP package was developed in 1972.

  1. High sensitive FBG load cell for icing of overhead transmission lines

    NASA Astrophysics Data System (ADS)

    Mao, Naiqiang; Ma, Guoming; Li, Chengrong; Li, Yabo; Shi, Cheng; Du, Yue

    2017-04-01

    Heavy ice coating of overhead transmission lines created the serious threat on the safe operation of power grid. The measurement of conductor icing had been an effective and reliable methods to prevent potential risks, such as conductor breakage, insulator flashover and tower collapse. Because of the advantages of immunity to electromagnetic interference and no demand for power supply in site, the optical load cell has been widely applied in monitoring the ice coating of overhead transmission lines. In this paper, we have adopted the shearing structure with additional grooves as elastic element of load cell to detect the eccentric load. Then, two welding package fiber Bragg gratings (FBGs) were mounted onto the grooves of elastic element with a direction deviation of 90° to eliminate temperature effects on strain measurement without extra FBG. After that, to avoid the occurrence of load cell breakage in heavy load measurement, the protection part has been proposed and added to the elastic element. The results of tension experiments indicate that the resolution of the load cell is 7.78 N in the conventional measuring range (0-10 kN). And in addition, the load cell proposed in this paper also has a good performance in actual experiment in which the load and temperature change simultaneously.

  2. Method of forming a package for MEMS-based fuel cell

    DOEpatents

    Morse, Jeffrey D; Jankowski, Alan F

    2013-05-21

    A MEMS-based fuel cell package and method thereof is disclosed. The fuel cell package comprises seven layers: (1) a sub-package fuel reservoir interface layer, (2) an anode manifold support layer, (3) a fuel/anode manifold and resistive heater layer, (4) a Thick Film Microporous Flow Host Structure layer containing a fuel cell, (5) an air manifold layer, (6) a cathode manifold support structure layer, and (7) a cap. Fuel cell packages with more than one fuel cell are formed by positioning stacks of these layers in series and/or parallel. The fuel cell package materials such as a molded plastic or a ceramic green tape material can be patterned, aligned and stacked to form three dimensional microfluidic channels that provide electrical feedthroughs from various layers which are bonded together and mechanically support a MEMS-based miniature fuel cell. The package incorporates resistive heating elements to control the temperature of the fuel cell stack. The package is fired to form a bond between the layers and one or more microporous flow host structures containing fuel cells are inserted within the Thick Film Microporous Flow Host Structure layer of the package.

  3. Method of forming a package for mems-based fuel cell

    DOEpatents

    Morse, Jeffrey D.; Jankowski, Alan F.

    2004-11-23

    A MEMS-based fuel cell package and method thereof is disclosed. The fuel cell package comprises seven layers: (1) a sub-package fuel reservoir interface layer, (2) an anode manifold support layer, (3) a fuel/anode manifold and resistive heater layer, (4) a Thick Film Microporous Flow Host Structure layer containing a fuel cell, (5) an air manifold layer, (6) a cathode manifold support structure layer, and (7) a cap. Fuel cell packages with more than one fuel cell are formed by positioning stacks of these layers in series and/or parallel. The fuel cell package materials such as a molded plastic or a ceramic green tape material can be patterned, aligned and stacked to form three dimensional microfluidic channels that provide electrical feedthroughs from various layers which are bonded together and mechanically support a MEMOS-based miniature fuel cell. The package incorporates resistive heating elements to control the temperature of the fuel cell stack. The package is fired to form a bond between the layers and one or more microporous flow host structures containing fuel cells are inserted within the Thick Film Microporous Flow Host Structure layer of the package.

  4. Parameterizable Library Components for SAW Devices

    NASA Technical Reports Server (NTRS)

    Wilson, William C.; Atkinson, Gary M.

    2006-01-01

    To facilitate quick fabrication of Surface Acoustic Wave (SAW) sensors we have found it necessary to develop a library of parameterizable components. This library is the first module in our strategy towards a design tool that is integrated into existing Electronic Design Automation (EDA) tools. This library is similar to the standard cell libraries found in digital design packages. The library cells allow the user to input the design parameters which automatically generate a detailed layout of the SAW component. This paper presents the results of our development of parameterizable cells for an InterDigitated Transducer (IDT), reflector, SAW delay line, and both one and two port resonators.

  5. Line drawing Scientific Instrument Module and lunar orbital science package

    NASA Technical Reports Server (NTRS)

    1970-01-01

    A line drawing of the Scientific Instrument Module (SIM) with its lunar orbital science package. The SIM will be mounted in a previously vacant sector of the Apollo Service Module. It will carry specialized cameras and instrumentation for gathering lunar orbit scientific data.

  6. Recombinant Human Parvovirus B19 Vectors: Erythroid Cell-Specific Delivery and Expression of Transduced Genes

    PubMed Central

    Ponnazhagan, Selvarangan; Weigel, Kirsten A.; Raikwar, Sudhanshu P.; Mukherjee, Pinku; Yoder, Mervin C.; Srivastava, Arun

    1998-01-01

    A novel packaging strategy combining the salient features of two human parvoviruses, namely the pathogenic parvovirus B19 and the nonpathogenic adeno-associated virus type 2 (AAV), was developed to achieve erythroid cell-specific delivery as well as expression of the transduced gene. The development of such a chimeric vector system was accomplished by packaging heterologous DNA sequences cloned within the inverted terminal repeats of AAV and subsequently packaging the DNA inside the capsid structure of B19 virus. Recombinant B19 virus particles were assembled, as evidenced by electron microscopy as well as DNA slot blot analyses. The hybrid vector failed to transduce nonerythroid human cells, such as 293 cells, as expected. However, MB-02 cells, a human megakaryocytic leukemia cell line which can be infected by B19 virus following erythroid differentiation with erythropoietin (N. C. Munshi, S. Z. Zhou, M. J. Woody, D. A. Morgan, and A. Srivastava, J. Virol. 67:562–566, 1993) but lacks the putative receptor for AAV (S. Ponnazhagan, X.-S. Wang, M. J. Woody, F. Luo, L. Y. Kang, M. L. Nallari, N. C. Munshi, S. Z. Zhou, and A. Srivastava, J. Gen. Virol. 77:1111–1122, 1996), were readily transduced by this vector. The hybrid vector was also found to specifically target the erythroid population in primary human bone marrow cells as well as more immature hematopoietic progenitor cells following erythroid differentiation, as evidenced by selective expression of the transduced gene in these target cells. Preincubation with anticapsid antibodies against B19 virus, but not anticapsid antibodies against AAV, inhibited transduction of primary human erythroid cells. The efficiency of transduction of primary human erythroid cells by the recombinant B19 virus vector was significantly higher than that by the recombinant AAV vector. Further development of the AAV-B19 virus hybrid vector system should prove beneficial in gene therapy protocols aimed at the correction of inherited and acquired human diseases affecting cells of erythroid lineage. PMID:9573295

  7. Technology and design of an active-matrix OLED on crystalline silicon direct-view display for a wristwatch computer

    NASA Astrophysics Data System (ADS)

    Sanford, James L.; Schlig, Eugene S.; Prache, Olivier; Dove, Derek B.; Ali, Tariq A.; Howard, Webster E.

    2002-02-01

    The IBM Research Division and eMagin Corp. jointly have developed a low-power VGA direct view active matrix OLED display, fabricated on a crystalline silicon CMOS chip. The display is incorporated in IBM prototype wristwatch computers running the Linus operating system. IBM designed the silicon chip and eMagin developed the organic stack and performed the back-end-of line processing and packaging. Each pixel is driven by a constant current source controlled by a CMOS RAM cell, and the display receives its data from the processor memory bus. This paper describes the OLED technology and packaging, and outlines the design of the pixel and display electronics and the processor interface. Experimental results are presented.

  8. Networking of differentially expressed genes in human cancer cells resistant to methotrexate

    PubMed Central

    2009-01-01

    Background The need for an integrated view of data obtained from high-throughput technologies gave rise to network analyses. These are especially useful to rationalize how external perturbations propagate through the expression of genes. To address this issue in the case of drug resistance, we constructed biological association networks of genes differentially expressed in cell lines resistant to methotrexate (MTX). Methods Seven cell lines representative of different types of cancer, including colon cancer (HT29 and Caco2), breast cancer (MCF-7 and MDA-MB-468), pancreatic cancer (MIA PaCa-2), erythroblastic leukemia (K562) and osteosarcoma (Saos-2), were used. The differential expression pattern between sensitive and MTX-resistant cells was determined by whole human genome microarrays and analyzed with the GeneSpring GX software package. Genes deregulated in common between the different cancer cell lines served to generate biological association networks using the Pathway Architect software. Results Dikkopf homolog-1 (DKK1) is a highly interconnected node in the network generated with genes in common between the two colon cancer cell lines, and functional validations of this target using small interfering RNAs (siRNAs) showed a chemosensitization toward MTX. Members of the UDP-glucuronosyltransferase 1A (UGT1A) family formed a network of genes differentially expressed in the two breast cancer cell lines. siRNA treatment against UGT1A also showed an increase in MTX sensitivity. Eukaryotic translation elongation factor 1 alpha 1 (EEF1A1) was overexpressed among the pancreatic cancer, leukemia and osteosarcoma cell lines, and siRNA treatment against EEF1A1 produced a chemosensitization toward MTX. Conclusions Biological association networks identified DKK1, UGT1As and EEF1A1 as important gene nodes in MTX-resistance. Treatments using siRNA technology against these three genes showed chemosensitization toward MTX. PMID:19732436

  9. chemalot and chemalot_knime: Command line programs as workflow tools for drug discovery.

    PubMed

    Lee, Man-Ling; Aliagas, Ignacio; Feng, Jianwen A; Gabriel, Thomas; O'Donnell, T J; Sellers, Benjamin D; Wiswedel, Bernd; Gobbi, Alberto

    2017-06-12

    Analyzing files containing chemical information is at the core of cheminformatics. Each analysis may require a unique workflow. This paper describes the chemalot and chemalot_knime open source packages. Chemalot is a set of command line programs with a wide range of functionalities for cheminformatics. The chemalot_knime package allows command line programs that read and write SD files from stdin and to stdout to be wrapped into KNIME nodes. The combination of chemalot and chemalot_knime not only facilitates the compilation and maintenance of sequences of command line programs but also allows KNIME workflows to take advantage of the compute power of a LINUX cluster. Use of the command line programs is demonstrated in three different workflow examples: (1) A workflow to create a data file with project-relevant data for structure-activity or property analysis and other type of investigations, (2) The creation of a quantitative structure-property-relationship model using the command line programs via KNIME nodes, and (3) The analysis of strain energy in small molecule ligand conformations from the Protein Data Bank database. The chemalot and chemalot_knime packages provide lightweight and powerful tools for many tasks in cheminformatics. They are easily integrated with other open source and commercial command line tools and can be combined to build new and even more powerful tools. The chemalot_knime package facilitates the generation and maintenance of user-defined command line workflows, taking advantage of the graphical design capabilities in KNIME. Graphical abstract Example KNIME workflow with chemalot nodes and the corresponding command line pipe.

  10. Reliability evaluation of hermetic dual in-line flat microcircuit packages

    NASA Technical Reports Server (NTRS)

    Johnson, G. M.; Conaway, L. K.

    1977-01-01

    The relative strengths and weaknesses of 35 commonly used hermetic flat and dual in-line packages were determined and used to rank each of the packages according to a numerical weighting scheme for package attributes. The list of attributes included desirable features in five major areas: lead and lead seal, body construction, body materials, lid and lid seal, and marking. The metal flat pack and multilayer integral ceramic flat pack and DIP received the highest rankings, and the soft glass Cerdip and Cerpak types received the lowest rankings. Loss of package hermeticity due to lead and lid seal problems was found to be the predominant failure mode from the literature/data search. However, environmental test results showed that lead and lid seal failures due to thermal stressing was only a problem with the hard glass (Ceramic) body DIP utilizing a metal lid and/or bottom. Insufficient failure data were generated for the other package types tested to correlate test results with the package ranking.

  11. Expression and processing of human preprogastrin in murine medullary thyroid carcinoma cells.

    PubMed

    Daugherty, D F; Dickinson, C J; Takeuchi, T; Bachwich, D; Yamada, T

    1991-05-01

    Gastrin, the primary hormonal mediator of postprandial gastric acid secretion, is produced from its precursor progastrin by a series of posttranslational processing reactions including dibasic residue cleavage and carboxyl-terminal alpha-amidation. Progastrin contains three dibasic cleavage signals, Arg57Arg58, Lys74Lys75, and Arg94Arg95, that appear to be cleaved differently in different tissues. Differential processing is a potential means by which the production of biologically active peptides may be regulated in a tissue-specific manner. To study these reactions further, we used the pZipNeo SV(X) retroviral vector to express human gastrin cDNA in a heterologous cell line (MTC 6-23) known to be capable of processing other peptide precursors. The psi 2 packaging cell line transfected with the gastrin cDNA-retroviral construct (pSVXgas) produced progastrin, but no substantial amounts of processed amidated gastrin were detected. amounts of processed amidated gastrin were detected. In contrast, MTC 6-23 cells infected with the viral stock obtained from the supernatant of pSVXgas-transfected psi 2 cells produced carboxyl-terminally amidated gastrin in all of its standard molecular forms, including sulfated and nonsulfated forms of tetratriacontagastrin (G-34), heptadecagastrin (G-17), and tetradecagastrin (G-14). These studies indicate that heterologous endocrine cell lines infected with a retroviral-peptide cDNA construct can serve as useful models for peptide hormone posttranslational processing.

  12. Tractor Mechanics: Maintaining and Servicing the Fuel System. Learning Activity Packages 20-33.

    ERIC Educational Resources Information Center

    Clemson Univ., SC. Vocational Education Media Center.

    Learning activity packages are presented for instruction in tractor mechanics. The packages deal with the duties involved in maintaining the fuel system. The following fourteen learning activity packages are included: servicing fuel and air filters, servicing fuel tanks and lines, adjusting a carburetor, servicing a carburetor, servicing the…

  13. Potential of Equine Herpesvirus 1 as a Vector for Immunization

    PubMed Central

    Trapp, Sascha; von Einem, Jens; Hofmann, Helga; Köstler, Josef; Wild, Jens; Wagner, Ralf; Beer, Martin; Osterrieder, Nikolaus

    2005-01-01

    Key problems using viral vectors for vaccination and gene therapy are antivector immunity, low transduction efficiencies, acute toxicity, and limited capacity to package foreign genetic information. It could be demonstrated that animal and human cells were efficiently transduced with equine herpesvirus 1 (EHV-1) reconstituted from viral DNA maintained and manipulated in Escherichia coli. Between 13 and 23% of primary human CD3+, CD4+, CD8+, CD11b+, and CD19+ cells and more than 70% of CD4+ MT4 cells or various human tumor cell lines (MeWo, Huh7, HeLa, 293T, or H1299) could be transduced with one infectious unit of EHV-1 per cell. After intranasal instillation of EHV-1 into mice, efficient transgene expression in lungs was detectable. Successful immunization using EHV-1 was shown after delivery of the human immunodeficiency virus type 1 Pr55gag precursor by the induction of a Gag-specific CD8+ immune response in mice. Because EHV-1 was not neutralized by human sera containing high titers of antibodies directed against human herpesviruses 1 to 5, it is concluded that this animal herpesvirus has enormous potential as a vaccine vector, because it is able to efficiently transduce a variety of animal and human cells, has high DNA packaging capacity, and can conveniently be maintained and manipulated in prokaryotic cells. PMID:15827159

  14. Safety evaluation for packaging (onsite) concrete-lined waste packaging

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Romano, T.

    1997-09-25

    The Pacific Northwest National Laboratory developed a package to ship Type A, non-transuranic, fissile excepted quantities of liquid or solid radioactive material and radioactive mixed waste to the Central Waste Complex for storage on the Hanford Site.

  15. Laser Frequency Stabilization for Coherent Lidar Applications using Novel All-Fiber Gas Reference Cell Fabrication Technique

    NASA Technical Reports Server (NTRS)

    Meras, Patrick, Jr.; Poberezhskiy, Ilya Y.; Chang, Daniel H.; Levin, Jason; Spiers, Gary D.

    2008-01-01

    Compact hollow-core photonic crystal fiber (HC-PCF)gas frequency reference cell was constructed using a novel packaging technique that relies on torch-sealing a quartz filling tube connected to a mechanical splice between regular and hollow-core fibers. The use of this gas cell for laser frequency stabilization was demonstrated by locking a tunable diode laser to the center of the P9 line from the (nu)1+(nu)3 band of acetylene with RMS frequency error of 2.06 MHz over 2 hours. This effort was performed in support of a task to miniaturize the laser frequency stabilization subsystem of JPL/LMCT Laser Absorption Spectrometer (LAS) instrument.

  16. The Human Immunodeficiency Virus Type 1 Vif Protein Reduces Intracellular Expression and Inhibits Packaging of APOBEC3G (CEM15), a Cellular Inhibitor of Virus Infectivity

    PubMed Central

    Kao, Sandra; Khan, Mohammad A.; Miyagi, Eri; Plishka, Ron; Buckler-White, Alicia; Strebel, Klaus

    2003-01-01

    Replication of human immunodeficiency virus type 1 (HIV-1) in most primary cells and some immortalized T-cell lines depends on the activity of the viral infectivity factor (Vif). Vif has the ability to counteract a cellular inhibitor, recently identified as CEM15, that blocks infectivity of Vif-defective HIV-1 variants. CEM15 is identical to APOBEC3G and belongs to a family of proteins involved in RNA and DNA deamination. We cloned APOBEC3G from a human kidney cDNA library and confirmed that the protein acts as a potent inhibitor of HIV replication and is sensitive to the activity of Vif. We found that wild-type Vif inhibits packaging of APOBEC3G into virus particles in a dose-dependent manner. In contrast, biologically inactive variants carrying in-frame deletions in various regions of Vif or mutation of two highly conserved cysteine residues did not inhibit packaging of APOBEC3G. Interestingly, expression of APOBEC3G in the presence of wild-type Vif not only affected viral packaging but also reduced its intracellular expression level. This effect was not seen in the presence of biologically inactive Vif variants. Pulse-chase analyses did not reveal a significant difference in the stability of APOBEC3G in the presence or absence of Vif. However, in the presence of Vif, the rate of synthesis of APOBEC3G was slightly reduced. The reduction of intracellular APOBEC3G in the presence of Vif does not fully account for the Vif-induced reduction of virus-associated APOBEC3G, suggesting that Vif may function at several levels to prevent packaging of APOBEC3G into virus particles. PMID:14557625

  17. Corneal epithelial cell biocompatibility to silicone hydrogel and conventional hydrogel contact lens packaging solutions

    PubMed Central

    Tanti, N.C.; Jones, L.; Sheardown, H.

    2010-01-01

    Purpose Although all contact lenses (CLs) are applied initially to the eye directly from a packaging solution, little is known about the effects of these solutions on human corneal epithelial cells (HCECs). Due to the porous nature of CL materials, they have the potential to sorb components of the packaging solution during storage, which could then be subsequently released upon insertion of the CL on the eye. The purpose of this study was to investigate the effect of various packaging solutions on HCECs, using an in vitro model. Methods An in vitro assay was developed whereby various silicone hydrogels and conventional, poly-2-hydroxyethylmethacrylate  (polyHEMA)-based lens materials were removed directly from their packaging and then incubated for up to 24 h with HCECs. The effect of the retained and released packaging solution components on HCECs was assessed by measuring cell viability, adhesion phenotype, and apoptosis. Results Incubation of HCECs with CLs stored in borate-buffered packaging solutions resulted in a significant reduction in cell viability. Adherent cells incubated with these CLs also exhibited reduced levels of β1 and α3 integrin. Soaking borate-buffered packaged CLs in PBS before cell incubation resolved viability and integrin expression in all cases, with the exception of galyfilcon A and balafilcon A, from which a 20% reduction in cell viability was still observed. In comparison, CLs stored in phosphate-buffered packaging solutions had cellular viability and expression of integrins similar to control cells (cells incubated in the absence of a lens). When incubated with cells at a 10% concentration in serum-free medium, borate-buffered packaging solutions and borate-containing saline (Unisol 4) significantly reduced cell viability and integrin expression. Neither caspase activation nor annexin V binding was observed on cells following exposure to borate buffer solution. However, a significant decrease in reactive oxygen species was observed at 24 h. These latter results suggest that in vitro exposure to low concentration of borate/boric acid results in cell dysfunction, leading to necrosis rather than apoptosis. Conclusions Borate-buffered packaging solutions were shown to adversely affect the viability and integrin expression of HCECs in vitro. When used in ophthalmic packaging solutions, the antimicrobial properties of borate buffer may be outweighed by its relatively cytotoxic effects on cells. PMID:20169012

  18. Corneal epithelial cell biocompatibility to silicone hydrogel and conventional hydrogel contact lens packaging solutions.

    PubMed

    Gorbet, M B; Tanti, N C; Jones, L; Sheardown, H

    2010-02-19

    Although all contact lenses (CLs) are applied initially to the eye directly from a packaging solution, little is known about the effects of these solutions on human corneal epithelial cells (HCECs). Due to the porous nature of CL materials, they have the potential to sorb components of the packaging solution during storage, which could then be subsequently released upon insertion of the CL on the eye. The purpose of this study was to investigate the effect of various packaging solutions on HCECs, using an in vitro model. An in vitro assay was developed whereby various silicone hydrogels and conventional, poly-2-hydroxyethylmethacrylate (polyHEMA)-based lens materials were removed directly from their packaging and then incubated for up to 24 h with HCECs. The effect of the retained and released packaging solution components on HCECs was assessed by measuring cell viability, adhesion phenotype, and apoptosis. Incubation of HCECs with CLs stored in borate-buffered packaging solutions resulted in a significant reduction in cell viability. Adherent cells incubated with these CLs also exhibited reduced levels of beta(1) and alpha(3) integrin. Soaking borate-buffered packaged CLs in PBS before cell incubation resolved viability and integrin expression in all cases, with the exception of galyfilcon A and balafilcon A, from which a 20% reduction in cell viability was still observed. In comparison, CLs stored in phosphate-buffered packaging solutions had cellular viability and expression of integrins similar to control cells (cells incubated in the absence of a lens). When incubated with cells at a 10% concentration in serum-free medium, borate-buffered packaging solutions and borate-containing saline (Unisol 4) significantly reduced cell viability and integrin expression. Neither caspase activation nor annexin V binding was observed on cells following exposure to borate buffer solution. However, a significant decrease in reactive oxygen species was observed at 24 h. These latter results suggest that in vitro exposure to low concentration of borate/boric acid results in cell dysfunction, leading to necrosis rather than apoptosis. Borate-buffered packaging solutions were shown to adversely affect the viability and integrin expression of HCECs in vitro. When used in ophthalmic packaging solutions, the antimicrobial properties of borate buffer may be outweighed by its relatively cytotoxic effects on cells.

  19. A novel paclitaxel-loaded poly(d,l-lactide-co-glycolide)-Tween 80 copolymer nanoparticle overcoming multidrug resistance for lung cancer treatment

    PubMed Central

    Yuan, Xun; Ji, Wenxiang; Chen, Si; Bao, Yuling; Tan, Songwei; Lu, Shun; Wu, Kongming; Chu, Qian

    2016-01-01

    Drug resistance has become a main obstacle for the effective treatment of lung cancer. To address this problem, a novel biocompatible nanoscale package, poly(d,l-lactide-co-glycolide)-Tween 80, was designed and synthesized to overcome paclitaxel (PTX) resistance in a PTX-resistant human lung cancer cell line. The poly(d,l-lactide-co-glycolide) (PLGA)-Tween 80 nanoparticles (NPs) could efficiently load PTX and release the drug gradually. There was an increased level of uptake of PLGA-Tween 80 in PTX-resistant lung cancer cell line A549/T, which achieved a significantly higher level of cytotoxicity than both PLGA NP formulation and Taxol®. The in vivo antitumor efficacy also showed that PLGA-Tween 80 NP was more effective than Taxol®, indicating that PLGA-Tween 80 copolymer was a promising carrier for PTX in resistant lung cancer. PMID:27307727

  20. Detection of osteoclastic cell-cell fusion through retroviral vector packaging.

    PubMed

    Kondo, Takako; Ikeda, Kyoji; Matsuo, Koichi

    2004-11-01

    Cell-cell fusion generates multinucleated cells such as osteoclasts in bone, myotubes in muscle, and trophoblasts in placenta. Molecular details governing these fusion processes are still largely unknown. As a step toward identification of fusogenic genes, we tested the concept that retroviral vectors can be packaged as a result of cell-cell fusion. First, we introduced replication-deficient retroviral vectors expressing mCAT-1, which mediates fusogenic interaction with the retroviral envelope protein Env, into Chinese hamster ovary (CHO) cells to generate vector cells. Plasmids expressing virion proteins Gag, Pol, and Env were introduced into a separate culture of CHO cells to generate packaging cells. Co-culturing vector and packaging cells resulted in production of infectious retroviruses carrying the mCAT-1 gene as a consequence of cell-cell fusion. Second, we introduced a retroviral vector into primary osteoclast precursors and co-cultured them with established osteoclast precursor RAW264.7 cells, which turned out to harbor packaging activity. Packaged retroviral vector was detected in culture supernatants only where the osteoclast differentiation factor receptor activator for NF-kappaB ligand (RANKL) induced fusion between these two cell types. These data suggest that retrovirus production can occur as a result of cell-cell fusion. This provides a novel approach for isolating and characterizing fusogenic genes using retroviral expression vectors.

  1. Low temperature co-fired ceramic packaging of CMOS capacitive sensor chip towards cell viability monitoring.

    PubMed

    Halonen, Niina; Kilpijärvi, Joni; Sobocinski, Maciej; Datta-Chaudhuri, Timir; Hassinen, Antti; Prakash, Someshekar B; Möller, Peter; Abshire, Pamela; Kellokumpu, Sakari; Lloyd Spetz, Anita

    2016-01-01

    Cell viability monitoring is an important part of biosafety evaluation for the detection of toxic effects on cells caused by nanomaterials, preferably by label-free, noninvasive, fast, and cost effective methods. These requirements can be met by monitoring cell viability with a capacitance-sensing integrated circuit (IC) microchip. The capacitance provides a measurement of the surface attachment of adherent cells as an indication of their health status. However, the moist, warm, and corrosive biological environment requires reliable packaging of the sensor chip. In this work, a second generation of low temperature co-fired ceramic (LTCC) technology was combined with flip-chip bonding to provide a durable package compatible with cell culture. The LTCC-packaged sensor chip was integrated with a printed circuit board, data acquisition device, and measurement-controlling software. The packaged sensor chip functioned well in the presence of cell medium and cells, with output voltages depending on the medium above the capacitors. Moreover, the manufacturing of microfluidic channels in the LTCC package was demonstrated.

  2. A fully wafer-level packaged RF MEMS switch with low actuation voltage using a piezoelectric actuator

    NASA Astrophysics Data System (ADS)

    Park, Jae-Hyoung; Lee, Hee-Chul; Park, Yong-Hee; Kim, Yong-Dae; Ji, Chang-Hyeon; Bu, Jonguk; Nam, Hyo-Jin

    2006-11-01

    In this paper, a fully wafer-level packaged RF MEMS switch has been demonstrated, which has low operation voltage, using a piezoelectric actuator. The piezoelectric actuator was designed to operate at low actuation voltage for application to advanced mobile handsets. The dc contact type RF switch was packaged using the wafer-level bonding process. The CPW transmission lines and piezoelectric actuators have been fabricated on separate wafers and assembled together by the wafer-level eutectic bonding process. A gold and tin composite was used for eutectic bonding at a low temperature of 300 °C. Via holes interconnecting the electrical contact pads through the wafer were filled completely with electroplated copper. The fully wafer-level packaged RF MEMS switch showed an insertion loss of 0.63 dB and an isolation of 26.4 dB at 5 GHz. The actuation voltage of the switch was 5 V. The resonant frequency of the piezoelectric actuator was 38.4 kHz and the spring constant of the actuator was calculated to be 9.6 N m-1. The size of the packaged SPST (single-pole single-through) switch was 1.2 mm × 1.2 mm including the packaging sealing rim. The effect of the proposed package structure on the RF performance was characterized with a device having CPW through lines and vertical feed lines excluding the RF switches. The measured packaging loss was 0.2 dB and the return loss was 33.6 dB at 5 GHz.

  3. NASA DOD Lead Free Electronics Project

    NASA Technical Reports Server (NTRS)

    Kessel, Kurt R.

    2008-01-01

    The primary'technical objective of this project is to undertake comprehensive testing to generate information on failure modes/criteria to better understand the reliability of: Packages (e.g., Thin Small Outline Package [TSOP], Ball Grid Array [BGA], Plastic Dual In-line Package [PDIPD assembled and reworked with lead-free alloys Packages (e.g., TSOP, BGA, PDIP) assembled and reworked with mixed (lead/lead-free) alloys.

  4. Biochemical and ultrastructural characterization of a novel cell structure associated with immunoglobulin secretion in B-lymphocytes.

    PubMed

    Mazzaferro, P K; Repasky, E A; Black, J; Kubo, R T; Bankert, R B

    1987-01-01

    In the companion paper, it was established that a secretory form of immunoglobulin, sIg, is present at or near the cell surface. This unexpected occurrence of sIg was postulated to be due to the labelling of sIg which remains temporarily associated with the cell packaged in a vesicle which appears to bud from the plasma membrane at a single pole of the cell. The question that is addressed in this report is whether or not this polar accumulation of sIg represents a common pathway for the exit of this protein which is used by antibody-producing cells. This question is important since, in spite of the fact that the intracellular events associated with immunoglobulin synthesis (processing and movement between subcellular compartments) have been defined, very little data exists on how or where immunoglobulin finally leaves the plasma cell. This question was approached here by first demonstrating that the polar immunoglobulin secretory vesicles (ISV) are associated with several sIg-producing cells including other hybridomas, B-cell lines, and mitogen-activated spleen cells. The second approach was to characterize the ISV on the cell ultrastructurally and to establish that these vesicles are released from the cell carrying with them sIg. Isolated vesicles released from biosynthetically labeled Ig-producing cells were analyzed by SDS-PAGE in order to confirm the presence of sIg and to determine the number of other proteins associated with the ISV, their molecular weights, and the degree of disulfide crosslinking of the molecules comprising this structure. Finally, the kinetics of sIg release was established by a pulse chase protocol for biosynthetically labeled cells, and by monitoring the release of radioactive Ig from surface iodinated cells. As was predicted from our biochemical studies of the ISV, we observed a very slow phase of sIg release as well as a rapid release phase. Our studies have established that at least one of the pathways for the release of Ig from hybridomas, B-cell lines, and normal splenic B-cells is via a polar multivesiculated structure that we have termed ISV, and that the sIg can be released either as a free form of the protein or packaged within a satellite vesicle which may release the sIg later and perhaps at considerable distance from the cell that produced it.

  5. Conductor backed and shielded multi-layer coplanar waveguide designs on LTCC for RF carrier boards for packaging PICs

    NASA Astrophysics Data System (ADS)

    Marraccini, Philip J.; Jezzini, Moises A.; Peters, Frank H.

    2016-05-01

    Designing photonic integrated circuits (PICs) with packaging in mind is important since this impacts the performance of the final product. In coherent optical communication applications there are a large number of DC and RF lines that need routed to connect the PIC to the outer packaging. These RF lines should be impedance matched to the devices, isolated from each other, low loss and protected against electromagnetic interference (EMI) over the frequency range of interest to achieve the performance required for the application. Multilevel low temperature co-fired ceramic (LTCC) boards can be used as a carrier board connecting the PIC to the packaging due to its good RF performance, machinability, compatibility with hermetic sealing, and ability to integrate drivers into the board. Flexibility with layer numbers enables additional layers for shielding against electromagnetic interference or increased space for routing electrical connections. In this paper the design, simulations, and measured results for a set of 4 phase matched transmission lines in LTCC that would be used with an IQ MZM are presented. The measured 3dB bandwidth for a set of four phase matched transmission lines for an IQ MZM was measured to be 19.8 GHz.

  6. Method for Fabricating and Packaging an M.Times.N Phased-Array Antenna

    NASA Technical Reports Server (NTRS)

    Xu, Xiaochuan (Inventor); Chen, Yihong (Inventor); Chen, Ray T. (Inventor); Subbaraman, Harish (Inventor)

    2017-01-01

    A method for fabricating an M.times.N, P-bit phased-array antenna on a flexible substrate is disclosed. The method comprising ink jet printing and hardening alignment marks, antenna elements, transmission lines, switches, an RF coupler, and multilayer interconnections onto the flexible substrate. The substrate of the M.times.N, P-bit phased-array antenna may comprise an integrated control circuit of printed electronic components such as, photovoltaic cells, batteries, resistors, capacitors, etc. Other embodiments are described and claimed.

  7. 75 FR 81943 - Appliance Labeling Rule

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-29

    ....\\5\\ This substantial increase--from as many as 1,500 packaging styles per full- line bulb manufacturer in 1994 to as many as 3,500 packaging styles today--greatly increases the burden on manufacturers.... Specifically, the much larger number of packaging styles involved than in 1994, the difficulties posed by...

  8. NASA-DoD Lead-Free Electronics Project

    NASA Technical Reports Server (NTRS)

    Kessel, Kurt R.

    2009-01-01

    The primary technical objective of this project is to undertake comprehensive testing to generate information on failure modes/criteria to better understand the reliability of: (1) Packages (e.g., Thin Small Outline Package [TSOP], Ball Grid Array [BGA], Plastic Dual In-line Package [PDIP]) assembled and reworked with lead-free alloys, (2) Packages (e.g., TSOP, BGA, PDIP) assembled and reworked with mixed (lead/lead-free) alloys.

  9. Characteristics of Minimally Oversized Adeno-Associated Virus Vectors Encoding Human Factor VIII Generated Using Producer Cell Lines and Triple Transfection.

    PubMed

    Nambiar, Bindu; Cornell Sookdeo, Cathleen; Berthelette, Patricia; Jackson, Robert; Piraino, Susan; Burnham, Brenda; Nass, Shelley; Souza, David; O'Riordan, Catherine R; Vincent, Karen A; Cheng, Seng H; Armentano, Donna; Kyostio-Moore, Sirkka

    2017-02-01

    Several ongoing clinical studies are evaluating recombinant adeno-associated virus (rAAV) vectors as gene delivery vehicles for a variety of diseases. However, the production of vectors with genomes >4.7 kb is challenging, with vector preparations frequently containing truncated genomes. To determine whether the generation of oversized rAAVs can be improved using a producer cell-line (PCL) process, HeLaS3-cell lines harboring either a 5.1 or 5.4 kb rAAV vector genome encoding codon-optimized cDNA for human B-domain deleted Factor VIII (FVIII) were isolated. High-producing "masterwells" (MWs), defined as producing >50,000 vg/cell, were identified for each oversized vector. These MWs provided stable vector production for >20 passages. The quality and potency of the AAVrh8R/FVIII-5.1 and AAVrh8R/FVIII-5.4 vectors generated by the PCL method were then compared to those prepared via transient transfection (TXN). Southern and dot blot analyses demonstrated that both production methods resulted in packaging of heterogeneously sized genomes. However, the PCL-derived rAAV vector preparations contained some genomes >4.7 kb, whereas the majority of genomes generated by the TXN method were ≤4.7 kb. The PCL process reduced packaging of non-vector DNA for both the AAVrh8R/FVIII-5.1 and the AAVrh8R/FVIII-5.4 kb vector preparations. Furthermore, more DNA-containing viral particles were obtained for the AAVrh8R/FVIII-5.1 vector. In a mouse model of hemophilia A, animals administered a PCL-derived rAAV vector exhibited twofold higher plasma FVIII activity and increased levels of vector genomes in the liver than mice treated with vector produced via TXN did. Hence, the quality of oversized vectors prepared using the PCL method is greater than that of vectors generated using the TXN process, and importantly this improvement translates to enhanced performance in vivo.

  10. CalQuo: automated, simultaneous single-cell and population-level quantification of global intracellular Ca2+ responses.

    PubMed

    Fritzsche, Marco; Fernandes, Ricardo A; Colin-York, Huw; Santos, Ana M; Lee, Steven F; Lagerholm, B Christoffer; Davis, Simon J; Eggeling, Christian

    2015-11-13

    Detecting intracellular calcium signaling with fluorescent calcium indicator dyes is often coupled with microscopy techniques to follow the activation state of non-excitable cells, including lymphocytes. However, the analysis of global intracellular calcium responses both at the single-cell level and in large ensembles simultaneously has yet to be automated. Here, we present a new software package, CalQuo (Calcium Quantification), which allows the automated analysis and simultaneous monitoring of global fluorescent calcium reporter-based signaling responses in up to 1000 single cells per experiment, at temporal resolutions of sub-seconds to seconds. CalQuo quantifies the number and fraction of responding cells, the temporal dependence of calcium signaling and provides global and individual calcium-reporter fluorescence intensity profiles. We demonstrate the utility of the new method by comparing the calcium-based signaling responses of genetically manipulated human lymphocytic cell lines.

  11. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Claus, Claudia; Tzeng, W.-P.; Liebert, Uwe Gerd

    During serial passaging of rubella virus (RUB) in cell culture, the dominant species of defective-interfering RNA (DI) generated contains an in-frame deletion between the capsid protein (C) gene and E1 glycoprotein gene resulting in production of a C-E1 fusion protein that is necessary for the maintenance of the DI [Tzeng, W.P., Frey, T.K. (2006). C-E1 fusion protein synthesized by rubella virus DI RNAs maintained during serial passage. Virology 356 198-207.]. A BHK cell line stably expressing the RUB structural proteins was established which was used to package DIs into virus particles following transfection with in vitro transcripts from DI infectiousmore » cDNA constructs. Packaging of a DI encoding an in-frame C-GFP-E1 reporter fusion protein corresponding to the C-E1 fusion protein expressed in a native DI was only marginally more efficient than packaging of a DI encoding GFP, indicating that the C-E1 fusion protein did not function by enhancing packaging. However, infection with the DI encoding the C-GFP-E1 fusion protein (in the absence of wt RUB helper virus) resulted in formation of clusters of GFP-positive cells and the percentage of GFP-positive cells in the culture following infection remained relatively constant. In contrast, a DI encoding GFP did not form GFP-positive clusters and the percentage of GFP-positive cells declined by roughly half from 2 to 4 days post-infection. Cluster formation and sustaining the percentage of infected (GFP-positive) cells required the C part of the fusion protein, including the downstream but not the upstream of two arginine clusters (both of which are associated with RNA binding and association with mitochondrial p32 protein) and the E1 part through the transmembrane sequence, but not the C-terminal cytoplasmic tail. Among a collection of mutant DI constructs, cluster formation and sustaining infected cell percentage correlated with maintenance during serial passage with wt RUB. We hypothesize that cluster formation and sustaining infected cell percentage increase the likelihood of co-infection by a DI and wt RUB during serial passage thus enhancing maintenance of the DI. Cluster formation and sustaining infected cell percentage were found to be due to a combination of attenuated cytopathogenicity of DIs that express the C-E1 fusion protein and cell-to-cell movement of the DI. In infected cells, the C-GFP-E1 fusion protein was localized to potentially novel vesicular structures that appear to originate from ER-Golgi transport vacuoles. This species of DI expressing a C-E1 fusion protein that exhibits attenuated cytopathogenicity and the ability to increase the number of infected cells through cell-to-cell movement could be the basis for development of an attractive vaccine vector.« less

  12. Hermetic Packages For Millimeter-Wave Circuits

    NASA Technical Reports Server (NTRS)

    Herman, Martin I.; Lee, Karen A.; Lowry, Lynn E.; Carpenter, Alain; Wamhof, Paul

    1994-01-01

    Advanced hermetic packages developed to house electronic circuits operating at frequencies from 1 to 100 gigahertz and beyond. Signals coupled into and out of packages electromagnetically. Provides circuit packages small, lightweight, rugged, and inexpensive in mass production. Packages embedded in planar microstrip and coplanar waveguide circuits, in waveguide-to-planar and planar-to-waveguide circuitry, in waveguide-to-waveguide circuitry, between radiating (antenna) elements, and between planar transmission lines and radiating elements. Other applications in automotive, communication, radar, remote sensing, and biomedical electronic systems foreseen.

  13. An Examination of Superintendent Salaries and Compensation Packages in Kentucky

    ERIC Educational Resources Information Center

    Carroll, Lisa M.

    2014-01-01

    The salaries and compensation packages of women in the United States fall short of those to men holding similar employment positions. This study will look specifically at the salaries and compensation packages of current Kentucky school superintendents and investigate whether or not there exists discrepencies among them along gender lines. The…

  14. SMALL DIAMETER CEMENT LINING FROM STAIRWAY. United States Pipe ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    SMALL DIAMETER CEMENT LINING FROM STAIRWAY. - United States Pipe & Foundry Company Plant, Coating, Painting, Lining & Packaging Building, 2023 St. Louis Avenue at I-20/59, Bessemer, Jefferson County, AL

  15. Retractable pin dual in-line package test clip

    DOEpatents

    Bandzuch, Gregory S.; Kosslow, William J.

    1996-01-01

    This invention is a Dual In-Line Package (DIP) test clip for use when troubleshooting circuits containing DIP integrated circuits. This test clip is a significant improvement over existing DIP test clips in that it has retractable pins which will permit troubleshooting without risk of accidentally shorting adjacent pins together when moving probes to different pins on energized circuits or when the probe is accidentally bumped while taking measurements.

  16. HB-Line Plutonium Oxide Data Collection Strategy

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Watkins, R.; Varble, J.; Jordan, J.

    2015-05-26

    HB-Line and H-Canyon will handle and process plutonium material to produce plutonium oxide for feed to the Mixed Oxide Fuel Fabrication Facility (MFFF). However, the plutonium oxide product will not be transferred to the MFFF directly from HB-Line until it is packaged into a qualified DOE-STD-3013-2012 container. In the interim, HB-Line will load plutonium oxide into an inner, filtered can. The inner can will be placed in a filtered bag, which will be loaded into a filtered outer can. The outer can will be loaded into a certified 9975 with getter assembly in compliance with onsite transportation requirement, for subsequentmore » storage and transfer to the K-Area Complex (KAC). After DOE-STD-3013-2012 container packaging capabilities are established, the product will be returned to HB-Line to be packaged into a qualified DOE-STD-3013-2012 container. To support the transfer of plutonium oxide to KAC and then eventually to MFFF, various material and packaging data will have to be collected and retained. In addition, data from initial HB-Line processing operations will be needed to support future DOE-STD-3013-2012 qualification as amended by the HB-Line DOE Standard equivalency. As production increases, the volume of data to collect will increase. The HB-Line data collected will be in the form of paper copies and electronic media. Paper copy data will, at a minimum, consist of facility procedures, nonconformance reports (NCRs), and DCS print outs. Electronic data will be in the form of Adobe portable document formats (PDFs). Collecting all the required data for each plutonium oxide can will be no small effort for HB-Line, and will become more challenging once the maximum annual oxide production throughput is achieved due to the sheer volume of data to be collected. The majority of the data collected will be in the form of facility procedures, DCS print outs, and laboratory results. To facilitate complete collection of this data, a traveler form will be developed which identifies the required facility procedures, DCS print outs, and laboratory results needed to assemble a final data package for each HB-Line plutonium oxide interim oxide can. The data traveler may identify the specific values (data) required to be extracted from the collected facility procedures and DCS print outs. The data traveler may also identify associated criteria to be checked. Inevitably there will be procedure anomalies during the course of the HB-Line plutonium oxide campaign that will have to be addressed in a timely manner.« less

  17. SMALL DIAMETER PRECEMENT LINING FROM CATWALK ABOVE. United States ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    SMALL DIAMETER PRE-CEMENT LINING FROM CATWALK ABOVE. - United States Pipe & Foundry Company Plant, Coating, Painting, Lining & Packaging Building, 2023 St. Louis Avenue at I-20/59, Bessemer, Jefferson County, AL

  18. A fast hidden line algorithm with contour option. M.S. Thesis

    NASA Technical Reports Server (NTRS)

    Thue, R. E.

    1984-01-01

    The JonesD algorithm was modified to allow the processing of N-sided elements and implemented in conjunction with a 3-D contour generation algorithm. The total hidden line and contour subsystem is implemented in the MOVIE.BYU Display package, and is compared to the subsystems already existing in the MOVIE.BYU package. The comparison reveals that the modified JonesD hidden line and contour subsystem yields substantial processing time savings, when processing moderate sized models comprised of 1000 elements or less. There are, however, some limitations to the modified JonesD subsystem.

  19. Open-source Software for Exoplanet Atmospheric Modeling

    NASA Astrophysics Data System (ADS)

    Cubillos, Patricio; Blecic, Jasmina; Harrington, Joseph

    2018-01-01

    I will present a suite of self-standing open-source tools to model and retrieve exoplanet spectra implemented for Python. These include: (1) a Bayesian-statistical package to run Levenberg-Marquardt optimization and Markov-chain Monte Carlo posterior sampling, (2) a package to compress line-transition data from HITRAN or Exomol without loss of information, (3) a package to compute partition functions for HITRAN molecules, (4) a package to compute collision-induced absorption, and (5) a package to produce radiative-transfer spectra of transit and eclipse exoplanet observations and atmospheric retrievals.

  20. Packaging of silicon photonic devices: from prototypes to production

    NASA Astrophysics Data System (ADS)

    Morrissey, Padraic E.; Gradkowski, Kamil; Carroll, Lee; O'Brien, Peter

    2018-02-01

    The challenges associated with the photonic packaging of silicon devices is often underestimated and remains technically challenging. In this paper, we review some key enabling technologies that will allow us to overcome the current bottleneck in silicon photonic packaging; while also describing the recent developments in standardisation, including the establishment of PIXAPP as the worlds first open-access PIC packaging and assembly Pilot Line. These developments will allow the community to move from low volume prototype photonic packaged devices to large scale volume manufacturing, where the full commercialisation of PIC technology can be realised.

  1. TCGA Workflow: Analyze cancer genomics and epigenomics data using Bioconductor packages

    PubMed Central

    Bontempi, Gianluca; Ceccarelli, Michele; Noushmehr, Houtan

    2016-01-01

    Biotechnological advances in sequencing have led to an explosion of publicly available data via large international consortia such as The Cancer Genome Atlas (TCGA), The Encyclopedia of DNA Elements (ENCODE), and The NIH Roadmap Epigenomics Mapping Consortium (Roadmap). These projects have provided unprecedented opportunities to interrogate the epigenome of cultured cancer cell lines as well as normal and tumor tissues with high genomic resolution. The Bioconductor project offers more than 1,000 open-source software and statistical packages to analyze high-throughput genomic data. However, most packages are designed for specific data types (e.g. expression, epigenetics, genomics) and there is no one comprehensive tool that provides a complete integrative analysis of the resources and data provided by all three public projects. A need to create an integration of these different analyses was recently proposed. In this workflow, we provide a series of biologically focused integrative analyses of different molecular data. We describe how to download, process and prepare TCGA data and by harnessing several key Bioconductor packages, we describe how to extract biologically meaningful genomic and epigenomic data. Using Roadmap and ENCODE data, we provide a work plan to identify biologically relevant functional epigenomic elements associated with cancer. To illustrate our workflow, we analyzed two types of brain tumors: low-grade glioma (LGG) versus high-grade glioma (glioblastoma multiform or GBM). This workflow introduces the following Bioconductor packages: AnnotationHub, ChIPSeeker, ComplexHeatmap, pathview, ELMER, GAIA, MINET, RTCGAToolbox,  TCGAbiolinks. PMID:28232861

  2. TCGA Workflow: Analyze cancer genomics and epigenomics data using Bioconductor packages.

    PubMed

    Silva, Tiago C; Colaprico, Antonio; Olsen, Catharina; D'Angelo, Fulvio; Bontempi, Gianluca; Ceccarelli, Michele; Noushmehr, Houtan

    2016-01-01

    Biotechnological advances in sequencing have led to an explosion of publicly available data via large international consortia such as The Cancer Genome Atlas (TCGA), The Encyclopedia of DNA Elements (ENCODE), and The NIH Roadmap Epigenomics Mapping Consortium (Roadmap). These projects have provided unprecedented opportunities to interrogate the epigenome of cultured cancer cell lines as well as normal and tumor tissues with high genomic resolution. The Bioconductor project offers more than 1,000 open-source software and statistical packages to analyze high-throughput genomic data. However, most packages are designed for specific data types (e.g. expression, epigenetics, genomics) and there is no one comprehensive tool that provides a complete integrative analysis of the resources and data provided by all three public projects. A need to create an integration of these different analyses was recently proposed. In this workflow, we provide a series of biologically focused integrative analyses of different molecular data. We describe how to download, process and prepare TCGA data and by harnessing several key Bioconductor packages, we describe how to extract biologically meaningful genomic and epigenomic data. Using Roadmap and ENCODE data, we provide a work plan to identify biologically relevant functional epigenomic elements associated with cancer. To illustrate our workflow, we analyzed two types of brain tumors: low-grade glioma (LGG) versus high-grade glioma (glioblastoma multiform or GBM). This workflow introduces the following Bioconductor packages: AnnotationHub, ChIPSeeker, ComplexHeatmap, pathview, ELMER, GAIA, MINET, RTCGAToolbox,  TCGAbiolinks.

  3. Adenovirus Vector Pseudotyping in Fiber-Expressing Cell Lines: Improved Transduction of Epstein-Barr Virus-Transformed B Cells

    PubMed Central

    Von Seggern, Dan J.; Huang, Shuang; Fleck, Shonna Kaye; Stevenson, Susan C.; Nemerow, Glen R.

    2000-01-01

    While adenovirus (Ad) gene delivery vectors are useful in many gene therapy applications, their broad tropism means that they cannot be directed to a specific target cell. There are also a number of cell types involved in human disease which are not transducible with standard Ad vectors, such as Epstein-Barr virus (EBV)-transformed B lymphocytes. Adenovirus binds to host cells via the viral fiber protein, and Ad vectors have previously been retargeted by modifying the fiber gene on the viral chromosome. This requires that the modified fiber be able to bind to the cell in which the vector is grown, which prevents truly specific vector targeting. We previously reported a gene delivery system based on a fiber gene-deleted Ad type 5 (Ad5) vector (Ad5.βgal.ΔF) and packaging cells that express the viral fiber protein. Expression of different fibers in packaging cells will allow Ad retargeting without modifying the viral chromosome. Importantly, fiber proteins which can no longer bind to the producer cells can also be used. Using this approach, we generated for the first time pseudotyped Ad5.βgal.ΔF particles containing either the wild-type Ad5 fiber protein or a chimeric fiber with the receptor-binding knob domain of the Ad3 fiber. Particles equipped with the chimeric fiber bound to the Ad3 receptor rather than the coxsackievirus-adenovirus receptor protein used by Ad5. EBV-transformed B lymphocytes were infected efficiently by the Ad3-pseudotyped particles but poorly by virus containing the Ad5 fiber protein. The strategy described here represents a broadly applicable method for targeting gene delivery to specific cell types. PMID:10590124

  4. Production of recombinant adenovirus containing human interlukin-4 gene.

    PubMed

    Mojarrad, Majid; Abdolazimi, Yassan; Hajati, Jamshid; Modarressi, Mohammad Hossein

    2011-11-01

    Recombinant adenoviruses are currently used for a variety of purposes, including in vitro gene transfer, in vivo vaccination, and gene therapy. Ability to infect many cell types, high efficiency in gene transfer, entering both dividing and non dividing cells, and growing to high titers make this virus a good choice for using in various experiments. In the present experiment, a recombinant adenovirus containing human IL-4 coding sequence was made. IL-4 has several characteristics that made it a good choice for using in cancer gene therapy, controlling inflammatory diseases, and studies on autoimmune diseases. In brief, IL-4 coding sequence was amplified by and cloned in pAd-Track-CMV. Then, by means of homologous recombination between recombinant pAd-Track-CMV and Adeasy-1 plasmid in bacteria, recombinant adenovirus complete genome was made and IL-4 containing shuttle vector was incorporated into the viral backbone. After linearization, for virus packaging, viral genome was transfected into HEK-293 cell line. Viral production was conveniently followed with the aid of green fluorescent protein. Recombinant adenovirus produced here, was capable to infecting cell lines and express interlukin-4 in cell. This system can be used as a powerful, easy, and cost benefit tool in various studies on cancer gene therapy and also studies on immunogenetics.

  5. CellTree: an R/bioconductor package to infer the hierarchical structure of cell populations from single-cell RNA-seq data.

    PubMed

    duVerle, David A; Yotsukura, Sohiya; Nomura, Seitaro; Aburatani, Hiroyuki; Tsuda, Koji

    2016-09-13

    Single-cell RNA sequencing is fast becoming one the standard method for gene expression measurement, providing unique insights into cellular processes. A number of methods, based on general dimensionality reduction techniques, have been suggested to help infer and visualise the underlying structure of cell populations from single-cell expression levels, yet their models generally lack proper biological grounding and struggle at identifying complex differentiation paths. Here we introduce cellTree: an R/Bioconductor package that uses a novel statistical approach, based on document analysis techniques, to produce tree structures outlining the hierarchical relationship between single-cell samples, while identifying latent groups of genes that can provide biological insights. With cellTree, we provide experimentalists with an easy-to-use tool, based on statistically and biologically-sound algorithms, to efficiently explore and visualise single-cell RNA data. The cellTree package is publicly available in the online Bionconductor repository at: http://bioconductor.org/packages/cellTree/ .

  6. Profiling of Glycan Receptors for Minute Virus of Mice in Permissive Cell Lines Towards Understanding the Mechanism of Cell Recognition

    PubMed Central

    Halder, Sujata; Cotmore, Susan; Heimburg-Molinaro, Jamie; Smith, David F.; Cummings, Richard D.; Chen, Xi; Trollope, Alana J.; North, Simon J.; Haslam, Stuart M.; Dell, Anne; Tattersall, Peter; McKenna, Robert; Agbandje-McKenna, Mavis

    2014-01-01

    The recognition of sialic acids by two strains of minute virus of mice (MVM), MVMp (prototype) and MVMi (immunosuppressive), is an essential requirement for successful infection. To understand the potential for recognition of different modifications of sialic acid by MVM, three types of capsids, virus-like particles, wild type empty (no DNA) capsids, and DNA packaged virions, were screened on a sialylated glycan microarray (SGM). Both viruses demonstrated a preference for binding to 9-O-methylated sialic acid derivatives, while MVMp showed additional binding to 9-O-acetylated and 9-O-lactoylated sialic acid derivatives, indicating recognition differences. The glycans recognized contained a type-2 Galβ1-4GlcNAc motif (Neu5Acα2-3Galβ1-4GlcNAc or 3′SIA-LN) and were biantennary complex-type N-glycans with the exception of one. To correlate the recognition of the 3′SIA-LN glycan motif as well as the biantennary structures to their natural expression in cell lines permissive for MVMp, MVMi, or both strains, the N- and O-glycans, and polar glycolipids present in three cell lines used for in vitro studies, A9 fibroblasts, EL4 T lymphocytes, and the SV40 transformed NB324K cells, were analyzed by MALDI-TOF/TOF mass spectrometry. The cells showed an abundance of the sialylated glycan motifs recognized by the viruses in the SGM and previous glycan microarrays supporting their role in cellular recognition by MVM. Significantly, the NB324K showed fucosylation at the non-reducing end of their biantennary glycans, suggesting that recognition of these cells is possibly mediated by the Lewis X motif as in 3′SIA-LeX identified in a previous glycan microarray screen. PMID:24475195

  7. Lentiviral Delivery of HIV-1 Vpr Protein Induces Apoptosis in Transformed Cells

    NASA Astrophysics Data System (ADS)

    Stewart, Sheila A.; Poon, Betty; Jowett, Jeremy B. M.; Xie, Yiming; Chen, Irvin S. Y.

    1999-10-01

    Most current anticancer therapies act by inducing tumor cell stasis followed by apoptosis. HIV-1 Vpr effectively induces apoptosis of T cells after arrest of cells at a G2/M checkpoint. Here, we investigated whether this property of Vpr could be exploited for use as a potential anticancer agent. As a potentially safer alternative to transfer of genes encoding Vpr, we developed a method to efficiently introduce Vpr protein directly into cells. Vpr packaged into HIV-1 virions lacking a genome induced efficient cell cycle arrest and apoptosis. Introduction of Vpr into tumor cell lines of various tissue origin, including those bearing predisposing mutations in p53, XPA, and hMLH1, induced cell cycle arrest and apoptosis with high efficiency. Significantly, apoptosis mediated by virion-associated Vpr was more effective on rapidly dividing cells compared with slow-growing cells, thus, in concept, providing a potential differential effect between some types of tumor cells and surrounding normal cells. This model system provides a rationale and proof of concept for the development of potential cancer therapeutic agents based on the growth-arresting and apoptotic properties of Vpr.

  8. Effect of packaging material on enological parameters and volatile compounds of dry white wine.

    PubMed

    Revi, M; Badeka, A; Kontakos, S; Kontominas, M G

    2014-01-01

    The enological parameters and volatile compounds of white wine packaged in dark coloured glass and two commercial bag-in-box (BIB) pouches (low density polyethylene - LDPE and ethylene vinyl acetate - EVA lined) were determined for a period of 6 months at 20 °C. Parameters monitored included: titratable acidity, volatile acidity, pH, total SO2, free SO2, colour, volatile compounds and sensory attributes. The BIB packaging materials affected the titratable acidity, total and free SO2 and colour of wine. A substantial portion of the wine aroma compounds was adsorbed by the plastic materials or lost to the environment through leakage of the valve fitment. Between the two plastics, the LDPE lined pouch showed a considerably higher aroma sorption as compared to EVA. Wine packaged in glass retained the largest portion of its aroma compounds. Sensory evaluation showed that white wine packaged in both plastics was of acceptable quality for 3 months vs. at least 6 months for that in glass bottles. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Frequency stabilization of a 2.05 μm laser using hollow-core fiber CO2 frequency reference cell

    NASA Astrophysics Data System (ADS)

    Meras, Patrick; Poberezhskiy, Ilya Y.; Chang, Daniel H.; Spiers, Gary D.

    2010-04-01

    We have designed and built a hollow-core fiber frequency reference cell, filled it with CO2, and used it to demonstrate frequency stabilization of a 2.05 μm Tm:Ho:YLF laser using frequency modulation (FM) spectroscopy technique. The frequency reference cell is housed in a compact and robust hermetic package that contains a several meter long hollow-core photonic crystal fiber optically coupled to index-guiding fibers with a fusion splice on one end and a mechanical splice on the other end. The package has connectorized fiber pigtails and a valve used to evacuate, refill it, or adjust the gas pressure. We have demonstrated laser frequency standard deviation decreasing from >450MHz (free-running) to <2.4MHz (stabilized). The 2.05 μm laser wavelength is of particular interest for spectroscopic instruments due to the presence of many CO2 and H20 absorption lines in its vicinity. To our knowledge, this is the first reported demonstration of laser frequency stabilization at this wavelength using a hollow-core fiber reference cell. This approach enables all-fiber implementation of the optical portion of laser frequency stabilization system, thus making it dramatically more lightweight, compact, and robust than the traditional free-space version that utilizes glass or metal gas cells. It can also provide much longer interaction length of light with gas and does not require any alignment. The demonstrated frequency reference cell is particularly attractive for use in aircraft and space coherent lidar instruments for measuring atmospheric CO2 profile.

  10. Mechanical-Electrochemical-Thermal Simulation of Lithium-Ion Cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Santhanagopalan, Shriram; Zhang, Chao; Sprague, Michael A.

    2016-06-01

    Models capture the force response for single-cell and cell-string levels to within 15%-20% accuracy and predict the location for the origin of failure based on the deformation data from the experiments. At the module level, there is some discrepancy due to poor mechanical characterization of the packaging material between the cells. The thermal response (location and value of maximum temperature) agrees qualitatively with experimental data. In general, the X-plane results agree with model predictions to within 20% (pending faulty thermocouples, etc.); the Z-plane results show a bigger variability both between the models and test-results, as well as among multiple repeatsmore » of the tests. The models are able to capture the timing and sequence in voltage drop observed in the multi-cell experiments; the shapes of the current and temperature profiles need more work to better characterize propagation. The cells within packaging experience about 60% less force under identical impact test conditions, so the packaging on the test articles is robust. However, under slow-crush simulations, the maximum deformation of the cell strings with packaging is about twice that of cell strings without packaging.« less

  11. Combination screening in vitro identifies synergistically acting KP372-1 and cytarabine against acute myeloid leukemia.

    PubMed

    Österroos, A; Kashif, M; Haglund, C; Blom, K; Höglund, M; Andersson, C; Gustafsson, M G; Eriksson, A; Larsson, R

    2016-10-15

    Cytogenetic lesions often alter kinase signaling in acute myeloid leukemia (AML) and the addition of kinase inhibitors to the treatment arsenal is of interest. We have screened a kinase inhibitor library and performed combination testing to find promising drug-combinations for synergistic killing of AML cells. Cytotoxicity of 160 compounds in the library InhibitorSelect™ 384-Well Protein Kinase Inhibitor I was measured using the fluorometric microculture cytotoxicity assay (FMCA) in three AML cell lines. The 15 most potent substances were evaluated for dose-response. The 6 most cytotoxic compounds underwent combination synergy analysis based on the FMCA readouts after either simultaneous or sequential drug addition in AML cell lines. The 4 combinations showing the highest level of synergy were evaluated in 5 primary AML samples. Synergistic calculations were performed using the combination interaction analysis package COMBIA, written in R, using the Bliss independence model. Based on obtained results, an iterative combination search was performed using the therapeutic algorithmic combinatorial screen (TACS) algorithm. Of 160 substances, cell survival was ⩽50% at <0.5μM for Cdk/Crk inhibitor, KP372-1, synthetic fascaplysin, herbimycin A, PDGF receptor tyrosine kinase inhibitor IV and reference-drug cytarabine. KP372-1, synthetic fascaplysin or herbimycin A obtained synergy when combined with cytarabine in AML cell lines MV4-11 and HL-60. KP372-1 added 24h before cytarabine gave similar results in patient cells. The iterative search gave further improved synergy between cytarabine and KP372-1. In conclusion, our in vitro studies suggest that combining KP372-1 and cytarabine is a potent and synergistic drug combination in AML. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. [The effect of Foxc2 overexpression on the osteogenic properties of C3H10T1/2 cells].

    PubMed

    Wang, Min-Jiao; Si, Jia-Wen; Li, Hong-Liang; Ouyang, Ning-Juan; Shen, Guo-Fang

    2016-08-01

    To investigate the effect of Foxc2 overexpression on osteogenic and adipogenic differentiation of C3H10T1/2 cells. C3H10T1/2 cells were transfected with plenti-Foxc2 and selected with puromycin for stable clones. The expression of Foxc2 was determined by real-time PCR and Western blot. Cell proliferation was detected by CCK-8 kit. Cell cycle and apoptosis were detected by flow cytometry. The level of osteogenic biomarkers Runx2, OPN, OCN and adipogenic biomarker PPARγ were quantified by real-time PCR and Western blot. Alkaline phosphatase (ALP) staining and oil red staining were conducted to evaluate the effect of Foxc2 overexpression on osteogenic and adipogenic differentiation. Statistical analysis was performed using SPSS 17.0 software package. C3H10T1/2-Foxc2 cell line was successfully constructed and verified by direct sequencing and Foxc2 overexpression in vitro. Cell proliferation was reduced and cell cycle was blocked in G1/G0 phase. Enhanced ALP staining and reduced oil red staining were observed in C3H10T1/2-Foxc2 cells as compared with the control. Foxc2 overexpression up-regulated Runx2, OPN, OCN during osteogenic differentiation and down-regulated PPARγduring adipogenic differentiation. C3H10T1/2 cell line stably expressing Foxc2 gene was successfully established, cell proliferation was reduced, osteogenesis biomarkers were up-regulated during the osteogenesis by overexpression Foxc2, PPARγwas down-regulated during adipogenesis.

  13. Intracellular Crosslinking of Filoviral Nucleoproteins with Xintrabodies Restricts Viral Packaging

    PubMed Central

    Darling, Tamarand Lee; Sherwood, Laura Jo; Hayhurst, Andrew

    2017-01-01

    Viruses assemble large macromolecular repeat structures that become part of the infectious particles or virions. Ribonucleocapsids (RNCs) of negative strand RNA viruses are a prime example where repetition of nucleoprotein (NP) along the genome creates a core polymeric helical scaffold that accommodates other nucleocapsid proteins including viral polymerase. The RNCs are transported through the cytosol for packaging into virions through association with viral matrix proteins at cell membranes. We hypothesized that RNC would be ideal targets for crosslinkers engineered to promote aberrant protein–protein interactions, thereby blocking their orderly transport and packaging. Previously, we had generated single-domain antibodies (sdAbs) against Filoviruses that have all targeted highly conserved C-terminal regions of NP known to be repetitively exposed along the length of the RNCs of Marburgvirus (MARV) and Ebolavirus (EBOV). Our crosslinker design consisted of dimeric sdAb expressed intracellularly, which we call Xintrabodies (X- for crosslinking). Electron microscopy of purified NP polymers incubated with purified sdAb constructs showed NP aggregation occurred in a genus-specific manner with dimeric and not monomeric sdAb. A virus-like particle (VLP) assay was used for initial evaluation where we found that dimeric sdAb inhibited NP incorporation into VP40-based VLPs whereas monomeric sdAb did not. Inhibition of NP packaging was genus specific. Confocal microscopy revealed dimeric sdAb was diffuse when expressed alone but focused on pools of NP when the two were coexpressed, while monomeric sdAb showed ambivalent partition. Infection of stable Vero cell lines expressing dimeric sdAb specific for either MARV or EBOV NP resulted in smaller plaques and reduced progeny of cognate virus relative to wild-type Vero cells. Though the impact was marginal at later time-points, the collective data suggest that viral replication can be reduced by crosslinking intracellular NP using relatively small amounts of dimeric sdAb to restrict NP packaging. The stoichiometry and ease of application of the approach would likely benefit from transitioning away from intracellular expression of crosslinking sdAb to exogenous delivery of antibody. By retuning sdAb specificity, the approach of crosslinking highly conserved regions of assembly critical proteins may well be applicable to inhibiting replication processes of a broad spectrum of viruses. PMID:29021793

  14. SuperSegger: robust image segmentation, analysis and lineage tracking of bacterial cells.

    PubMed

    Stylianidou, Stella; Brennan, Connor; Nissen, Silas B; Kuwada, Nathan J; Wiggins, Paul A

    2016-11-01

    Many quantitative cell biology questions require fast yet reliable automated image segmentation to identify and link cells from frame-to-frame, and characterize the cell morphology and fluorescence. We present SuperSegger, an automated MATLAB-based image processing package well-suited to quantitative analysis of high-throughput live-cell fluorescence microscopy of bacterial cells. SuperSegger incorporates machine-learning algorithms to optimize cellular boundaries and automated error resolution to reliably link cells from frame-to-frame. Unlike existing packages, it can reliably segment microcolonies with many cells, facilitating the analysis of cell-cycle dynamics in bacteria as well as cell-contact mediated phenomena. This package has a range of built-in capabilities for characterizing bacterial cells, including the identification of cell division events, mother, daughter and neighbouring cells, and computing statistics on cellular fluorescence, the location and intensity of fluorescent foci. SuperSegger provides a variety of postprocessing data visualization tools for single cell and population level analysis, such as histograms, kymographs, frame mosaics, movies and consensus images. Finally, we demonstrate the power of the package by analyzing lag phase growth with single cell resolution. © 2016 John Wiley & Sons Ltd.

  15. Ultra high speed image processing techniques. [electronic packaging techniques

    NASA Technical Reports Server (NTRS)

    Anthony, T.; Hoeschele, D. F.; Connery, R.; Ehland, J.; Billings, J.

    1981-01-01

    Packaging techniques for ultra high speed image processing were developed. These techniques involve the development of a signal feedthrough technique through LSI/VLSI sapphire substrates. This allows the stacking of LSI/VLSI circuit substrates in a 3 dimensional package with greatly reduced length of interconnecting lines between the LSI/VLSI circuits. The reduced parasitic capacitances results in higher LSI/VLSI computational speeds at significantly reduced power consumption levels.

  16. Virus-Like Particles Derived from HIV-1 for Delivery of Nuclear Proteins: Improvement of Production and Activity by Protein Engineering.

    PubMed

    Robert, Marc-André; Lytvyn, Viktoria; Deforet, Francis; Gilbert, Rénald; Gaillet, Bruno

    2017-01-01

    Virus-like particles (VLPs) derived from retroviruses and lentiviruses can be used to deliver recombinant proteins without the fear of causing insertional mutagenesis to the host cell genome. In this study we evaluate the potential of an inducible lentiviral vector packaging cell line for VLP production. The Gag gene from HIV-1 was fused to a gene encoding a selected protein and it was transfected into the packaging cells. Three proteins served as model: the green fluorescent protein and two transcription factors-the cumate transactivator (cTA) of the inducible CR5 promoter and the human Krüppel-like factor 4 (KLF4). The sizes of the VLPs were 120-150 nm in diameter and they were resistant to freeze/thaw cycles. Protein delivery by the VLPs reached up to 100% efficacy in human cells and was well tolerated. Gag-cTA triggered up to 1100-fold gene activation of the reporter gene in comparison to the negative control. Protein engineering was required to detect Gag-KLF4 activity. Thus, insertion of the VP16 transactivation domain increased the activity of the VLPs by eightfold. An additional 2.4-fold enhancement was obtained by inserting nuclear export signal. In conclusion, our platform produced VLPs capable of efficient protein transfer, and it was shown that protein engineering can be used to improve the activity of the delivered proteins as well as VLP production.

  17. Solving Differential Equations in R: Package deSolve

    EPA Science Inventory

    In this paper we present the R package deSolve to solve initial value problems (IVP) written as ordinary differential equations (ODE), differential algebraic equations (DAE) of index 0 or 1 and partial differential equations (PDE), the latter solved using the method of lines appr...

  18. Implementation of interconnect simulation tools in spice

    NASA Technical Reports Server (NTRS)

    Satsangi, H.; Schutt-Aine, J. E.

    1993-01-01

    Accurate computer simulation of high speed digital computer circuits and communication circuits requires a multimode approach to simulate both the devices and the interconnects between devices. Classical circuit analysis algorithms (lumped parameter) are needed for circuit devices and the network formed by the interconnected devices. The interconnects, however, have to be modeled as transmission lines which incorporate electromagnetic field analysis. An approach to writing a multimode simulator is to take an existing software package which performs either lumped parameter analysis or field analysis and add the missing type of analysis routines to the package. In this work a traditionally lumped parameter simulator, SPICE, is modified so that it will perform lossy transmission line analysis using a different model approach. Modifying SPICE3E2 or any other large software package is not a trivial task. An understanding of the programming conventions used, simulation software, and simulation algorithms is required. This thesis was written to clarify the procedure for installing a device into SPICE3E2. The installation of three devices is documented and the installations of the first two provide a foundation for installation of the lossy line which is the third device. The details of discussions are specific to SPICE, but the concepts will be helpful when performing installations into other circuit analysis packages.

  19. The graphics and data acquisition software package

    NASA Technical Reports Server (NTRS)

    Crosier, W. G.

    1981-01-01

    A software package was developed for use with micro and minicomputers, particularly the LSI-11/DPD-11 series. The package has a number of Fortran-callable subroutines which perform a variety of frequently needed tasks for biomedical applications. All routines are well documented, flexible, easy to use and modify, and require minimal programmer knowledge of peripheral hardware. The package is also economical of memory and CPU time. A single subroutine call can perform any one of the following functions: (1) plot an array of integer values from sampled A/D data, (2) plot an array of Y values versus an array of X values; (3) draw horizontal and/or vertical grid lines of selectable type; (4) annotate grid lines with user units; (5) get coordinates of user controlled crosshairs from the terminal for interactive graphics; (6) sample any analog channel with program selectable gain; (7) wait a specified time interval, and (8) perform random access I/O of one or more blocks of a sequential disk file. Several miscellaneous functions are also provided.

  20. FABRICATION AND DEPLOYMENT OF THE 9979 TYPE AF RADIOACTIVE WASTE PACKAGING FOR THE DEPARTMENT OF ENERGY

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Blanton, P.; Eberl, K.

    2013-10-10

    This paper summarizes the development, testing, and certification of the 9979 Type A Fissile Packaging that replaces the UN1A2 Specification Shipping Package eliminated from Department of Transportation (DOT) 49 CFR 173. The DOT Specification Package was used for many decades by the U.S. nuclear industry as a fissile waste container until its removal as an authorized container by DOT. This paper will discuss stream lining procurement of high volume radioactive material packaging manufacturing, such as the 9979, to minimize packaging production costs without sacrificing Quality Assurance. The authorized content envelope (combustible and non-combustible) as well as planned content envelope expansionmore » will be discussed.« less

  1. Development of a functional cell-based assay that probes the specific interaction between influenza A virus NP and its packaging signal sequence RNA.

    PubMed

    Woo, Jiwon; Yu, Kyung Lee; Lee, Sun Hee; You, Ji Chang

    2015-02-06

    Although cis-acting packaging signal RNA sequences for the influenza virus NP encoding vRNA have been identified recently though genetic studies, little is known about the interaction between NP and the vRNA packaging signals either in vivo or in vitro. Here, we provide evidence that NP is able to interact specifically with the vRNA packaging sequence RNA within living cells and that the specific RNA binding activity of NP in vivo requires both the N-terminal and central region of the protein. This assay established would be a valuable tool for further detailed studies of the NP-packaging signal RNA interaction in living cells. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Production of Recombinant Adenovirus Containing Human Interlukin-4 Gene

    PubMed Central

    Mojarrad, Majid; Abdolazimi, Yassan; Hajati, Jamshid; Modarressi, Mohammad Hossein

    2011-01-01

    Objective(s) Recombinant adenoviruses are currently used for a variety of purposes, including in vitro gene transfer, in vivo vaccination, and gene therapy. Ability to infect many cell types, high efficiency in gene transfer, entering both dividing and non dividing cells, and growing to high titers make this virus a good choice for using in various experiments. In the present experiment, a recombinant adenovirus containing human IL-4 coding sequence was made. IL-4 has several characteristics that made it a good choice for using in cancer gene therapy, controlling inflammatory diseases, and studies on autoimmune diseases. Materials and Methods In brief, IL-4 coding sequence was amplified by and cloned in pAd-Track-CMV. Then, by means of homologous recombination between recombinant pAd-Track-CMV and Adeasy-1 plasmid in bacteria, recombinant adenovirus complete genome was made and IL-4 containing shuttle vector was incorporated into the viral backbone. After linearization, for virus packaging, viral genome was transfected into HEK-293 cell line. Viral production was conveniently followed with the aid of green fluorescent protein. Results Recombinant adenovirus produced here, was capable to infecting cell lines and express interlukin-4 in cell. Conclusion This system can be used as a powerful, easy, and cost benefit tool in various studies on cancer gene therapy and also studies on immunogenetics. PMID:23493491

  3. Raman Spectroscopy of DNA Packaging in Individual Human Sperm Cells distinguishes Normal from Abnormal Cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Huser, T; Orme, C; Hollars, C

    Healthy human males produce sperm cells of which about 25-40% have abnormal head shapes. Increases in the percentage of sperm exhibiting aberrant sperm head morphologies have been correlated with male infertility, and biochemical studies of pooled sperm have suggested that sperm with abnormal shape may contain DNA that has not been properly repackaged by protamine during spermatid development. We have used micro-Raman spectroscopy to obtain Raman spectra from individual human sperm cells and examined how differences in the Raman spectra of sperm chromatin correlate with cell shape. We show that Raman spectra of individual sperm cells contain vibrational marker modesmore » that can be used to assess the efficiency of DNA-packaging for each cell. Raman spectra obtained from sperm cells with normal shape provide evidence that DNA in these sperm is very efficiently packaged. We find, however, that the relative protein content per cell and DNA packaging efficiencies are distributed over a relatively wide range for sperm cells with both normal and abnormal shape. These findings indicate that single cell Raman spectroscopy should be a valuable tool in assessing the quality of sperm cells for in-vitro fertilization.« less

  4. Seismic Line Location Map Hot Pot Project, Humboldt County, Nevada 2010

    DOE Data Explorer

    Lane, Michael

    2010-01-01

    Seismic Line Location Map Hot Pot Project, Humboldt County, Nevada 2010. ArcGIS map package containing topographic base map, Township and Range layer, Oski BLM and private leases at time of survey, and locations, with selected shot points, of the five seismic lines.

  5. Independent Orbiter Assessment (IOA): Analysis of the electrical power generation/power reactant storage and distribution subsystem

    NASA Technical Reports Server (NTRS)

    Gotch, S. M.

    1986-01-01

    The results of the Independent Orbiter Assessment (IOA) of the Failure Modes and Effects Analysis (FMEA) and Critical Items List (CIL) are presented. The IOA approach features a top-down analysis of the hardware to determine failure modes, criticality, and potential critical items. To preserve independence, this analysis was accomplished without reliance upon the results contained within the NAA FMEA/CIL documentation. The independent analysis results corresponding to the Orbiter Electrical Power Generation (EPG)/Power Reactants Storage and Distribution (PRSD) System Hardware is documented. The EPG/PRSD hardware is required for performing critical functions of cryogenic hydrogen and oxygen storage and distribution to the Fuel Cell Powerplants (FCP) and Atmospheric Revitalization Pressure Control Subsystem (ARPCS). Specifically, the EPG/PRSD hardware consists of the following: Hydryogen (H2) tanks; Oxygen (O2) tanks; H2 Relief Valve/Filter Packages (HRVFP); O2 Relief Valve/Filter Packages (ORVFP); H2 Valve Modules (HVM); O2 Valve Modules (OVM); and O2 and H2 lines, components, and fittings.

  6. Proposed Ancestors of Phage Nucleic Acid Packaging Motors (and Cells)

    PubMed Central

    Serwer, Philip

    2011-01-01

    I present a hypothesis that begins with the proposal that abiotic ancestors of phage RNA and DNA packaging systems (and cells) include mobile shells with an internal, molecule-transporting cavity. The foundations of this hypothesis include the conjecture that current nucleic acid packaging systems have imprints from abiotic ancestors. The abiotic shells (1) initially imbibe and later also bind and transport organic molecules, thereby providing a means for producing molecular interactions that are links in the chain of events that produces ancestors to the first molecules that are both information carrying and enzymatically active, and (2) are subsequently scaffolds on which proteins assemble to form ancestors common to both shells of viral capsids and cell membranes. Emergence of cells occurs via aggregation and merger of shells and internal contents. The hypothesis continues by using proposed imprints of abiotic and biotic ancestors to deduce an ancestral thermal ratchet-based DNA packaging motor that subsequently evolves to integrate a DNA packaging ATPase that provides a power stroke. PMID:21994778

  7. Effects of retail style or food service style packaging type and storage time on sensory characteristics of bacon manufactured from commercially sourced pork bellies.

    PubMed

    Lowe, B K; Bohrer, B M; Holmer, S F; Boler, D D; Dilger, A C

    2014-06-01

    Objectives were to characterize differences in pork bellies that were stored frozen for different durations prior to processing and characterize sensory properties of the bacon derived from those bellies when stored in either retail or food service style packaging. Bellies (n = 102) were collected from 4 different time periods, fresh bellies (never frozen) and bellies frozen for 2, 5, or 7 mo, and manufactured into bacon under commercial conditions. Food service bacon was packaged in oxygen-permeable polyvinyl lined boxes layered on wax-covered lined paper and blast frozen (-33 °C) for 45 or 90 d after slicing. Retail bacon was vacuum-packaged in retail packages and refrigerated (2 °C) in the dark for 60 or 120 d after slicing. At the end of respective storage times after slicing, bacon was analyzed for sensory attributes and lipid oxidation. Off-flavor and oxidized odor of bacon increased (P < 0.01) with increasing storage time in both packaging types. Lipid oxidation increased (P < 0.01) as storage time increased from day 0 to day 45 in food service packaged bacon from frozen bellies, but was unchanged (P ≥ 0.07) with time in food service packaged bacon from fresh bellies. Lipid oxidation was also unchanged (P ≥ 0.21) over time in retail packaged bacon, with the exception of bellies frozen for 5 mo, which was increased from day 0 to day 90. Overall, off-flavor, oxidized odor, and lipid oxidation increased as storage time after processing increased. Freezing bellies before processing may exacerbate lipid oxidation as storage time after processing was extended. Bacon can be packaged and managed several different ways before it reaches the consumer. This research simulated food service (frozen) and retail packaged (refrigerated) bacon over a range of storage times after slicing. Off-flavor and oxidized odor increased as storage time after processing increased in both packaging types. Lipid oxidation increased as storage time after slicing increased to a greater extent in food service packaging. © 2014 Institute of Food Technologists®

  8. 76 FR 51257 - First-Class Package Service

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-18

    ... Ground, and Bound Printed Matter prices. * * * * * * * * 2.0 Additional Physical Standards by Class of... ``PRSRT'') First-Class Package'' (or ``PKG'') must be printed as part of; directly below; or to the left... follows:] b. * * * labeling: * * * * * 2. Line 2: ``FC PARCELS 3D.'' [Revise item 4.4c2 by changing ``FCM...

  9. 49 CFR Appendix E to Part 178 - Flame Penetration Resistance Test

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) SPECIFICATIONS FOR PACKAGINGS Pt...) At least three specimens of the outer packaging materials must be tested; (2) Each test must be... of cargo compartment lining materials to resist flame penetration with a 2 gallon per hour (GPH) #2...

  10. 49 CFR Appendix E to Part 178 - Flame Penetration Resistance Test

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) SPECIFICATIONS FOR PACKAGINGS Pt...) At least three specimens of the outer packaging materials must be tested; (2) Each test must be... of cargo compartment lining materials to resist flame penetration with a 2 gallon per hour (GPH) #2...

  11. 49 CFR Appendix E to Part 178 - Flame Penetration Resistance Test

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) SPECIFICATIONS FOR PACKAGINGS Pt...) At least three specimens of the outer packaging materials must be tested; (2) Each test must be... of cargo compartment lining materials to resist flame penetration with a 2 gallon per hour (GPH) #2...

  12. 49 CFR Appendix E to Part 178 - Flame Penetration Resistance Test

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) SPECIFICATIONS FOR PACKAGINGS Pt...) At least three specimens of the outer packaging materials must be tested; (2) Each test must be... of cargo compartment lining materials to resist flame penetration with a 2 gallon per hour (GPH) #2...

  13. 49 CFR 178.507 - Standards for plywood drums.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... PACKAGINGS Non-bulk Performance-Oriented Packaging Standards § 178.507 Standards for plywood drums. (a) The... effectiveness of the drum for the purpose intended. A material other than plywood, of at least equivalent..., lids must be lined with kraft paper or some other equivalent material which must be securely fastened...

  14. Encapsulants for protecting MEMS devices during post-packaging release etch

    DOEpatents

    Peterson, Kenneth A.

    2005-10-18

    The present invention relates to methods to protect a MEMS or microsensor device through one or more release or activation steps in a "package first, release later" manufacturing scheme: This method of fabrication permits wirebonds, other interconnects, packaging materials, lines, bond pads, and other structures on the die to be protected from physical, chemical, or electrical damage during the release etch(es) or other packaging steps. Metallic structures (e.g., gold, aluminum, copper) on the device are also protected from galvanic attack because they are protected from contact with HF or HCL-bearing solutions.

  15. Algebraic grid generation for coolant passages of turbine blades with serpentine channels and pin fins

    NASA Technical Reports Server (NTRS)

    Shih, T. I.-P.; Roelke, R. J.; Steinthorsson, E.

    1991-01-01

    In order to study numerically details of the flow and heat transfer within coolant passages of turbine blades, a method must first be developed to generate grid systems within the very complicated geometries involved. In this study, a grid generation package was developed that is capable of generating the required grid systems. The package developed is based on an algebraic grid generation technique that permits the user considerable control over how grid points are to be distributed in a very explicit way. These controls include orthogonality of grid lines next to boundary surfaces and ability to cluster about arbitrary points, lines, and surfaces. This paper describes that grid generation package and shows how it can be used to generate grid systems within complicated-shaped coolant passages via an example.

  16. User's Guide for the MapImage Reprojection Software Package, Version 1.01

    USGS Publications Warehouse

    Finn, Michael P.; Trent, Jason R.

    2004-01-01

    Scientists routinely accomplish small-scale geospatial modeling in the raster domain, using high-resolution datasets (such as 30-m data) for large parts of continents and low-resolution to high-resolution datasets for the entire globe. Recently, Usery and others (2003a) expanded on the previously limited empirical work with real geographic data by compiling and tabulating the accuracy of categorical areas in projected raster datasets of global extent. Geographers and applications programmers at the U.S. Geological Survey's (USGS) Mid-Continent Mapping Center (MCMC) undertook an effort to expand and evolve an internal USGS software package, MapImage, or mapimg, for raster map projection transformation (Usery and others, 2003a). Daniel R. Steinwand of Science Applications International Corporation, Earth Resources Observation Systems Data Center in Sioux Falls, S. Dak., originally developed mapimg for the USGS, basing it on the USGS's General Cartographic Transformation Package (GCTP). It operated as a command line program on the Unix operating system. Through efforts at MCMC, and in coordination with Mr. Steinwand, this program has been transformed from an application based on a command line into a software package based on a graphic user interface for Windows, Linux, and Unix machines. Usery and others (2003b) pointed out that many commercial software packages do not use exact projection equations and that even when exact projection equations are used, the software often results in error and sometimes does not complete the transformation for specific projections, at specific resampling resolutions, and for specific singularities. Direct implementation of point-to-point transformation with appropriate functions yields the variety of projections available in these software packages, but implementation with data other than points requires specific adaptation of the equations or prior preparation of the data to allow the transformation to succeed. Additional constraints apply to global raster data. It appears that some packages use the USGS's GCTP or similar point transformations without adaptation to the specific characteristics of raster data (Usery and others, 2003b). It is most common for programs to compute transformations of raster data in an inverse fashion. Such mapping can result in an erroneous position and replicate data or create pixels not in the original space. As Usery and others (2003a) indicated, mapimg performs a corresponding forward transformation to ensure the same location results from both methods. The primary benefit of this function is to mask cells outside the domain. MapImage 1.01 is now on the Web. You can download the User's Guide, source, and binaries from the following site: http://mcmcweb.er.usgs.gov/carto_research/projection/acc_proj_data.html

  17. HIV-1 adaptation studies reveal a novel Env-mediated homeostasis mechanism for evading lethal hypermutation by APOBEC3G

    PubMed Central

    Ikeda, Terumasa; Albin, John S.; Li, Ming; Thali, Markus

    2018-01-01

    HIV-1 replication normally requires Vif-mediated neutralization of APOBEC3 antiviral enzymes. Viruses lacking Vif succumb to deamination-dependent and -independent restriction processes. Here, HIV-1 adaptation studies were leveraged to ask whether viruses with an irreparable vif deletion could develop resistance to restrictive levels of APOBEC3G. Several resistant viruses were recovered with multiple amino acid substitutions in Env, and these changes alone are sufficient to protect Vif-null viruses from APOBEC3G-dependent restriction in T cell lines. Env adaptations cause decreased fusogenicity, which results in higher levels of Gag-Pol packaging. Increased concentrations of packaged Pol in turn enable faster virus DNA replication and protection from APOBEC3G-mediated hypermutation of viral replication intermediates. Taken together, these studies reveal that a moderate decrease in one essential viral activity, namely Env-mediated fusogenicity, enables the virus to change other activities, here, Gag-Pol packaging during particle production, and thereby escape restriction by the antiviral factor APOBEC3G. We propose a new paradigm in which alterations in viral homeostasis, through compensatory small changes, constitute a general mechanism used by HIV-1 and other viral pathogens to escape innate antiviral responses and other inhibitions including antiviral drugs. PMID:29677220

  18. Electromagnetic interference reduction using electromagnetic bandgap structures in packages, enclosures, cavities, and antennas

    NASA Astrophysics Data System (ADS)

    Mohajer Iravani, Baharak

    Electromagnetic interference (EMI) is a source of noise problems in electronic devices. The EMI is attributed to coupling between sources of radiation and components placed in the same media such as package or chassis. This coupling can be either through conducting currents or through radiation. The radiation of electromagnetic (EM) fields is supported by surface currents. Thus, minimizing these surface currents is considered a major and critical step to suppress EMI. In this work, we present novel strategies to confine surface currents in different applications including packages, enclosures, cavities, and antennas. The efficiency of present methods of EM noise suppression is limited due to different drawbacks. For example, the traditional use of lossy materials and absorbers suffers from considerable disadvantages including mechanical and thermal reliability leading to limited life time, cost, volume, and weight. In this work, we consider the use of Electromagnetic Band Gap (EBG) structures. These structures are suitable for suppressing surface currents within a frequency band denoted as the bandgap. Their design is straight forward, they are inexpensive to implement, and they do not suffer from the limitations of the previous methods. A new method of EM noise suppression in enclosures and cavity-backed antennas using mushroom-type EBG structures is introduced. The effectiveness of the EBG as an EMI suppresser is demonstrated using numerical simulations and experimental measurements. To allow integration of EBGs in printed circuit boards and packages, novel miniaturized simple planar EBG structures based on use of high-k dielectric material (epsilonr > 100) are proposed. The design consists of meander lines and patches. The inductive meander lines serve to provide current continuity bridges between the capacitive patches. The high-k dielectric material increases the effective capacitive load substantially in comparison to commonly used material with much lower dielectric constant. Meander lines can increase the effective inductive load which pushes down the lower edge of bandgap, thus resulting in a wider bandgap. Simulation results are included to show that the proposed EBG structures provide very wide bandgap (˜10GHz) covering the multiple harmonics of of currently available microprocessors and its harmonics. To speed up the design procedure, a model based on combination of lumped elements and transmission lines is proposed. The derived model predicts accurately the starting edge of bandgap. This result is verified with full-wave analysis. Finally, another novel compact wide band mushroom-type EBG structure using magneto-dielectric materials is designed. Numerical simulations show that the proposed EBG structure provides in-phase reflection bandgap which is several times greater than the one obtained from a conventional EBG operating at the same frequency while its cell size is smaller. This type of EBG structure can be used efficiently as a ground plane for low-profile wideband antennas.

  19. Airborne Sensor Thermal Management Solution

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ng, K. K.

    2015-06-03

    The customer wants to outfit aircraft (de Havilland Twin Otter) with optical sensors. In previous product generations the sensor line-of-sight direction was fixed – the sensor’s direction relied on the orientation of the aircraft. The next generation sensor will be packaged in a rotatable turret so that the line-of-sight is reasonably independent of the aircraft’s orientation. This turret will be mounted on a boom protruding from the side of the aircraft. The customer wants to outfit aircraft (de Havilland Twin Otter) with optical sensors. In previous product generations the sensor line-of-sight direction was fixed – the sensor’s direction relied onmore » the orientation of the aircraft. The next generation sensor will be packaged in a rotatable turret so that the line-of-sight is reasonably independent of the aircraft’s orientation. This turret will be mounted on a boom protruding from the side of the aircraft.« less

  20. Passing in Command Line Arguments and Parallel Cluster/Multicore Batching in R with batch.

    PubMed

    Hoffmann, Thomas J

    2011-03-01

    It is often useful to rerun a command line R script with some slight change in the parameters used to run it - a new set of parameters for a simulation, a different dataset to process, etc. The R package batch provides a means to pass in multiple command line options, including vectors of values in the usual R format, easily into R. The same script can be setup to run things in parallel via different command line arguments. The R package batch also provides a means to simplify this parallel batching by allowing one to use R and an R-like syntax for arguments to spread a script across a cluster or local multicore/multiprocessor computer, with automated syntax for several popular cluster types. Finally it provides a means to aggregate the results together of multiple processes run on a cluster.

  1. An adaptive optics package designed for astronomical use with a laser guide star tuned to an absorption line of atomic sodium

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Salmon, J.T.; Avicola, K.; Brase, J.M.

    1994-04-11

    We present the design and implementation of a very compact adaptive optic system that senses the return light from a sodium guide-star and controls a deformable mirror and a pointing mirror to compensate atmospheric perturbations in the wavefront. The deformable mirror has 19 electrostrictive actuators and triangular subapertures. The wavefront sensor is a Hartmann sensor with lenslets on triangular centers. The high-bandwidth steering mirror assembly incorporates an analog controller that samples the tilt with an avalanche photodiode quad cell. An {line_integral}/25 imaging leg focuses the light into a science camera that can either obtain long-exposure images or speckle data. Inmore » laboratory tests overall Strehl ratios were improved by a factor of 3 when a mylar sheet was used as an aberrator. The crossover frequency at unity gain is 30 Hz.« less

  2. Practical Issues in Estimating Classification Accuracy and Consistency with R Package cacIRT

    ERIC Educational Resources Information Center

    Lathrop, Quinn N.

    2015-01-01

    There are two main lines of research in estimating classification accuracy (CA) and classification consistency (CC) under Item Response Theory (IRT). The R package cacIRT provides computer implementations of both approaches in an accessible and unified framework. Even with available implementations, there remains decisions a researcher faces when…

  3. Expression of human argininosuccinate synthetase after retroviral-mediated gene transfer.

    PubMed

    Wood, P A; Partridge, C A; O'Brien, W E; Beaudet, A L

    1986-09-01

    The cDNA sequence for human argininosuccinate synthetase (AS) was introduced into plasmid expression vectors with an SV40 promoter or Rous sarcoma virus promoter to construct pSV2-AS and pRSV-AS, respectively, and human enzyme was synthesized after gene transfer into Chinese hamster cells. The functional cDNA was inserted into the retroviral vectors pZIP-NeoSV(X) and pZIP-NeoSV(B). Ecotropic AS retrovirus was produced after calcium-phosphate-mediated gene transfer of these constructions into the packaging cell line psi-2, and viral titers up to 10(5) CFU/ml were obtained. Recombinant AS retrovirus was evaluated by detecting G-418-resistant colonies after infection of the rodent cells, XC, NRK, and 3T3. Colonies were also obtained when infected XC cells were selected in citrulline medium for expression of AS activity. Southern blot analysis of infected cells demonstrated that the recombinant retroviral genome was not altered grossly after infecting some rodent cells, while other cells showed evidence of rearrangement. A rapid assay for detecting AS retrovirus was developed based on the incorporation of [14C]citrulline into protein by intact 3T3 cells or XC cells.

  4. Targeting of phage particles towards endothelial cells by antibodies selected through a multi-parameter selection strategy.

    PubMed

    Mandrup, Ole A; Lykkemark, Simon; Kristensen, Peter

    2017-02-10

    One of the hallmarks of cancer is sustained angiogenesis. Here, normal endothelial cells are activated, and their formation of new blood vessels leads to continued tumour growth. An improved patient condition is often observed when angiogenesis is prevented or normalized through targeting of these genomically stable endothelial cells. However, intracellular targets constitute a challenge in therapy, as the agents modulating these targets have to be delivered and internalized specifically to the endothelial cells. Selection of antibodies binding specifically to certain cell types is well established. It is nonetheless a challenge to ensure that the binding of antibodies to the target cell will mediate internalization. Previously selection of such antibodies has been performed targeting cancer cell lines; most often using either monovalent display or polyvalent display. In this article, we describe selections that isolate internalizing antibodies by sequential combining monovalent and polyvalent display using two types of helper phages, one which increases display valence and one which reduces background. One of the selected antibodies was found to mediate internalization into human endothelial cells, although our results confirms that the single stranded nature of the DNA packaged into phage particles may limit applications aimed at targeting nucleic acids in mammalian cells.

  5. Elimination of endogenous aberrant kappa chain transcripts from sp2/0-derived hybridoma cells by specific ribozyme cleavage: utility in genetic therapy of HIV-1 infections.

    PubMed Central

    Duan, L; Pomerantz, R J

    1994-01-01

    The pooled degenerate-primer polymerase chain reaction (PCR) technology is now widely used in the amplification and cloning of murine hybridoma-specific immunoglobulin gene cDNAs. The design of primers is mainly based on the highly conserved 5' terminus of immunoglobulin gene variable regions and the constant region in the 3' terminus. Of note, most murine hybridoma cell lines are derived from the Sp2/0 cell line, which is demonstrated to express endogenous aberrant kappa chains (abV kappa). This high-level endogenous abV kappa mixes with specific kappa chains in the hybridomas and interferes with the efficiency of the reverse transcriptase (RT)-PCR cloning strategy. In this report, during the cloning of murine anti-human immunodeficiency virus type I (HIV-1) hybridoma immunoglobulin cDNAs, a specific primer-PCR screening system was developed, based on the abV kappa complementarity-defining region (CDR), to eliminate abV kappa-carrying plasmids. Furthermore, an abV kappa sequence-specific derived ribozyme was developed and packaged in a retroviral expression vector system. This abV kappa ribozyme can be transduced into different murine hybridomas, and expressed intracellularly to potently eliminate endogenous abV kappa RNA. Images PMID:7816635

  6. Plant Equipment Package Modernization Program. Volume 4-1. Model Lines. Shell, HE, M483/M107-155MM Case, Cartridge, M115B1, M148A1B1, M150B1-105MM Shell, HEAT-T, M456A1-105MM Fuze, PD, M739

    DTIC Science & Technology

    1976-04-01

    Cartridge, M115B1, M148A1B1, M15#1B1-15MM J .. Shell, HEAT-T, M456A1-105MM Fuze, PD, M739 # prepared for Project Manager Munitions Production Base...ENGINEERS PLANT EQUIPMENT PACKAGE MODERNIZATION PROGRAM Volume 4-1 Report No. 75-86-R-4- MODEL LINE DEVELOPMENT FUZE,PD, M739 prepared for Project...In preparing the model line for the manufacture of piece parts for the M739 fuze, a number of facts became obvious and affect the detailed de- [ sign

  7. MOPEX: a software package for astronomical image processing and visualization

    NASA Astrophysics Data System (ADS)

    Makovoz, David; Roby, Trey; Khan, Iffat; Booth, Hartley

    2006-06-01

    We present MOPEX - a software package for astronomical image processing and display. The package is a combination of command-line driven image processing software written in C/C++ with a Java-based GUI. The main image processing capabilities include creating mosaic images, image registration, background matching, point source extraction, as well as a number of minor image processing tasks. The combination of the image processing and display capabilities allows for much more intuitive and efficient way of performing image processing. The GUI allows for the control over the image processing and display to be closely intertwined. Parameter setting, validation, and specific processing options are entered by the user through a set of intuitive dialog boxes. Visualization feeds back into further processing by providing a prompt feedback of the processing results. The GUI also allows for further analysis by accessing and displaying data from existing image and catalog servers using a virtual observatory approach. Even though originally designed for the Spitzer Space Telescope mission, a lot of functionalities are of general usefulness and can be used for working with existing astronomical data and for new missions. The software used in the package has undergone intensive testing and benefited greatly from effective software reuse. The visualization part has been used for observation planning for both the Spitzer and Herschel Space Telescopes as part the tool Spot. The visualization capabilities of Spot have been enhanced and integrated with the image processing functionality of the command-line driven MOPEX. The image processing software is used in the Spitzer automated pipeline processing, which has been in operation for nearly 3 years. The image processing capabilities have also been tested in off-line processing by numerous astronomers at various institutions around the world. The package is multi-platform and includes automatic update capabilities. The software package has been developed by a small group of software developers and scientists at the Spitzer Science Center. It is available for distribution at the Spitzer Science Center web page.

  8. User’s Manual for the Modular Analysis-Package Libraries ANAPAC and TRANL

    DTIC Science & Technology

    1977-09-01

    number) Computer software Fourier transforms Computer software library Interpolation software Digitized data...disregarded to give the user a simplified plot. (b) The last digit of ISPACE determines the type of line to be drawn, provided KODE is not...negative. If the last digit of ISPACE is 0 a solid line is drawn 1 a dashed line is drawn - - - 2 a dotted line is drawn .... 3 a dash-dot line is

  9. Documentation for the MODFLOW 6 Groundwater Flow Model

    USGS Publications Warehouse

    Langevin, Christian D.; Hughes, Joseph D.; Banta, Edward R.; Niswonger, Richard G.; Panday, Sorab; Provost, Alden M.

    2017-08-10

    This report documents the Groundwater Flow (GWF) Model for a new version of MODFLOW called MODFLOW 6. The GWF Model for MODFLOW 6 is based on a generalized control-volume finite-difference approach in which a cell can be hydraulically connected to any number of surrounding cells. Users can define the model grid using one of three discretization packages, including (1) a structured discretization package for defining regular MODFLOW grids consisting of layers, rows, and columns, (2) a discretization by ver­tices package for defining layered unstructured grids consisting of layers and cells, and (3) a general unstruc­tured discretization package for defining flexible grids comprised of cells and their connection properties. For layered grids, a new capability is available for removing thin cells and vertically connecting cells overlying and underlying the thin cells. For complex problems involving water-table conditions, an optional Newton-Raphson formulation, based on the formulations in MODFLOW-NWT and MODFLOW-USG, can be acti­vated. Use of the Newton-Raphson formulation will often improve model convergence and allow solutions to be obtained for difficult problems that cannot be solved using the traditional wetting and drying approach. The GWF Model is divided into “packages,” as was done in previous MODFLOW versions. A package is the part of the model that deals with a single aspect of simulation. Packages included with the GWF Model include those related to internal calculations of groundwater flow (discretization, initial conditions, hydraulic conduc­tance, and storage), stress packages (constant heads, wells, recharge, rivers, general head boundaries, drains, and evapotranspiration), and advanced stress packages (streamflow routing, lakes, multi-aquifer wells, and unsaturated zone flow). An additional package is also available for moving water available in one package into the individual features of the advanced stress packages. The GWF Model also has packages for obtaining and controlling output from the model. This report includes detailed explanations of physical and mathematical concepts on which the GWF Model and its packages are based.Like its predecessors, MODFLOW 6 is based on a highly modular structure; however, this structure has been extended into an object-oriented framework. The framework includes a robust and generalized numeri­cal solution object, which can be used to solve many different types of models. The numerical solution object has several different matrix preconditioning options as well as several methods for solving the linear system of equations. In this new framework, the GWF Model itself is an object as are each of the GWF Model packages. A benefit of the object-oriented structure is that multiple objects of the same type can be used in a single sim­ulation. Thus, a single forward run with MODFLOW 6 may contain multiple GWF Models. GWF Models can be hydraulically connected using GWF-GWF Exchange objects. Connecting GWF models in different ways permits the user to utilize a local grid refinement strategy consisting of parent and child models or to couple adjacent GWF Models. An advantage of the approach implemented in MODFLOW 6 is that multiple models and their exchanges can be incorporated into a single numerical solution object. With this design, models can be tightly coupled at the matrix level.

  10. Capable Copper Electrodeposition Process for Integrated Circuit - substrate Packaging Manufacturing

    NASA Astrophysics Data System (ADS)

    Ghanbari, Nasrin

    This work demonstrates a capable reverse pulse deposition methodology to influence gap fill behavior inside microvia along with a uniform deposit in the fine line patterned regions for substrate packaging applications. Interconnect circuitry in IC substrate packages comprises of stacked microvia that varies in depth from 20microm to 100microm with an aspect ratio of 0.5 to 1.5 and fine line patterns defined by photolithography. Photolithography defined pattern regions incorporate a wide variety of feature sizes including large circular pad structures with diameter of 20microm - 200microm, fine traces with varying widths of 3microm - 30microm and additional planar regions to define a IC substrate package. Electrodeposition of copper is performed to establish the desired circuit. Electrodeposition of copper in IC substrate applications holds certain unique challenges in that they require a low cost manufacturing process that enables a void-free gap fill inside the microvia along with uniform deposition of copper on exposed patterned regions. Deposition time scales to establish the desired metal thickness for such packages could range from several minutes to few hours. This work showcases a reverse pulse electrodeposition methodology that achieves void-free gap fill inside the microvia and uniform plating in FLS (Fine Lines and Spaces) regions with significantly higher deposition rates than traditional approaches. In order to achieve this capability, systematic experimental and simulation studies were performed. A strong correlation of independent parameters that govern the electrodeposition process such as bath temperature, reverse pulse plating parameters and the ratio of electrolyte concentrations is shown to the deposition kinetics and deposition uniformity in fine patterned regions and gap fill rate inside the microvia. Additionally, insight into the physics of via fill process is presented with secondary and tertiary current simulation efforts. Such efforts lead to show "smart" control of deposition rate at the top and bottom of via to avoid void formation. Finally, a parametric effect on grain size and the ensuing copper metallurgical characteristics of bulk copper is also shown to enable high reliability substrate packages for the IC packaging industry.

  11. High-performance packaging for monolithic microwave and millimeter-wave integrated circuits

    NASA Technical Reports Server (NTRS)

    Shalkhauser, K. A.; Li, K.; Shih, Y. C.

    1992-01-01

    Packaging schemes were developed that provide low-loss, hermetic enclosure for advanced monolithic microwave and millimeter-wave integrated circuits (MMICs). The package designs are based on a fused quartz substrate material that offers improved radio frequency (RF) performance through 44 gigahertz (GHz). The small size and weight of the packages make them appropriate for a variety of applications, including phased array antenna systems. Packages were designed in two forms; one for housing a single MMIC chip, the second in the form of a multi-chip phased array module. The single chip array module was developed in three separate sizes, for chips of different geometry and frequency requirements. The phased array module was developed to address packaging directly for antenna applications, and includes transmission line and interconnect structures to support multi-element operation. All packages are fabricated using fused quartz substrate materials. As part of the packaging effort, a test fixture was developed to interface the single chip packages to conventional laboratory instrumentation for characterization of the packaged devices. The package and test fixture designs were both developed in a generic sense, optimizing performance for a wide range of possible applications and devices.

  12. In-vitro antitumor activity evaluation of hyperforin derivatives.

    PubMed

    Sun, Feng; Liu, Jin-Yun; He, Feng; Liu, Zhong; Wang, Rui; Wang, Dong-Mei; Wang, Yi-Fei; Yang, De-Po

    2011-08-01

    The derivatives of hyperforin, namely hyperforin acetate (2), 17,18,22,23,27,28,32,33-octahydrohyperforin acetate (3), and N,N-dicyclohexylamine salt of hyperforin (4), have been investigated for their antitumor properties. In-vitro studies demonstrated that 2 and 4 were active against HeLa (human cervical cancer), A375 (human malignant melanoma), HepG2 (human hepatocellular carcinoma), MCF-7 (human breast cancer), A549 (human nonsmall cell lung cancer), K562 (human chronic myeloid leukemia), and K562/ADR (human adriamycin-resistant K562) cell lines with IC(50) values in the range of 3.2-64.1 μM. The energy differences between highest occupied molecular orbital and lowest unoccupied molecular orbital of 2-4 were calculated to be 0.39778, 0.43106, and 0.30900 a.u., respectively, using the Gaussian 03 software package and ab initio method with the HF/6-311 G* basis set. The result indicated that the biological activity of 4 might be the strongest and that of 3 might be the weakest, which was in accordance with their corresponding antiproliferative effects against the tested tumor cell lines. Compound 4 caused cell cycle arrest at G2/M phase in flow cytometry experiment and induced apoptosis by 4',6-diamidino-2-phenylindole staining and Annexin V-FITC/PI (propidium iodide) double-labeled staining in HepG2 cells. The results indicated a potential for N,N-dicyclohexylamine salt of hyperforin as a new antitumor drug.

  13. ANALYSIS/PLOT: a graphics package for use with the SORT/ANALYSIS data bases

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sady, C.A.

    1983-08-01

    This report describes a graphics package that is used with the SORT/ANALYSIS data bases. The data listed by the SORT/ANALYSIS program can be presented in pie, bar, line, or Gantt chart form. Instructions for the use of the plotting program and descriptions of the subroutines are given in the report.

  14. A Comparison of Authoring Software for Developing Mathematics Self-Learning Software Packages.

    ERIC Educational Resources Information Center

    Suen, Che-yin; Pok, Yang-ming

    Four years ago, the authors started to develop a self-paced mathematics learning software called NPMaths by using an authoring package called Tencore. However, NPMaths had some weak points. A development team was hence formed to develop similar software called Mathematics On Line. This time the team used another development language called…

  15. Granatum: a graphical single-cell RNA-Seq analysis pipeline for genomics scientists.

    PubMed

    Zhu, Xun; Wolfgruber, Thomas K; Tasato, Austin; Arisdakessian, Cédric; Garmire, David G; Garmire, Lana X

    2017-12-05

    Single-cell RNA sequencing (scRNA-Seq) is an increasingly popular platform to study heterogeneity at the single-cell level. Computational methods to process scRNA-Seq data are not very accessible to bench scientists as they require a significant amount of bioinformatic skills. We have developed Granatum, a web-based scRNA-Seq analysis pipeline to make analysis more broadly accessible to researchers. Without a single line of programming code, users can click through the pipeline, setting parameters and visualizing results via the interactive graphical interface. Granatum conveniently walks users through various steps of scRNA-Seq analysis. It has a comprehensive list of modules, including plate merging and batch-effect removal, outlier-sample removal, gene-expression normalization, imputation, gene filtering, cell clustering, differential gene expression analysis, pathway/ontology enrichment analysis, protein network interaction visualization, and pseudo-time cell series construction. Granatum enables broad adoption of scRNA-Seq technology by empowering bench scientists with an easy-to-use graphical interface for scRNA-Seq data analysis. The package is freely available for research use at http://garmiregroup.org/granatum/app.

  16. Uptake of DNA by cancer cells without a transfection reagent.

    PubMed

    Kong, Yanping; Zhang, Xianbo; Zhao, Yongliang; Xue, Yanfang; Zhang, Ye

    2017-01-21

    Cancer cells exhibit elevated levels of glucose uptake and may obtain pre-formed, diet-derived fatty acids from the bloodstream to boost their rapid growth; they may also use nucleic acid from their microenvironment. The study of processing nucleic acid by cancer cells will help improve the understanding of the metabolism of cancer. DNA is commonly packaged into a viral or lipid particle to be transferred into cells; this process is called transfection in laboratory. Cancer cells are known for having gene mutations and the evolving ability of endocytosis. Their uptake of DNAs might be different from normal cells; they may take in DNAs directly from the environment. In this report, we studied the uptake of DNAs in cancer cells without a transfection reagent. A group of DNA fragments were prepared with PCR and labeled with isotope phosphorous-32 to test their uptake by Huh 7 (liver cancer) and THLE3 (normal liver cells) after incubation overnight by counting radioactivity of the cells' genomic DNA. Multiple cell lines including breast cancer and lung cancer were tested with the same method. DNA molecules were also labeled with fluorescence to test the location in the cells using a kit of "label it fluorescence in situ hybridization (FISH)" from Mirus (USA). The data demonstrated that hepatocellular carcinoma cells possess the ability to take in large DNA fragments directly without a transfection reagent whereas normal liver cells cannot. Huh7 and MDA-MB231 cells displayed a significantly higher Rhodamine density in the cytoplasmic phagosomes and this suggests that the mechanism of uptake of large DNA by cancer cells is likely endocytosis. The efficacy of uptake is related to the DNA's size. Some cell lines of lung cancer and breast cancer also showed similar uptake of DNA. In the present study, we have revealed the evidence that some cancer cells, but not nontumorigenic cells, can take DNA fragments directly from the environment without the aid of the transfecting reagent.

  17. Soluble soybean polysaccharide/TiO2 bionanocomposite film for food application.

    PubMed

    Salarbashi, Davoud; Tafaghodi, Mohsen; Bazzaz, Bibi Sedigheh Fazly

    2018-04-15

    In the current study, a set of biodegradable soybean polysaccharide (SSPS) nanocomposites containing different ratios of TiO 2 nanoparticles was characterized as new packaging system. X-ray diffraction (XRD) measurement showed that the crystalline structure of the TiO 2 nanoparticles remained intact in the polysaccharide matrix and the surface of nanocomposites containing 1-3% TiO 2 was observed morphologically uniform under scanning electron microscopy (SEM). Dynamic mechanical thermal analysis revealed that the magnitude of storage modulus was 3.62-fold higher in SPSS/TiO 2 nanocomposites containing 7 wt.% of TiO 2 than control SSPS indicating improvement in the physical properties of the film supposed to be utilized for food packaging. With respect to the concern over the safety of these nanocomposites, inductively coupled plasma-optical emission spectroscopy (ICP-OES) showed that no TiO 2 was detected in bread samples covered by SSPS/TiO 2 film and stored for 6 months. Similarly, the nanocomposite films only released a minuscule amount (21.05 ± 0.054 ppm) of TiO 2 in water. TiO 2 nanoparticles were found in the plasma membrane of epithelial cell line after long-term exposure (10-day) of these cells to large amounts of the free nanoparticles. SSPS/TiO 2 nanocomposites showed excellent antimicrobial activity against Staphylococcus aureus PTCC 1431 (ATCC 25923), while neither anti-cancerous nor pro-cancerous activity was observed for these nanocomposites denoting their neutrality with respect to cancer suppression or progression in gastrointestinal tract. In conclusion, SSPS/TiO 2 nanocomposites could be a promising packaging system for food industries' objective regarding their physical characteristics, low rate of Ti transition, and low health risk. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. Uv-Light Stabilization Additive Package For Solar Cell Module And Laminated Glass Applications

    DOEpatents

    Hanoka, Jack I.; Klemchuk, Peter P.

    2002-03-05

    An ultraviolet light stabilization additive package is used in an encapsulant material that may be used in solar cell modules, laminated glass and a variety of other applications. The ultraviolet light stabilization additive package comprises a first hindered amine light stabilizer and a second hindered amine light stabilizer. The first hindered amine light stabilizer provides thermal oxidative stabilization, and the second hindered amine light stabilizer providing photo-oxidative stabilization.

  19. From Cells to Virus Particles: Quantitative Methods to Monitor RNA Packaging

    PubMed Central

    Ferrer, Mireia; Henriet, Simon; Chamontin, Célia; Lainé, Sébastien; Mougel, Marylène

    2016-01-01

    In cells, positive strand RNA viruses, such as Retroviridae, must selectively recognize their full-length RNA genome among abundant cellular RNAs to assemble and release particles. How viruses coordinate the intracellular trafficking of both RNA and protein components to the assembly sites of infectious particles at the cell surface remains a long-standing question. The mechanisms ensuring packaging of genomic RNA are essential for viral infectivity. Since RNA packaging impacts on several essential functions of retroviral replication such as RNA dimerization, translation and recombination events, there are many studies that require the determination of RNA packaging efficiency and/or RNA packaging ability. Studies of RNA encapsidation rely upon techniques for the identification and quantification of RNA species packaged by the virus. This review focuses on the different approaches available to monitor RNA packaging: Northern blot analysis, ribonuclease protection assay and quantitative reverse transcriptase-coupled polymerase chain reaction as well as the most recent RNA imaging and sequencing technologies. Advantages, disadvantages and limitations of these approaches will be discussed in order to help the investigator to choose the most appropriate technique. Although the review was written with the prototypic simple murine leukemia virus (MLV) and complex human immunodeficiency virus type 1 (HIV-1) in mind, the techniques were described in order to benefit to a larger community. PMID:27556480

  20. On-Line Retrieval II.

    ERIC Educational Resources Information Center

    Kurtz, Peter; And Others

    This report is concerned with the implementation of two interrelated computer systems: an automatic document analysis and classification package, and an on-line interactive information retrieval system which utilizes the information gathered during the automatic classification phase. Well-known techniques developed by Salton and Dennis have been…

  1. PLOT3D- DRAWING THREE DIMENSIONAL SURFACES

    NASA Technical Reports Server (NTRS)

    Canright, R. B.

    1994-01-01

    PLOT3D is a package of programs to draw three-dimensional surfaces of the form z = f(x,y). The function f and the boundary values for x and y are the input to PLOT3D. The surface thus defined may be drawn after arbitrary rotations. However, it is designed to draw only functions in rectangular coordinates expressed explicitly in the above form. It cannot, for example, draw a sphere. Output is by off-line incremental plotter or online microfilm recorder. This package, unlike other packages, will plot any function of the form z = f(x,y) and portrays continuous and bounded functions of two independent variables. With curve fitting; however, it can draw experimental data and pictures which cannot be expressed in the above form. The method used is division into a uniform rectangular grid of the given x and y ranges. The values of the supplied function at the grid points (x, y) are calculated and stored; this defines the surface. The surface is portrayed by connecting successive (y,z) points with straight-line segments for each x value on the grid and, in turn, connecting successive (x,z) points for each fixed y value on the grid. These lines are then projected by parallel projection onto the fixed yz-plane for plotting. This program has been implemented on the IBM 360/67 with on-line CDC microfilm recorder.

  2. Protecting enzymatic function through directed packaging into bacterial outer membrane vesicles

    PubMed Central

    Alves, Nathan J.; Turner, Kendrick B.; Medintz, Igor L.; Walper, Scott A.

    2016-01-01

    Bacteria possess innate machinery to transport extracellular cargo between cells as well as package virulence factors to infect host cells by secreting outer membrane vesicles (OMVs) that contain small molecules, proteins, and genetic material. These robust proteoliposomes have evolved naturally to be resistant to degradation and provide a supportive environment to extend the activity of encapsulated cargo. In this study, we sought to exploit bacterial OMV formation to package and maintain the activity of an enzyme, phosphotriesterase (PTE), under challenging storage conditions encountered for real world applications. Here we show that OMV packaged PTE maintains activity over free PTE when subjected to elevated temperatures (>100-fold more activity after 14 days at 37 °C), iterative freeze-thaw cycles (3.4-fold post four-cycles), and lyophilization (43-fold). We also demonstrate how lyophilized OMV packaged PTE can be utilized as a cell free reagent for long term environmental remediation of pesticide/chemical warfare contaminated areas. PMID:27117743

  3. Fully Packaged Carbon Nanotube Supercapacitors by Direct Ink Writing on Flexible Substrates.

    PubMed

    Chen, Bolin; Jiang, Yizhou; Tang, Xiaohui; Pan, Yayue; Hu, Shan

    2017-08-30

    The ability to print fully packaged integrated energy storage components (e.g., supercapacitors) is of critical importance for practical applications of printed electronics. Due to the limited variety of printable materials, most studies on printed supercapacitors focus on printing the electrode materials but rarely the full-packaged cell. This work presents for the first time the printing of a fully packaged single-wall carbon nanotube-based supercapacitor with direct ink writing (DIW) technology. Enabled by the developed ink formula, DIW setup, and cell architecture, the whole printing process is mask free, transfer free, and alignment free with precise and repeatable control on the spatial distribution of all constituent materials. Studies on cell design show that a wider electrode pattern and narrower gap distance between electrodes lead to higher specific capacitance. The as-printed fully packaged supercapacitors have energy and power performances that are among the best in recently reported planar carbon-based supercapacitors that are only partially printed or nonprinted.

  4. BIB mixers

    NASA Technical Reports Server (NTRS)

    1995-01-01

    We have determined that the multi-pin 'microprocessor style' packages in which current Blocked Impurity Band (BIB) devices are mounted will not meet our IF bandwidth spec of greater than 2 GHz for a practical mixer. Hence we have started to repackage the Ga:Ge BIB devices in new microwave compatible packages. The smaller size of the microwave package mount necessitates cutting the BIB array down to include only the 3 smallest detectors: 0.2, 0.4, and 0.6 mm sq. A FIR beam incident at f/1.5 can be focussed on the smallest element for wavelengths shorter than 100 microns. A more typical (easier) beam convergence of f/3 will require 0.4 mm elements at 100 microns and 0.6 mm elements at 170 microns wavelength. Since the device capacitance (parasitic loss) scales with detector size, there is a tradeoff of speed of response and optical convenience. Our existing optics produce only the slower convergence beam, so we need to redesign the optical layout and are looking at long focal length all-reflective microscope objectives. BIB detectors and the edge-coupled microbolometers have restricted IF bandwidths, an order of magnitude less than what is possible with the Schottky-diode mixers we currently use for astronomical observations. Consequently the frequencies of the FIR laser lines must be close to the astronomical line of interest to be an effective Local Oscillator (LO). We have therefore begun a coordinated effort to discover and measure new FIR laser transition lines in close frequency coincidence with important astrophysical lines. Most of this effort involves pumping isotopic variants of known good laser molecules with laser lines from isotopic variants of CO2. We have been most successful in detecting new FIR lines in deuterated ammonia. One line in particular is very close to the frequency of HD rotational line at 2675 GHz.

  5. Hypertext-based computer vision teaching packages

    NASA Astrophysics Data System (ADS)

    Marshall, A. David

    1994-10-01

    The World Wide Web Initiative has provided a means for providing hypertext and multimedia based information across the whole INTERNET. Many applications have been developed on such http servers. At Cardiff we have developed a http hypertext based multimedia server, the Cardiff Information Server, using the widely available Mosaic system. The server provides a variety of information ranging from the provision of teaching modules, on- line documentation, timetables for departmental activities to more light hearted hobby interests. One important and novel development to the server has been the development of courseware facilities. This ranges from the provision of on-line lecture notes, exercises and their solutions to more interactive teaching packages. A variety of disciplines have benefitted notably Computer Vision, and Image Processing but also C programming, X Windows, Computer Graphics and Parallel Computing. This paper will address the issues of the implementation of the Computer Vision and Image Processing packages, the advantages gained from using a hypertext based system and also will relate practical experiences of using the packages in a class environment. The paper addresses issues of how best to provide information in such a hypertext based system and how interactive image processing packages can be developed and integrated into courseware. The suite of tools developed facilitates a flexible and powerful courseware package that has proved popular in the classroom and over the Internet. The paper will also detail many future developments we see possible. One of the key points raised in the paper is that Mosaic's hypertext language (html) is extremely powerful and yet relatively straightforward to use. It is also possible to link in Unix calls so that programs and shells can be executed. This provides a powerful suite of utilities that can be exploited to develop many packages.

  6. MeV+R: using MeV as a graphical user interface for Bioconductor applications in microarray analysis

    PubMed Central

    Chu, Vu T; Gottardo, Raphael; Raftery, Adrian E; Bumgarner, Roger E; Yeung, Ka Yee

    2008-01-01

    We present MeV+R, an integration of the JAVA MultiExperiment Viewer program with Bioconductor packages. This integration of MultiExperiment Viewer and R is easily extensible to other R packages and provides users with point and click access to traditionally command line driven tools written in R. We demonstrate the ability to use MultiExperiment Viewer as a graphical user interface for Bioconductor applications in microarray data analysis by incorporating three Bioconductor packages, RAMA, BRIDGE and iterativeBMA. PMID:18652698

  7. ELSA: An integrated, semi-automated nebular abundance package

    NASA Astrophysics Data System (ADS)

    Johnson, Matthew D.; Levitt, Jesse S.; Henry, Richard B. C.; Kwitter, Karen B.

    We present ELSA, a new modular software package, written in C, to analyze and manage spectroscopic data from emission-line objects. In addition to calculating plasma diagnostics and abundances from nebular emission lines, the software provides a number of convenient features including the ability to ingest logs produced by IRAF's splot task, to semi-automatically merge spectra in different wavelength ranges, and to automatically generate various data tables in machine-readable or LaTeX format. ELSA features a highly sophisticated interstellar reddening correction scheme that takes into account temperature and density effects as well as He II contamination of the hydrogen Balmer lines. Abundance calculations are performed using a 5-level atom approximation with recent atomic data, based on R. Henry's ABUN program. Downloading and detailed documentation for all aspects of ELSA are available at the following URL:

  8. In cell mutational interference mapping experiment (in cell MIME) identifies the 5' polyadenylation signal as a dual regulator of HIV-1 genomic RNA production and packaging.

    PubMed

    Smyth, Redmond P; Smith, Maureen R; Jousset, Anne-Caroline; Despons, Laurence; Laumond, Géraldine; Decoville, Thomas; Cattenoz, Pierre; Moog, Christiane; Jossinet, Fabrice; Mougel, Marylène; Paillart, Jean-Christophe; von Kleist, Max; Marquet, Roland

    2018-05-18

    Non-coding RNA regulatory elements are important for viral replication, making them promising targets for therapeutic intervention. However, regulatory RNA is challenging to detect and characterise using classical structure-function assays. Here, we present in cell Mutational Interference Mapping Experiment (in cell MIME) as a way to define RNA regulatory landscapes at single nucleotide resolution under native conditions. In cell MIME is based on (i) random mutation of an RNA target, (ii) expression of mutated RNA in cells, (iii) physical separation of RNA into functional and non-functional populations, and (iv) high-throughput sequencing to identify mutations affecting function. We used in cell MIME to define RNA elements within the 5' region of the HIV-1 genomic RNA (gRNA) that are important for viral replication in cells. We identified three distinct RNA motifs controlling intracellular gRNA production, and two distinct motifs required for gRNA packaging into virions. Our analysis reveals the 73AAUAAA78 polyadenylation motif within the 5' PolyA domain as a dual regulator of gRNA production and gRNA packaging, and demonstrates that a functional polyadenylation signal is required for viral packaging even though it negatively affects gRNA production.

  9. In cell mutational interference mapping experiment (in cell MIME) identifies the 5′ polyadenylation signal as a dual regulator of HIV-1 genomic RNA production and packaging

    PubMed Central

    Smith, Maureen R; Jousset, Anne-Caroline; Despons, Laurence; Laumond, Géraldine; Decoville, Thomas; Cattenoz, Pierre; Moog, Christiane; Jossinet, Fabrice; Mougel, Marylène; Paillart, Jean-Christophe

    2018-01-01

    Abstract Non-coding RNA regulatory elements are important for viral replication, making them promising targets for therapeutic intervention. However, regulatory RNA is challenging to detect and characterise using classical structure-function assays. Here, we present in cell Mutational Interference Mapping Experiment (in cell MIME) as a way to define RNA regulatory landscapes at single nucleotide resolution under native conditions. In cell MIME is based on (i) random mutation of an RNA target, (ii) expression of mutated RNA in cells, (iii) physical separation of RNA into functional and non-functional populations, and (iv) high-throughput sequencing to identify mutations affecting function. We used in cell MIME to define RNA elements within the 5′ region of the HIV-1 genomic RNA (gRNA) that are important for viral replication in cells. We identified three distinct RNA motifs controlling intracellular gRNA production, and two distinct motifs required for gRNA packaging into virions. Our analysis reveals the 73AAUAAA78 polyadenylation motif within the 5′ PolyA domain as a dual regulator of gRNA production and gRNA packaging, and demonstrates that a functional polyadenylation signal is required for viral packaging even though it negatively affects gRNA production. PMID:29514260

  10. A Python library for FAIRer access and deposition to the Metabolomics Workbench Data Repository.

    PubMed

    Smelter, Andrey; Moseley, Hunter N B

    2018-01-01

    The Metabolomics Workbench Data Repository is a public repository of mass spectrometry and nuclear magnetic resonance data and metadata derived from a wide variety of metabolomics studies. The data and metadata for each study is deposited, stored, and accessed via files in the domain-specific 'mwTab' flat file format. In order to improve the accessibility, reusability, and interoperability of the data and metadata stored in 'mwTab' formatted files, we implemented a Python library and package. This Python package, named 'mwtab', is a parser for the domain-specific 'mwTab' flat file format, which provides facilities for reading, accessing, and writing 'mwTab' formatted files. Furthermore, the package provides facilities to validate both the format and required metadata elements of a given 'mwTab' formatted file. In order to develop the 'mwtab' package we used the official 'mwTab' format specification. We used Git version control along with Python unit-testing framework as well as continuous integration service to run those tests on multiple versions of Python. Package documentation was developed using sphinx documentation generator. The 'mwtab' package provides both Python programmatic library interfaces and command-line interfaces for reading, writing, and validating 'mwTab' formatted files. Data and associated metadata are stored within Python dictionary- and list-based data structures, enabling straightforward, 'pythonic' access and manipulation of data and metadata. Also, the package provides facilities to convert 'mwTab' files into a JSON formatted equivalent, enabling easy reusability of the data by all modern programming languages that implement JSON parsers. The 'mwtab' package implements its metadata validation functionality based on a pre-defined JSON schema that can be easily specialized for specific types of metabolomics studies. The library also provides a command-line interface for interconversion between 'mwTab' and JSONized formats in raw text and a variety of compressed binary file formats. The 'mwtab' package is an easy-to-use Python package that provides FAIRer utilization of the Metabolomics Workbench Data Repository. The source code is freely available on GitHub and via the Python Package Index. Documentation includes a 'User Guide', 'Tutorial', and 'API Reference'. The GitHub repository also provides 'mwtab' package unit-tests via a continuous integration service.

  11. 49 CFR 172.315 - Packages containing limited quantities.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... applicable, for the entry as shown in the § 172.101 Table, and placed within a square-on-point border in... to the package as to be readily visible. The width of line forming the square-on-point must be at... square-on-points bearing a single ID number, or a single square-on-point large enough to include each...

  12. MGGPOD: a Monte Carlo Suite for Modeling Instrumental Line and Continuum Backgrounds in Gamma-Ray Astronomy

    NASA Technical Reports Server (NTRS)

    Weidenspointner, G.; Harris, M. J.; Sturner, S.; Teegarden, B. J.; Ferguson, C.

    2004-01-01

    Intense and complex instrumental backgrounds, against which the much smaller signals from celestial sources have to be discerned, are a notorious problem for low and intermediate energy gamma-ray astronomy (approximately 50 keV - 10 MeV). Therefore a detailed qualitative and quantitative understanding of instrumental line and continuum backgrounds is crucial for most stages of gamma-ray astronomy missions, ranging from the design and development of new instrumentation through performance prediction to data reduction. We have developed MGGPOD, a user-friendly suite of Monte Carlo codes built around the widely used GEANT (Version 3.21) package, to simulate ab initio the physical processes relevant for the production of instrumental backgrounds. These include the build-up and delayed decay of radioactive isotopes as well as the prompt de-excitation of excited nuclei, both of which give rise to a plethora of instrumental gamma-ray background lines in addition t o continuum backgrounds. The MGGPOD package and documentation are publicly available for download. We demonstrate the capabilities of the MGGPOD suite by modeling high resolution gamma-ray spectra recorded by the Transient Gamma-Ray Spectrometer (TGRS) on board Wind during 1995. The TGRS is a Ge spectrometer operating in the 40 keV to 8 MeV range. Due to its fine energy resolution, these spectra reveal the complex instrumental background in formidable detail, particularly the many prompt and delayed gamma-ray lines. We evaluate the successes and failures of the MGGPOD package in reproducing TGRS data, and provide identifications for the numerous instrumental lines.

  13. Creation of Rift Valley Fever Viruses with Four-Segmented Genomes Reveals Flexibility in Bunyavirus Genome Packaging

    PubMed Central

    Oreshkova, Nadia; Moormann, Rob J. M.; Kortekaas, Jeroen

    2014-01-01

    ABSTRACT Bunyavirus genomes comprise a small (S), a medium (M), and a large (L) RNA segment of negative polarity. Although the untranslated regions have been shown to comprise signals required for transcription, replication, and encapsidation, the mechanisms that drive the packaging of at least one S, M, and L segment into a single virion to generate infectious virus are largely unknown. One of the most important members of the Bunyaviridae family that causes devastating disease in ruminants and occasionally humans is the Rift Valley fever virus (RVFV). We studied the flexibility of RVFV genome packaging by splitting the glycoprotein precursor gene, encoding the (NSm)GnGc polyprotein, into two individual genes encoding either (NSm)Gn or Gc. Using reverse genetics, six viruses with a segmented glycoprotein precursor gene were rescued, varying from a virus comprising two S-type segments in the absence of an M-type segment to a virus consisting of four segments (RVFV-4s), of which three are M-type. Despite that all virus variants were able to grow in mammalian cell lines, they were unable to spread efficiently in cells of mosquito origin. Moreover, in vivo studies demonstrated that RVFV-4s is unable to cause disseminated infection and disease in mice, even in the presence of the main virulence factor NSs, but induced a protective immune response against a lethal challenge with wild-type virus. In summary, splitting bunyavirus glycoprotein precursor genes provides new opportunities to study bunyavirus genome packaging and offers new methods to develop next-generation live-attenuated bunyavirus vaccines. IMPORTANCE Rift Valley fever virus (RVFV) causes devastating disease in ruminants and occasionally humans. Virions capable of productive infection comprise at least one copy of the small (S), medium (M), and large (L) RNA genome segments. The M segment encodes a glycoprotein precursor (GPC) protein that is cotranslationally cleaved into Gn and Gc, which are required for virus entry and fusion. We studied the flexibility of RVFV genome packaging and developed experimental live-attenuated vaccines by applying a unique strategy based on the splitting of the GnGc open reading frame. Several RVFV variants, varying from viruses comprising two S-type segments to viruses consisting of four segments (RVFV-4s), of which three are M-type, could be rescued and were shown to induce a rapid protective immune response. Altogether, the segmentation of bunyavirus GPCs provides a new method for studying bunyavirus genome packaging and facilitates the development of novel live-attenuated bunyavirus vaccines. PMID:25008937

  14. Creation of Rift Valley fever viruses with four-segmented genomes reveals flexibility in bunyavirus genome packaging.

    PubMed

    Wichgers Schreur, Paul J; Oreshkova, Nadia; Moormann, Rob J M; Kortekaas, Jeroen

    2014-09-01

    Bunyavirus genomes comprise a small (S), a medium (M), and a large (L) RNA segment of negative polarity. Although the untranslated regions have been shown to comprise signals required for transcription, replication, and encapsidation, the mechanisms that drive the packaging of at least one S, M, and L segment into a single virion to generate infectious virus are largely unknown. One of the most important members of the Bunyaviridae family that causes devastating disease in ruminants and occasionally humans is the Rift Valley fever virus (RVFV). We studied the flexibility of RVFV genome packaging by splitting the glycoprotein precursor gene, encoding the (NSm)GnGc polyprotein, into two individual genes encoding either (NSm)Gn or Gc. Using reverse genetics, six viruses with a segmented glycoprotein precursor gene were rescued, varying from a virus comprising two S-type segments in the absence of an M-type segment to a virus consisting of four segments (RVFV-4s), of which three are M-type. Despite that all virus variants were able to grow in mammalian cell lines, they were unable to spread efficiently in cells of mosquito origin. Moreover, in vivo studies demonstrated that RVFV-4s is unable to cause disseminated infection and disease in mice, even in the presence of the main virulence factor NSs, but induced a protective immune response against a lethal challenge with wild-type virus. In summary, splitting bunyavirus glycoprotein precursor genes provides new opportunities to study bunyavirus genome packaging and offers new methods to develop next-generation live-attenuated bunyavirus vaccines. Rift Valley fever virus (RVFV) causes devastating disease in ruminants and occasionally humans. Virions capable of productive infection comprise at least one copy of the small (S), medium (M), and large (L) RNA genome segments. The M segment encodes a glycoprotein precursor (GPC) protein that is cotranslationally cleaved into Gn and Gc, which are required for virus entry and fusion. We studied the flexibility of RVFV genome packaging and developed experimental live-attenuated vaccines by applying a unique strategy based on the splitting of the GnGc open reading frame. Several RVFV variants, varying from viruses comprising two S-type segments to viruses consisting of four segments (RVFV-4s), of which three are M-type, could be rescued and were shown to induce a rapid protective immune response. Altogether, the segmentation of bunyavirus GPCs provides a new method for studying bunyavirus genome packaging and facilitates the development of novel live-attenuated bunyavirus vaccines. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  15. Use of optical technique for inspection of warpage of IC packages

    NASA Astrophysics Data System (ADS)

    Toh, Siew-Lok; Chau, Fook S.; Ong, Sim Heng

    2001-06-01

    The packaging of IC packages has changed over the years, form dual-in-line, wire-bond, and pin-through-hole in printed wiring board technologies in the 1970s to ball grid array, chip scale and surface mount technologies in the 1990s. Reliability has been a big problem for manufacturers for some moisture-sensitive packages. One of the potential problems in plastic IC packages is moisture-induced popcorn effect which can arise during the reflow process. Shearography is a non-destructive inspection technique that may be used to detect the delamination and warpage of IC packages. It is non-contacting and permits a full-field observation of surface displacement derivatives. Another advantage of this technique is that it is able to give the real-time formation of the fringes which indicate flaws in the IC package under real-time simulation condition of Surface Mount Technology (SMT) IR reflow profile. It is extremely fast and convenient to study the true behavior of the packaging deformation during the SMT process. It can be concluded that shearography has the potential for the real- time detection, in situ and non-destructive inspection of IC packages during the surface mount process.

  16. 7 CFR 993.106 - In-line inspection.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false In-line inspection. 993.106 Section 993.106 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing... means inspection of prunes where samples are drawn from a flow of prunes prior to packaging. Effective...

  17. 7 CFR 993.504 - In-line inspection.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false In-line inspection. 993.504 Section 993.504 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing... inspection of prunes where samples are drawn from a flow of prunes prior to packaging. Effective Date Note...

  18. A high molecular weight-melanoma associated antigen-specific chimeric antigen receptor redirects lymphocytes to target human melanomas

    PubMed Central

    Burns, William R.; Zhao, Yangbing; Frankel, Timothy L.; Hinrichs, Christian S.; Zheng, Zhili; Xu, Hui; Feldman, Steven A.; Ferrone, Soldano; Rosenberg, Steven A.; Morgan, Richard A.

    2011-01-01

    Immunotherapy, particularly the adoptive cell transfer (ACT) of tumor infiltrating lymphocytes (TIL), is a very promising therapy for metastatic melanoma. Some patients unable to receive TIL have been successfully treated with autologous peripheral blood lymphocytes (PBL), genetically modified to express HLA class I antigen restricted, melanoma antigen-reactive T-cell receptors; however, substantial numbers of patients remain ineligible due to the lack of expression of the restricting HLA class I allele. We sought to overcome this limitation by designing a non-MHC-restricted, chimeric antigen receptor (CAR) targeting the high molecular weight-melanoma associated antigen (HMW-MAA), which is highly expressed on over 90% of human melanomas but has a restricted distribution in normal tissues. HMW-MAA-specific CARs containing an antigen recognition domain based on variations of the HMW-MAA-specific monoclonal antibody (mAb) 225.28S and a T-cell activation domain based on combinations of CD28, 4-1BB, and CD3ζ activation motifs were constructed within a retroviral vector to allow stable gene transfer into cells and their progeny. Following optimization of the HMW-MAA-specific CAR for expression and function in human PBL, these gene-modified T cells secreted cytokines, were cytolytic, and proliferated in response to HMW-MAA expressing cell lines. Furthermore, the receptor functioned in both CD4+ and CD8+ cells, was non-MHC-restricted, and reacted against explanted human melanomas. To evaluate this HMW-MAA-specific CAR in patients with metastatic melanoma, we developed a clinical-grade retroviral packaging line. This may represent a novel means to treat the majority of patients with advanced melanoma, most notably those unable to receive current ACT therapies. PMID:20395199

  19. Transcriptome changes induced in vitro by alcohol-containing mouthwashes in normal and dysplastic oral keratinocytes.

    PubMed

    Fox, Simon A; Currie, Sean S; Dalley, Andrew J; Farah, Camile S

    2018-05-01

    The role of alcohol-containing mouthwash as a risk factor for the development of oral cancer is a subject of conflicting epidemiological evidence in the literature despite alcohol being a recognised carcinogen. The aim of this study was to use in vitro models to investigate mechanistic and global gene expression effects of exposure to alcohol-containing mouthwash. Two brands of alcohol-containing mouthwash and their alcohol-free counterparts were used to treat two oral cell lines derived from normal (OKF6-TERT) and dysplastic (DOK) tissues. Genotoxicity was determined by Comet assay. RNA-seq was performed using the Ion Torrent platform. Bioinformatics analysis used R/Bioconductor packages with differential expression using DEseq2. Pathway enrichment analysis used EnrichR with the WikiPathways and Kegg databases. Both cell lines displayed dose-dependent DNA damage in response to acute exposure to ethanol and alcohol-containing mouthwashes as well as alcohol-free mouthwashes reconstituted with ethanol as shown by Comet assay. The transcriptomic effects of alcohol-containing mouthwash exposure were more complex with significant differential gene expression ranging from >2000 genes in dysplastic (DOK) cells to <100 genes in normal (OKF6-TERT) cells. Pathway enrichment analysis in DOK cells revealed alcohol-containing mouthwashes showed common features between the two brands used including DNA damage response as well as cancer-associated pathways. In OKF6-TERT cells, the most significantly enriched pathways involved inflammatory signalling. Alcohol-containing mouthwashes are genotoxic in vitro to normal and dysplastic oral keratinocytes and induce widespread changes in gene expression. Dysplastic cells are more susceptible to the transcriptomic effects of mouthwash. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  20. Thermal enclosures for electronically scanned pressure modules operating in cryogenic environments

    NASA Technical Reports Server (NTRS)

    Mitchell, Michael; Sealey, Bradley S.

    1989-01-01

    Specific guidelines to design, construct, and test ESP thermal enclosures for applications at cryogenic temperatures are given. The enclosures maintain the ESP modules at a constant temperature (10 C plus or minus 1 C) to minimize thermal zero and sensitivity shifts, to minimize the frequency of expensive on-line calibrations, and to avoid adverse effects on tunnel and model boundary layers. The enclosures are constructed of a rigid closed-cell foam and are capable of withstanding the stagnation pressures to 932kPa (135 psia) without reduction in thermal insulation properties. This construction procedure has been used to construct several thermal packages which have been successfully used in National Transonic Facility.

  1. Construction and production of oncotropic vectors, derived from MVM(p), that share reduced sequence homology with helper plasmids.

    PubMed

    Clément, Nathalie; Velu, Thierry; Brandenburger, Annick

    2002-09-01

    The production of currently available vectors derived from autonomous parvoviruses requires the expression of capsid proteins in trans, from helper sequences. Cotransfection of a helper plasmid always generates significant amounts of replication-competent virus (RCV) that can be reduced by the integration of helper sequences into a packaging cell line. Although stocks of minute virus of mice (MVM)-based vectors with no detectable RCV could be produced by transfection into packaging cells; the latter appear after one or two rounds of replication, precluding further amplification of the vector stock. Indeed, once RCVs become detectable, they are efficiently amplified and rapidly take over the culture. Theoretically RCV-free vector stocks could be produced if all homology between vector and helper DNA is eliminated, thus preventing homologous recombination. We constructed new vectors based on the structure of spontaneously occurring defective particles of MVM. Based on published observations related to the size of vectors and the sequence of the viral origin of replication, these vectors were modified by the insertion of foreign DNA sequences downstream of the transgene and by the introduction of a consensus NS-1 nick site near the origin of replication to optimize their production. In one of the vectors the inserted fragment of mouse genomic DNA had a synergistic effect with the modified origin of replication in increasing vector production.

  2. New insights into the nucleolar localization of a plant RNA virus-encoded protein that acts in both RNA packaging and RNA silencing suppression: involvement of importins alpha and relevance for viral infection.

    PubMed

    Pérez-Cañamás, Miryam; Hernández, Carmen

    2018-05-21

    Despite replication of plus strand RNA viruses takes place in the cytoplasm of host cells, different proteins encoded by these infectious agents have been shown to localize in the nucleus, with high accumulation at the nucleolus. In most cases, the molecular determinants and/or biological significance of such subcellular localization remain elusive. Recently, we reported that protein p37 encoded by Pelargonium line pattern virus (family Tombusviridae) acts in both RNA packaging and RNA silencing suppression. Connsistently with these functions, p37 was detected in the cytoplasm of plant cells though it was also present in the nucleus and, particularly, in the nucleolus. Here, we have aimed to gain further insights into factors influencing p37 nucleolar localization and into its potential relevance for viral infection. Besides mapping the protein region containing the nucleolar localization signal, we have found that p37 interacts with distinct members of the importin alpha family -main cellular transporters for nucleo-cytoplasmic traffic of proteins-, and that these interactions are crucial for nucleolar targeting of p37. Impairment of p37 nucleolar localization through down-regulation of importin alpha expression resulted in a reduction of viral accumulation, suggesting that sorting of the protein to the major subnuclear compartment is advantageous for the infection process.

  3. Statistical issues in signal extraction from microarrays

    NASA Astrophysics Data System (ADS)

    Bergemann, Tracy; Quiaoit, Filemon; Delrow, Jeffrey J.; Zhao, Lue Ping

    2001-06-01

    Microarray technologies are increasingly used in biomedical research to study genome-wide expression profiles in the post genomic era. Their popularity is largely due to their high throughput and economical affordability. For example, microarrays have been applied to studies of cell cycle, regulatory circuitry, cancer cell lines, tumor tissues, and drug discoveries. One obstacle facing the continued success of applying microarray technologies, however, is the random variaton present on microarrays: within signal spots, between spots and among chips. In addition, signals extracted by available software packages seem to vary significantly. Despite a variety of software packages, it appears that there are two major approaches to signal extraction. One approach is to focus on the identification of signal regions and hence estimation of signal levels above background levels. The other approach is to use the distribution of intensity values as a way of identifying relevant signals. Building upon both approaches, the objective of our work is to develop a method that is statistically rigorous and also efficient and robust. Statistical issues to be considered here include: (1) how to refine grid alignment so that the overall variation is minimized, (2) how to estimate the signal levels relative to the local background levels as well as the variance of this estimate, and (3) how to integrate red and green channel signals so that the ratio of interest is stable, simultaneously relaxing distributional assumptions.

  4. Concise classification of the genomic porcine endogenous retroviral gamma1 load to defined lineages.

    PubMed

    Klymiuk, Nikolai; Wolf, Eckhard; Aigner, Bernhard

    2008-02-05

    We investigated the infection history of porcine endogenous retroviruses (PERV) gamma1 by analyzing published env and LTR sequences. PERV sequences from various breeds, porcine cell lines and infected human primary cells were included in the study. We identified a considerable number of retroviral lineages indicating multiple independent colonization events of the porcine genome. A recent boost of the proviral load in an isolated pig herd and exclusive occurrence of distinct lineages in single studies indicated the ongoing colonization of the porcine genome with endogenous retroviruses. Retroviral recombination between co-packaged genomes was a general factor for PERV gamma1 diversity which indicated the simultaneous expression of different proviral loci over a period of time. In total, our detailed description of endogenous retroviral lineages is the prerequisite for breeding approaches to minimize the infectious potential of porcine tissues for the subsequent use in xenotransplantation.

  5. Plectin-1 Targeted AAV Vector for the Molecular Imaging of Pancreatic Cancer

    PubMed Central

    Konkalmatt, Prasad R.; Deng, Defeng; Thomas, Stephanie; Wu, Michael T.; Logsdon, Craig D.; French, Brent A.; Kelly, Kimberly A.

    2013-01-01

    Pancreatic ductal adenocarcinoma (PDAC) is highly malignant disease that is the fourth leading cause of cancer-related death in the US. Gene therapy using AAV vectors to selectively deliver genes to PDAC cells is an attractive treatment option for pancreatic cancer. However, most AAV serotypes display a broad spectrum of tissue tropism and none of the existing serotypes specifically target PDAC cells. This study tests the hypothesis that AAV2 can be genetically re-engineered to specifically target PDAC cells by modifying the capsid surface to display a peptide that has previously been shown to bind plectin-1. Toward this end, a Plectin-1 Targeting Peptide (PTP) was inserted into the loop IV region of the AAV2 capsid, and the resulting capsid (AAV-PTP) was used in a series of in vitro and in vivo experiments. In vitro, AAV-PTP was found to target all five human PDAC cell lines tested (PANC-1, MIA PaCa-2, HPAC, MPanc-96, and BxPC-3) preferentially over two non-neoplastic human pancreatic cell lines (human pancreatic ductal epithelial and human pancreatic stellate cells). In vivo, mice bearing subcutaneous tumor xenografts were generated using the PANC-1 cell line. Once tumors reached a size of ∼1–2 mm in diameter, the mice were injected intravenously with luciferase reporter vectors packaged in the either AAV-PTP or wild type AAV2 capsids. Luciferase expression was then monitored by bioluminescence imaging on days 3, 7, and 14 after vector injection. The results indicate that the AAV-PTP capsid displays a 37-fold preference for PANC-1 tumor xenographs over liver and other tissues; whereas the wild type AAV2 capsid displays a complementary preference for liver over tumors and other tissues. Together, these results establish proof-of-principle for the ability of PTP-modified AAV capsids to selectively target gene delivery to PDAC cells in vivo, which opens promising new avenues for the early detection, diagnosis, and treatment of pancreatic cancer. PMID:23616947

  6. Studies of the earth-limb absorptions in the near ultraviolet. [atmospheric chemistry

    NASA Technical Reports Server (NTRS)

    Murcray, D. G.

    1980-01-01

    The use of solar occultation techniques as a means of obtaining data on the photochemistry of the ozone layer was investigated. A spectrometer equipped with a photomultiplier detector and associated electronics necessary to amplify the output of the detector up to a level suitable for on-board recording and also for telemetering to the ground, was packaged with a solar telescope and flown by balloon in order to obtain solar spectra in the region between 3080 A and 3100 A with a resolution of close to 0.05 A. Data reduction techniques required to remove solar OH lines from atmospheric OH line spectra are described as well as the instrument package.

  7. Herpes Simplex Virus DNA Packaging without Measurable DNA Synthesis

    PubMed Central

    Church, Geoffrey A.; Dasgupta, Anindya; Wilson, Duncan W.

    1998-01-01

    Herpes simplex virus (HSV) type 1 DNA synthesis and packaging occur within the nuclei of infected cells; however, the extent to which the two processes are coupled remains unclear. Correct packaging is thought to be dependent upon DNA debranching or other repair processes, and such events commonly involve new DNA synthesis. Furthermore, the HSV UL15 gene product, essential for packaging, nevertheless localizes to sites of active DNA replication and may link the two events. It has previously been difficult to determine whether packaging requires concomitant DNA synthesis due to the complexity of these processes and of the viral life cycle; however, we have recently described a model system which simplifies the study of HSV assembly. Cells infected with HSV strain tsProt.A accumulate unpackaged capsids at the nonpermissive temperature of 39°C. Following release of the temperature block, these capsids proceed to package viral DNA in a single, synchronous wave. Here we report that, when DNA replication was inhibited prior to release of the temperature block, DNA packaging and later events in viral assembly nevertheless occurred at near-normal levels. We conclude that, under our conditions, HSV DNA packaging does not require detectable levels of DNA synthesis. PMID:9525593

  8. AuNP-DG: deoxyglucose-labeled gold nanoparticles as X-ray computed tomography contrast agents for cancer imaging.

    PubMed

    Aydogan, Bulent; Li, Ji; Rajh, Tijana; Chaudhary, Ahmed; Chmura, Steven J; Pelizzari, Charles; Wietholt, Christian; Kurtoglu, Metin; Redmond, Peter

    2010-10-01

    To study the feasibility of using 2-deoxy-D-glucose (2-DG)-labeled gold nanoparticle (AuNP-DG) as a computed tomography (CT) contrast agent with tumor targeting capability through in vitro experiments. Gold nanoparticles (AuNP) were fabricated and were conjugated with 2-deoxy-D-glucose. The human alveolar epithelial cancer cell line, A-549, was chosen for the in vitro cellular uptake assay. Two groups of cell samples were incubated with the AuNP-DG and the unlabeled AuNP, respectively. Following the incubation, the cells were washed with sterile PBS to remove the excess gold nanoparticles and spun to cell pellets using a centrifuge. The cell pellets were imaged using a microCT scanner immediately after the centrifugation. The reconstructed CT images were analyzed using a commercial software package. Significant contrast enhancement in the cell samples incubated with the AuNP-DG with respect to the cell samples incubated with the unlabeled AuNP was observed in multiple CT slices. Results from this study demonstrate enhanced uptake of 2-DG-labeled gold nanoparticle by cancer cells in vitro and warrant further experiments to study the exact molecular mechanism by which the AuNP-DG is internalized and retained in the tumor cells.

  9. 77 FR 14729 - Certain Lined Paper Products From India: Notice of Final Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-13

    ... DEPARTMENT OF COMMERCE International Trade Administration [A-533-843] Certain Lined Paper Products... Administration, International Trade Administration, Department of Commerce. SUMMARY: On October 7, 2011, the... & Vials Mfg. Co. Ltd.; AR Printing & Packaging (India) Pvt.; Bafna Exports; Cello International Pvt. Ltd...

  10. 78 FR 13016 - Certain Lined Paper Products From India: Notice of Partial Rescission of Countervailing Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-26

    ... From India: Notice of Partial Rescission of Countervailing Duty Administrative Review; 2011 AGENCY... an administrative review of the countervailing duty order on certain lined paper products from India..., including its requests for both Navneet Publications (India) Ltd. (Navneet) and A.R. Printing & Packaging...

  11. 78 FR 62584 - Certain Lined Paper Products From India: Preliminary Results of Countervailing Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-22

    ... From India: Preliminary Results of Countervailing Duty Administrative Review; Calendar Year 2011 AGENCY... (CVD) order on certain lined paper products from India. The period of review (POR) is January 1, 2011.... Printing & Packaging India Pvt. Ltd. (AR Printing). We have preliminarily determined that AR Printing...

  12. An Ada/SQL (Structured Query Language) Application Scanner.

    DTIC Science & Technology

    1988-03-01

    Digital ...8217 (" DIGITS "), 46 new STRING’ ("DO"), new STRING’ ("ELSE"), new STRING’ ("ELSIF"), new STRING’ ("END"), new STRING’ ("ENTRY"), new STRING’ ("EXCEPTION...INTEGERPRINT; generic type NUM is digits <>; package FLOATPRINT is package txtprts.ada 18 prcdr PR (FL inFL %YE LINE n LINTYPE UNCLASSIFIED procedure

  13. Abnormality in catalase import into peroxisomes leads to severe neurological disorder

    PubMed Central

    Sheikh, Faruk G.; Pahan, Kalipada; Khan, Mushfiquddin; Barbosa, Ernest; Singh, Inderjit

    1998-01-01

    Peroxisomal disorders are lethal inherited diseases caused by either defects in peroxisome assembly or dysfunction of single or multiple enzymatic function(s). The peroxisomal matrix proteins are targeted to peroxisomes via the interaction of peroxisomal targeting signal sequences 1 and 2 (PTS1 or PTS2) with their respective cytosolic receptors. We have studied human skin fibroblast cell lines that have multiple peroxisomal dysfunctions with normal packaging of PTS1 and PTS2 signal-containing proteins but lack catalase in peroxisomes. To understand the defect in targeting of catalase to peroxisomes and the loss of multiple enzyme activities, we transfected the mutant cells with normal catalase modified to contain either PTS1 or PTS2 signal sequence. We demonstrate the integrity of these pathways by targeting catalase into peroxisomes via PTS1 or PTS2 pathways. Furthermore, restoration of peroxisomal functions by targeting catalase-SKL protein (a catalase fused to the PTS1 sequence) to peroxisomes indicates that loss of multiple functions may be due to their inactivation by H2O2 or other oxygen species in these catalase-negative peroxisomes. In addition to enzyme activities, targeting of catalase-SKL chimera to peroxisomes also corrected the in situ levels of fatty acids and plasmalogens in these mutant cell lines. In normal fibroblasts treated with aminotriazole to inhibit catalase, we found that peroxisomal functions were inhibited to the level found in mutant cells, an observation that supports the conclusion that multiple peroxisomal enzyme defects in these patients are caused by H2O2 toxicity in catalase-negative peroxisomes. Moreover, targeting of catalase to peroxisomes via PTS1 and PTS2 pathways in these mutant cell lines suggests that there is another pathway for catalase import into peroxisomes and that an abnormality in this pathway manifests as a peroxisomal disease. PMID:9501198

  14. Meso-scale controlled motion for a microfluidic drop ejector.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Galambos, Paul C.; Givler, Richard C.; Pohl, Kenneth Roy

    2004-12-01

    The objective of this LDRD was to develop a uniquely capable, novel droplet solution based manufacturing system built around a new MEMS drop ejector. The development all the working subsystems required was completed, leaving the integration of these subsystems into a working prototype still left to accomplish. This LDRD report will focus on the three main subsystems: (1) MEMS drop ejector--the MEMS ''sideshooter'' effectively ejected 0.25 pl drops at 10 m/s, (2) packaging--a compact ejector package based on a modified EMDIP (Electro-Microfluidic Dual In-line Package--SAND2002-1941) was fabricated, and (3) a vision/stage system allowing precise ejector package positioning in 3 dimensionsmore » above a target was developed.« less

  15. Primary lithium cell life studies

    NASA Technical Reports Server (NTRS)

    Capulli, John; Donley, Sam; Deligiannis, Frank; Shen, David

    1990-01-01

    One solution for providing a truly independent power source is to package, within the critical subsystem element, a primary battery that can remain dormant for time periods as long as the mission life, which can be 10-15 years, maximum. When primary power from the spacecraft solar array/battery system is interrupted, the backup battery system, which is connected through a diode to the power input line, would automatically support the load to avoid a power interruption to the critical load for a time period long enough to ensure that ground control could access the satellite and correct the anomaly by sending appropriate commands to the spacecraft. Critical subsystems identified for the application are telemetry and command circuits, volatile computer memory, attitude control circuits, and some critical payloads. Due to volume packaging and weight restrictions that exist on most spacecraft, coupled with the long storage periods required, lithium cell technology was selected for the backup power source. Because of the high energy density (200-400 Wh/kg), long shelf life, and load capability, soluble cathode primary lithium technology was chosen. The most important lithium cell properties that require detail characterization for this application are capacity loss, shelf life, and the voltage delay mechanism. These are functions of storage time and temperature. During storage, a passive film builds up on the lithium electrode. The film protects the lithium electrode from progressive capacity decay but requires time to break down when a load is applied. This phenomenon results in a depressed voltage during the period of film breakdown which can last from fractions of a second to minutes.

  16. STILTS -- Starlink Tables Infrastructure Library Tool Set

    NASA Astrophysics Data System (ADS)

    Taylor, Mark

    STILTS is a set of command-line tools for processing tabular data. It has been designed for, but is not restricted to, use on astronomical data such as source catalogues. It contains both generic (format-independent) table processing tools and tools for processing VOTable documents. Facilities offered include crossmatching, format conversion, format validation, column calculation and rearrangement, row selection, sorting, plotting, statistical calculations and metadata display. Calculations on cell data can be performed using a powerful and extensible expression language. The package is written in pure Java and based on STIL, the Starlink Tables Infrastructure Library. This gives it high portability, support for many data formats (including FITS, VOTable, text-based formats and SQL databases), extensibility and scalability. Where possible the tools are written to accept streamed data so the size of tables which can be processed is not limited by available memory. As well as the tutorial and reference information in this document, detailed on-line help is available from the tools themselves. STILTS is available under the GNU General Public Licence.

  17. [Suppression of VEGF protein expression by arctigenin in oral squamous cell carcinoma].

    PubMed

    Pu, Guang-rui; Liu, Fa-yu; Wang, Bo

    2015-08-01

    To observe arctigenin's inhibitory effect on oral squamous cell carcinoma, and explore the possible mechanism. The expression of VEGF in 32 cases of oral squamous cell cancer and 20 adjacent tissue specimen were detected with immunohistochemistry. Human nude mouse transplantation tumor model of oral squamous cell cancer was prepared with HSC-3 cells line. Transplanted tumor growth and VEGF expression in transplanted tumor tissues were assayed after treatment with arctigenin. One-way ANOVA was used for comparison between groups with SPSS 16.0 software package. Compared with the adjacent tissue, immunohistochemical staining score of VEGF was significantly higher (P<0.01) in oral squamous cell carcinoma tissues. After treatment with arctigenin, the growth of oral squamous cell transplanted tumors in nude mouse was inhibited (P<0.05), and decreased weight in end point of observation was noted (P<0.05). There were significant differences between high dose group and low dose group (P<0.05). Compared with the nude mouse model group, the optical density of VEGF staining was significantly lower in arctigenin group (P<0.05). There were significant differences between high dose group and low dose group (P<0.05). Arctigenin can dose-dependently inhibit the growth of oral squamous cell carcinomas, and this effect may be related to down regulation of VEGF expression.

  18. Versatile Software Package For Near Real-Time Analysis of Experimental Data

    NASA Technical Reports Server (NTRS)

    Wieseman, Carol D.; Hoadley, Sherwood T.

    1998-01-01

    This paper provides an overview of a versatile software package developed for time- and frequency-domain analyses of experimental wind-tunnel data. This package, originally developed for analyzing data in the NASA Langley Transonic Dynamics Tunnel (TDT), is applicable for analyzing any time-domain data. A Matlab-based software package, TDT-analyzer, provides a compendium of commonly-required dynamic analysis functions in a user-friendly interactive and batch processing environment. TDT-analyzer has been used extensively to provide on-line near real-time and post-test examination and reduction of measured data acquired during wind tunnel tests of aeroelastically-scaled models of aircraft and rotorcraft as well as a flight test of the NASA High Alpha Research Vehicle (HARV) F-18. The package provides near real-time results in an informative and timely manner far exceeding prior methods of data reduction at the TDT.

  19. 49 CFR 173.4a - Excepted quantities.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... of withstanding without leakage the pressure differential specified in § 173.27(c) of this part. (b... ice), and lithium batteries and cells. (c) Inner packaging limits. The maximum quantity of hazardous..., rigid outer packaging. (5) Placement of the material in the package or packing different materials in...

  20. Prolonged shelf life and reduced drip loss of chicken filets by the use of carbon dioxide emitters and modified atmosphere packaging.

    PubMed

    Holck, Askild L; Pettersen, Marit K; Moen, Marie H; Sørheim, Oddvin

    2014-07-01

    Modified atmosphere packaging containing CO2 is widely used for extending the shelf life of chicken meat. Active packaging by adding CO2 emitter sachets to packages of meat is an alternative to traditional modified atmosphere packaging. The purpose of the study was to investigate the shelf life of chicken filets under different CO2 concentrations at 4°C storage. The inhibition of microbial growth was proportional to the CO2 concentration. Storage in 100% CO2 both with and without a CO2 emitter sachet gave a microbiological shelf-life extension of 7 days compared with 60% CO2. Carnobacterium divergens, Carnobacterium sp., and Lactococcus sp. were the dominating species at the end of the storage period. During storage in pure CO2, the carbon dioxide dissolved in the meat and caused the collapse of the packages. The resulting squeeze of the meat lead to a severe increase in drip loss. The drip loss was reduced profoundly by using the CO2 emitting sachet in the packages. The addition of CO2 emitters can easily be implemented at industrial packaging lines without reduction in production efficiency.

  1. Employing online quantum random number generators for generating truly random quantum states in Mathematica

    NASA Astrophysics Data System (ADS)

    Miszczak, Jarosław Adam

    2013-01-01

    The presented package for the Mathematica computing system allows the harnessing of quantum random number generators (QRNG) for investigating the statistical properties of quantum states. The described package implements a number of functions for generating random states. The new version of the package adds the ability to use the on-line quantum random number generator service and implements new functions for retrieving lists of random numbers. Thanks to the introduced improvements, the new version provides faster access to high-quality sources of random numbers and can be used in simulations requiring large amount of random data. New version program summaryProgram title: TRQS Catalogue identifier: AEKA_v2_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/AEKA_v2_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: Standard CPC licence, http://cpc.cs.qub.ac.uk/licence/licence.html No. of lines in distributed program, including test data, etc.: 18 134 No. of bytes in distributed program, including test data, etc.: 2 520 49 Distribution format: tar.gz Programming language: Mathematica, C. Computer: Any supporting Mathematica in version 7 or higher. Operating system: Any platform supporting Mathematica; tested with GNU/Linux (32 and 64 bit). RAM: Case-dependent Supplementary material: Fig. 1 mentioned below can be downloaded. Classification: 4.15. External routines: Quantis software library (http://www.idquantique.com/support/quantis-trng.html) Catalogue identifier of previous version: AEKA_v1_0 Journal reference of previous version: Comput. Phys. Comm. 183(2012)118 Does the new version supersede the previous version?: Yes Nature of problem: Generation of random density matrices and utilization of high-quality random numbers for the purpose of computer simulation. Solution method: Use of a physical quantum random number generator and an on-line service providing access to the source of true random numbers generated by quantum real number generator. Reasons for new version: Added support for the high-speed on-line quantum random number generator and improved methods for retrieving lists of random numbers. Summary of revisions: The presented version provides two signicant improvements. The first one is the ability to use the on-line Quantum Random Number Generation service developed by PicoQuant GmbH and the Nano-Optics groups at the Department of Physics of Humboldt University. The on-line service supported in the version 2.0 of the TRQS package provides faster access to true randomness sources constructed using the laws of quantum physics. The service is freely available at https://qrng.physik.hu-berlin.de/. The use of this service allows using the presented package with the need of a physical quantum random number generator. The second improvement introduced in this version is the ability to retrieve arrays of random data directly for the used source. This increases the speed of the random number generation, especially in the case of an on-line service, where it reduces the time necessary to establish the connection. Thanks to the speed improvement of the presented version, the package can now be used in simulations requiring larger amounts of random data. Moreover, the functions for generating random numbers provided by the current version of the package more closely follow the pattern of functions for generating pseudo- random numbers provided in Mathematica. Additional comments: Speed comparison: The implementation of the support for the QRNG on-line service provides a noticeable improvement in the speed of random number generation. For the samples of real numbers of size 101; 102,…,107 the times required to generate these samples using Quantis USB device and QRNG service are compared in Fig. 1. The presented results show that the use of the on-line service provides faster access to random numbers. One should note, however, that the speed gain can increase or decrease depending on the connection speed between the computer and the server providing random numbers. Running time: Depends on the used source of randomness and the amount of random data used in the experiment. References: [1] M. Wahl, M. Leifgen, M. Berlin, T. Röhlicke, H.-J. Rahn, O. Benson., An ultrafast quantum random number generator with provably bounded output bias based on photon arrival time measurements, Applied Physics Letters, Vol. 098, 171105 (2011). http://dx.doi.org/10.1063/1.3578456.

  2. Adaptation of retrovirus producer cells to serum deprivation: Implications in lipid biosynthesis and vector production.

    PubMed

    Rodrigues, A F; Amaral, A I; Veríssimo, V; Alves, P M; Coroadinha, A S

    2012-05-01

    The manufacture of enveloped virus, particularly retroviral (RV) and lentiviral (LV) vectors, faces the challenge of low titers that are aggravated under serum deprivation culture conditions. Also, the scarce knowledge on the biochemical pathways related with virus production is still limiting the design of rational strategies for improved production yields. This work describes the adaptation to serum deprivation of two human RV packaging cell lines, 293 FLEX and Te Fly and its effects on lipid biosynthetic pathways and infectious vector production. Total lipid content as well as cellular cholesterol were quantified and lipid biosynthesis was assessed by (13)C-NMR spectroscopy; changes in gene expression of lipid biosynthetic enzymes were also evaluated. The effects of adaptation to serum deprivation in lipid biosynthesis were cell line specific and directly correlated with infectious virus titers: 293 FLEX cells faced severe lipid starvation-up to 50% reduction in total lipid content-along with a 68-fold reduction in infectious vector titers; contrarily, Te Fly cells were able to maintain identical levels of total lipid content by rising de novo lipid biosynthesis, particularly for cholesterol-50-fold increase-with the consequent recovery of infectious vector productivities. Gene expression analysis of lipid biosynthetic enzymes further confirmed cholesterol production pathway to be prominently up-regulated under serum deprivation conditions for Te Fly cells, providing a genotype-phenotype validation for enhanced cholesterol synthesis. These results highlight lipid metabolism dynamics and the ability to activate lipid biosynthesis under serum deprivation as an important feature for high retroviral titers. Mechanisms underlying virus production and its relationship with lipid biosynthesis, with special focus on cholesterol, are discussed as potential targets for cellular metabolic engineering. Copyright © 2011 Wiley Periodicals, Inc.

  3. soilphysics: An R package to determine soil preconsolidation pressure

    NASA Astrophysics Data System (ADS)

    da Silva, Anderson Rodrigo; de Lima, Renato Paiva

    2015-11-01

    Preconsolidation pressure is a parameter obtained from the soil compression curve and has been used as an indicator of load-bearing capacity of soil, as well as to characterize the impacts suffered by the use of machines. Despite its importance in soil physics, there is a few software or computational routines to support its determination. In this paper we present a computational package in R language, the package soilphysics, which contains implementations of the main methods for determining preconsolidation pressure, such as the method of Casagrande, Pacheco Silva, regression methods and the method of the virgin compression line intercept. There is still a consensus that Casagrande is the standard method, although the method of Pacheco Silva has shown similar values. The method of the virgin compression line intercept can be used when trying to be more conservative on the value (smaller) of preconsolidation pressure. Furthermore, Casagrande could be replaced by a regression method when the compression curve is obtained from saturated soils. The theory behind each method is presented and the algorithms are thoroughly described. We also give some support on how to use the R functions. Examples are used to illustrate the capabilities of the package, and the results are briefly discussed. The latter were validated using a recently published VBA. With soilphysics, the user has all the graphical and statistical power of R to determine preconsolidation pressure using different methods. The package is distribution free (under the GPL-2|3) and is currently available from the Comprehensive R Archive Network.

  4. destiny: diffusion maps for large-scale single-cell data in R.

    PubMed

    Angerer, Philipp; Haghverdi, Laleh; Büttner, Maren; Theis, Fabian J; Marr, Carsten; Buettner, Florian

    2016-04-15

    : Diffusion maps are a spectral method for non-linear dimension reduction and have recently been adapted for the visualization of single-cell expression data. Here we present destiny, an efficient R implementation of the diffusion map algorithm. Our package includes a single-cell specific noise model allowing for missing and censored values. In contrast to previous implementations, we further present an efficient nearest-neighbour approximation that allows for the processing of hundreds of thousands of cells and a functionality for projecting new data on existing diffusion maps. We exemplarily apply destiny to a recent time-resolved mass cytometry dataset of cellular reprogramming. destiny is an open-source R/Bioconductor package "bioconductor.org/packages/destiny" also available at www.helmholtz-muenchen.de/icb/destiny A detailed vignette describing functions and workflows is provided with the package. carsten.marr@helmholtz-muenchen.de or f.buettner@helmholtz-muenchen.de Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  5. Ada (Trademark) Case Studies II.

    DTIC Science & Technology

    1984-01-01

    number ( CSN ) in that character set’s "transmission identifier". The array is named START CSN , but a better name would be STARTOFCSN. The word of...protocols), CSN (Channel Sequence Number), EON (End of Message), LMF (Line Media Format), MCB (Message Control Block), RI (Routing Indicator), and TI...component names should end with -PART, - FIELD , or COMPONENT.El - Package names should end with PACKAGE. - Enumeration literals should have suffixes

  6. 1986 Petroleum Software Directory. [800 mini, micro and mainframe computer software packages

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1985-01-01

    Pennwell's 1986 Petroleum Software Directory is a complete listing of software created specifically for the petroleum industry. Details are provided on over 800 mini, micro and mainframe computer software packages from more than 250 different companies. An accountant can locate programs to automate bookkeeping functions in large oil and gas production firms. A pipeline engineer will find programs designed to calculate line flow and wellbore pressure drop.

  7. Oufti: An integrated software package for high-accuracy, high-throughput quantitative microscopy analysis

    PubMed Central

    Paintdakhi, Ahmad; Parry, Bradley; Campos, Manuel; Irnov, Irnov; Elf, Johan; Surovtsev, Ivan; Jacobs-Wagner, Christine

    2016-01-01

    Summary With the realization that bacteria display phenotypic variability among cells and exhibit complex subcellular organization critical for cellular function and behavior, microscopy has re-emerged as a primary tool in bacterial research during the last decade. However, the bottleneck in today’s single-cell studies is quantitative image analysis of cells and fluorescent signals. Here, we address current limitations through the development of Oufti, a stand-alone, open-source software package for automated measurements of microbial cells and fluorescence signals from microscopy images. Oufti provides computational solutions for tracking touching cells in confluent samples, handles various cell morphologies, offers algorithms for quantitative analysis of both diffraction and non-diffraction-limited fluorescence signals, and is scalable for high-throughput analysis of massive datasets, all with subpixel precision. All functionalities are integrated in a single package. The graphical user interface, which includes interactive modules for segmentation, image analysis, and post-processing analysis, makes the software broadly accessible to users irrespective of their computational skills. PMID:26538279

  8. Truck circuits diagnosis for railway lines equipped with an automatic block signalling system

    NASA Astrophysics Data System (ADS)

    Spunei, E.; Piroi, I.; Muscai, C.; Răduca, E.; Piroi, F.

    2018-01-01

    This work presents a diagnosis method for detecting track circuits failures on a railway traffic line equipped with an Automatic Block Signalling installation. The diagnosis method uses the installation’s electrical schemas, based on which a series of diagnosis charts have been created. Further, the diagnosis charts were used to develop a software package, CDCBla, which substantially contributes to reducing the diagnosis time and human error during failure remedies. The proposed method can also be used as a training package for the maintenance staff. Since the diagnosis method here does not need signal or measurement inputs, using it does not necessitate additional IT knowledge and can be deployed on a mobile computing device (tablet, smart phone).

  9. Gene therapy for human ovarian cancer cells using efficient expression of Fas gene combined with γδT cells.

    PubMed

    Lin, Jiajing; Zeng, Dingyuan; He, Hongying; Tan, Guangping; Lan, Ying; Jiang, Fuyan; Sheng, Shuting

    2017-10-01

    Low tissue specificity and efficiency of exogenous gene expression are the two major obstacles in tumor‑targeted gene therapy. The Fas cell surface death receptor (Fas)/Fas ligand pathway is one of the primary pathways responsible for the regulation of cell apoptosis. The aim of the present study was to explore whether the regulation of tumor specific promoters and a two‑step transcriptional amplification system (TSTA) assured efficient, targeted expression of their downstream Fas gene in human ovarian cancer cells, and to assess the killing effect of γδT cells on these cells with high Fas expression. Three shuttle plasmids containing different control elements of the human telomerase reverse transcriptase (hTERT) promoter and/or TSTA were constructed and packaged into adenovirus 5 (Ad5) vectors for the expression of exogenous Fas gene. The human ovarian cancer cell line SKOV3 and a control human embryonic lung fibroblast cell line were transfected with Ad5‑hTERT‑Fas or Ad5‑hTERT‑TSTA‑Fas. Fas mRNA and protein expression were examined by reverse transcription‑quantitative polymerase chain reaction and western blot analysis. γδT lymphocytes were isolated, cultured and mixed at different ratios with SKOV3 cells with Fas expression in order to assess the killing effect of γδT cells. hTERT promoter induced the specific expression of FAS gene in SKOV3 cells, and the TSTA strategy increased FAS expression by 14.2‑fold. The killing effect of γδT cells increased with the expression level of Fas and the effector‑target cell ratio. The killing rate for SKOV3 cells with high FAS expression was 72.5% at an effector‑target cell ratio of 40:1. The regulators of hTERT promoter and TSTA assure the efficient and targeted expression of their downstream Fas gene in SKOV3 cells. The killing effect of γδT cells for ovarian cancer cells with relatively high Fas expression was improved.

  10. Pathogen inactivation treatment of plasma and platelet concentrates and their predicted functionality in massive transfusion protocols.

    PubMed

    Arbaeen, Ahmad F; Schubert, Peter; Serrano, Katherine; Carter, Cedric J; Culibrk, Brankica; Devine, Dana V

    2017-05-01

    Trauma transfusion packages for hemorrhage control consist of red blood cells, plasma, and platelets at a set ratio. Although pathogen reduction improves the transfusion safety of platelet and plasma units, there is an associated reduction in quality. This study aimed to investigate the impact of riboflavin/ultraviolet light-treated plasma or platelets in transfusion trauma packages composed of red blood cell, plasma, and platelet units in a ratio of 1:1:1 in vitro by modeling transfusion scenarios for trauma patients and assessing function by rotational thromboelastometry. Pathogen-reduced or untreated plasma and buffy coat platelet concentrate units produced in plasma were used in different combinations with red blood cells in trauma transfusion packages. After reconstitution of these packages with hemodiluted blood, the hemostatic functionality was analyzed by rotational thromboelastometry. Hemostatic profiles of pathogen-inactivated buffy coat platelet concentrate and plasma indicated decreased activity compared with their respective controls. Reconstitution of hemodiluted blood (hematocrit = 20%) with packages that contained treated or nontreated components resulted in increased alpha and maximum clot firmness and enhanced clot-formation time. Simulating transfusion scenarios based on 30% blood replacement with a transfusion trauma package resulted in a nonsignificant difference in rotational thromboelastometry parameters between packages containing treated and nontreated blood components (p ≥ 0.05). Effects of pathogen inactivation treatment were evident when the trauma package percentage was 50% or greater and contained both pathogen inactivation-treated plasma and buffy coat platelet concentrate. Rotational thromboelastometry investigations suggest that there is relatively little impact of pathogen inactivation treatment on whole blood clot formation unless large amounts of treated components are used. © 2017 AABB.

  11. The Product of the Herpes Simplex Virus Type 1 UL25 Gene Is Required for Encapsidation but Not for Cleavage of Replicated Viral DNA

    PubMed Central

    McNab, Alistair R.; Desai, Prashant; Person, Stan; Roof, Lori L.; Thomsen, Darrell R.; Newcomb, William W.; Brown, Jay C.; Homa, Fred L.

    1998-01-01

    The herpes simplex virus type 1 (HSV-1) UL25 gene contains a 580-amino-acid open reading frame that codes for an essential protein. Previous studies have shown that the UL25 gene product is a virion component (M. A. Ali et al., Virology 216:278–283, 1996) involved in virus penetration and capsid assembly (C. Addison et al., Virology 138:246–259, 1984). In this study, we describe the isolation of a UL25 mutant (KUL25NS) that was constructed by insertion of an in-frame stop codon in the UL25 open reading frame and propagated on a complementing cell line. Although the mutant was capable of synthesis of viral DNA, it did not form plaques or produce infectious virus in noncomplementing cells. Antibodies specific for the UL25 protein were used to demonstrate that KUL25NS-infected Vero cells did not express the UL25 protein. Western immunoblotting showed that the UL25 protein was associated with purified, wild-type HSV A, B, and C capsids. Transmission electron microscopy indicated that the nucleus of Vero cells infected with KUL25NS contained large numbers of both A and B capsids but no C capsids. Analysis of infected cells by sucrose gradient sedimentation analysis confirmed that the ratio of A to B capsids was elevated in KUL25NS-infected Vero cells. Following restriction enzyme digestion, specific terminal fragments were observed in DNA isolated from KUL25NS-infected Vero cells, indicating that the UL25 gene was not required for cleavage of replicated viral DNA. The latter result was confirmed by pulsed-field gel electrophoresis (PFGE), which showed the presence of genome-size viral DNA in KUL25NS-infected Vero cells. DNase I treatment prior to PFGE demonstrated that monomeric HSV DNA was not packaged in the absence of the UL25 protein. Our results indicate that the product of the UL25 gene is required for packaging but not cleavage of replicated viral DNA. PMID:9445000

  12. [Effects of 5-aza-2-deoxycytidine on methylation status of RECK gene and cancer cell invasion in tongue cancer SCC-4 cells].

    PubMed

    Jiang, Xv

    2014-10-01

    To investigate the effects of 5-aza-2-deoxycytidine on methylation status and invasion ability of RECK gene in tongue cancer SCC-4 cells. Tongue cancer cell line SCC-4 cells were treated with 5-aza-dC at different concentrations for 72 h. Methylation status of RECK gene of SCC-4 cells was detected by methylation specific PCR (MSP), the expression of RECK gene mRNA was detected by real-time quantitative PCR. The expression of RECK protein was detected by Western blot, and the invasion ability of SCC-4 cell was examined by Transwell assay. SPSS13.0 software package was used for statistical analysis. RECK gene of SCC-4 cells was in high methylation status in untreated group, abnormal methylation was effectively reversed by 5-aza-dC treatment. After treatment with different concentration of 5-aza-dC for 72 h, relative mRNA expression level increased gradually (P<0.05). The relative expression level of RECK protein in 5-aza-dC treated group was significantly higher than that in the control group,the invasion ability of SCC-4 cell was decreased gradually. 5-aza-dC treatment for tongue cancer SCC-4 cells can successfully reverse high methylation status of RECK gene and restore the expression of RECK gene mRNA and protein, and reduced the invasion ability.

  13. Glyceraldehyde 3-phosphate dehydrogenase negatively regulates human immunodeficiency virus type 1 infection

    PubMed Central

    2012-01-01

    Background Host proteins are incorporated inside human immunodeficiency virus type 1 (HIV-1) virions during assembly and can either positively or negatively regulate HIV-1 infection. Although the identification efficiency of host proteins is improved by mass spectrometry, how those host proteins affect HIV-1 replication has not yet been fully clarified. Results In this study, we show that virion-associated glyceraldehyde 3-phosphate dehydrogenase (GAPDH) does not allosterically inactivate HIV-1 reverse transcriptase (RT) but decreases the efficiency of reverse transcription reactions by decreasing the packaging efficiency of lysyl-tRNA synthetase (LysRS) and tRNALys3 into HIV-1 virions. Two-dimensional (2D) gel electrophoresis demonstrated that some isozymes of GAPDH with different isoelectric points were expressed in HIV-1-producing CEM/LAV-1 cells, and a proportion of GAPDH was selectively incorporated into the virions. Suppression of GAPDH expression by RNA interference in CEM/LAV-1 cells resulted in decreased GAPDH packaging inside the virions, and the GAPDH-packaging-defective virus maintained at least control levels of viral production but increased the infectivity. Quantitative analysis of reverse transcription products indicated that the levels of early cDNA products of the GAPDH-packaging-defective virus were higher than those of the control virus owing to the higher packaging efficiencies of LysRS and tRNALys3 into the virions rather than the GAPDH-dependent negative allosteric modulation for RT. Furthermore, immunoprecipitation assay using an anti-GAPDH antibody showed that GAPDH directly interacted with Pr55gag and p160gag-pol and the overexpression of LysRS in HIV-1-producing cells resulted in a decrease in the efficiency of GAPDH packaging in HIV particles. In contrast, the viruses produced from cells expressing a high level of GAPDH showed decreased infectivity in TZM-bl cells and reverse transcription efficiency in TZM-bl cells and peripheral blood mononuclear cells (PBMCs). Conclusions These findings indicate that GAPDH negatively regulates HIV-1 infection and provide insights into a novel function of GAPDH in the HIV-1 life cycle and a new host defense mechanism against HIV-1 infection. PMID:23237566

  14. Leadless Chip Carrier Packaging and CAD/CAM-Supported Wire Wrap Interconnect Technology for Subnanosecond ECL.

    DTIC Science & Technology

    1981-11-01

    Showing Wire . 99 Impregnanted Silicone Rubber Contacts, Chip Carrier, ard Lid 35. Technit Connector For 68-Pad JEDEC Type A Leadless . . 100 Chip Carrier...Points of Various . . . . 124 Solders 4. Composition of Alloys Employed in Dual-In-Line . . . . 128 Package Pins and Plating by Mass Spectrographic...swings, and subnanosecond gate delays and risetimes. Presently, emitter coupled logic (ECL) and current mode logic (CML), both fabricated with silicon tech

  15. Power Line Technician. Second Edition. Module B. Instructor's Manual [and] Student Workbook.

    ERIC Educational Resources Information Center

    Oklahoma State Dept. of Vocational and Technical Education, Stillwater. Curriculum and Instructional Materials Center.

    This module is the second in a series that is designed to help students achieve greater professional and personal success as power line technicians. Each module represents one or more complete units of instruction with components organized into one instructor and one student "package": the instructor's manual and the student workbook.…

  16. Power Line Technician. Second Edition. Module A. Instructor's Manual [and] Student Workbook.

    ERIC Educational Resources Information Center

    Oklahoma State Dept. of Vocational and Technical Education, Stillwater. Curriculum and Instructional Materials Center.

    This module is the first in a series that is designed to help students achieve greater professional and personal success as power line technicians. Each module represents one or more complete units of instruction with components organized into one instructor and one student "package": the instructor's manual and the student workbook. The…

  17. 9 CFR 381.222 - States designated under paragraph 5(c) of the Act; application of regulations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... that separation of toilet soil lines from house drainage lines to a point outside the buildings will... mechanical printing device or a self-destructive pressure sensitive sticker, and provided the label shows the... by the formula and details of preparation and packaging for each product. Within 90 days after...

  18. 40 CFR 63.3300 - Which of my emission sources are affected by this subpart?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... CATEGORIES National Emission Standards for Hazardous Air Pollutants: Paper and Other Web Coating What This... subject to this subpart is the collection of all web coating lines at your facility. This includes web coating lines engaged in the coating of metal webs that are used in flexible packaging, and web coating...

  19. Electrical Experiments. VT-214-12-2. Part II. A-C Across the Line Control.

    ERIC Educational Resources Information Center

    Connecticut State Dept. of Education, Hartford. Div. of Vocational Education.

    Designed for high school electronics students, this second document in a series of six electrical learning activity packages focuses on alternating current across-the-line control. An introductory section gives the objective for the activities, an introduction, and an outline of the content. The remainder of the activity book is comprised of…

  20. 40 CFR 63.3300 - Which of my emission sources are affected by this subpart?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... CATEGORIES (CONTINUED) National Emission Standards for Hazardous Air Pollutants: Paper and Other Web Coating... affected source subject to this subpart is the collection of all web coating lines at your facility. This includes web coating lines engaged in the coating of metal webs that are used in flexible packaging, and...

  1. 40 CFR 63.3300 - Which of my emission sources are affected by this subpart?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... CATEGORIES (CONTINUED) National Emission Standards for Hazardous Air Pollutants: Paper and Other Web Coating... affected source subject to this subpart is the collection of all web coating lines at your facility. This includes web coating lines engaged in the coating of metal webs that are used in flexible packaging, and...

  2. 40 CFR 63.3300 - Which of my emission sources are affected by this subpart?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... CATEGORIES (CONTINUED) National Emission Standards for Hazardous Air Pollutants: Paper and Other Web Coating... affected source subject to this subpart is the collection of all web coating lines at your facility. This includes web coating lines engaged in the coating of metal webs that are used in flexible packaging, and...

  3. 40 CFR 63.3300 - Which of my emission sources are affected by this subpart?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... CATEGORIES National Emission Standards for Hazardous Air Pollutants: Paper and Other Web Coating What This... subject to this subpart is the collection of all web coating lines at your facility. This includes web coating lines engaged in the coating of metal webs that are used in flexible packaging, and web coating...

  4. Impact of policy changes on infant feeding decisions among low-income women participating in the Special Supplemental Nutrition Program for Women, Infants, and Children.

    PubMed

    Whaley, Shannon E; Koleilat, Maria; Whaley, Mike; Gomez, Judy; Meehan, Karen; Saluja, Kiran

    2012-12-01

    We present infant feeding data before and after the 2009 Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) food package change that supported and incentivized breastfeeding. We describe the key role of California WIC staff in supporting these policy changes. We analyzed WIC data on more than 180,000 infants in Southern California. We employed the analysis of variance and Tukey (honestly significant difference) tests to compare issuance rates of postpartum and infant food packages before and after the changes. We used analysis of covariance to adjust for poverty status changes as a potential confounder. Issuance rates of the "fully breastfeeding" package at infant WIC enrollment increased by 86% with the package changes. Rates also increased significantly for 2- and 6-month-old infants. Issuance rates of packages that included formula decreased significantly. All outcomes remained highly significant in the adjusted model. Policy changes, training of front-line WIC staff, and participant education influenced issuance rates of WIC food packages. In California, the issuance rates of packages that include formula have significantly decreased and the rate for those that include no formula has significantly increased.

  5. Documentation of the seawater intrusion (SWI2) package for MODFLOW

    USGS Publications Warehouse

    Bakker, Mark; Schaars, Frans; Hughes, Joseph D.; Langevin, Christian D.; Dausman, Alyssa M.

    2013-01-01

    The SWI2 Package is the latest release of the Seawater Intrusion (SWI) Package for MODFLOW. The SWI2 Package allows three-dimensional vertically integrated variable-density groundwater flow and seawater intrusion in coastal multiaquifer systems to be simulated using MODFLOW-2005. Vertically integrated variable-density groundwater flow is based on the Dupuit approximation in which an aquifer is vertically discretized into zones of differing densities, separated from each other by defined surfaces representing interfaces or density isosurfaces. The numerical approach used in the SWI2 Package does not account for diffusion and dispersion and should not be used where these processes are important. The resulting differential equations are equivalent in form to the groundwater flow equation for uniform-density flow. The approach implemented in the SWI2 Package allows density effects to be incorporated into MODFLOW-2005 through the addition of pseudo-source terms to the groundwater flow equation without the need to solve a separate advective-dispersive transport equation. Vertical and horizontal movement of defined density surfaces is calculated separately using a combination of fluxes calculated through solution of the groundwater flow equation and a simple tip and toe tracking algorithm. Use of the SWI2 Package in MODFLOW-2005 only requires the addition of a single additional input file and modification of boundary heads to freshwater heads referenced to the top of the aquifer. Fluid density within model layers can be represented using zones of constant density (stratified flow) or continuously varying density (piecewise linear in the vertical direction) in the SWI2 Package. The main advantage of using the SWI2 Package instead of variable-density groundwater flow and dispersive solute transport codes, such as SEAWAT and SUTRA, is that fewer model cells are required for simulations using the SWI2 Package because every aquifer can be represented by a single layer of cells. This reduction in number of required model cells and the elimination of the need to solve the advective-dispersive transport equation results in substantial model run-time savings, which can be large for regional aquifers. The accuracy and use of the SWI2 Package is demonstrated through comparison with existing exact solutions and numerical solutions with SEAWAT. Results for an unconfined aquifer are also presented to demonstrate application of the SWI2 Package to a large-scale regional problem.

  6. Profound Amplification of Pathogenic Murine Polytropic Retrovirus Release from Coinfected Cells

    PubMed Central

    Rosenke, Kyle; Lavignon, Marc; Malik, Frank; Kolokithas, Angelo; Hendrick, Duncan; Virtaneva, Kimmo; Peterson, Karin

    2012-01-01

    Previous studies indicate that mice infected with mixtures of mouse retroviruses (murine leukemia viruses [MuLVs]) exhibit dramatically altered pathology compared to mice infected with individual viruses of the mixture. Coinoculation of the ecotropic virus Friend MuLV (F-MuLV) with Fr98, a polytropic MuLV, induced a rapidly fatal neurological disease that was not observed in infections with either virus alone. The polytropic virus load in coinoculated mice was markedly enhanced, while the ecotropic F-MuLV load was unchanged. Furthermore, pseudotyping of the polytropic MuLV genome within ecotropic virions was nearly complete in coinoculated mice. In an effort to better understand these phenomena, we examined mixed retrovirus infections by utilizing in vitro cell lines. Similar to in vivo mixed infections, the polytropic MuLV genome was extensively pseudotyped within ecotropic virions; polytropic virus release was profoundly elevated in coinfected cells, and the ecotropic virus release was unchanged. A reduced level of polytropic SU protein on the surfaces of coinfected cells was observed and correlated with a reduced level of nonpseudotyped polytropic virion release. Marked amplification and pseudotyping of the polytropic MuLV were also observed in mixed Fr98–F-MuLV infections of cell lines derived from the central nervous system (CNS), the target for Fr98 pathogenesis. Additional experiments indicated that pseudotyping contributed to the elevated polytropic virus titer by increasing the efficiency of packaging and release of the polytropic genomes within ecotropic virions. Mixed infections are the rule rather than the exception in retroviral infection, and the ability to examine them in vitro should facilitate a more thorough understanding of retroviral interactions in general. PMID:22514353

  7. WASTE PACKAGE REMEDIATION SYSTEM DESCRIPTION DOCUMENT

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    N.D. Sudan

    2000-06-22

    The Waste Package Remediation System remediates waste packages (WPs) and disposal containers (DCs) in one of two ways: preparation of rejected DC closure welds for repair or opening of the DC/WP. DCs are brought to the Waste Package Remediation System for preparation of rejected closure welds if testing of the closure weld by the Disposal Container Handling System indicates an unacceptable, but repairable, welding flaw. DC preparation of rejected closure welds will require removal of the weld in such a way that the Disposal Container Handling System may resume and complete the closure welding process. DCs/WPs are brought to themore » Waste Package Remediation System for opening if the Disposal Container Handling System testing of the DC closure weld indicates an unrepairable welding flaw, or if a WP is recovered from the subsurface repository because suspected damage to the WP or failure of the WP has occurred. DC/WP opening will require cutting of the DC/WP such that a temporary seal may be installed and the waste inside the DC/WP removed by another system. The system operates in a Waste Package Remediation System hot cell located in the Waste Handling Building that has direct access to the Disposal Container Handling System. One DC/WP at a time can be handled in the hot cell. The DC/WP arrives on a transfer cart, is positioned within the cell for system operations, and exits the cell without being removed from the cart. The system includes a wide variety of remotely operated components including a manipulator with hoist and/or jib crane, viewing systems, machine tools for opening WPs, and equipment used to perform pressure and gas composition sampling. Remotely operated equipment is designed to facilitate DC/WP decontamination and hot cell equipment maintenance, and interchangeable components are provided where appropriate. The Waste Package Remediation System interfaces with the Disposal Container Handling System for the receipt and transport of WPs and DCs. The Waste Handling Building System houses the system, and provides the facility, safety, and auxiliary systems required to support operations. The system receives power from the Waste Handling Building Electrical System. The system also interfaces with the various DC systems.« less

  8. Supervision: Substance and Style

    ERIC Educational Resources Information Center

    Gellerman, Saul W.

    1976-01-01

    Argues that managerial style and substance are inextricably intertwined, illustrating the discussion with excerpts from an extensive study and job analysis of first-line supervisors in a food packaging plant. (JG)

  9. BatTool: an R package with GUI for assessing the effect of White-nose syndrome and other take events on Myotis spp. of bats

    PubMed Central

    2014-01-01

    Background Myotis species of bats such as the Indiana Bat and Little Brown Bat are facing population declines because of White-nose syndrome (WNS). These species also face threats from anthropogenic activities such as wind energy development. Population models may be used to provide insights into threats facing these species. We developed a population model, BatTool, as an R package to help decision makers and natural resource managers examine factors influencing the dynamics of these species. The R package includes two components: 1) a deterministic and stochastic model that are accessible from the command line and 2) a graphical user interface (GUI). Results BatTool is an R package allowing natural resource managers and decision makers to understand Myotis spp. population dynamics. Through the use of a GUI, the model allows users to understand how WNS and other take events may affect the population. The results are saved both graphically and as data files. Additionally, R-savvy users may access the population functions through the command line and reuse the code as part of future research. This R package could also be used as part of a population dynamics or wildlife management course. Conclusions BatTool provides access to a Myotis spp. population model. This tool can help natural resource managers and decision makers with the Endangered Species Act deliberations for these species and with issuing take permits as part of regulatory decision making. The tool is available online as part of this publication. PMID:24955110

  10. BatTool: an R package with GUI for assessing the effect of White-nose syndrome and other take events on Myotis spp. of bats

    USGS Publications Warehouse

    Erickson, Richard A.; Thogmartin, Wayne E.; Szymanski, Jennifer A.

    2014-01-01

    Background: Myotis species of bats such as the Indiana Bat and Little Brown Bat are facing population declines because of White-nose syndrome (WNS). These species also face threats from anthropogenic activities such as wind energy development. Population models may be used to provide insights into threats facing these species. We developed a population model, BatTool, as an R package to help decision makers and natural resource managers examine factors influencing the dynamics of these species. The R package includes two components: 1) a deterministic and stochastic model that are accessible from the command line and 2) a graphical user interface (GUI). Results: BatTool is an R package allowing natural resource managers and decision makers to understand Myotis spp. population dynamics. Through the use of a GUI, the model allows users to understand how WNS and other take events may affect the population. The results are saved both graphically and as data files. Additionally, R-savvy users may access the population functions through the command line and reuse the code as part of future research. This R package could also be used as part of a population dynamics or wildlife management course. Conclusions: BatTool provides access to a Myotis spp. population model. This tool can help natural resource managers and decision makers with the Endangered Species Act deliberations for these species and with issuing take permits as part of regulatory decision making. The tool is available online as part of this publication.

  11. BatTool: an R package with GUI for assessing the effect of White-nose syndrome and other take events on Myotis spp. of bats.

    PubMed

    Erickson, Richard A; Thogmartin, Wayne E; Szymanski, Jennifer A

    2014-01-01

    Myotis species of bats such as the Indiana Bat and Little Brown Bat are facing population declines because of White-nose syndrome (WNS). These species also face threats from anthropogenic activities such as wind energy development. Population models may be used to provide insights into threats facing these species. We developed a population model, BatTool, as an R package to help decision makers and natural resource managers examine factors influencing the dynamics of these species. The R package includes two components: 1) a deterministic and stochastic model that are accessible from the command line and 2) a graphical user interface (GUI). BatTool is an R package allowing natural resource managers and decision makers to understand Myotis spp. population dynamics. Through the use of a GUI, the model allows users to understand how WNS and other take events may affect the population. The results are saved both graphically and as data files. Additionally, R-savvy users may access the population functions through the command line and reuse the code as part of future research. This R package could also be used as part of a population dynamics or wildlife management course. BatTool provides access to a Myotis spp. population model. This tool can help natural resource managers and decision makers with the Endangered Species Act deliberations for these species and with issuing take permits as part of regulatory decision making. The tool is available online as part of this publication.

  12. Circadian Behavioral Responses to Light and Optic Chiasm-Evoked Glutamatergic EPSCs in the Suprachiasmatic Nucleus of ipRGC Conditional vGlut2 Knock-Out Mice

    PubMed Central

    2018-01-01

    Abstract Intrinsically photosensitive retinal ganglion cells (ipRGCs) innervate the hypothalamic suprachiasmatic nucleus (SCN), a circadian oscillator that functions as a biological clock. ipRGCs use vesicular glutamate transporter 2 (vGlut2) to package glutamate into synaptic vesicles and light-evoked resetting of the SCN circadian clock is widely attributed to ipRGC glutamatergic neurotransmission. Pituitary adenylate cyclase-activating polypeptide (PACAP) is also packaged into vesicles in ipRGCs and PACAP may be coreleased with glutamate in the SCN. vGlut2 has been conditionally deleted in ipRGCs in mice [conditional knock-outs (cKOs)] and their aberrant photoentrainment and residual attenuated light responses have been ascribed to ipRGC PACAP release. However, there is no direct evidence that all ipRGC glutamatergic neurotransmission is eliminated in vGlut2 cKOs. Here, we examined two lines of ipRGC vGlut2 cKO mice for SCN-mediated behavioral responses under several lighting conditions and for ipRGC glutamatergic neurotransmission in the SCN. Circadian behavioral responses varied from a very limited response to light to near normal photoentrainment. After collecting behavioral data, hypothalamic slices were prepared and evoked EPSCs (eEPSCs) were recorded from SCN neurons by stimulating the optic chiasm. In cKOs, glutamatergic eEPSCs were recorded and all eEPSC parameters examined (stimulus threshold, amplitude, rise time or time-to-peak and stimulus strength to evoke a maximal response) were similar to controls. We conclude that a variable number but functionally significant percentage of ipRGCs in two vGlut2 cKO mouse lines continue to release glutamate. Thus, the residual SCN-mediated light responses in these cKO mouse lines cannot be attributed solely to ipRGC PACAP release. PMID:29756029

  13. The Impact of Chromatin Dynamics on Cas9-Mediated Genome Editing in Human Cells.

    PubMed

    Daer, René M; Cutts, Josh P; Brafman, David A; Haynes, Karmella A

    2017-03-17

    In order to efficiently edit eukaryotic genomes, it is critical to test the impact of chromatin dynamics on CRISPR/Cas9 function and develop strategies to adapt the system to eukaryotic contexts. So far, research has extensively characterized the relationship between the CRISPR endonuclease Cas9 and the composition of the RNA-DNA duplex that mediates the system's precision. Evidence suggests that chromatin modifications and DNA packaging can block eukaryotic genome editing by custom-built DNA endonucleases like Cas9; however, the underlying mechanism of Cas9 inhibition is unclear. Here, we demonstrate that closed, gene-silencing-associated chromatin is a mechanism for the interference of Cas9-mediated DNA editing. Our assays use a transgenic cell line with a drug-inducible switch to control chromatin states (open and closed) at a single genomic locus. We show that closed chromatin inhibits binding and editing at specific target sites and that artificial reversal of the silenced state restores editing efficiency. These results provide new insights to improve Cas9-mediated editing in human and other mammalian cells.

  14. Mass decomposition of galaxies using DECA software package

    NASA Astrophysics Data System (ADS)

    Mosenkov, A. V.

    2014-01-01

    The new DECA software package, which is designed to perform photometric analysis of the images of disk and elliptical galaxies having a regular structure, is presented. DECA is written in Python interpreted language and combines the capabilities of several widely used packages for astronomical data processing such as IRAF, SExtractor, and the GALFIT code used to perform two-dimensional decomposition of galaxy images into several photometric components (bulge+disk). DECA has the advantage that it can be applied to large samples of galaxies with different orientations with respect to the line of sight (including edge-on galaxies) and requires minimum human intervention. Examples of using the package to study a sample of simulated galaxy images and a sample of real objects are shown to demonstrate that DECA can be a reliable tool for the study of the structure of galaxies.

  15. Parasitic heat loss reduction in AMTEC cells by heat shield optimization

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Borkowski, C.A.; Svedberg, R.C.; Hendricks, T.J.

    1997-12-31

    Alkali metal thermal to electric conversion (AMTEC) cell performance can be increased by the proper design of thermal radiative shielding internal to the AMTEC cell. These heat shields essentially lower the radiative heat transfer between the heat input zone of the cell and the heat rejection zone of the cell. In addition to lowering the radiative heat transfer between the heat input and heat rejection surfaces of the cell, the shields raise the AMTEC cell performance by increasing the temperature of the beta alumina solid electrolyte (BASE). This increase in temperature of the BASE tube allows the evaporator temperature tomore » be increased without sodium condensing within the BASE tubes. Experimental testing and theoretical analysis have been performed to compare the relative merits of two candidate heat shield packages: (1) chevron, and (2) cylindrical heat shields. These two heat shield packages were compared to each other and a baseline cell which had no heat shields installed. For the two heat shield packages, the reduction in total heat transfer is between 17--27% for the heat input surface temperature varying from 700 C, 750 C, and 800 C with the heat rejection surface temperature kept at 300 C.« less

  16. Calculation Package for the Analysis of Performance of Cells 1-6, with Underdrain, of the Environmental Management Waste Management Facility Oak Ridge, Tennessee

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gonzales D.

    2010-03-30

    This calculation package presents the results of an assessment of the performance of the 6 cell design of the Environmental Management Waste Management Facility (EMWMF). The calculations show that the new cell 6 design at the EMWMF meets the current WAC requirement. QA/QC steps were taken to verify the input/output data for the risk model and data transfer from modeling output files to tables and calculation.

  17. Apparel Manufacture

    NASA Technical Reports Server (NTRS)

    1995-01-01

    Marshall Space Flight Center teamed with the University of Alabama in Huntsville (UAH) in 1989 on a program involving development of advanced simulation software. Concurrently, the State of Alabama chartered UAH to conduct a technology advancement program in support of the state's apparel manufacturers. In 1992, under contract to Marshall, UAH developed an apparel-specific software package that allows manufacturers to design and analyze modules without making an actual investment -- it functions on ordinary PC equipment. By 1995, Marshall had responded to requests for the package from more than 400 companies in 36 states; some of which reported savings up to $2 million. The National Garment Company of Missouri, for example, uses the system to design and balance a modular line before committing to expensive hardware; for setting up sewing lines; and for determining the composition of a new team.

  18. Solvation Structure and Thermodynamic Mapping (SSTMap): An Open-Source, Flexible Package for the Analysis of Water in Molecular Dynamics Trajectories.

    PubMed

    Haider, Kamran; Cruz, Anthony; Ramsey, Steven; Gilson, Michael K; Kurtzman, Tom

    2018-01-09

    We have developed SSTMap, a software package for mapping structural and thermodynamic water properties in molecular dynamics trajectories. The package introduces automated analysis and mapping of local measures of frustration and enhancement of water structure. The thermodynamic calculations are based on Inhomogeneous Fluid Solvation Theory (IST), which is implemented using both site-based and grid-based approaches. The package also extends the applicability of solvation analysis calculations to multiple molecular dynamics (MD) simulation programs by using existing cross-platform tools for parsing MD parameter and trajectory files. SSTMap is implemented in Python and contains both command-line tools and a Python module to facilitate flexibility in setting up calculations and for automated generation of large data sets involving analysis of multiple solutes. Output is generated in formats compatible with popular Python data science packages. This tool will be used by the molecular modeling community for computational analysis of water in problems of biophysical interest such as ligand binding and protein function.

  19. Trends in the use of natural antioxidants in active food packaging: a review.

    PubMed

    Sanches-Silva, Ana; Costa, Denise; Albuquerque, Tânia G; Buonocore, Giovanna Giuliana; Ramos, Fernando; Castilho, Maria Conceição; Machado, Ana Vera; Costa, Helena S

    2014-01-01

    The demand for natural antioxidant active packaging is increasing due to its unquestionable advantages compared with the addition of antioxidants directly to the food. Therefore, the search for antioxidants perceived as natural, namely those that naturally occur in herbs and spices, is a field attracting great interest. In line with this, in the last few years, natural antioxidants such as α-tocopherol, caffeic acid, catechin, quercetin, carvacrol and plant extracts (e.g. rosemary extract) have been incorporated into food packaging. On the other hand, consumers and the food industry are also interested in active biodegradable/compostable packaging and edible films to reduce environmental impact, minimise food loss and minimise contaminants from industrial production and reutilisation by-products. The present review focuses on the natural antioxidants already applied in active food packaging, and it reviews the methods used to determine the oxidation protection effect of antioxidant active films and the methods used to quantify natural antioxidants in food matrices or food simulants. Lastly consumers' demands and industry trends are also addressed.

  20. A geometry package for generation of input data for a three-dimensional potential-flow program

    NASA Technical Reports Server (NTRS)

    Halsey, N. D.; Hess, J. L.

    1978-01-01

    The preparation of geometric data for input to three-dimensional potential flow programs was automated and simplified by a geometry package incorporated into the NASA Langley version of the 3-D lifting potential flow program. Input to the computer program for the geometry package consists of a very sparse set of coordinate data, often with an order of magnitude of fewer points than required for the actual potential flow calculations. Isolated components, such as wings, fuselages, etc. are paneled automatically, using one of several possible element distribution algorithms. Curves of intersection between components are calculated, using a hybrid curve-fit/surface-fit approach. Intersecting components are repaneled so that adjacent elements on either side of the intersection curves line up in a satisfactory manner for the potential-flow calculations. Many cases may be run completely (from input, through the geometry package, and through the flow calculations) without interruption. Use of the package significantly reduces the time and expense involved in making three-dimensional potential flow calculations.

  1. Easing access to R using 'shiny' to create graphical user interfaces: An example for the R package 'Luminescence'

    NASA Astrophysics Data System (ADS)

    Burow, Christoph; Kreutzer, Sebastian; Dietze, Michael; Fuchs, Margret C.; Schmidt, Christoph; Fischer, Manfred; Brückner, Helmut

    2017-04-01

    Since the release of the R package 'Luminescence' (Kreutzer et al., 2012) the functionality of the package has been greatly enhanced by implementing further functions for measurement data processing, statistical analysis and graphical output. Despite its capabilities for complex and non-standard analysis of luminescence data, working with the command-line interface (CLI) of R can be tedious at best and overwhelming at worst, especially for users without experience in programming languages. Even though much work is put into simplifying the usage of the package to continuously lower the entry threshold, at least basic knowledge of R will always be required. Thus, the potential user base of the package cannot be exhausted, at least as long as the CLI is the only means of utilising the 'Luminescence' package. But even experienced users may find it tedious to iteratively run a function until a satisfying results is produced. For example, plotting data is also at least partly subject to personal aesthetic tastes in accordance with the information it is supposed to convey and iterating through all the possible options in the R CLI can be a time-consuming task. An alternative approach to the CLI is the graphical user interface (GUI), which allows direct, interactive manipulation and interaction with the underlying software. For users with little or no experience with command-lines a GUI offers intuitive access that counteracts the perceived steep learning curve of a CLI. Even though R lacks native support for GUI functions, its capabilities of linking it to other programming languages allows to utilise external frameworks to build graphical user interfaces. A recent attempt to provide a GUI toolkit for R was the introduction of the 'shiny' package (Chang et al., 2016), which allows automatic construction of HTML, CSS and JavaScript based user interfaces straight from R. Here, we give (1) a brief introduction to the 'shiny' framework for R, before we (2) present a GUI for the R package 'Luminescence' in the form of interactive web applications. These applications can be accessed online so that a user is not even required to have a local installation of R and which provide access to most of the plotting functions of the R package 'Luminescence'. These functionalities will be demonstrated live during the PICO session. References Chang, W., Cheng, J., Allaire, JJ., Xie, Y., McPherson, J., 2016. shiny: Web Application Framework for R. R package version 0.13.2. https://CRAN.R-project.org/package=shiny Kreutzer, S., Schmidt, C., Fuchs, M.C., Dietze, M., Fischer, M., Fuchs, M., 2012. Introducing an R package for luminescence dating analysis. Ancient TL, 30: 1-8, 2012.

  2. The R package 'icosa' for coarse resolution global triangular and penta-hexagonal gridding

    NASA Astrophysics Data System (ADS)

    Kocsis, Adam T.

    2017-04-01

    With the development of the internet and the computational power of personal computers, open source programming environments have become indispensable for science in the past decade. This includes the increase of the GIS capacity of the free R environment, which was originally developed for statistical analyses. The flexibility of R made it a preferred programming tool in a multitude of disciplines from the area of the biological and geological sciences. Many of these subdisciplines operate with incidence (occurrence) data that are in a large number of cases to be grained before further analyses can be conducted. This graining is executed mostly by gridding data to cells of a Gaussian grid of various resolutions to increase the density of data in a single unit of the analyses. This method has obvious shortcomings despite the ease of its application: well-known systematic biases are induced to cell sizes and shapes that can interfere with the results of statistical procedures, especially if the number of incidence points influences the metrics in question. The 'icosa' package employs a common method to overcome this obstacle by implementing grids with roughly equal cell sizes and shapes that are based on tessellated icosahedra. These grid objects are essentially polyhedra with xyz Cartesian vertex data that are linked to tables of faces and edges. At its current developmental stage, the package uses a single method of tessellation which balances grid cell size and shape distortions, but its structure allows the implementation of various other types of tessellation algorithms. The resolution of the grids can be set by the number of breakpoints inserted into a segment forming an edge of the original icosahedron. Both the triangular and their inverted penta-hexagonal grids are available for creation with the package. The package also incorporates functions to look up coordinates in the grid very effectively and data containers to link data to the grid structure. The classes defined in the package are communicating with classes of the 'sp' and 'raster' packages and functions are supplied that allow resolution change and type conversions. Three-dimensional rendering is made available with the 'rgl' package and two-dimensional projections can be calculated using 'sp' and 'rgdal'. The package was developed as part of a project funded by the Deutsche Forschungsgemeinschaft (KO - 5382/1-1).

  3. Trip Report - June 1989 Swallow Float Deployment with RUM

    DTIC Science & Technology

    1990-12-01

    Float 1. with its external geophone package resting on the sediment, and float 3, equipped with an infra - sonic hydrophone and tethered to the bottom...an external, triaxial geophone package resting on the ocean bottom and the other equippd with an infrasonic hydrophone and bottom-tethered by a 0.5... infrasonic hydrophone and bottom-tethered by a 0.5-meter line, are presented in this report Introduction An experiment designed to compare the ambient sound

  4. Materials for High-Density Electronic Packaging and Interconnection

    DTIC Science & Technology

    1990-04-10

    play a prominent role in the future. Glass and Porcelain The earliest use of electronic ceramics was as insulators for carrying telegraph lines...Administration 61L & CORES , (Ot. stem. SAI WCJm 76. LOISS (C". SUMt *oW WVCf B’%2101 Constitution Avenue. N W Washington, D.C. 20418 Washington. D.C. 20301 G...Density Packaging 84 Tape Automated Bonding 87 Diamond 88 Superconductors 88 Composites 89 Materials for Very-High-Frequency Digital Systems 91

  5. Use of CellNetAnalyzer in biotechnology and metabolic engineering.

    PubMed

    von Kamp, Axel; Thiele, Sven; Hädicke, Oliver; Klamt, Steffen

    2017-11-10

    Mathematical models of the cellular metabolism have become an essential tool for the optimization of biotechnological processes. They help to obtain a systemic understanding of the metabolic processes in the used microorganisms and to find suitable genetic modifications maximizing the production performance. In particular, methods of stoichiometric and constraint-based modeling are frequently used in the context of metabolic and bioprocess engineering. Since metabolic networks can be complex and comprise hundreds or even thousands of metabolites and reactions, dedicated software tools are required for an efficient analysis. One such software suite is CellNetAnalyzer, a MATLAB package providing, among others, various methods for analyzing stoichiometric and constraint-based metabolic models. CellNetAnalyzer can be used via command-line based operations or via a graphical user interface with embedded network visualizations. Herein we will present key functionalities of CellNetAnalyzer for applications in biotechnology and metabolic engineering and thereby review constraint-based modeling techniques such as metabolic flux analysis, flux balance analysis, flux variability analysis, metabolic pathway analysis (elementary flux modes) and methods for computational strain design. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

  6. Development of an in vitro PIG-A gene mutation assay in human cells

    PubMed Central

    Rees, Benjamin J; Tate, Matthew; Lynch, Anthony M; Thornton, Catherine A; Jenkins, Gareth J; Walmsley, Richard M; Johnson, George E

    2017-01-01

    Abstract Mutagens can be carcinogens, and traditionally, they have been identified in vitro using the Salmonella ‘Ames’ reverse mutation assay. However, prokaryotic DNA packaging, replication and repair systems are mechanistically very different to those in the humans we inevitably seek to protect. Therefore, for many years, mammalian cell line genotoxicity assays that can detect eukaryotic mutagens as well as clastogens and aneugens have been used. The apparent lack of specificity in these largely rodent systems, due partly to their mutant p53 status, has contributed to the use of animal studies to resolve data conflicts. Recently, silencing mutations at the PIG-A locus have been demonstrated to prevent glycophosphatidylinositol (GPI) anchor synthesis and consequentially result in loss of GPI-anchored proteins from the cell’s extracellular surface. The successful exploitation of this mutant phenotype in animal studies has triggered interest in the development of an analogous in vitro PIG-A mutation screening assay. This article describes the development of a robust assay design using metabolically active human cells. The assay includes viability and cell membrane integrity assessment and conforms to the future ideas of the 21st-century toxicology testing. PMID:28057708

  7. Comparison of porcine endogenous retroviruses infectious potential in supernatants of producer cells and in cocultures.

    PubMed

    Costa, Michael Rodrigues; Fischer, Nicole; Gulich, Barbara; Tönjes, Ralf R

    2014-01-01

    Porcine endogenous retroviruses (PERV) pose a zoonotic risk potential in pig-to-human xenotransplantation given that PERV capacity to infect different human cell lines in vitro has been clearly shown in the past. However, PERV infectious potential for human peripheral blood mononuclear cells (huPBMC) has been also demonstrated, albeit with controversial results. As productive PERV infection of huPBMC involves immune suppression that may attract opportunistic pathogens as shown for other retroviruses, it is crucial to ascertain unequivocally huPBMC susceptibility for PERV. To address this question, we first investigated in vitro infectivity of PERV for huPBMC using supernatants containing highly infectious PERV-A/C. Second, huPBMC were cocultivated with PERV-A/C producer cells to come a step closer to the in vivo situation of xenotransplantation. In addition, cocultivation of huPBMC with porcine PBMC (poPBMC) isolated from German landrace pigs was performed to distinguish PERV replication competence when they were constitutively produced by immortalized cells or by primary poPBMC. Supernatants containing recombinant highly infectious PERV-A/C were used to infect PHA-activated huPBMC in the presence or absence of polybrene. Next, PERV-producing cell lines such as human 293/5° and primary mitogenically activated poPBMC of three German landrace pigs were cocultivated with huPBMC as well as with susceptible human and porcine cell lines as controls. PERV infection was monitored by using three test approaches. The presence of provirus DNA in putatively infected cells was detected via sensitive nested PCR. Viral expression was determined by screening for the activity of gammaretroviral reverse transcriptase (RT) in cell-free supernatants of infected cells. Virus release was monitored by counting the number of packaged RNA particles in supernatants via PERV-specific quantitative one-step real-time reverse transcriptase PCR. Porcine endogenous retroviruses-A/C in supernatants of human producer 293/5° cells was not able to infect huPBMC. Neither RT activity nor PERV copies were detected. Even provirus could not be detected displaying the inability of PERV-A/C to induce a productive infection in huPBMC. In cocultivation experiments only non-productive infection of huPBMC with PERV derived from 293/5° cell line and from PHA-activated poPBMC was observed by detection of provirus DNA in infected cells. Recombinant PERV-A/C in supernatants of producer cells failed to infect huPBMC, whereas coculture experiments with producer cell lines lead to non-productive infection of huPBMC. PERV in supernatants seem to have not sufficient infectious potential for huPBMC. However, extensive PERV exposure to huPBMC via cocultivation enabled at least virus cell entry as provirus was detected by nested PCR. Furthermore, results presented support previous data showing German landrace pigs as low producers with negligible infectious potential due to the absence of replication-competent PERV in the genome. The low PERV expression profile and the lack of significant replication competence of German landrace pigs raise hope for considering these animals as putative donor animals in future pig-to-human xenotransplantation. Nonetheless, data imply that PERV still represent a virological risk in the course of xenotransplantation, as the presence of PERV provirus in host cells may lead to a provirus integration resulting in insertional mutagenesis and chromosomal rearrangements. © 2014 John Wiley & Sons A/S.

  8. Program Package for the Analysis of High Resolution High Signal-To-Noise Stellar Spectra

    NASA Astrophysics Data System (ADS)

    Piskunov, N.; Ryabchikova, T.; Pakhomov, Yu.; Sitnova, T.; Alekseeva, S.; Mashonkina, L.; Nordlander, T.

    2017-06-01

    The program package SME (Spectroscopy Made Easy), designed to perform an analysis of stellar spectra using spectral fitting techniques, was updated due to adding new functions (isotopic and hyperfine splittins) in VALD and including grids of NLTE calculations for energy levels of few chemical elements. SME allows to derive automatically stellar atmospheric parameters: effective temperature, surface gravity, chemical abundances, radial and rotational velocities, turbulent velocities, taking into account all the effects defining spectral line formation. SME package uses the best grids of stellar atmospheres that allows us to perform spectral analysis with the similar accuracy in wide range of stellar parameters and metallicities - from dwarfs to giants of BAFGK spectral classes.

  9. WannierTools: An open-source software package for novel topological materials

    NASA Astrophysics Data System (ADS)

    Wu, QuanSheng; Zhang, ShengNan; Song, Hai-Feng; Troyer, Matthias; Soluyanov, Alexey A.

    2018-03-01

    We present an open-source software package WannierTools, a tool for investigation of novel topological materials. This code works in the tight-binding framework, which can be generated by another software package Wannier90 (Mostofi et al., 2008). It can help to classify the topological phase of a given material by calculating the Wilson loop, and can get the surface state spectrum, which is detected by angle resolved photoemission (ARPES) and in scanning tunneling microscopy (STM) experiments. It also identifies positions of Weyl/Dirac points and nodal line structures, calculates the Berry phase around a closed momentum loop and Berry curvature in a part of the Brillouin zone (BZ).

  10. IceBridge Provides Novel Evidence for Thick Units of Basal Freeze-on Ice Along Petermann Glacier, Greenland

    NASA Astrophysics Data System (ADS)

    Bell, R. E.; Tinto, K. J.; Wolovick, M.; Block, A. E.; Frearson, N.; Das, I.; Abdi, A.; Creyts, T. T.; Cochran, J. R.; Csatho, B. M.; Babonis, G. S.

    2011-12-01

    The Petermann Glacier, one of the major outlet glaciers in Greenland, drains six percent of the Greenland ice from a basin largely below sea level. Petermann Glacier and its large ice shelf may be susceptible to increased change as the waters along the Greenland margin warm. The 2010 and 2011 Operation IceBridge mission, acquired a comprehensive aerogeophysical data set over the Petermann Glacier that provides insights into the ice sheet structure. This analysis employs most of the data streams acquired by the Icebridge platform including ice-penetrating radar, laser altimetry, gravity and magnetics. An orthogonal 10 km grid extends from 60 km upstream of the grounding line to 240 km inland. The ice velocities in the region range from <50m/yr to >200m/yr. On the interior lines the internal layers are pulled down over 2-3 km wide regions. Up to 400m of ice from the base of the ice sheet appears to be absent in these regions. We interpret these pulled down regions as basal melt. These melt regions are mainly located along the upstream side of a 80 km wide east-west trending topographic ridge that separates the interior ice from the Petermann Fjord. The IceBridge magnetic data indicates that this broad flat ridge is the boundary between the Franklinian Basins and the Ellsmerian Foldbelt to the north. Downstream of these pull-down layers we have identified 4 distinct packages of ice that thicken downstream and are characterized by a strong upper reflector. These packages develop at the base of the ice sheet and reach thicknesses of 500-700m over distances of 10-20 km. These basal packages can be traced for 30-100 km following the direction of flow, and may be present close to the grounding line. These basal reflectors deflect the overlying internal layers upward indicating the addition of ice to the base of the ice sheet. The IceBridge gravity data indicates that these features are probably not off-nadir topography since these would show up as around 30mGal anomalies in the gravity data, and no such signature exists. We interpret these basal features in the radar data as ice. The geometry of these packages is very similar to the frozen-on ice packages imaged along the margins of the Gamburtsev Mountains, East Antarctica. We have interpreted these packages as bodies of accreted ice sourced from the upstream melting. Similar packages of basal ice up to 1200m thick are found throughout the margins of the Greenland ice sheet north of 70°N. The accretion process is modifying the base of the ice sheet in the onset regions of Petermann and other outlet glaciers in Greenland. Any change in rheology between basal and meteoric ice is likely to influence ice flow fostering enhanced melting and deformation of the basal ice. These frozen-on basal ice packages may be more susceptible to melt when ice sheet goes afloat and may be the origin of the elongate melt channels in the ice shelf. Accretion may influence both ice flow in outlet glaciers and melt rates at the grounding line and the adjacent ice shelves.

  11. Pizza.py Toolkit

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Plimpton, Steve; Jones, Matt; Crozier, Paul

    2006-01-01

    Pizza.py is a loosely integrated collection of tools, many of which provide support for the LAMMPS molecular dynamics and ChemCell cell modeling packages. There are tools to create input files. convert between file formats, process log and dump files, create plots, and visualize and animate simulation snapshots. Software packages that are wrapped by Pizza.py. so they can invoked from within Python, include GnuPlot, MatLab, Raster3d. and RasMol. Pizza.py is written in Python and runs on any platform that supports Python. Pizza.py enhances the standard Python interpreter in a few simple ways. Its tools are Python modules which can be invokedmore » interactively, from scripts, or from GUIs when appropriate. Some of the tools require additional Python packages to be installed as part of the users Python. Others are wrappers on software packages (as listed above) which must be available on the users system. It is easy to modify or extend Pizza.py with new functionality or new tools, which need not have anything to do with LAMMPS or ChemCell.« less

  12. The Ocean Observatories Initiative: Data Acquisition Functions and Its Built-In Automated Python Modules

    NASA Astrophysics Data System (ADS)

    Smith, M. J.; Vardaro, M.; Crowley, M. F.; Glenn, S. M.; Schofield, O.; Belabbassi, L.; Garzio, L. M.; Knuth, F.; Fram, J. P.; Kerfoot, J.

    2016-02-01

    The Ocean Observatories Initiative (OOI), funded by the National Science Foundation, provides users with access to long-term datasets from a variety of oceanographic sensors. The Endurance Array in the Pacific Ocean consists of two separate lines off the coasts of Oregon and Washington. The Oregon line consists of 7 moorings, two cabled benthic experiment packages and 6 underwater gliders. The Washington line comprises 6 moorings and 6 gliders. Each mooring is outfitted with a variety of instrument packages. The raw data from these instruments are sent to shore via satellite communication and in some cases, via fiber optic cable. Raw data is then sent to the cyberinfrastructure (CI) group at Rutgers where it is aggregated, parsed into thousands of different data streams, and integrated into a software package called uFrame. The OOI CI delivers the data to the general public via a web interface that outputs data into commonly used scientific data file formats such as JSON, netCDF, and CSV. The Rutgers data management team has developed a series of command-line Python tools that streamline data acquisition in order to facilitate the QA/QC review process. The first step in the process is querying the uFrame database for a list of all available platforms. From this list, a user can choose a specific platform and automatically download all available datasets from the specified platform. The downloaded dataset is plotted using a generalized Python netcdf plotting routine that utilizes a data visualization toolbox called matplotlib. This routine loads each netCDF file separately and outputs plots by each available parameter. These Python tools have been uploaded to a Github repository that is openly available to help facilitate OOI data access and visualization.

  13. Analysis and characterizations of planar transmission structures and components for superconducting and monolithic integrated circuits

    NASA Technical Reports Server (NTRS)

    Itoh, Tatsuo

    1991-01-01

    The analysis and modeling of superconducting planar transmission lines were performed. Theoretically, the highest possible Q values of superconducting microstrip line was calculated and, as a result, it provided the Q value that the experiment can aim for. As an effort to search for a proper superconducting transmission line structure, the superconducting microstrip line and coplanar waveguide were compared in terms of loss characteristics and their design aspects. Also, the research was expanded to a superconducting coplanar waveguide family in the microwave packaging environment. Theoretically, it was pointed out that the substrate loss is critical in the superconducting transmission line structures.

  14. Ground-Based GPS Sensing of Azimuthal Variations in Precipitable Water Vapor

    NASA Technical Reports Server (NTRS)

    Kroger, P. M.; Bar-Sever, Y. E.

    1997-01-01

    Current models for troposphere delay employed by GPS software packages map the total zenith delay to the line-of-sight delay of the individual satellite-receiver link under the assumption of azimuthal homogeneity. This could be a poor approximation for many sites, in particular, those located at an ocean front or next to a mountain range. We have modified the GIPSY-OASIS II software package to include a simple non-symmetric mapping function (MacMillan, 1995) which introduces two new parameters.

  15. Reflective Packaging

    NASA Technical Reports Server (NTRS)

    1994-01-01

    The aluminized polymer film used in spacecraft as a radiation barrier to protect both astronauts and delicate instruments has led to a number of spinoff applications. Among them are aluminized shipping bags, food cart covers and medical bags. Radiant Technologies purchases component materials and assembles a barrier made of layers of aluminized foil. The packaging reflects outside heat away from the product inside the container. The company is developing new aluminized lines, express mailers, large shipping bags, gel packs and insulated panels for the building industry.

  16. TMAP: Tübingen NLTE Model-Atmosphere Package

    NASA Astrophysics Data System (ADS)

    Werner, Klaus; Dreizler, Stefan; Rauch, Thomas

    2012-12-01

    The Tübingen NLTE Model-Atmosphere Package (TMAP) is a tool to calculate stellar atmospheres in spherical or plane-parallel geometry in hydrostatic and radiative equilibrium allowing departures from local thermodynamic equilibrium (LTE) for the population of atomic levels. It is based on the Accelerated Lambda Iteration (ALI) method and is able to account for line blanketing by metals. All elements from hydrogen to nickel may be included in the calculation with model atoms which are tailored for the aims of the user.

  17. NASA-DoD Lead-Free Electronics Project

    NASA Technical Reports Server (NTRS)

    Kessel, Kurt

    2009-01-01

    In response to concerns about risks from lead-free induced faults to high reliability products, NASA has initiated a multi-year project to provide manufacturers and users with data to clarify the risks of lead-free materials in their products. The project will also be of interest to component manufacturers supplying to high reliability markets. The project was launched in November 2006. The primary technical objective of the project is to undertake comprehensive testing to generate information on failure modes/criteria to better understand the reliability of: (1) Packages (e.g., Thin Small Outline Package [TSOP], Ball Grid Array [BGA], Plastic Dual In-line Package [PDIP]) assembled and reworked with solder interconnects consisting of lead-free alloys (2) Packages (e.g., TSOP, BGA, PDIP) assembled and reworked with solder interconnects consisting of mixed alloys, lead component finish/lead-free solder and lead-free component finish/SnPb solder

  18. Delivery of chemotherapeutic drugs in tumour cell-derived microparticles.

    PubMed

    Tang, Ke; Zhang, Yi; Zhang, Huafeng; Xu, Pingwei; Liu, Jing; Ma, Jingwei; Lv, Meng; Li, Dapeng; Katirai, Foad; Shen, Guan-Xin; Zhang, Guimei; Feng, Zuo-Hua; Ye, Duyun; Huang, Bo

    2012-01-01

    Cellular microparticles are vesicular plasma membrane fragments with a diameter of 100-1,000 nanometres that are shed by cells in response to various physiological and artificial stimuli. Here we demonstrate that tumour cell-derived microparticles can be used as vectors to deliver chemotherapeutic drugs. We show that tumour cells incubated with chemotherapeutic drugs package these drugs into microparticles, which can be collected and used to effectively kill tumour cells in murine tumour models without typical side effects. We describe several mechanisms involved in this process, including uptake of drug-containing microparticles by tumour cells, synthesis of additional drug-packaging microparticles by these cells that contribute to the cytotoxic effect and the inhibition of drug efflux from tumour cells. This study highlights a novel drug delivery strategy with potential clinical application.

  19. Impact of Policy Changes on Infant Feeding Decisions Among Low-Income Women Participating in the Special Supplemental Nutrition Program for Women, Infants, and Children

    PubMed Central

    Koleilat, Maria; Whaley, Mike; Gomez, Judy; Meehan, Karen; Saluja, Kiran

    2012-01-01

    Objectives. We present infant feeding data before and after the 2009 Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) food package change that supported and incentivized breastfeeding. We describe the key role of California WIC staff in supporting these policy changes. Methods. We analyzed WIC data on more than 180 000 infants in Southern California. We employed the analysis of variance and Tukey (honestly significant difference) tests to compare issuance rates of postpartum and infant food packages before and after the changes. We used analysis of covariance to adjust for poverty status changes as a potential confounder. Results. Issuance rates of the “fully breastfeeding” package at infant WIC enrollment increased by 86% with the package changes. Rates also increased significantly for 2- and 6-month-old infants. Issuance rates of packages that included formula decreased significantly. All outcomes remained highly significant in the adjusted model. Conclusions. Policy changes, training of front-line WIC staff, and participant education influenced issuance rates of WIC food packages. In California, the issuance rates of packages that include formula have significantly decreased and the rate for those that include no formula has significantly increased. PMID:23078467

  20. Generation of an Adenovirus-Parvovirus Chimera with Enhanced Oncolytic Potential

    PubMed Central

    El-Andaloussi, Nazim; Bonifati, Serena; Kaufmann, Johanna K.; Mailly, Laurent; Daeffler, Laurent; Deryckère, François; Nettelbeck, Dirk M.; Rommelaere, Jean

    2012-01-01

    In this study, our goal was to generate a chimeric adenovirus-parvovirus (Ad-PV) vector that combines the high-titer and efficient gene transfer of adenovirus with the anticancer potential of rodent parvovirus. To this end, the entire oncolytic PV genome was inserted into a replication-defective E1- and E3-deleted Ad5 vector genome. As we found that parvoviral NS expression inhibited Ad-PV chimera production, we engineered the parvoviral P4 early promoter, which governs NS expression, by inserting into its sequence tetracycline operator elements. As a result of these modifications, P4-driven expression was blocked in the packaging T-REx-293 cells, which constitutively express the tetracycline repressor, allowing high-yield chimera production. The chimera effectively delivered the PV genome into cancer cells, from which fully infectious replication-competent parvovirus particles were generated. Remarkably, the Ad-PV chimera exerted stronger cytotoxic activities against various cancer cell lines, compared with the PV and Ad parental viruses, while being still innocuous to a panel of tested healthy primary human cells. This Ad-PV chimera represents a novel versatile anticancer agent which can be subjected to further genetic manipulations in order to reinforce its enhanced oncolytic capacity through arming with transgenes or retargeting into tumor cells. PMID:22787235

  1. Generation of an adenovirus-parvovirus chimera with enhanced oncolytic potential.

    PubMed

    El-Andaloussi, Nazim; Bonifati, Serena; Kaufmann, Johanna K; Mailly, Laurent; Daeffler, Laurent; Deryckère, François; Nettelbeck, Dirk M; Rommelaere, Jean; Marchini, Antonio

    2012-10-01

    In this study, our goal was to generate a chimeric adenovirus-parvovirus (Ad-PV) vector that combines the high-titer and efficient gene transfer of adenovirus with the anticancer potential of rodent parvovirus. To this end, the entire oncolytic PV genome was inserted into a replication-defective E1- and E3-deleted Ad5 vector genome. As we found that parvoviral NS expression inhibited Ad-PV chimera production, we engineered the parvoviral P4 early promoter, which governs NS expression, by inserting into its sequence tetracycline operator elements. As a result of these modifications, P4-driven expression was blocked in the packaging T-REx-293 cells, which constitutively express the tetracycline repressor, allowing high-yield chimera production. The chimera effectively delivered the PV genome into cancer cells, from which fully infectious replication-competent parvovirus particles were generated. Remarkably, the Ad-PV chimera exerted stronger cytotoxic activities against various cancer cell lines, compared with the PV and Ad parental viruses, while being still innocuous to a panel of tested healthy primary human cells. This Ad-PV chimera represents a novel versatile anticancer agent which can be subjected to further genetic manipulations in order to reinforce its enhanced oncolytic capacity through arming with transgenes or retargeting into tumor cells.

  2. Tobacco packaging and labeling policies under the U.S. Tobacco Control Act: research needs and priorities.

    PubMed

    Hammond, David

    2012-01-01

    The Family Smoking Prevention and Tobacco Control Act (the "Act"), enacted in June 2009, gave the U.S. Food and Drug Administration authority to regulate tobacco products. The current paper reviews the provisions for packaging and labeling, including the existing evidence and research priorities. Narrative review using electronic literature search of published and unpublished sources in 3 primary areas: health warnings, constituent labeling, and prohibitions on the promotional elements of packaging. The Act requires 9 pictorial health warnings covering half of cigarette packages and 4 text warnings covering 30% of smokeless tobacco packages. The Act also prohibits potentially misleading information on packaging, including the terms "light" and "mild," and provides a mandate to require disclosure of chemical constituents on packages. Many of the specific regulatory provisions are based on the extent to which they promote "greater public understanding of the risks of tobacco." As a result, research on consumer perceptions has the potential to shape the design and renewal of health warnings and to determine what, if any, information on product constituents should appear on packages. Research on consumer perceptions of existing and novel tobacco products will also be critical to help identify potentially misleading information that should be restricted under the Act. Packaging and labeling regulations required under the Act will bring the United States in line with international standards. There is an immediate need for research to evaluate these measures to guide future regulatory action.

  3. Interstellar lines in high resolution IUE spectra. Part 1: Groningen data reduction package and technical results

    NASA Astrophysics Data System (ADS)

    Gilra, D. P.; Pwa, T. H.; Arnal, E. M.; de Vries, J.

    1982-06-01

    In order to process and analyze high resolution IUE data on a large number of interstellar lines in a large number of images for a large number of stars, computer programs were developed for 115 lines in the short wavelength range and 40 in the long wavelength range. Programs include extraction, processing, plotting, averaging, and profile fitting. Wavelength calibration in high resolution spectra, fixed pattern noise, instrument profile and resolution, and the background problem in the region where orders are crowding are discussed. All the expected lines are detected in at least one spectrum.

  4. Spontaneous Packaging and Hypothermic Storage of Mammalian Cells with a Cell-Membrane-Mimetic Polymer Hydrogel in a Microchip.

    PubMed

    Xu, Yan; Mawatari, Kazuma; Konno, Tomohiro; Kitamori, Takehiko; Ishihara, Kazuhiko

    2015-10-21

    Currently, continuous culture/passage and cryopreservation are two major, well-established methods to provide cultivated mammalian cells for experiments in laboratories. Due to the lack of flexibility, however, both laboratory-oriented methods are unable to meet the need for rapidly growing cell-based applications, which require cell supply in a variety of occasions outside of laboratories. Herein, we report spontaneous packaging and hypothermic storage of mammalian cells under refrigerated (4 °C) and ambient conditions (25 °C) using a cell-membrane-mimetic methacryloyloxyethyl phosphorylcholine (MPC) polymer hydrogel incorporated within a glass microchip. Its capability for hypothermic storage of cells was comparatively evaluated over 16 days. The results reveal that the cytocompatible MPC polymer hydrogel, in combination with the microchip structure, enabled hypothermic storage of cells with quite high viability, high intracellular esterase activity, maintained cell membrane integrity, and small morphological change for more than 1 week at 4 °C and at least 4 days at 25 °C. Furthermore, the stored cells could be released from the hydrogel and exhibited the ability to adhere to a surface and achieve confluence under standard cell culture conditions. Both hypothermic storage conditions are ordinary flexible conditions which can be easily established in places outside of laboratories. Therefore, cell packaging and storage using the hydrogel incorporated within the microchip would be a promising miniature and portable solution for flexible supply and delivery of small amounts of cells from bench to bedside.

  5. funtooNorm: an R package for normalization of DNA methylation data when there are multiple cell or tissue types.

    PubMed

    Oros Klein, Kathleen; Grinek, Stepan; Bernatsky, Sasha; Bouchard, Luigi; Ciampi, Antonio; Colmegna, Ines; Fortin, Jean-Philippe; Gao, Long; Hivert, Marie-France; Hudson, Marie; Kobor, Michael S; Labbe, Aurelie; MacIsaac, Julia L; Meaney, Michael J; Morin, Alexander M; O'Donnell, Kieran J; Pastinen, Tomi; Van Ijzendoorn, Marinus H; Voisin, Gregory; Greenwood, Celia M T

    2016-02-15

    DNA methylation patterns are well known to vary substantially across cell types or tissues. Hence, existing normalization methods may not be optimal if they do not take this into account. We therefore present a new R package for normalization of data from the Illumina Infinium Human Methylation450 BeadChip (Illumina 450 K) built on the concepts in the recently published funNorm method, and introducing cell-type or tissue-type flexibility. funtooNorm is relevant for data sets containing samples from two or more cell or tissue types. A visual display of cross-validated errors informs the choice of the optimal number of components in the normalization. Benefits of cell (tissue)-specific normalization are demonstrated in three data sets. Improvement can be substantial; it is strikingly better on chromosome X, where methylation patterns have unique inter-tissue variability. An R package is available at https://github.com/GreenwoodLab/funtooNorm, and has been submitted to Bioconductor at http://bioconductor.org. © The Author 2015. Published by Oxford University Press.

  6. Trans-packaging of human immunodeficiency virus type 1 genome into Gag virus-like particles in Saccharomyces cerevisiae.

    PubMed

    Tomo, Naoki; Goto, Toshiyuki; Morikawa, Yuko

    2013-03-26

    Yeast is recognized as a generally safe microorganism and is utilized for the production of pharmaceutical products, including vaccines. We previously showed that expression of human immunodeficiency virus type 1 (HIV-1) Gag protein in Saccharomyces cerevisiae spheroplasts released Gag virus-like particles (VLPs) extracellularly, suggesting that the production system could be used in vaccine development. In this study, we further establish HIV-1 genome packaging into Gag VLPs in a yeast cell system. The nearly full-length HIV-1 genome containing the entire 5' long terminal repeat, U3-R-U5, did not transcribe gag mRNA in yeast. Co-expression of HIV-1 Tat, a transcription activator, did not support the transcription. When the HIV-1 promoter U3 was replaced with the promoter for the yeast glyceraldehyde-3-phosphate dehydrogenase gene, gag mRNA transcription was restored, but no Gag protein expression was observed. Co-expression of HIV-1 Rev, a factor that facilitates nuclear export of gag mRNA, did not support the protein synthesis. Progressive deletions of R-U5 and its downstream stem-loop-rich region (SL) to the gag start ATG codon restored Gag protein expression, suggesting that a highly structured noncoding RNA generated from the R-U5-SL region had an inhibitory effect on gag mRNA translation. When a plasmid containing the HIV-1 genome with the R-U5-SL region was coexpressed with an expression plasmid for Gag protein, the HIV-1 genomic RNA was transcribed and incorporated into Gag VLPs formed by Gag protein assembly, indicative of the trans-packaging of HIV-1 genomic RNA into Gag VLPs in a yeast cell system. The concentration of HIV-1 genomic RNA in Gag VLPs released from yeast was approximately 500-fold higher than that in yeast cytoplasm. The deletion of R-U5 to the gag gene resulted in the failure of HIV-1 RNA packaging into Gag VLPs, indicating that the packaging signal of HIV-1 genomic RNA present in the R-U5 to gag region functions similarly in yeast cells. Our data indicate that selective trans-packaging of HIV-1 genomic RNA into Gag VLPs occurs in a yeast cell system, analogous to a mammalian cell system, suggesting that yeast may provide an alternative packaging system for lentiviral RNA.

  7. A Mutation in UL15 of Herpes Simplex Virus 1 That Reduces Packaging of Cleaved Genomes▿

    PubMed Central

    Yang, Kui; Wills, Elizabeth G.; Baines, Joel D.

    2011-01-01

    Herpesvirus genomic DNA is cleaved from concatemers that accumulate in infected cell nuclei. Genomic DNA is inserted into preassembled capsids through a unique portal vertex. Extensive analyses of viral mutants have indicated that intact capsids, the portal vertex, and all components of a tripartite terminase enzyme are required to both cleave and package viral DNA, suggesting that DNA cleavage and packaging are inextricably linked. Because the processes have not been functionally separable, it has been difficult to parse the roles of individual proteins in the DNA cleavage/packaging reaction. In the present study, a virus bearing the deletion of codons 400 to 420 of UL15, encoding a terminase component, was analyzed. This virus, designated vJB27, failed to replicate on noncomplementing cells but cleaved concatemeric DNA to ca. 35 to 98% of wild-type levels. No DNA cleavage was detected in cells infected with a UL15-null virus or a virus lacking UL15 codons 383 to 385, comprising a motif proposed to couple ATP hydrolysis to DNA translocation. The amount of vJB27 DNA protected from DNase I digestion was reduced compared to the wild-type virus by 6.5- to 200-fold, depending on the DNA fragment analyzed, thus indicating a profound defect in DNA packaging. Capsids containing viral DNA were not detected in vJB27-infected cells, as determined by electron microscopy. These data suggest that pUL15 plays an essential role in DNA translocation into the capsid and indicate that this function is separable from its role in DNA cleavage. PMID:21880766

  8. Randomization and In Vivo Selection Reveal a GGRG Motif Essential for Packaging Human Immunodeficiency Virus Type 2 RNA ▿ †

    PubMed Central

    Baig, Tayyba T.; Lanchy, Jean-Marc; Lodmell, J. Stephen

    2009-01-01

    The packaging signal (ψ) of human immunodeficiency virus type 2 (HIV-2) is present in the 5′ noncoding region of RNA and contains a 10-nucleotide palindrome (pal; 5′-392-GGAGUGCUCC) located upstream of the dimerization signal stem-loop 1 (SL1). pal has been shown to be functionally important in vitro and in vivo. We previously showed that the 3′ side of pal (GCUCC-3′) is involved in base-pairing interactions with a sequence downstream of SL1 to make an extended SL1, which is important for replication in vivo and the regulation of dimerization in vitro. However, the role of the 5′ side of pal (5′-GGAGU) was less clear. Here, we characterized this role using an in vivo SELEX approach. We produced a population of HIV-2 DNA genomes with random sequences within the 5′ side of pal and transfected these into COS-7 cells. Viruses from COS-7 cells were used to infect C8166 permissive cells. After several weeks of serial passage in C8166 cells, surviving viruses were sequenced. On the 5′ side of pal there was a striking convergence toward a GGRGN consensus sequence. Individual clones with consensus and nonconsensus sequences were tested in infectivity and packaging assays. Analysis of individuals that diverged from the consensus sequence showed normal viral RNA and protein synthesis but had replication defects and impaired RNA packaging. These findings clearly indicate that the GGRG motif is essential for viral replication and genomic RNA packaging. PMID:18971263

  9. A Study of Direct Digital Manufactured RF/Microwave Packaging

    NASA Astrophysics Data System (ADS)

    Stratton, John W. I.

    Various facets of direct digital manufactured (DDM) microwave packages are studied. The rippled surface inherent in fused deposition modeling (FDM) fabricated geometries is modeled in Ansoft HFSS, and its effect on the performance of microstrip transmission lines is assessed via simulation and measurement. The thermal response of DDM microstrip transmission lines is analyzed over a range of RF input powers, and linearity is confirmed over that range. Two IC packages are embedded into DDM printed circuit boards, and their performance is analyzed. The first is a low power RF switch, and the second is an RF front end device that includes a low noise amplifier (LNA) and a power amplifier (PA). The RF switch is shown to perform well, as compared to a layout designed for a Rogers 4003C microwave laminate substrate. The LNA performs within datasheet specifications. The power amplifier generates substantial heat, so a thermal management attempt is described. Finally, a capacitively loaded 6dB Wilkinson power divider is designed and fabricated using DDM techniques and materials. Its performance is analyzed and compared to simulation. The device is shown to compare favorably to a similar device fabricated on a Rogers 4003C microwave laminate using traditional printed circuit board techniques.

  10. The SCUBA map reduction cookbook

    NASA Astrophysics Data System (ADS)

    Sandell, G.; Jessop, N.; Jenness, T.

    This cookbook tells you how to reduce and analyze maps obtained with SCUBA using the off-line SCUBA reduction package, SURF, and the Starlink KAPPA, Figaro, GAIA and CONVERT applications. The easiest way of using these packages is to run-up ORAC-DR, a general purpose pipeline for reducing data from any telescope. A set of data reduction recipes are available to ORAC-DR for use when working with scuba maps, these recipes utilize the SURF and KAPPA packages. This cookbook makes no attempts to explain why and how, for that there is a comprehensive Starlink User Note 216 which properly documents all the software tasks in SURF, which should be consulted for those who need to know details of a task, or how the task really works.

  11. Development of program package for investigation and modeling of carbon nanostructures in diamond like carbon films with the help of Raman scattering and infrared absorption spectra line resolving

    NASA Astrophysics Data System (ADS)

    Hayrapetyan, David B.; Hovhannisyan, Levon; Mantashyan, Paytsar A.

    2013-04-01

    The analysis of complex spectra is an actual problem for modern science. The work is devoted to the creation of a software package, which analyzes spectrum in the different formats, possesses by dynamic knowledge database and self-study mechanism, performs automated analysis of the spectra compound based on knowledge database by application of certain algorithms. In the software package as searching systems, hyper-spherical random search algorithms, gradient algorithms and genetic searching algorithms were used. The analysis of Raman and IR spectrum of diamond-like carbon (DLC) samples were performed by elaborated program. After processing the data, the program immediately displays all the calculated parameters of DLC.

  12. Novel high-density packaging of solid state diode pumped eye-safe laser for LIBS

    NASA Astrophysics Data System (ADS)

    Bares, Kim; Torgerson, Justin; McNeil, Laine; Maine, Patrick; Patterson, Steve

    2018-02-01

    Laser-Induced Breakdown Spectroscopy (LIBS) has proven to be a useful research tool for material analysis for decades. However, because of the amount of energy required in a few nanosecond pulse to generate a stable and reliable LIBS signal, the lasers are often large and inefficient, relegating their implementation to research facilities, factory floors, and assembly lines. Small portable LIBS systems are now possible without having to compromise on energy needs by leveraging off of advances in high-density packaging of electronics, opto-mechanics, and highly efficient laser resonator architecture. This paper explores the integration of these techniques to achieve a mJ class eye-safe LIBS laser source, while retaining a small, light-weight package suitable for handheld systems.

  13. HapMap filter 1.0: a tool to preprocess the HapMap genotypic data for association studies.

    PubMed

    Zhang, Wei; Duan, Shiwei; Dolan, M Eileen

    2008-05-13

    The International HapMap Project provides a resource of genotypic data on single nucleotide polymorphisms (SNPs), which can be used in various association studies to identify the genetic determinants for phenotypic variations. Prior to the association studies, the HapMap dataset should be preprocessed in order to reduce the computation time and control the multiple testing problem. The less informative SNPs including those with very low genotyping rate and SNPs with rare minor allele frequencies to some extent in one or more population are removed. Some research designs only use SNPs in a subset of HapMap cell lines. Although the HapMap website and other association software packages have provided some basic tools for optimizing these datasets, a fast and user-friendly program to generate the output for filtered genotypic data would be beneficial for association studies. Here, we present a flexible, straight-forward bioinformatics program that can be useful in preparing the HapMap genotypic data for association studies by specifying cell lines and two common filtering criteria: minor allele frequencies and genotyping rate. The software was developed for Microsoft Windows and written in C++. The Windows executable and source code in Microsoft Visual C++ are available at Google Code (http://hapmap-filter-v1.googlecode.com/) or upon request. Their distribution is subject to GNU General Public License v3.

  14. Enhancing the Breadth and Efficacy of Therapeutic Vaccines for Breast Cancer

    DTIC Science & Technology

    2014-10-01

    sequence data produced by the Slansky team following their single-cell emulsion RT-PCR technique; however, it can be packaged and shared for use...cell emulsion RT-PCR. Additional modifications were made to our epitope discovery workflow to increase efficacy of transcript and neoantigen candidate...the MiTCR [8] open source software package developed by MiLaboratory. MiTCR is a highly efficient and fast approach to CDR3 extraction, clonotype

  15. 49 CFR 171.16 - Detailed hazardous materials incident reports.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ..., explosion or dangerous evolution of heat (i.e., an amount of heat sufficient to be dangerous to packaging or... seal, pump, compressor, or valve; or (iii) Connection or disconnection of loading or unloading lines...

  16. Packaging - Materials review

    NASA Astrophysics Data System (ADS)

    Herrmann, Matthias

    2014-06-01

    Nowadays, a large number of different electrochemical energy storage systems are known. In the last two decades the development was strongly driven by a continuously growing market of portable electronic devices (e.g. cellular phones, lap top computers, camcorders, cameras, tools). Current intensive efforts are under way to develop systems for automotive industry within the framework of electrically propelled mobility (e.g. hybrid electric vehicles, plug-in hybrid electric vehicles, full electric vehicles) and also for the energy storage market (e.g. electrical grid stability, renewable energies). Besides the different systems (cell chemistries), electrochemical cells and batteries were developed and are offered in many shapes, sizes and designs, in order to meet performance and design requirements of the widespread applications. Proper packaging is thereby one important technological step for designing optimum, reliable and safe batteries for operation. In this contribution, current packaging approaches of cells and batteries together with the corresponding materials are discussed. The focus is laid on rechargeable systems for industrial applications (i.e. alkaline systems, lithium-ion, lead-acid). In principle, four different cell types (shapes) can be identified - button, cylindrical, prismatic and pouch. Cell size can be either in accordance with international (e.g. International Electrotechnical Commission, IEC) or other standards or can meet application-specific dimensions. Since cell housing or container, terminals and, if necessary, safety installations as inactive (non-reactive) materials reduce energy density of the battery, the development of low-weight packages is a challenging task. In addition to that, other requirements have to be fulfilled: mechanical stability and durability, sealing (e.g. high permeation barrier against humidity for lithium-ion technology), high packing efficiency, possible installation of safety devices (current interrupt device, valve, etc.), chemical inertness, cost issues, and others. Finally, proper cell design has to be considered for effective thermal management (i.e. cooling and heating) of battery packs.

  17. Solving PDEs with Intrepid

    DOE PAGES

    Bochev, P.; Edwards, H. C.; Kirby, R. C.; ...

    2012-01-01

    Intrepid is a Trilinos package for advanced discretizations of Partial Differential Equations (PDEs). The package provides a comprehensive set of tools for local, cell-based construction of a wide range of numerical methods for PDEs. This paper describes the mathematical ideas and software design principles incorporated in the package. We also provide representative examples showcasing the use of Intrepid both in the context of numerical PDEs and the more general context of data analysis.

  18. Savannah River Site Eastern Transportation Hub: A Concept For a DOE Eastern Packaging, Staging and Maintenance Center - 13143

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    England, Jeffery L.; Adams, Karen; Maxted, Maxcine

    2013-07-01

    The Department of Energy (DOE) is working to de-inventory sites and consolidate hazardous materials for processing and disposal. The DOE administers a wide range of certified shipping packages for the transport of hazardous materials to include Special Nuclear Material (SNM), radioactive materials, sealed sources and radioactive wastes. A critical element to successful and safe transportation of these materials is the availability of certified shipping packages. There are over seven thousand certified packagings (i.e., Type B/Type AF) utilized within the DOE for current missions. The synergistic effects of consolidated maintenance, refurbishment, testing, certification, and costing of these services would allow formore » efficient management of the packagings inventory and to support anticipated future in-commerce shipping needs. The Savannah River Site (SRS) receives and ships radioactive materials (including SNM) and waste on a regular basis for critical missions such as consolidated storage, stabilization, purification, or disposition using H-Canyon and HB-Line. The Savannah River National Laboratory (SRNL) has the technical capability and equipment for all aspects of packaging management. SRS has the only active material processing facility in the DOE complex and is one of the sites of choice for nuclear material consolidation. SRS is a logical location to perform maintenance and periodic testing of the DOE fleet of certified packagings. This initiative envisions a DOE Eastern Packaging Staging and Maintenance Center (PSMC) at the SRS and a western hub at the Nevada National Security Site (NNSS), an active DOE Regional Disposal Site. The PSMC's would be the first place DOE would go to meet their radioactive packaging needs and the primary locations projects would go to disposition excess packaging for beneficial reuse. These two hubs would provide the centralized management of a packaging fleet rather than the current approach to design, procure, maintain and dispose of packagings on a project-by-project basis. This initiative provides significant savings in packaging costs and acceleration of project schedules. In addition to certified packaging, the PSMC would be well suited for select designs of 7A Type A packaging and Industrial Packaging. (authors)« less

  19. New Argentine Central-West line taps rich Neuquen gas field

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Watts, J.

    1982-02-01

    Argentina's new Central-West gas pipeline consists of 697 miles of 30-in. line and 451 miles of smaller gathering and distribution lines that link the rich Neuquen gas field with cities to the north. A financing package drawn up by 21 banks in the US and Europe allowed Cogasco S.A. to build the line for Gas del Estado across the roadless pampas east of the Andes. Primarily an agricultural country, Argentina had to import all the equipment and materials for the project. Site work began in July, 1980 with 800 workers employed on three spreads; the line was commissioned in November,more » 1981, 15 months ahead of the contract schedule.« less

  20. On-line data analysis and monitoring for H1 drift chambers

    NASA Astrophysics Data System (ADS)

    Düllmann, Dirk

    1992-05-01

    The on-line monitoring, slow control and calibration of the H1 central jet chamber uses a VME multiprocessor system to perform the analysis and a connected Macintosh computer as graphical interface to the operator on shift. Task of this system are: - analysis of event data including on-line track search, - on-line calibration from normal events and testpulse events, - control of the high voltage and monitoring of settings and currents, - monitoring of temperature, pressure and mixture of the chambergas. A program package is described which controls the dataflow between data aquisition, differnt VME CPUs and Macintosh. It allows to run off-line style programs for the different tasks.

  1. Production and first-in-man use of T cells engineered to express a HSVTK-CD34 sort-suicide gene.

    PubMed

    Zhan, Hong; Gilmour, Kimberly; Chan, Lucas; Farzaneh, Farzin; McNicol, Anne Marie; Xu, Jin-Hua; Adams, Stuart; Fehse, Boris; Veys, Paul; Thrasher, Adrian; Gaspar, Hubert; Qasim, Waseem

    2013-01-01

    Suicide gene modified donor T cells can improve immune reconstitution after allogeneic haematopoietic stem cell transplantation (SCT), but can be eliminated in the event of graft versus host disease (GVHD) through the administration of prodrug. Here we report the production and first-in-man use of mismatched donor T cells modified with a gamma-retroviral vector expressing a herpes simplex thymidine kinase (HSVTK):truncated CD34 (tCD34) suicide gene/magnetic selection marker protein. A stable packaging cell line was established to produce clinical grade vector pseudotyped with the Gibbon Ape Leukaemia Virus (GALV). T cells were transduced in a closed bag system following activation with anti-CD3/CD28 beads, and enriched on the basis of CD34 expression. Engineered cells were administered in two escalating doses to three children receiving T-depleted, CD34 stem cell selected, mismatched allogeneic grafts. All patients had pre-existing viral infections and received chemotherapy conditioning without serotherapy. In all three subjects cell therapy was tolerated without acute toxicity or the development of acute GVHD. Circulating gene modified T cells were detectable by flow cytometry and by molecular tracking in all three subjects. There was resolution of virus infections, concordant with detectable antigen-specific T cell responses and gene modified cells persisted for over 12 months. These findings highlight the suitability of tCD34 as a GMP compliant selection marker and demonstrate the feasibility, safety and immunological potential of HSVTK-tCD34 suicide gene modified donor T cells. ClinicalTrials.gov NCT01204502

  2. The analysis of novel microRNA mimic sequences in cancer cells reveals lack of specificity in stem-loop RT-qPCR-based microRNA detection.

    PubMed

    Winata, Patrick; Williams, Marissa; McGowan, Eileen; Nassif, Najah; van Zandwijk, Nico; Reid, Glen

    2017-11-17

    MicroRNAs are frequently downregulated in cancer, and restoring expression has tumour suppressive activity in tumour cells. Our recent phase I clinical trial investigated microRNA-based therapy in patients with malignant pleural mesothelioma. Treatment with TargomiRs, microRNA mimics with novel sequence packaged in EGFR antibody-targeted bacterial minicells, revealed clear signs of clinical activity. In order to detect delivery of microRNA mimics to tumour cells in future clinical trials, we tested hydrolysis probe-based assays specific for the sequence of the novel mimics in transfected mesothelioma cell lines using RT-qPCR. The custom assays efficiently and specifically amplified the consensus mimics. However, we found that these assays gave a signal when total RNA from untransfected and control mimic-transfected cells were used as templates. Further investigation revealed that the reverse transcription step using stem-loop primers appeared to introduce substantial non-specific amplification with either total RNA or synthetic RNA templates. This suggests that reverse transcription using stem-loop primers suffers from an intrinsic lack of specificity for the detection of highly similar microRNAs in the same family, especially when analysing total RNA. These results suggest that RT-qPCR is unlikely to be an effective means to detect delivery of microRNA mimic-based drugs to tumour cells in patients.

  3. The Reusable Load Cell with Protection Applied for Online Monitoring of Overhead Transmission Lines Based on Fiber Bragg Grating

    PubMed Central

    Ma, Guoming; Mao, Naiqiang; Li, Yabo; Jiang, Jun; Zhou, Hongyang; Li, Chengrong

    2016-01-01

    Heavy ice coating of high–voltage overhead transmission lines may lead to conductor breakage and tower collapse causing the unexpected interrupt of power supply. The optical load cell applied in ice monitoring systems is immune to electromagnetic interference and has no need of a power supply on site. Therefore, it has become a hot research topic in China and other countries. In this paper, to solve the problem of eccentric load in measurement, we adopt the shearing structure with additional grooves to improve the strain distribution and acquire good repeatability. Then, the fiber Bragg grating (FBG) with a permanent weldable package are mounted onto the front/rear groove of the elastic element by spot welding, the direction deviation of FBGs is 90° from each other to achieve temperature compensation without an extra FBG. After that, protection parts are designed to guarantee high sensitivity for a light load condition and industrial safety under a heavy load up to 65 kN. The results of tension experiments indicate that the sensitivity and resolution of the load cell is 0.1285 pm/N and 7.782 N in the conventional measuring range (0–10 kN). Heavy load tension experiments prove that the protection structure works and the sensitivity and resolution are not changed after several high load (65 kN) cycles. In addition, the experiment shows that the resolution of the sensor is 87.79 N in the large load range, allowing the parameter to be used in heavy icing monitoring. PMID:27338403

  4. Off-line robot programming and graphical verification of path planning

    NASA Technical Reports Server (NTRS)

    Tonkay, Gregory L.

    1989-01-01

    The objective of this project was to develop or specify an integrated environment for off-line programming, graphical path verification, and debugging for robotic systems. Two alternatives were compared. The first was the integration of the ASEA Off-line Programming package with ROBSIM, a robotic simulation program. The second alternative was the purchase of the commercial product IGRIP. The needs of the RADL (Robotics Applications Development Laboratory) were explored and the alternatives were evaluated based on these needs. As a result, IGRIP was proposed as the best solution to the problem.

  5. Tobacco Packaging and Labeling Policies Under the U.S. Tobacco Control Act: Research Needs and Priorities

    PubMed Central

    2012-01-01

    Introduction: The Family Smoking Prevention and Tobacco Control Act (the “Act”), enacted in June 2009, gave the U.S. Food and Drug Administration authority to regulate tobacco products. The current paper reviews the provisions for packaging and labeling, including the existing evidence and research priorities. Methods: Narrative review using electronic literature search of published and unpublished sources in 3 primary areas: health warnings, constituent labeling, and prohibitions on the promotional elements of packaging. Results: The Act requires 9 pictorial health warnings covering half of cigarette packages and 4 text warnings covering 30% of smokeless tobacco packages. The Act also prohibits potentially misleading information on packaging, including the terms “light” and “mild,” and provides a mandate to require disclosure of chemical constituents on packages. Many of the specific regulatory provisions are based on the extent to which they promote “greater public understanding of the risks of tobacco.” As a result, research on consumer perceptions has the potential to shape the design and renewal of health warnings and to determine what, if any, information on product constituents should appear on packages. Research on consumer perceptions of existing and novel tobacco products will also be critical to help identify potentially misleading information that should be restricted under the Act. Conclusion: Packaging and labeling regulations required under the Act will bring the United States in line with international standards. There is an immediate need for research to evaluate these measures to guide future regulatory action. PMID:22039072

  6. Multilingual Speech and Language Processing

    DTIC Science & Technology

    2003-04-01

    client software handles the user end of the transaction. Historically, four clients were provided: e-mail, web, FrameMaker , and command line. By...command-line client and an API. The API allows integration of CyberTrans into a number of processes including word processing packages ( FrameMaker ...preservation and logging, and others. The available clients remain e-mail, Web and FrameMaker . Platforms include both Unix and PC for clients, with

  7. Ada (tradename) Compiler Validation Summary Report. VAX Ada V1.3, Version 1.8 of the Ada Compiler Validation Capability (ACVC).

    DTIC Science & Technology

    1986-11-07

    and must apply to a function declared by an earlier declarative item of the same declarativ ,2 part or package specificatic.i. In the case of a...34IS" is misspelled at line 45. C34018A: The call of function G at line 114 is ambiguous in the presence of implicit conversions and inconsistente

  8. Asynchronous Discrete Control of Continuous Processes.

    DTIC Science & Technology

    1986-02-24

    Input. When 40-lead dual-in-line ceramic packages (D suffix) and in the mode input is high IMOOE=1), the CDPIS54A is 40-lead dual-in-line plastic ...DYNAIC U ThIi. CA~ YRi5iC5 TACDPI8541 CDPI854C DYNAMC LECRICL CARA ERITIC atTA 40 o +UC. DD M W O 1110 VlH’& 7 VDD. VIL, 3 . LIIMTS ’.- CARACTERISTIC

  9. Micro packaged MEMS pressure sensor for intracranial pressure measurement

    NASA Astrophysics Data System (ADS)

    Xiong, Liu; Yan, Yao; Jiahao, Ma; Yanhang, Zhang; Qian, Wang; Zhaohua, Zhang; Tianling, Ren

    2015-06-01

    This paper presents a micro packaged MEMS pressure sensor for intracranial pressure measurement which belongs to BioMEMS. It can be used in lumbar puncture surgery to measure intracranial pressure. Miniaturization is key for lumbar puncture surgery because the sensor must be small enough to allow it be placed in the reagent chamber of the lumbar puncture needle. The size of the sensor is decided by the size of the sensor chip and package. Our sensor chip is based on silicon piezoresistive effect and the size is 400 × 400 μm2. It is much smaller than the reported polymer intracranial pressure sensors such as liquid crystal polymer sensors. In terms of package, the traditional dual in-line package obviously could not match the size need, the minimal size of recently reported MEMS-based intracranial pressure sensors after packaging is 10 × 10 mm2. In this work, we are the first to introduce a quad flat no-lead package as the package form of piezoresistive intracranial pressure sensors, the whole size of the sensor is minimized to only 3 × 3 mm2. Considering the liquid measurement environment, the sensor is gummed and waterproof performance is tested; the sensitivity of the sensor is 0.9 × 10-2 mV/kPa. Project supported by the National Natural Science Foundation of China (Nos. 61025021, 61434001), and the ‘Thousands Talents’ Program for Pioneer Researchers and Its Innovation Team, China.

  10. An Object-Oriented Serial DSMC Simulation Package

    NASA Astrophysics Data System (ADS)

    Liu, Hongli; Cai, Chunpei

    2011-05-01

    A newly developed three-dimensional direct simulation Monte Carlo (DSMC) simulation package, named GRASP ("Generalized Rarefied gAs Simulation Package"), is reported in this paper. This package utilizes the concept of simulation engine, many C++ features and software design patterns. The package has an open architecture which can benefit further development and maintenance of the code. In order to reduce the engineering time for three-dimensional models, a hybrid grid scheme, combined with a flexible data structure compiled by C++ language, are implemented in this package. This scheme utilizes a local data structure based on the computational cell to achieve high performance on workstation processors. This data structure allows the DSMC algorithm to be very efficiently parallelized with domain decomposition and it provides much flexibility in terms of grid types. This package can utilize traditional structured, unstructured or hybrid grids within the framework of a single code to model arbitrarily complex geometries and to simulate rarefied gas flows. Benchmark test cases indicate that this package has satisfactory accuracy for complex rarefied gas flows.

  11. INTERIOR VIEW, LOOKING WEST, WITH PIPE BEING SPRAYED WITH ZINC ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    INTERIOR VIEW, LOOKING WEST, WITH PIPE BEING SPRAYED WITH ZINC COATING. - United States Pipe & Foundry Company Plant, Coating, Painting, Lining & Packaging Building, 2023 St. Louis Avenue at I-20/59, Bessemer, Jefferson County, AL

  12. 16 CFR 307.6 - Requirements for disclosure on the label.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ..., plug of chewing tobacco, or dispenser of individual packages of smokeless tobacco that may be purchased... plug of chewing tobacco—Eight point type, provided that if the warning statement is printed in one line...

  13. Privacy Impact Assessment for the Superfund Cost Recovery Package Imaging and On-Line System

    EPA Pesticide Factsheets

    This system collects financial data, associated documents, including travel, payroll and voucher data. Learn about how this data is collected, used, accessed, and the record retention policies for this data.

  14. fluff: exploratory analysis and visualization of high-throughput sequencing data

    PubMed Central

    Georgiou, Georgios

    2016-01-01

    Summary. In this article we describe fluff, a software package that allows for simple exploration, clustering and visualization of high-throughput sequencing data mapped to a reference genome. The package contains three command-line tools to generate publication-quality figures in an uncomplicated manner using sensible defaults. Genome-wide data can be aggregated, clustered and visualized in a heatmap, according to different clustering methods. This includes a predefined setting to identify dynamic clusters between different conditions or developmental stages. Alternatively, clustered data can be visualized in a bandplot. Finally, fluff includes a tool to generate genomic profiles. As command-line tools, the fluff programs can easily be integrated into standard analysis pipelines. The installation is straightforward and documentation is available at http://fluff.readthedocs.org. Availability. fluff is implemented in Python and runs on Linux. The source code is freely available for download at https://github.com/simonvh/fluff. PMID:27547532

  15. A computer program (MACPUMP) for interactive aquifer-test analysis

    USGS Publications Warehouse

    Day-Lewis, F. D.; Person, M.A.; Konikow, Leonard F.

    1995-01-01

    This report introduces MACPUMP (Version 1.0), an aquifer-test-analysis package for use with Macintosh4 computers. The report outlines the input- data format, describes the solutions encoded in the program, explains the menu-items, and offers a tutorial illustrating the use of the program. The package reads list-directed aquifer-test data from a file, plots the data to the screen, generates and plots type curves for several different test conditions, and allows mouse-controlled curve matching. MACPUMP features pull-down menus, a simple text viewer for displaying data-files, and optional on-line help windows. This version includes the analytical solutions for nonleaky and leaky confined aquifers, using both type curves and straight-line methods, and for the analysis of single-well slug tests using type curves. An executable version of the code and sample input data sets are included on an accompanying floppy disk.

  16. An Interpolation Method for Obtaining Thermodynamic Properties Near Saturated Liquid and Saturated Vapor Lines

    NASA Technical Reports Server (NTRS)

    Nguyen, Huy H.; Martin, Michael A.

    2004-01-01

    The two most common approaches used to formulate thermodynamic properties of pure substances are fundamental (or characteristic) equations of state (Helmholtz and Gibbs functions) and a piecemeal approach that is described in Adebiyi and Russell (1992). This paper neither presents a different method to formulate thermodynamic properties of pure substances nor validates the aforementioned approaches. Rather its purpose is to present a method to generate property tables from existing property packages and a method to facilitate the accurate interpretation of fluid thermodynamic property data from those tables. There are two parts to this paper. The first part of the paper shows how efficient and usable property tables were generated, with the minimum number of data points, using an aerospace industry standard property package. The second part describes an innovative interpolation technique that has been developed to properly obtain thermodynamic properties near the saturated liquid and saturated vapor lines.

  17. A comprehensive approach to addiction medicine as an appropriate response to the HIV epidemic among drug users.

    PubMed

    Gerra, Gilberto

    2013-12-01

    The services for drug-dependence treatment and care, particularly in low-income countries, should not be fragmented and uncoordinated. A basic package of interventions should be provided in the same place and managed by the same team, with a one-stop shop approach. The services for substance use disorders should be appealing, accessible, voluntary-based, and science-based. They should also, like efforts to fight other diseases, be included in the community and the public health systems; that is, those who are affected by drug use and those who serve them should not face discrimination. The first-line assistance and the second-line essential elements of the comprehensive package will be described. The work of the United Nations Office on Drugs and Crime (UNODC) and World Health Organization (WHO) to promote science-based and voluntary-based ethical treatment in Asia will be illustrated.

  18. Corporate ergonomics programme at BCM Airdrie. Boots Contract Manufacturing.

    PubMed

    Smyth, Joanne

    2003-01-01

    The production processes at the BCM Airdrie site range from manual loading tasks in the manufacturing areas to high frequency packaging assembly tasks on the production lines. Both are jobs that are known to carry risk to musculoskeletal health, so an ergonomist was appointed to design and co-ordinate an ergonomics programme for the site to control these risks. This paper details the programme that has evolved to proactively manage musculoskeletal risks in the design of both new and existing equipment and processes. The ergonomics procedures described primarily involve the engineers from all areas of the factory, and the process for ergonomics involvement with engineering design projects is described. Shop-floor personnel involvement is considered to be an essential part of the programme and 'Ergonomics Champions' are being trained on the packing lines to monitor the risks that are sometimes introduced with the different designs of product packaging.

  19. Coronal Magnetism and Forward Solarsoft Idl Package

    NASA Astrophysics Data System (ADS)

    Gibson, S. E.

    2014-12-01

    The FORWARD suite of Solar Soft IDL codes is a community resource for model-data comparison, with a particular emphasis on analyzing coronal magnetic fields. FORWARD may be used both to synthesize a broad range of coronal observables, and to access and compare to existing data. FORWARD works with numerical model datacubes, interfaces with the web-served Predictive Science Inc MAS simulation datacubes and the Solar Soft IDL Potential Field Source Surface (PFSS) package, and also includes several analytic models (more can be added). It connects to the Virtual Solar Observatory and other web-served observations to download data in a format directly comparable to model predictions. It utilizes the CHIANTI database in modeling UV/EUV lines, and links to the CLE polarimetry synthesis code for forbidden coronal lines. FORWARD enables "forward-fitting" of specific observations, and helps to build intuition into how the physical properties of coronal magnetic structures translate to observable properties.

  20. A passive and active microwave-vector radiative transfer (PAM-VRT) model

    NASA Astrophysics Data System (ADS)

    Yang, Jun; Min, Qilong

    2015-11-01

    A passive and active microwave vector radiative transfer (PAM-VRT) package has been developed. This fast and accurate forward microwave model, with flexible and versatile input and output components, self-consistently and realistically simulates measurements/radiation of passive and active microwave sensors. The core PAM-VRT, microwave radiative transfer model, consists of five modules: gas absorption (two line-by-line databases and four fast models); hydrometeor property of water droplets and ice (spherical and nonspherical) particles; surface emissivity (from Community Radiative Transfer Model (CRTM)); vector radiative transfer of successive order of scattering (VSOS); and passive and active microwave simulation. The PAM-VRT package has been validated against other existing models, demonstrating good accuracy. The PAM-VRT not only can be used to simulate or assimilate measurements of existing microwave sensors, but also can be used to simulate observation results at some new microwave sensors.

  1. GOplot: an R package for visually combining expression data with functional analysis.

    PubMed

    Walter, Wencke; Sánchez-Cabo, Fátima; Ricote, Mercedes

    2015-09-01

    Despite the plethora of methods available for the functional analysis of omics data, obtaining comprehensive-yet detailed understanding of the results remains challenging. This is mainly due to the lack of publicly available tools for the visualization of this type of information. Here we present an R package called GOplot, based on ggplot2, for enhanced graphical representation. Our package takes the output of any general enrichment analysis and generates plots at different levels of detail: from a general overview to identify the most enriched categories (bar plot, bubble plot) to a more detailed view displaying different types of information for molecules in a given set of categories (circle plot, chord plot, cluster plot). The package provides a deeper insight into omics data and allows scientists to generate insightful plots with only a few lines of code to easily communicate the findings. The R package GOplot is available via CRAN-The Comprehensive R Archive Network: http://cran.r-project.org/web/packages/GOplot. The shiny web application of the Venn diagram can be found at: https://wwalter.shinyapps.io/Venn/. A detailed manual of the package with sample figures can be found at https://wencke.github.io/ fscabo@cnic.es or mricote@cnic.es. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  2. Antimicrobial food packaging: potential and pitfalls

    PubMed Central

    Malhotra, Bhanu; Keshwani, Anu; Kharkwal, Harsha

    2015-01-01

    Nowadays food preservation, quality maintenance, and safety are major growing concerns of the food industry. It is evident that over time consumers’ demand for natural and safe food products with stringent regulations to prevent food-borne infectious diseases. Antimicrobial packaging which is thought to be a subset of active packaging and controlled release packaging is one such promising technology which effectively impregnates the antimicrobial into the food packaging film material and subsequently delivers it over the stipulated period of time to kill the pathogenic microorganisms affecting food products thereby increasing the shelf life to severe folds. This paper presents a picture of the recent research on antimicrobial agents that are aimed at enhancing and improving food quality and safety by reduction of pathogen growth and extension of shelf life, in a form of a comprehensive review. Examination of the available antimicrobial packaging technologies is also presented along with their significant impact on food safety. This article entails various antimicrobial agents for commercial applications, as well as the difference between the use of antimicrobials under laboratory scale and real time applications. Development of resistance amongst microorganisms is considered as a future implication of antimicrobials with an aim to come up with actual efficacies in extension of shelf life as well as reduction in bacterial growth through the upcoming and promising use of antimicrobials in food packaging for the forthcoming research down the line. PMID:26136740

  3. Using Cell-ID 1.4 with R for Microscope-Based Cytometry

    PubMed Central

    Bush, Alan; Chernomoretz, Ariel; Yu, Richard; Gordon, Andrew

    2012-01-01

    This unit describes a method for quantifying various cellular features (e.g., volume, total and subcellular fluorescence localization) from sets of microscope images of individual cells. It includes procedures for tracking cells over time. One purposefully defocused transmission image (sometimes referred to as bright-field or BF) is acquired to segment the image and locate each cell. Fluorescent images (one for each of the color channels to be analyzed) are then acquired by conventional wide-field epifluorescence or confocal microscopy. This method uses the image processing capabilities of Cell-ID (Gordon et al., 2007, as updated here) and data analysis by the statistical programming framework R (R-Development-Team, 2008), which we have supplemented with a package of routines for analyzing Cell-ID output. Both Cell-ID and the analysis package are open-source. PMID:23026908

  4. Versatile Cosmid Vectors for the Isolation, Expression, and Rescue of Gene Sequences: Studies with the Human α -globin Gene Cluster

    NASA Astrophysics Data System (ADS)

    Lau, Yun-Fai; Kan, Yuet Wai

    1983-09-01

    We have developed a series of cosmids that can be used as vectors for genomic recombinant DNA library preparations, as expression vectors in mammalian cells for both transient and stable transformations, and as shuttle vectors between bacteria and mammalian cells. These cosmids were constructed by inserting one of the SV2-derived selectable gene markers-SV2-gpt, SV2-DHFR, and SV2-neo-in cosmid pJB8. High efficiency of genomic cloning was obtained with these cosmids and the size of the inserts was 30-42 kilobases. We isolated recombinant cosmids containing the human α -globin gene cluster from these genomic libraries. The simian virus 40 DNA in these selectable gene markers provides the origin of replication and enhancer sequences necessary for replication in permissive cells such as COS 7 cells and thereby allows transient expression of α -globin genes in these cells. These cosmids and their recombinants could also be stably transformed into mammalian cells by using the respective selection systems. Both of the adult α -globin genes were more actively expressed than the embryonic zeta -globin genes in these transformed cell lines. Because of the presence of the cohesive ends of the Charon 4A phage in the cosmids, the transforming DNA sequences could readily be rescued from these stably transformed cells into bacteria by in vitro packaging of total cellular DNA. Thus, these cosmid vectors are potentially useful for direct isolation of structural genes.

  5. Experimental Methods Using Photogrammetric Techniques for Parachute Canopy Shape Measurements

    NASA Technical Reports Server (NTRS)

    Jones, Thomas W.; Downey, James M.; Lunsford, Charles B.; Desabrais, Kenneth J.; Noetscher, Gregory

    2007-01-01

    NASA Langley Research Center in partnership with the U.S. Army Natick Soldier Center has collaborated on the development of a payload instrumentation package to record the physical parameters observed during parachute air drop tests. The instrumentation package records a variety of parameters including canopy shape, suspension line loads, payload 3-axis acceleration, and payload velocity. This report discusses the instrumentation design and development process, as well as the photogrammetric measurement technique used to provide shape measurements. The scaled model tests were conducted in the NASA Glenn Plum Brook Space Propulsion Facility, OH.

  6. Advanced fingerprint verification software

    NASA Astrophysics Data System (ADS)

    Baradarani, A.; Taylor, J. R. B.; Severin, F.; Maev, R. Gr.

    2016-05-01

    We have developed a fingerprint software package that can be used in a wide range of applications from law enforcement to public and private security systems, and to personal devices such as laptops, vehicles, and door- locks. The software and processing units are a unique implementation of new and sophisticated algorithms that compete with the current best systems in the world. Development of the software package has been in line with the third generation of our ultrasonic fingerprinting machine1. Solid and robust performance is achieved in the presence of misplaced and low quality fingerprints.

  7. A graphics package for meteorological data, version 1.5

    NASA Technical Reports Server (NTRS)

    Moorthi, Shrinivas; Suarez, Max; Phillips, Bill; Schemm, Jae-Kyung; Schubert, Siegfried

    1989-01-01

    A plotting package has been developed to simplify the task of plotting meteorological data. The calling sequences and examples of high level yet flexible routines which allow contouring, vectors and shading of cylindrical, polar, orthographic and Mollweide (egg) projections are given. Routines are also included for contouring pressure-latitude and pressure-longitude fields with linear or log scales in pressure (interpolation to fixed grid interval is done automatically). Also included is a fairly general line plotting routine. The present version (1.5) produces plots on WMS laser printers and uses graphics primitives from WOLFPLOT.

  8. Survival of Spoilage and Pathogenic Microorganisms on Cardboard and Plastic Packaging Materials

    PubMed Central

    Siroli, Lorenzo; Patrignani, Francesca; Serrazanetti, Diana I.; Chiavari, Cristiana; Benevelli, Marzia; Grazia, Luigi; Lanciotti, Rosalba

    2017-01-01

    The aim of this work was to study the interaction of corrugated and plastic materials with pathogenic and spoiling microorganisms frequently associated to fresh produce. The effect of the two packaging materials on the survival during the storage of microorganisms belonging to the species Escherichia coli, Listeria monocytogenes, Salmonella enteritidis, Saccharomyces cerevisiae, Lactobacillus plantarum, Pseudomonas fluorescens, and Aspergillus flavus was studied through traditional plate counting and scanning electron microscopy (SEM). The results obtained showed that cardboard materials, if correctly stored, reduced the potential of packaging to cross-contaminate food due to a faster viability loss by spoilage and pathogenic microorganisms compared to the plastic ones. In fact, the cell loads of the pathogenic species considered decreased over time independently on the inoculation level and packaging material used. However, the superficial viability losses were significantly faster in cardboard compared to plastic materials. The same behavior was observed for the spoilage microorganisms considered. The SEM microphotographs indicate that the reduction of superficial contamination on cardboard surfaces was due to the entrapping of the microbial cells within the fibers and the pores of this material. In addition, SEM data showed that the entrapped cells were subjected to more or less rapid lyses, depending on the species, due to the absence of water and nutrients, with the exception of molds. The latter spoilers were able to proliferate inside the cardboard fibers only when the absorption of water was not prevented during the storage. In conclusion, the findings of this work showed the reduction of cross-contamination potential of corrugated compared to plastic packaging materials used in fruit and vegetable sector. However, the findings outlined the importance of hygiene and low humidity during cardboard storage to prevent the mold growth on packaging. PMID:29312271

  9. Nef exosomes isolated from the plasma of individuals with HIV-associated dementia (HAD) can induce Aβ(1-42) secretion in SH-SY5Y neural cells.

    PubMed

    Khan, Mahfuz B; Lang, Michelle J; Huang, Ming-Bo; Raymond, Andrea; Bond, Vincent C; Shiramizu, Bruce; Powell, Michael D

    2016-04-01

    In the era of combined antiretroviral therapy (CART), many of the complications due to HIV-1 infection have diminished. One exception is HIV-associated neurocognitive disorder (HAND). HAND is a spectrum of disorders in cognitive function that ranges from asymptomatic disease to severe dementia (HAD). The milder form of HAND has actually remained the same or slightly increased in prevalence in the CART era. Even in individuals who have maintained undetectable HIV RNA loads, viral proteins such as Nef and Tat can continue to be expressed. In this report, we show that Nef protein and nef messenger RNA (mRNA) are packaged into exosomes that remain in circulation in patients with HAD. Plasma-derived Nef exosomes from patients with HAD have the ability to interact with the neuroblastoma cell line SH-SY5Y and deliver nef mRNA. The mRNA can induce expression of Nef in target cells and subsequently increase expression and secretion of beta-amyloid (Aβ) and Aβ peptides. Increase secretion of amyloid peptide could contribute to cognitive impairment seen in HAND.

  10. Measurement value analysis overall equipment effectiveness (OEE) packaging process in line 2 (Case Study of PT. MBI Tbk)

    NASA Astrophysics Data System (ADS)

    Rimawan, Erry; Kholil, Muhammad; Hendri

    2018-03-01

    PT. MBI Tbk is engaged in the manufacture of beverage industry, where the company’s production is based on the magnitude of customer demand that is marketing offices that had been scattered in various regions of Indonesia. In the packaging process steps in PT.MBI through the line 3 lines including racking, canning line, bottling line. In the canning process to existing packing on Line 2 (canning line), there are some machines that are used continuously, among other Depalletizer machine, filler machine, can seamer machine, pasteurizer machine, machine FLD, Wrap Around engine, engine Shrink Wrap. Due to the large demand from customers that is relentless, therefore the calculation of overall equipment effectiveness (OEE) as a whole on line 2 (canning line) is needed in order to make improvements continuously (Continuous Improvement) at line 2 (canning line). This study aims to determine the value of overall equipment effectiveness (OEE) and Losses of the most influential of the big six OEE Losses focused on equipment or machinery as a whole into a single unit that is on the line 2, which will then be known root cause of the losses that occur from the research over the field. From the calculation of overall equipment effectiveness (OEE), there are two ratios are still poor and under world-class standards, while the ratio of the availability of 88.85% of the world-class standards by 90% and the performance ratio of 78.51% of the standard world class by 95%, whereas for quality ratio has entered the world-class standard that is equal to 99.90%. Thus the value of OEE on Line 2 line is below world class standards. In this study there were only five losses, which can be identified, and while the losses were very influential, namely the Speed Reduced Losses, losses, these losses accounted for the largest percentage of the value of the rate of 19.12%, of the results of this study losses occurred due to poor surveillance systems (less good) that causes the employee or operator does not perform the work in accordance with a predetermined.

  11. ATLAS software configuration and build tool optimisation

    NASA Astrophysics Data System (ADS)

    Rybkin, Grigory; Atlas Collaboration

    2014-06-01

    ATLAS software code base is over 6 million lines organised in about 2000 packages. It makes use of some 100 external software packages, is developed by more than 400 developers and used by more than 2500 physicists from over 200 universities and laboratories in 6 continents. To meet the challenge of configuration and building of this software, the Configuration Management Tool (CMT) is used. CMT expects each package to describe its build targets, build and environment setup parameters, dependencies on other packages in a text file called requirements, and each project (group of packages) to describe its policies and dependencies on other projects in a text project file. Based on the effective set of configuration parameters read from the requirements files of dependent packages and project files, CMT commands build the packages, generate the environment for their use, or query the packages. The main focus was on build time performance that was optimised within several approaches: reduction of the number of reads of requirements files that are now read once per package by a CMT build command that generates cached requirements files for subsequent CMT build commands; introduction of more fine-grained build parallelism at package task level, i.e., dependent applications and libraries are compiled in parallel; code optimisation of CMT commands used for build; introduction of package level build parallelism, i. e., parallelise the build of independent packages. By default, CMT launches NUMBER-OF-PROCESSORS build commands in parallel. The other focus was on CMT commands optimisation in general that made them approximately 2 times faster. CMT can generate a cached requirements file for the environment setup command, which is especially useful for deployment on distributed file systems like AFS or CERN VMFS. The use of parallelism, caching and code optimisation significantly-by several times-reduced software build time, environment setup time, increased the efficiency of multi-core computing resources utilisation, and considerably improved software developer and user experience.

  12. 49 CFR 171.12 - North American Shipments.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...) Lithium metal cells and batteries. Lithium metal cells and batteries (UN3090) are forbidden for transport aboard passenger-carrying aircraft. The outside of each package that contains lithium cells or batteries...

  13. Interactions between HIV-1 Gag and Viral RNA Genome Enhance Virion Assembly.

    PubMed

    Dilley, Kari A; Nikolaitchik, Olga A; Galli, Andrea; Burdick, Ryan C; Levine, Louis; Li, Kelvin; Rein, Alan; Pathak, Vinay K; Hu, Wei-Shau

    2017-08-15

    Most HIV-1 virions contain two copies of full-length viral RNA, indicating that genome packaging is efficient and tightly regulated. However, the structural protein Gag is the only component required for the assembly of noninfectious viruslike particles, and the viral RNA is dispensable in this process. The mechanism that allows HIV-1 to achieve such high efficiency of genome packaging when a packageable viral RNA is not required for virus assembly is currently unknown. In this report, we examined the role of HIV-1 RNA in virus assembly and found that packageable HIV-1 RNA enhances particle production when Gag is expressed at levels similar to those in cells containing one provirus. However, such enhancement is diminished when Gag is overexpressed, suggesting that the effects of viral RNA can be replaced by increased Gag concentration in cells. We also showed that the specific interactions between Gag and viral RNA are required for the enhancement of particle production. Taken together, these studies are consistent with our previous hypothesis that specific dimeric viral RNA-Gag interactions are the nucleation event of infectious virion assembly, ensuring that one RNA dimer is packaged into each nascent virion. These studies shed light on the mechanism by which HIV-1 achieves efficient genome packaging during virus assembly. IMPORTANCE Retrovirus assembly is a well-choreographed event, during which many viral and cellular components come together to generate infectious virions. The viral RNA genome carries the genetic information to new host cells, providing instructions to generate new virions, and therefore is essential for virion infectivity. In this report, we show that the specific interaction of the viral RNA genome with the structural protein Gag facilitates virion assembly and particle production. These findings resolve the conundrum that HIV-1 RNA is selectively packaged into virions with high efficiency despite being dispensable for virion assembly. Understanding the mechanism used by HIV-1 to ensure genome packaging provides significant insights into viral assembly and replication. Copyright © 2017 American Society for Microbiology.

  14. Bayesian Atmospheric Radiative Transfer (BART): Model, Statistics Driver, and Application to HD 209458b

    NASA Astrophysics Data System (ADS)

    Cubillos, Patricio; Harrington, Joseph; Blecic, Jasmina; Stemm, Madison M.; Lust, Nate B.; Foster, Andrew S.; Rojo, Patricio M.; Loredo, Thomas J.

    2014-11-01

    Multi-wavelength secondary-eclipse and transit depths probe the thermo-chemical properties of exoplanets. In recent years, several research groups have developed retrieval codes to analyze the existing data and study the prospects of future facilities. However, the scientific community has limited access to these packages. Here we premiere the open-source Bayesian Atmospheric Radiative Transfer (BART) code. We discuss the key aspects of the radiative-transfer algorithm and the statistical package. The radiation code includes line databases for all HITRAN molecules, high-temperature H2O, TiO, and VO, and includes a preprocessor for adding additional line databases without recompiling the radiation code. Collision-induced absorption lines are available for H2-H2 and H2-He. The parameterized thermal and molecular abundance profiles can be modified arbitrarily without recompilation. The generated spectra are integrated over arbitrary bandpasses for comparison to data. BART's statistical package, Multi-core Markov-chain Monte Carlo (MC3), is a general-purpose MCMC module. MC3 implements the Differental-evolution Markov-chain Monte Carlo algorithm (ter Braak 2006, 2009). MC3 converges 20-400 times faster than the usual Metropolis-Hastings MCMC algorithm, and in addition uses the Message Passing Interface (MPI) to parallelize the MCMC chains. We apply the BART retrieval code to the HD 209458b data set to estimate the planet's temperature profile and molecular abundances. This work was supported by NASA Planetary Atmospheres grant NNX12AI69G and NASA Astrophysics Data Analysis Program grant NNX13AF38G. JB holds a NASA Earth and Space Science Fellowship.

  15. GKS. Minimal Graphical Kernel System C Binding

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Simons, R.W.

    1985-10-01

    GKS (the Graphical Kernel System) is both an American National Standard (ANS) and an ISO international standard graphics package. It conforms to ANS X3.124-1985 and to the May 1985 draft proposal for the GKS C Language Binding standard under development by the X3H3 Technical Committee. This implementation includes level ma (the lowest level of the ANS) and some routines from level mb. The following graphics capabilities are supported: two-dimensional lines, markers, text, and filled areas; control over color, line type, and character height and alignment; multiple simultaneous workstations and multiple transformations; and locator and choice input. Tektronix 4014 and 4115more » terminals are supported, and support for other devices may be added. Since this implementation was developed under UNIX, it uses makefiles, C shell scripts, the ar library maintainer, editor scripts, and other UNIX utilities. Therefore, implementing it under another operating system may require considerable effort. Also included with GKS is the small plot package (SPP), a direct descendant of the WEASEL plot package developed at Sandia. SPP is built on the GKS; therefore, all of the capabilities of GKS are available. It is not necessary to use GKS functions, since entire plots can be produced using only SPP functions, but the addition of GKS will give the programmer added power and flexibility. SPP provides single-call plot commands, linear and logarithmic axis commands, control for optional plotting of tick marks and tick mark labels, and permits plotting of data with or without markers and connecting lines.« less

  16. An automated approach to Litchfield and Wilcoxon's evaluation of dose–effect experiments using the R package LW1949

    USGS Publications Warehouse

    Adams, Jean V.; Slaght, Karen; Boogaard, Michael A.

    2016-01-01

    The authors developed a package, LW1949, for use with the statistical software R to automatically carry out the manual steps of Litchfield and Wilcoxon's method of evaluating dose–effect experiments. The LW1949 package consistently finds the best fitting dose–effect relation by minimizing the chi-squared statistic of the observed and expected number of affected individuals and substantially speeds up the line-fitting process and other calculations that Litchfield and Wilcoxon originally carried out by hand. Environ Toxicol Chem 2016;9999:1–4. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America.

  17. GenomeDiagram: a python package for the visualization of large-scale genomic data.

    PubMed

    Pritchard, Leighton; White, Jennifer A; Birch, Paul R J; Toth, Ian K

    2006-03-01

    We present GenomeDiagram, a flexible, open-source Python module for the visualization of large-scale genomic, comparative genomic and other data with reference to a single chromosome or other biological sequence. GenomeDiagram may be used to generate publication-quality vector graphics, rastered images and in-line streamed graphics for webpages. The package integrates with datatypes from the BioPython project, and is available for Windows, Linux and Mac OS X systems. GenomeDiagram is freely available as source code (under GNU Public License) at http://bioinf.scri.ac.uk/lp/programs.html, and requires Python 2.3 or higher, and recent versions of the ReportLab and BioPython packages. A user manual, example code and images are available at http://bioinf.scri.ac.uk/lp/programs.html.

  18. Enhanced Electric Power Transmission by Hybrid Compensation Technique

    NASA Astrophysics Data System (ADS)

    Palanichamy, C.; Kiu, G. Q.

    2015-04-01

    In today's competitive environment, new power system engineers are likely to contribute immediately to the task, without years of seasoning via on-the-job training, mentoring, and rotation assignments. At the same time it is becoming obligatory to train power system engineering graduates for an increasingly quality-minded corporate environment. In order to achieve this, there is a need to make available better-quality tools for educating and training power system engineering students and in-service system engineers too. As a result of the swift advances in computer hardware and software, many windows-based computer software packages were developed for the purpose of educating and training. In line with those packages, a simulation package called Hybrid Series-Shunt Compensators (HSSC) has been developed and presented in this paper for educational purposes.

  19. Scater: pre-processing, quality control, normalization and visualization of single-cell RNA-seq data in R.

    PubMed

    McCarthy, Davis J; Campbell, Kieran R; Lun, Aaron T L; Wills, Quin F

    2017-04-15

    Single-cell RNA sequencing (scRNA-seq) is increasingly used to study gene expression at the level of individual cells. However, preparing raw sequence data for further analysis is not a straightforward process. Biases, artifacts and other sources of unwanted variation are present in the data, requiring substantial time and effort to be spent on pre-processing, quality control (QC) and normalization. We have developed the R/Bioconductor package scater to facilitate rigorous pre-processing, quality control, normalization and visualization of scRNA-seq data. The package provides a convenient, flexible workflow to process raw sequencing reads into a high-quality expression dataset ready for downstream analysis. scater provides a rich suite of plotting tools for single-cell data and a flexible data structure that is compatible with existing tools and can be used as infrastructure for future software development. The open-source code, along with installation instructions, vignettes and case studies, is available through Bioconductor at http://bioconductor.org/packages/scater . davis@ebi.ac.uk. Supplementary data are available at Bioinformatics online. © The Author 2017. Published by Oxford University Press.

  20. MICA: Multiple interval-based curve alignment

    NASA Astrophysics Data System (ADS)

    Mann, Martin; Kahle, Hans-Peter; Beck, Matthias; Bender, Bela Johannes; Spiecker, Heinrich; Backofen, Rolf

    2018-01-01

    MICA enables the automatic synchronization of discrete data curves. To this end, characteristic points of the curves' shapes are identified. These landmarks are used within a heuristic curve registration approach to align profile pairs by mapping similar characteristics onto each other. In combination with a progressive alignment scheme, this enables the computation of multiple curve alignments. Multiple curve alignments are needed to derive meaningful representative consensus data of measured time or data series. MICA was already successfully applied to generate representative profiles of tree growth data based on intra-annual wood density profiles or cell formation data. The MICA package provides a command-line and graphical user interface. The R interface enables the direct embedding of multiple curve alignment computation into larger analyses pipelines. Source code, binaries and documentation are freely available at https://github.com/BackofenLab/MICA

  1. GenomeGraphs: integrated genomic data visualization with R.

    PubMed

    Durinck, Steffen; Bullard, James; Spellman, Paul T; Dudoit, Sandrine

    2009-01-06

    Biological studies involve a growing number of distinct high-throughput experiments to characterize samples of interest. There is a lack of methods to visualize these different genomic datasets in a versatile manner. In addition, genomic data analysis requires integrated visualization of experimental data along with constantly changing genomic annotation and statistical analyses. We developed GenomeGraphs, as an add-on software package for the statistical programming environment R, to facilitate integrated visualization of genomic datasets. GenomeGraphs uses the biomaRt package to perform on-line annotation queries to Ensembl and translates these to gene/transcript structures in viewports of the grid graphics package. This allows genomic annotation to be plotted together with experimental data. GenomeGraphs can also be used to plot custom annotation tracks in combination with different experimental data types together in one plot using the same genomic coordinate system. GenomeGraphs is a flexible and extensible software package which can be used to visualize a multitude of genomic datasets within the statistical programming environment R.

  2. Thermal shock testing for assuring reliability of glass-sealed microelectronic packages

    NASA Technical Reports Server (NTRS)

    Thomas, Walter B., III; Lewis, Michael D.

    1991-01-01

    Tests were performed to determine if thermal shocking is destructive to glass-to-metal seal microelectronic packages and if thermal shock step stressing can compare package reliabilities. Thermal shocking was shown to be not destructive to highly reliable glass seals. Pin-pull tests used to compare the interfacial pin glass strengths showed no differences between thermal shocked and not-thermal shocked headers. A 'critical stress resistance temperature' was not exhibited by the 14 pin Dual In-line Package (DIP) headers evaluated. Headers manufactured in cryogenic nitrogen based and exothermically generated atmospheres showed differences in as-received leak rates, residual oxide depths and pin glass interfacial strengths; these were caused by the different manufacturing methods, in particular, by the chemically etched pins used by one manufacturer. Both header types passed thermal shock tests to temperature differentials of 646 C. The sensitivity of helium leak rate measurements was improved up to 70 percent by baking headers for two hours at 200 C after thermal shocking.

  3. Smart sign enhancement, phase 2 : final report, September 2007.

    DOT National Transportation Integrated Search

    2007-09-01

    An on line ordering system, called Smart Sign Ordering System (SSOS), was : developed by The University of Akron for Ohio Department of Transportation (ODOT) : in 2004. Driven by the demand of managing planning, fabrication, packaging and : delivery,...

  4. Py4CAtS - Python tools for line-by-line modelling of infrared atmospheric radiative transfer

    NASA Astrophysics Data System (ADS)

    Schreier, Franz; García, Sebastián Gimeno

    2013-05-01

    Py4CAtS — Python scripts for Computational ATmospheric Spectroscopy is a Python re-implementation of the Fortran infrared radiative transfer code GARLIC, where compute-intensive code sections utilize the Numeric/Scientific Python modules for highly optimized array-processing. The individual steps of an infrared or microwave radiative transfer computation are implemented in separate scripts to extract lines of relevant molecules in the spectral range of interest, to compute line-by-line cross sections for given pressure(s) and temperature(s), to combine cross sections to absorption coefficients and optical depths, and to integrate along the line-of-sight to transmission and radiance/intensity. The basic design of the package, numerical and computational aspects relevant for optimization, and a sketch of the typical workflow are presented.

  5. Evaluation of tools for highly variable gene discovery from single-cell RNA-seq data.

    PubMed

    Yip, Shun H; Sham, Pak Chung; Wang, Junwen

    2018-02-21

    Traditional RNA sequencing (RNA-seq) allows the detection of gene expression variations between two or more cell populations through differentially expressed gene (DEG) analysis. However, genes that contribute to cell-to-cell differences are not discoverable with RNA-seq because RNA-seq samples are obtained from a mixture of cells. Single-cell RNA-seq (scRNA-seq) allows the detection of gene expression in each cell. With scRNA-seq, highly variable gene (HVG) discovery allows the detection of genes that contribute strongly to cell-to-cell variation within a homogeneous cell population, such as a population of embryonic stem cells. This analysis is implemented in many software packages. In this study, we compare seven HVG methods from six software packages, including BASiCS, Brennecke, scLVM, scran, scVEGs and Seurat. Our results demonstrate that reproducibility in HVG analysis requires a larger sample size than DEG analysis. Discrepancies between methods and potential issues in these tools are discussed and recommendations are made.

  6. Expedient Measures in Postattack Industrial Recovery: Petroleum Refining

    DTIC Science & Technology

    1980-12-01

    type of damage chAZge from superficial damage of [Fagile equipiivtt (e.g., iv- strtments with glass covers) vero buckling of heavy steel framing andj t...Pressure vessels, cylindrical A. horizontal, glass -lined 1 B. horizontal, unlined 48 1 C. horizontal, near ground 2 D. vertical 75 2 30. Package...Pressure vessels, cylindrical A. horizontal, glass -lined 320 350 30 B. horizontal, unlined 260 290 48 C. horizontal, near ground 230 250 48 D

  7. MODFLOW-NWT, A Newton formulation for MODFLOW-2005

    USGS Publications Warehouse

    Niswonger, Richard G.; Panday, Sorab; Ibaraki, Motomu

    2011-01-01

    This report documents a Newton formulation of MODFLOW-2005, called MODFLOW-NWT. MODFLOW-NWT is a standalone program that is intended for solving problems involving drying and rewetting nonlinearities of the unconfined groundwater-flow equation. MODFLOW-NWT must be used with the Upstream-Weighting (UPW) Package for calculating intercell conductances in a different manner than is done in the Block-Centered Flow (BCF), Layer Property Flow (LPF), or Hydrogeologic-Unit Flow (HUF; Anderman and Hill, 2000) Packages. The UPW Package treats nonlinearities of cell drying and rewetting by use of a continuous function of groundwater head, rather than the discrete approach of drying and rewetting that is used by the BCF, LPF, and HUF Packages. This further enables application of the Newton formulation for unconfined groundwater-flow problems because conductance derivatives required by the Newton method are smooth over the full range of head for a model cell. The NWT linearization approach generates an asymmetric matrix, which is different from the standard MODFLOW formulation that generates a symmetric matrix. Because all linear solvers presently available for use with MODFLOW-2005 solve only symmetric matrices, MODFLOW-NWT includes two previously developed asymmetric matrix-solver options. The matrix-solver options include a generalized-minimum-residual (GMRES) Solver and an Orthomin / stabilized conjugate-gradient (CGSTAB) Solver. The GMRES Solver is documented in a previously published report, such that only a brief description and input instructions are provided in this report. However, the CGSTAB Solver (called XMD) is documented in this report. Flow-property input for the UPW Package is designed based on the LPF Package and material-property input is identical to that for the LPF Package except that the rewetting and vertical-conductance correction options of the LPF Package are not available with the UPW Package. Input files constructed for the LPF Package can be used with slight modification as input for the UPW Package. This report presents the theory and methods used by MODFLOW-NWT, including the UPW Package. Additionally, this report provides comparisons of the new methodology to analytical solutions of groundwater flow and to standard MODFLOW-2005 results by use of an unconfined aquifer MODFLOW example problem. The standard MODFLOW-2005 simulation uses the LPF Package with the wet/dry option active. A new example problem also is presented to demonstrate MODFLOW-NWT's ability to provide a solution for a difficult unconfined groundwater-flow problem.

  8. Virus Capsids as Targeted Nanoscale Delivery Vessels of Photoactive Compounds for Site-Specific Photodynamic Therapy

    NASA Astrophysics Data System (ADS)

    Cohen, Brian A.

    The research presented in this work details the use of a viral capsid as an addressable delivery vessel of photoactive compounds for use in photodynamic therapy. Photodynamic therapy is a treatment that involves the interaction of light with a photosensitizing molecule to create singlet oxygen, a reactive oxygen species. Overproduction of singlet oxygen in cells can cause oxidative damage leading to cytotoxicity and eventually cell death. Challenges with the current generation of FDA-approved photosensitizers for photodynamic therapy primarily stem from their lack of tissue specificity. This work describes the packaging of photoactive cationic porphyrins inside the MS2 bacteriophage capsid, followed by external modification of the capsid with cancer cell-targeting G-quadruplex DNA aptamers to generate a tumor-specific photosensitizing agent. First, a cationic porphyrin is loaded into the capsids via nucleotide-driven packaging, a process that involves charge interaction between the porphyrin and the RNA inside the capsid. Results show that over 250 porphyrin molecules associate with the RNA within each MS2 capsid. Removal of RNA from the capsid severely inhibits the packaging of the cationic porphyrins. Porphyrin-virus constructs were then shown to photogenerate singlet oxygen, and cytotoxicity in non-targeted photodynamic treatment experiments. Next, each porphyrin-loaded capsid is externally modified with approximately 60 targeting DNA aptamers by employing a heterobifunctional crosslinking agent. The targeting aptamer is known to bind the protein nucleolin, a ubiquitous protein that is overexpressed on the cell surface by many cancer cell types. MCF-7 human breast carcinoma cells and MCF-10A human mammary epithelial cells were selected as an in vitro model for breast cancer and normal tissue, respectively. Fluorescently tagged virus-aptamer constructs are shown to selectively target MCF-7 cells versus MCF-10A cells. Finally, results are shown in which porphyrin-virus-aptamer constructs selectively target and kill cancer cells versus non-cancer cells. Specifically, the results show that MS2 is a viable candidate as an addressable nanodelivery vessel of photoactive compounds, and the implications are that the nucleotide-driven packaging approach for modifying MS2 can be used to impart new functionalities for a host of diagnostic or therapeutic applications.

  9. A fast hidden line algorithm for plotting finite element models

    NASA Technical Reports Server (NTRS)

    Jones, G. K.

    1982-01-01

    Effective plotting of finite element models requires the use of fast hidden line plot techniques that provide interactive response. A high speed hidden line technique was developed to facilitate the plotting of NASTRAN finite element models. Based on testing using 14 different models, the new hidden line algorithm (JONES-D) appears to be very fast: its speed equals that for normal (all lines visible) plotting and when compared to other existing methods it appears to be substantially faster. It also appears to be very reliable: no plot errors were observed using the new method to plot NASTRAN models. The new algorithm was made part of the NPLOT NASTRAN plot package and was used by structural analysts for normal production tasks.

  10. CLO: The cell line ontology

    PubMed Central

    2014-01-01

    Background Cell lines have been widely used in biomedical research. The community-based Cell Line Ontology (CLO) is a member of the OBO Foundry library that covers the domain of cell lines. Since its publication two years ago, significant updates have been made, including new groups joining the CLO consortium, new cell line cells, upper level alignment with the Cell Ontology (CL) and the Ontology for Biomedical Investigation, and logical extensions. Construction and content Collaboration among the CLO, CL, and OBI has established consensus definitions of cell line-specific terms such as ‘cell line’, ‘cell line cell’, ‘cell line culturing’, and ‘mortal’ vs. ‘immortal cell line cell’. A cell line is a genetically stable cultured cell population that contains individual cell line cells. The hierarchical structure of the CLO is built based on the hierarchy of the in vivo cell types defined in CL and tissue types (from which cell line cells are derived) defined in the UBERON cross-species anatomy ontology. The new hierarchical structure makes it easier to browse, query, and perform automated classification. We have recently added classes representing more than 2,000 cell line cells from the RIKEN BRC Cell Bank to CLO. Overall, the CLO now contains ~38,000 classes of specific cell line cells derived from over 200 in vivo cell types from various organisms. Utility and discussion The CLO has been applied to different biomedical research studies. Example case studies include annotation and analysis of EBI ArrayExpress data, bioassays, and host-vaccine/pathogen interaction. CLO’s utility goes beyond a catalogue of cell line types. The alignment of the CLO with related ontologies combined with the use of ontological reasoners will support sophisticated inferencing to advance translational informatics development. PMID:25852852

  11. Curcumin may serve an anticancer role in human osteosarcoma cell line U-2 OS by targeting ITPR1.

    PubMed

    Luo, Zhanpeng; Li, Dawei; Luo, Xiaobo; Li, Litao; Gu, Suxi; Yu, Long; Ma, Yuanzheng

    2018-04-01

    The present study aimed to determine the mechanisms of action of curcumin in osteosarcoma. Human osteosarcoma U-2 OS cells was purchased from the Cell Bank of the Chinese Academy of Sciences. RNA sequencing analysis was performed for 2 curcumin-treated samples and 2 control samples using Illumina deep sequencing technology. The differentially expressed genes were identified using Cufflink software. Enrichment and protein-protein interaction network analyses were performed separately using cluster Profiler package and Cytoscape software to identify key genes. Then, the mRNA levels of key genes were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) in U-2 OS cells. Finally, cell apoptosis, proliferation, migration and invasion arrays were performed. In total, 201 DEGs were identified in the curcumin-treated group. EEF1A1 (degree=88), ATF7IP, HIF1A, SMAD7, CLTC, MCM10, ITPR1, ADAM15, WWP2 and ATP5C1, which were enriched in 'biological process', exhibited higher degrees than other genes in the PPI network. RT-qPCR demonstrated that treatment with curcumin was able to significantly increase the levels of CLTC and ITPR1 mRNA in curcumin-treated cells compared with control. In addition, targeting ITPR1 with curcumin significantly promoted apoptosis and suppressed proliferation, migration and invasion. Targeting ITPR1 via curcumin may serve an anticancer role by mediating apoptosis, proliferation, migration and invasion in U-2 OS cells.

  12. Improvement of COBRA-TF for modeling of PWR cold- and hot-legs during reactor transients

    NASA Astrophysics Data System (ADS)

    Salko, Robert K.

    COBRA-TF is a two-phase, three-field (liquid, vapor, droplets) thermal-hydraulic modeling tool that has been developed by the Pacific Northwest Laboratory under sponsorship of the NRC. The code was developed for Light Water Reactor analysis starting in the 1980s; however, its development has continued to this current time. COBRA-TF still finds wide-spread use throughout the nuclear engineering field, including nuclear-power vendors, academia, and research institutions. It has been proposed that extension of the COBRA-TF code-modeling region from vessel-only components to Pressurized Water Reactor (PWR) coolant-line regions can lead to improved Loss-of-Coolant Accident (LOCA) analysis. Improved modeling is anticipated due to COBRA-TF's capability to independently model the entrained-droplet flow-field behavior, which has been observed to impact delivery to the core region[1]. Because COBRA-TF was originally developed for vertically-dominated, in-vessel, sub-channel flow, extension of the COBRA-TF modeling region to the horizontal-pipe geometries of the coolant-lines required several code modifications, including: • Inclusion of the stratified flow regime into the COBRA-TF flow regime map, along with associated interfacial drag, wall drag and interfacial heat transfer correlations, • Inclusion of a horizontal-stratification force between adjacent mesh cells having unequal levels of stratified flow, and • Generation of a new code-input interface for the modeling of coolant-lines. The sheer number of COBRA-TF modifications that were required to complete this work turned this project into a code-development project as much as it was a study of thermal-hydraulics in reactor coolant-lines. The means for achieving these tasks shifted along the way, ultimately leading the development of a separate, nearly completely independent one-dimensional, two-phase-flow modeling code geared toward reactor coolant-line analysis. This developed code has been named CLAP, for Coolant-Line-Analysis Package. Versions were created that were both coupled to COBRA-TF and standalone, with the most recent version being a standalone code. This code performs a separate, simplified, 1-D solution of the conservation equations while making special considerations for coolant-line geometry and flow phenomena. The end of this project saw a functional code package that demonstrates a stable numerical solution and that has gone through a series of Validation and Verification tests using the Two-Phase Testing Facility (TPTF) experimental data[2]. The results indicate that CLAP is under-performing RELAP5-MOD3 in predicting the experimental void of the TPTF facility in some cases. There is no apparent pattern, however, to point to a consistent type of case that the code fails to predict properly (e.g., low-flow, high-flow, discharging to full vessel, or discharging to empty vessel). Pressure-profile predictions are sometimes unrealistic, which indicates that there may be a problem with test-case boundary conditions or with the coupling of continuity and momentum equations in the solution algorithm. The code does predict the flow regime correctly for all cases with the stratification-force model off. Turning the stratification model on can cause the low-flow case void profiles to over-react to the force and the flow regime to transition out of stratified flow. The code would benefit from an increased amount of Validation & Verification testing. The development of CLAP was significant, as it is a cleanly written, logical representation of the reactor coolant-line geometry. It is stable and capable of modeling basic flow physics in the reactor coolant-line. Code development and debugging required the temporary removal of the energy equation and mass-transfer terms in governing equations. The reintroduction of these terms will allow future coupling to RELAP and re-coupling with COBRA-TF. Adding in more applicable entrainment and de-entrainment models would allow the capture of more advanced physics in the coolant-line that can be expected during Loss-of-Coolant Accident. One of the package's benefits is its ability to be used as a platform for future coolant-line model development and implementation, including capturing of the important de-entrainment behavior in reactor hot-legs (steam-binding effect) and flow convection in the upper-plenum region of the vessel.

  13. msap: a tool for the statistical analysis of methylation-sensitive amplified polymorphism data.

    PubMed

    Pérez-Figueroa, A

    2013-05-01

    In this study msap, an R package which analyses methylation-sensitive amplified polymorphism (MSAP or MS-AFLP) data is presented. The program provides a deep analysis of epigenetic variation starting from a binary data matrix indicating the banding pattern between the isoesquizomeric endonucleases HpaII and MspI, with differential sensitivity to cytosine methylation. After comparing the restriction fragments, the program determines if each fragment is susceptible to methylation (representative of epigenetic variation) or if there is no evidence of methylation (representative of genetic variation). The package provides, in a user-friendly command line interface, a pipeline of different analyses of the variation (genetic and epigenetic) among user-defined groups of samples, as well as the classification of the methylation occurrences in those groups. Statistical testing provides support to the analyses. A comprehensive report of the analyses and several useful plots could help researchers to assess the epigenetic and genetic variation in their MSAP experiments. msap is downloadable from CRAN (http://cran.r-project.org/) and its own webpage (http://msap.r-forge.R-project.org/). The package is intended to be easy to use even for those people unfamiliar with the R command line environment. Advanced users may take advantage of the available source code to adapt msap to more complex analyses. © 2013 Blackwell Publishing Ltd.

  14. c-Src, Insulin-Like Growth Factor I Receptor, G-Protein-Coupled Receptor Kinases and Focal Adhesion Kinase are Enriched Into Prostate Cancer Cell Exosomes.

    PubMed

    DeRita, Rachel M; Zerlanko, Brad; Singh, Amrita; Lu, Huimin; Iozzo, Renato V; Benovic, Jeffrey L; Languino, Lucia R

    2017-01-01

    It is well known that Src tyrosine kinase, insulin-like growth factor 1 receptor (IGF-IR), and focal adhesion kinase (FAK) play important roles in prostate cancer (PrCa) development and progression. Src, which signals through FAK in response to integrin activation, has been implicated in many aspects of tumor biology, such as cell proliferation, metastasis, and angiogenesis. Furthermore, Src signaling is known to crosstalk with IGF-IR, which also promotes angiogenesis. In this study, we demonstrate that c-Src, IGF-IR, and FAK are packaged into exosomes (Exo), c-Src in particular being highly enriched in Exo from the androgen receptor (AR)-positive cell line C4-2B and AR-negative cell lines PC3 and DU145. Furthermore, we show that the active phosphorylated form of Src (Src pY416 ) is co-expressed in Exo with phosphorylated FAK (FAK pY861 ), a known target site of Src, which enhances proliferation and migration. We further demonstrate for the first time exosomal enrichment of G-protein-coupled receptor kinase (GRK) 5 and GRK6, both of which regulate Src and IGF-IR signaling and have been implicated in cancer. Finally, Src pY416 and c-Src are both expressed in Exo isolated from the plasma of prostate tumor-bearing TRAMP mice, and those same mice have higher levels of exosomal c-Src than their wild-type counterparts. In summary, we provide new evidence that active signaling molecules relevant to PrCa are enriched in Exo, and this suggests that the Src signaling network may provide useful biomarkers detectable by liquid biopsy, and may contribute to PrCa progression via Exo. J. Cell. Biochem. 118: 66-73, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  15. Localized heating and bonding technique for MEMS packaging

    NASA Astrophysics Data System (ADS)

    Cheng, Yu-Ting

    Localized heating and bonding techniques have been developed for hermetic and vacuum packaging of MEMS devices, including silicon-to-glass fusion, silicon-gold eutectic, and silicon-to-glass bonding using PSG, indium, aluminum, and aluminum/silicon alloy as the intermediate layer. Line shaped phosphorus-doped polysilicon or gold films are used as resistive microheaters to provide enough thermal energy for bonding. The bonding processes are conducted in the common environment of room temperature and atmospheric pressure and can achieve bonding strength comparable to the fracture toughness of bulk silicon in less than 10 minutes. About 5 watts of input power is needed for localized bonding which can seal a 500 x 500 mum2 area. The total input power is determined by the thermal properties of bonding materials, including the heat capacity and latent heat. Two important bonding results are obtained: (1) The surface step created by the electrical interconnect line can be planarized by reflowing the metal solder. (2) Small applied pressure, less than 1MPa, for intimate contact reduces mechanical damage to the device substrate. This new class of bonding technology has potential applications for MEMS fabrication and packaging that require low temperature processing at the wafer level, excellent bonding strength and hermetic sealing characteristics. A hermetic package based on localized aluminum/silicon-to-glass bonding has been successfully fabricated. Less than 0.2 MPa contact pressure with 46mA input current for two parallel 3.5mum wide polysilicon on-chip microheaters can create as high as 700°C bonding temperature and achieve a strong and reliable bond in 7.5 minutes. Accelerated testing in an autoclave shows some packages survive more than 450 hours under 3 atm, 100%RH and 128°C. Premature failure has been attributed to some unbonded regions on the failed samples. The bonding yield and reliability have been improved by increasing bonding time and applied pressure. Finally, vacuum encapsulation of folded-beam comb-drive mu-resonators used as pressure monitors has been demonstrated using localized aluminum/silicon-to-glass bonding. With 3.4 watt heating power, ˜0.2MPa applied contact pressure, and 90 minutes wait time before bonding, vacuum encapsulation can be achieved with the same vacuum level as the packaging environment which is about 25 mtorr. Metal coating used as diffusion barrier and a longer wait time before bonding are used to improve the vacuum level of the package. Long-term measurement of the Q of un-annealed vacuum-packaged mu-resonators, illustrates stable operation after 19 weeks.

  16. R software package based statistical optimization of process components to simultaneously enhance the bacterial growth, laccase production and textile dye decolorization with cytotoxicity study

    PubMed Central

    Dudhagara, Pravin; Tank, Shantilal

    2018-01-01

    The thermophilic bacterium, Bacillus licheniformis U1 is used for the optimization of bacterial growth (R1), laccase production (R2) and synthetic disperse blue DBR textile dye decolorization (R3) in the present study. Preliminary optimization has been performed by one variable at time (OVAT) approach using four media components viz., dye concentration, copper sulphate concentration, pH, and inoculum size. Based on OVAT result further statistical optimization of R1, R2 and R3 performed by Box–Behnken design (BBD) using response surface methodology (RSM) in R software with R Commander package. The total 29 experimental runs conducted in the experimental design study towards the construction of a quadratic model. The model indicated that dye concentration 110 ppm, copper sulphate 0.2 mM, pH 7.5 and inoculum size 6% v/v were found to be optimum to maximize the laccase production and bacterial growth. Whereas, maximum dye decolorization achieved in media containing dye concentration 110 ppm, copper sulphate 0.6 mM, pH 6 and inoculum size 6% v/v. R package predicted R2 of R1, R2 and R3 were 0.9917, 0.9831 and 0.9703 respectively; likened to Design-Expert (Stat-Ease) (DOE) predicted R2 of R1, R2, and R3 were 0.9893, 0.9822 and 0.8442 respectively. The values obtained by R software were more precise, reliable and reproducible, compared to the DOE model. The laccase production was 1.80 fold increased, and 2.24 fold enhancement in dye decolorization was achieved using optimized medium than initial experiments. Moreover, the laccase-treated sample demonstrated the less cytotoxic effect on L132 and MCF-7 cell lines compared to untreated sample using MTT assay. Higher cell viability and lower cytotoxicity observed in a laccase-treated sample suggest the impending application of bacterial laccase in the reduction of toxicity of dye to design rapid biodegradation process. PMID:29718934

  17. Thyroid cell lines in research on goitrogenesis.

    PubMed

    Gerber, H; Peter, H J; Asmis, L; Studer, H

    1991-12-01

    Thyroid cell lines have contributed a lot to the understanding of goitrogenesis. The cell lines mostly used in thyroid research are briefly discussed, namely the rat thyroid cell lines FRTL and FRTL-5, the porcine thyroid cell lines PORTHOS and ARTHOS, The sheep thyroid cell lines OVNIS 5H and 6H, the cat thyroid cell lines PETCAT 1 to 4 and ROMCAT, and the human thyroid cell lines FTC-133 and HTh 74. Chinese hamster ovary (CHO) cells and COS-7 cells, stably transfected with TSH receptor cDNA and expressing a functional TSH receptor, are discussed as examples for non-thyroidal cells, transfected with thyroid genes.

  18. Analysis of renal cancer cell lines from two major resources enables genomics-guided cell line selection

    NASA Astrophysics Data System (ADS)

    Sinha, Rileen; Winer, Andrew G.; Chevinsky, Michael; Jakubowski, Christopher; Chen, Ying-Bei; Dong, Yiyu; Tickoo, Satish K.; Reuter, Victor E.; Russo, Paul; Coleman, Jonathan A.; Sander, Chris; Hsieh, James J.; Hakimi, A. Ari

    2017-05-01

    The utility of cancer cell lines is affected by the similarity to endogenous tumour cells. Here we compare genomic data from 65 kidney-derived cell lines from the Cancer Cell Line Encyclopedia and the COSMIC Cell Lines Project to three renal cancer subtypes from The Cancer Genome Atlas: clear cell renal cell carcinoma (ccRCC, also known as kidney renal clear cell carcinoma), papillary (pRCC, also known as kidney papillary) and chromophobe (chRCC, also known as kidney chromophobe) renal cell carcinoma. Clustering copy number alterations shows that most cell lines resemble ccRCC, a few (including some often used as models of ccRCC) resemble pRCC, and none resemble chRCC. Human ccRCC tumours clustering with cell lines display clinical and genomic features of more aggressive disease, suggesting that cell lines best represent aggressive tumours. We stratify mutations and copy number alterations for important kidney cancer genes by the consistency between databases, and classify cell lines into established gene expression-based indolent and aggressive subtypes. Our results could aid investigators in analysing appropriate renal cancer cell lines.

  19. ARCHANGEL: Galaxy Photometry System

    NASA Astrophysics Data System (ADS)

    Schombert, James

    2011-07-01

    ARCHANGEL is a Unix-based package for the surface photometry of galaxies. While oriented for large angular size systems (i.e. many pixels), its tools can be applied to any imaging data of any size. The package core contains routines to perform the following critical galaxy photometry functions: sky determination; frame cleaning; ellipse fitting; profile fitting; and total and isophotal magnitudes. The goal of the package is to provide an automated, assembly-line type of reduction system for galaxy photometry of space-based or ground-based imaging data. The procedures outlined in the documentation are flux independent, thus, these routines can be used for non-optical data as well as typical imaging datasets. ARCHANGEL has been tested on several current OS's (RedHat Linux, Ubuntu Linux, Solaris, Mac OS X). A tarball for installation is available at the download page. The main routines are Python and FORTRAN based, therefore, a current installation of Python and a FORTRAN compiler are required. The ARCHANGEL package also contains Python hooks to the PGPLOT package, an XML processor and network tools which automatically link to data archives (i.e. NED, HST, 2MASS, etc) to download images in a non-interactive manner.

  20. Stitch-bond parallel-gap welding for IC circuits

    NASA Technical Reports Server (NTRS)

    Chvostal, P.; Tuttle, J.; Vanderpool, R.

    1980-01-01

    Stitch-bonded flatpacks are superior to soldered dual-in-lines where size, weight, and reliability are important. Results should interest designers of packaging for complex high-reliability electronics, such as that used in security systems, industrial process control, and vehicle electronics.

  1. Electronic ripple indicator

    NASA Technical Reports Server (NTRS)

    Davidson, J. K.; Houck, W. H.

    1971-01-01

    Electronic circuit for monitoring excessive ripple voltage on dc power lines senses voltage variations from few millivolts to maximum of 10 volts rms. Instrument is used wherever power supply fluctuations might endanger system operations or damage equipment. Device is inexpensive and easily packaged in small chassis.

  2. NASA Leads Demo for Drone Traffic Management Tech

    NASA Image and Video Library

    2017-06-30

    During the latest NASA-led demonstrations of technologies that could be part of an automated traffic management system for drones, pilots sent their vehicles beyond visual line-of-sight in simulated infrastructure inspections, search and rescue support, and package delivery.

  3. [Assessment of the probability of encountering staphylococcal enterotoxins in lactic acid cheese packaged in laminates].

    PubMed

    Steinka, Izabela

    2004-01-01

    Immunoassay methods were used to identify the presence of staphylococcal enterotoxins in lactic acid cheese vacuum and non-vacuum packed. There was assessed the probability of encountering staphylococcal enterotoxin in cheese dependent on different systems of packaging, count of staphylococcal cells, intensiveness of coagulase synthesis and tightness of packaging. The presence of enterotoxin was identified in 5% of researched samples of products stored for 14 days. The influence of packaging system and tightness on presence of enterotoxin was observed. The probability of presence of staphylococcal and enterotoxin in relation to researched factors was presented by the mathematical models.

  4. Flexible packaging for PV modules

    NASA Astrophysics Data System (ADS)

    Dhere, Neelkanth G.

    2008-08-01

    Economic, flexible packages that provide needed level of protection to organic and some other PV cells over >25-years have not yet been developed. However, flexible packaging is essential in niche large-scale applications. Typical configuration used in flexible photovoltaic (PV) module packaging is transparent frontsheet/encapsulant/PV cells/flexible substrate. Besides flexibility of various components, the solder bonds should also be flexible and resistant to fatigue due to cyclic loading. Flexible front sheets should provide optical transparency, mechanical protection, scratch resistance, dielectric isolation, water resistance, UV stability and adhesion to encapsulant. Examples are Tefzel, Tedlar and Silicone. Dirt can get embedded in soft layers such as silicone and obscure light. Water vapor transmittance rate (WVTR) of polymer films used in the food packaging industry as moisture barriers are ~0.05 g/(m2.day) under ambient conditions. In comparison, light emitting diodes employ packaging components that have WVTR of ~10-6 g/(m2.day). WVTR of polymer sheets can be improved by coating them with dense inorganic/organic multilayers. Ethylene vinyl acetate, an amorphous copolymer used predominantly by the PV industry has very high O2 and H2O diffusivity. Quaternary carbon chains (such as acetate) in a polymer lead to cleavage and loss of adhesional strength at relatively low exposures. Reactivity of PV module components increases in presence of O2 and H2O. Adhesional strength degrades due to the breakdown of structure of polymer by reactive, free radicals formed by high-energy radiation. Free radical formation in polymers is reduced when the aromatic rings are attached at regular intervals. This paper will review flexible packaging for PV modules.

  5. DEsingle for detecting three types of differential expression in single-cell RNA-seq data.

    PubMed

    Miao, Zhun; Deng, Ke; Wang, Xiaowo; Zhang, Xuegong

    2018-04-24

    The excessive amount of zeros in single-cell RNA-seq data include "real" zeros due to the on-off nature of gene transcription in single cells and "dropout" zeros due to technical reasons. Existing differential expression (DE) analysis methods cannot distinguish these two types of zeros. We developed an R package DEsingle which employed Zero-Inflated Negative Binomial model to estimate the proportion of real and dropout zeros and to define and detect 3 types of DE genes in single-cell RNA-seq data with higher accuracy. The R package DEsingle is freely available at https://github.com/miaozhun/DEsingle and is under Bioconductor's consideration now. zhangxg@tsinghua.edu.cn. Supplementary data are available at Bioinformatics online.

  6. Advances in rechargeable lithium molybdenum disulfide batteries

    NASA Technical Reports Server (NTRS)

    Brandt, K.; Stiles, J. A. R.

    1985-01-01

    The lithium molybdenum disulfide system as demonstrated in a C size cell, offers performance characteristics for applications where light weight and low volume are important. A gravimetric energy density of 90 watt hours per kilogram can be achieved in a C size cell package. The combination of charge retention capabilities, high energy density and a state of charge indicator in a rechargeable cell provides power package for a wide range of devices. The system overcomes the memory effect in Nicads where the full capacity of the battery cannot be utilized unless it was utilized on previous cycles. The development of cells with an advanced electrolyte formulation led to an improved rate capability especially at low temperatures and to a significantly improved life cycle.

  7. TLM-Tracker: software for cell segmentation, tracking and lineage analysis in time-lapse microscopy movies.

    PubMed

    Klein, Johannes; Leupold, Stefan; Biegler, Ilona; Biedendieck, Rebekka; Münch, Richard; Jahn, Dieter

    2012-09-01

    Time-lapse imaging in combination with fluorescence microscopy techniques enable the investigation of gene regulatory circuits and uncovered phenomena like culture heterogeneity. In this context, computational image processing for the analysis of single cell behaviour plays an increasing role in systems biology and mathematical modelling approaches. Consequently, we developed a software package with graphical user interface for the analysis of single bacterial cell behaviour. A new software called TLM-Tracker allows for the flexible and user-friendly interpretation for the segmentation, tracking and lineage analysis of microbial cells in time-lapse movies. The software package, including manual, tutorial video and examples, is available as Matlab code or executable binaries at http://www.tlmtracker.tu-bs.de.

  8. 49 CFR 173.4a - Excepted quantities.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... ice), and lithium batteries and cells. (c) Inner packaging limits. The maximum quantity of hazardous materials in each inner packaging is limited to: (1) For toxic material with a Division 6.1 primary or... excepted quantities must meet the following: (1) Each inner receptacle must be constructed of plastic, or...

  9. Disruption of Specific RNA-RNA Interactions in a Double-Stranded RNA Virus Inhibits Genome Packaging and Virus Infectivity

    PubMed Central

    Fajardo, Teodoro; Sung, Po-Yu; Roy, Polly

    2015-01-01

    Bluetongue virus (BTV) causes hemorrhagic disease in economically important livestock. The BTV genome is organized into ten discrete double-stranded RNA molecules (S1-S10) which have been suggested to follow a sequential packaging pathway from smallest to largest segment during virus capsid assembly. To substantiate and extend these studies, we have investigated the RNA sorting and packaging mechanisms with a new experimental approach using inhibitory oligonucleotides. Putative packaging signals present in the 3’untranslated regions of BTV segments were targeted by a number of nuclease resistant oligoribonucleotides (ORNs) and their effects on virus replication in cell culture were assessed. ORNs complementary to the 3’ UTR of BTV RNAs significantly inhibited virus replication without affecting protein synthesis. Same ORNs were found to inhibit complex formation when added to a novel RNA-RNA interaction assay which measured the formation of supramolecular complexes between and among different RNA segments. ORNs targeting the 3’UTR of BTV segment 10, the smallest RNA segment, were shown to be the most potent and deletions or substitution mutations of the targeted sequences diminished the RNA complexes and abolished the recovery of viable viruses using reverse genetics. Cell-free capsid assembly/RNA packaging assay also confirmed that the inhibitory ORNs could interfere with RNA packaging and further substitution mutations within the putative RNA packaging sequence have identified the recognition sequence concerned. Exchange of 3’UTR between segments have further demonstrated that RNA recognition was segment specific, most likely acting as part of the secondary structure of the entire genomic segment. Our data confirm that genome packaging in this segmented dsRNA virus occurs via the formation of supramolecular complexes formed by the interaction of specific sequences located in the 3’ UTRs. Additionally, the inhibition of packaging in-trans with inhibitory ORNs suggests this that interaction is a bona fide target for the design of compounds with antiviral activity. PMID:26646790

  10. Disruption of Specific RNA-RNA Interactions in a Double-Stranded RNA Virus Inhibits Genome Packaging and Virus Infectivity.

    PubMed

    Fajardo, Teodoro; Sung, Po-Yu; Roy, Polly

    2015-12-01

    Bluetongue virus (BTV) causes hemorrhagic disease in economically important livestock. The BTV genome is organized into ten discrete double-stranded RNA molecules (S1-S10) which have been suggested to follow a sequential packaging pathway from smallest to largest segment during virus capsid assembly. To substantiate and extend these studies, we have investigated the RNA sorting and packaging mechanisms with a new experimental approach using inhibitory oligonucleotides. Putative packaging signals present in the 3'untranslated regions of BTV segments were targeted by a number of nuclease resistant oligoribonucleotides (ORNs) and their effects on virus replication in cell culture were assessed. ORNs complementary to the 3' UTR of BTV RNAs significantly inhibited virus replication without affecting protein synthesis. Same ORNs were found to inhibit complex formation when added to a novel RNA-RNA interaction assay which measured the formation of supramolecular complexes between and among different RNA segments. ORNs targeting the 3'UTR of BTV segment 10, the smallest RNA segment, were shown to be the most potent and deletions or substitution mutations of the targeted sequences diminished the RNA complexes and abolished the recovery of viable viruses using reverse genetics. Cell-free capsid assembly/RNA packaging assay also confirmed that the inhibitory ORNs could interfere with RNA packaging and further substitution mutations within the putative RNA packaging sequence have identified the recognition sequence concerned. Exchange of 3'UTR between segments have further demonstrated that RNA recognition was segment specific, most likely acting as part of the secondary structure of the entire genomic segment. Our data confirm that genome packaging in this segmented dsRNA virus occurs via the formation of supramolecular complexes formed by the interaction of specific sequences located in the 3' UTRs. Additionally, the inhibition of packaging in-trans with inhibitory ORNs suggests this that interaction is a bona fide target for the design of compounds with antiviral activity.

  11. The New Web-Based Hera Data Processing System at the HEASARC

    NASA Technical Reports Server (NTRS)

    Pence, W.

    2011-01-01

    The HEASARC at NASA/GSFC has provide an on-line astronomical data processing system called Hera for several years. Hera provides a complete data processing environment, including installed software packages, local data storage, and the CPU resources needed to process the user's data. The original design of Hera, however, has 2 requirements that has limited it's usefulness for some users, namely, that 1) the user must download and install a small helper program on their own computer before using Hera, and 2) Hera requires that several computer ports/sockets be allowed to communicate through any local firewalls on the users machine. Both of these restrictions can be problematic for some users, therefore we are now migrating Hera into a purely Web based environment which only requires a standard Web browser. The first release of Web Hera is now publicly available at http://heasarc.gsfc.nasa.gov/webheara/. It currently provides a standard graphical interface for running hundreds of different data processing programs that are available in the HEASARC's ftools software package. Over the next year we to add more features to Web Hera, including an interactive command line interface, and more display and line capabilities.

  12. Viral nanoparticle-encapsidated enzyme and restructured DNA for cell delivery and gene expression

    PubMed Central

    Liu, Jinny L.; Dixit, Aparna Banerjee; Robertson, Kelly L.; Qiao, Eric; Black, Lindsay W.

    2014-01-01

    Packaging specific exogenous active proteins and DNAs together within a single viral-nanocontainer is challenging. The bacteriophage T4 capsid (100 × 70 nm) is well suited for this purpose, because it can hold a single long DNA or multiple short pieces of DNA up to 170 kb packed together with more than 1,000 protein molecules. Any linear DNA can be packaged in vitro into purified procapsids. The capsid-targeting sequence (CTS) directs virtually any protein into the procapsid. Procapsids are assembled with specific CTS-directed exogenous proteins that are encapsidated before the DNA. The capsid also can display on its surface high-affinity eukaryotic cell-binding peptides or proteins that are in fusion with small outer capsid and head outer capsid surface-decoration proteins that can be added in vivo or in vitro. In this study, we demonstrate that the site-specific recombinase cyclic recombination (Cre) targeted into the procapsid is enzymatically active within the procapsid and recircularizes linear plasmid DNA containing two terminal loxP recognition sites when packaged in vitro. mCherry expression driven by a cytomegalovirus promoter in the capsid containing Cre-circularized DNA is enhanced over linear DNA, as shown in recipient eukaryotic cells. The efficient and specific packaging into capsids and the unpackaging of both DNA and protein with release of the enzymatically altered protein–DNA complexes from the nanoparticles into cells have potential in numerous downstream drug and gene therapeutic applications. PMID:25161284

  13. Evaluating cell lines as tumour models by comparison of genomic profiles

    PubMed Central

    Domcke, Silvia; Sinha, Rileen; Levine, Douglas A.; Sander, Chris; Schultz, Nikolaus

    2013-01-01

    Cancer cell lines are frequently used as in vitro tumour models. Recent molecular profiles of hundreds of cell lines from The Cancer Cell Line Encyclopedia and thousands of tumour samples from the Cancer Genome Atlas now allow a systematic genomic comparison of cell lines and tumours. Here we analyse a panel of 47 ovarian cancer cell lines and identify those that have the highest genetic similarity to ovarian tumours. Our comparison of copy-number changes, mutations and mRNA expression profiles reveals pronounced differences in molecular profiles between commonly used ovarian cancer cell lines and high-grade serous ovarian cancer tumour samples. We identify several rarely used cell lines that more closely resemble cognate tumour profiles than commonly used cell lines, and we propose these lines as the most suitable models of ovarian cancer. Our results indicate that the gap between cell lines and tumours can be bridged by genomically informed choices of cell line models for all tumour types. PMID:23839242

  14. Isoform-level gene expression patterns in single-cell RNA-sequencing data.

    PubMed

    Vu, Trung Nghia; Wills, Quin F; Kalari, Krishna R; Niu, Nifang; Wang, Liewei; Pawitan, Yudi; Rantalainen, Mattias

    2018-02-27

    RNA sequencing of single cells enables characterization of transcriptional heterogeneity in seemingly homogeneous cell populations. Single-cell sequencing has been applied in a wide range of researches fields. However, few studies have focus on characterization of isoform-level expression patterns at the single-cell level. In this study we propose and apply a novel method, ISOform-Patterns (ISOP), based on mixture modeling, to characterize the expression patterns of isoform pairs from the same gene in single-cell isoform-level expression data. We define six principal patterns of isoform expression relationships and describe a method for differential-pattern analysis. We demonstrate ISOP through analysis of single-cell RNA-sequencing data from a breast cancer cell line, with replication in three independent datasets. We assigned the pattern types to each of 16,562 isoform-pairs from 4,929 genes. Among those, 26% of the discovered patterns were significant (p<0.05), while remaining patterns are possibly effects of transcriptional bursting, drop-out and stochastic biological heterogeneity. Furthermore, 32% of genes discovered through differential-pattern analysis were not detected by differential-expression analysis. The effect of drop-out events, mean expression level, and properties of the expression distribution on the performances of ISOP were also investigated through simulated datasets. To conclude, ISOP provides a novel approach for characterization of isoformlevel preference, commitment and heterogeneity in single-cell RNA-sequencing data. The ISOP method has been implemented as a R package and is available at https://github.com/nghiavtr/ISOP under a GPL-3 license. mattias.rantalainen@ki.se. Supplementary data are available at Bioinformatics online.

  15. Pharmacological mimicking of caloric restriction elicits epigenetic reprogramming of differentiated cells to stem-like self-renewal states.

    PubMed

    Oliveras-Ferraros, Cristina; Vazquez-Martin, Alejandro; Menendez, Javier A

    2010-10-01

    Networks of oncogenes and tumor suppressor genes that control cancer cell proliferation also regulate stem cell renewal and possibly stem cell aging. Because (de)differentiation processes might dictate tumor cells to retrogress to a more stem-like state in response to aging-relevant epigenetic and/or environmental players, we recently envisioned that cultured human cancer cells might be used as reliable models to test the ability of antiaging interventions for promoting the initiation and maintenance of self-renewing divisions. Cancer cell lines naturally bearing undetectable amounts of stem/progenitor-like cell populations were continuously cultured in the presence of the caloric restriction mimetic metformin for several months. Microarray technology was employed to profile expression of genes related to the identification, growth, and differentiation of stem cells. Detection of functionally related gene groups using a pathway analysis package provided annotated genetic signatures over- and underexpressed in response to pharmacological mimicking of caloric restriction. By following this methodological approach, we recently obtained data fitting a model in which, in response to chronic impairment of cellular bioenergetics imposed by metformin-induced mitochondrial uncoupling as assessed by the phosphorylation state of cAMP-response element binding protein (CREB), tumor cells can retrogress from a differentiated state to a more CD44(+) stem-like primitive state epigenetically governed by the Polycomb-group suppressor BMI1-a crucial "stemness" gene involved in the epigenetic maintenance of adult stem cells. These findings might provide a novel molecular avenue to investigate if antiaging benefits from caloric restriction mimetics might relate to their ability to epigenetically reprogram stemness while prolonging the capacity of stem-like cell states to proliferate, differentiate, and replace mature cells in adult aging tissues.

  16. simBio: a Java package for the development of detailed cell models.

    PubMed

    Sarai, Nobuaki; Matsuoka, Satoshi; Noma, Akinori

    2006-01-01

    Quantitative dynamic computer models, which integrate a variety of molecular functions into a cell model, provide a powerful tool to create and test working hypotheses. We have developed a new modeling tool, the simBio package (freely available from ), which can be used for constructing cell models, such as cardiac cells (the Kyoto model from Matsuoka et al., 2003, 2004 a, b, the LRd model from Faber and Rudy, 2000, and the Noble 98 model from Noble et al., 1998), epithelial cells (Strieter et al., 1990) and pancreatic beta cells (Magnus and Keizer, 1998). The simBio package is written in Java, uses XML and can solve ordinary differential equations. In an attempt to mimic biological functional structures, a cell model is, in simBio, composed of independent functional modules called Reactors, such as ion channels and the sarcoplasmic reticulum, and dynamic variables called Nodes, such as ion concentrations. The interactions between Reactors and Nodes are described by the graph theory and the resulting graph represents a blueprint of an intricate cellular system. Reactors are prepared in a hierarchical order, in analogy to the biological classification. Each Reactor can be composed or improved independently, and can easily be reused for different models. This way of building models, through the combination of various modules, is enabled through the use of object-oriented programming concepts. Thus, simBio is a straightforward system for the creation of a variety of cell models on a common database of functional modules.

  17. Continuous hematopoietic cell lines as model systems for leukemia-lymphoma research.

    PubMed

    Drexler, H G; Matsuo, A Y; MacLeod, R A

    2000-11-01

    Along with other improvements, the advent of continuous human leukemia-lymphoma (LL) cell lines as a rich resource of abundant, accessible and manipulable living cells has contributed significantly to a better understanding of the pathophysiology of hematopoietic tumors. The first LL cell lines, Burkitt's lymphoma-derived lines, were established in 1963. Since then, more than 1000 cell lines have been described, although not all of them in full detail. The major advantages of continuous cell lines is the unlimited supply and worldwide availability of identical cell material, and the infinite viable storability in liquid nitrogen. LL cell lines are characterized generally by monoclonal origin and differentiation arrest, sustained proliferation in vitro under preservation of most cellular features, and specific genetic alterations. The most practical classification of LL cell lines assigns them to one of the physiologically occurring cell lineages, based on their immunophenotype, genotype and functional features. Truly malignant cell lines must be discerned from Epstein-Barr virus (EBV)-immortalized normal cells, using various distinguishing parameters. However, the picture is not quite so straightforward, as some types of LL cell lines are indeed EBV+, and some EBV+ normal cell lines carry also genetic aberrations and may mimic malignancy-associated features. Apart from EBV and human T-cell leukemia virus in some lines, the majority of wild-type LL cell lines are virus-negative. The efficiency of cell line establishment is rather low and the deliberate establishment of new LL cell lines remains by and large an unpredictable random process. Difficulties in establishing continuous cell lines may be caused by the inappropriate selection of nutrients and growth factors for these cells. Clearly, a generally suitable microenvironment for hematopoietic cells, either malignant or normal, cannot yet be created in vitro. The characterization and publication of new LL cell lines should provide important and informative core data, attesting to their scientific significance. Large percentages of LL cell lines are contaminated with mycoplasma (about 30%) or are cross-contaminated with other cell lines (about 15-20%). Solutions to these problems are sensitive detection, effective elimination and rigorous prevention of mycoplasma infection, and proper, regular authentication of cell lines. The underlying cause, however, appears to be negligent cell culture practice. The willingness of investigators to make their LL cell lines available to others is all too often limited. There is a need in the scientific community for clean and authenticated high-quality LL cell lines to which every scientist has access. These are offered by various institutionalized public cell line banks. It has been argued that LL cell lines are genetically unstable (both cytogenetically and molecular genetically). For instance, cell lines are supposed to acquire numerical and structural chromosomal alterations and various types of mutations (e.g. point mutations) in vitro. We present evidence that while nearly 100% of all LL cell lines indeed carry genetic alterations, these alterations appear to be stable rather than unstable. As an example of the practical utility of LL cell lines, the recent advances in studies of classical and molecular cytogenetics, which in large part were made possible by cell lines, are highlighted. A list of the most useful, robust and publicly available reference cell lines that may be used for a variety of experimental purposes is proposed. Clearly, by opening new avenues for investigation, studies of LL cell lines have provided seminal insights into the biology of hematopoietic neoplasia. Over a period of nearly four decades, these initially rather exotic cell cultures, known only to a few specialists, have become ubiquitous powerful research tools that are available to every investigator.

  18. heatmaply: an R package for creating interactive cluster heatmaps for online publishing.

    PubMed

    Galili, Tal; O'Callaghan, Alan; Sidi, Jonathan; Sievert, Carson

    2018-05-01

    heatmaply is an R package for easily creating interactive cluster heatmaps that can be shared online as a stand-alone HTML file. Interactivity includes a tooltip display of values when hovering over cells, as well as the ability to zoom in to specific sections of the figure from the data matrix, the side dendrograms, or annotated labels. Thanks to the synergistic relationship between heatmaply and other R packages, the user is empowered by a refined control over the statistical and visual aspects of the heatmap layout. The heatmaply package is available under the GPL-2 Open Source license. It comes with a detailed vignette, and is freely available from: http://cran.r-project.org/package=heatmaply. tal.galili@math.tau.ac.il. Supplementary data are available at Bioinformatics online.

  19. Evaluation of a fast and flexible OPC package: OPTISSIMO

    NASA Astrophysics Data System (ADS)

    Maurer, Wilhelm; Waas, Thomas; Eisenmann, Hans

    1996-12-01

    It is out of question, that current state-of-the-art lithography--printing 350 nm structures with i-line tools or 250 nm structures with DUV tools--needs to correct for proximity effects (OPC). Otherwise, all the well-known effects like line-end shortening, linewidth variation as a function of adjacent patterns, linewidth non-linearity, etc. will produce a pattern, that is significantly different from the intended design. In this paper, we report first evaluation results of OPTISSIMO, a software package for automatic proximity correction. Besides the ability to handle full-chip designs by preserving as much as possible of the original data-hierarchy, there are significant options for the user. A large number of choices can be made to balance between the precision of the correction and the complexity of the corrected design. The main target of our evaluations was to check for full-chip OPC for the gate level of a state-of-the-art design. This corresponds to print either linewidths in the 350 nm to 400 nm range with i-line lithography or 250 nm/300 nm linewidth with DUV lithography. Taking 400 nm i-line lithography as an example, 3% precision OPC which has been demonstrated. By using hierarchical data handling, it was shown, that even the data complexity of a 256 M DRAM can be managed within reasonable time.

  20. Cyrface: An interface from Cytoscape to R that provides a user interface to R packages.

    PubMed

    Gonçalves, Emanuel; Mirlach, Franz; Saez-Rodriguez, Julio

    2013-01-01

    There is an increasing number of software packages to analyse biological experimental data in the R environment. In particular, Bioconductor, a repository of curated R packages, is one of the most comprehensive resources for bioinformatics and biostatistics. The use of these packages is increasing, but it requires a basic understanding of the R language, as well as the syntax of the specific package used. The availability of user graphical interfaces for these packages would decrease the learning curve and broaden their application. Here, we present a Cytoscape app termed Cyrface that allows Cytoscape apps to connect to any function and package developed in R. Cyrface can be used to run R packages from within the Cytoscape environment making use of a graphical user interface. Moreover, it can link R packages with the capabilities of Cytoscape and its apps, in particular network visualization and analysis. Cyrface's utility has been demonstrated for two Bioconductor packages ( CellNOptR and DrugVsDisease), and here we further illustrate its usage by implementing a workflow of data analysis and visualization. Download links, installation instructions and user guides can be accessed from the Cyrface's homepage ( http://www.ebi.ac.uk/saezrodriguez/cyrface/) and from the Cytoscape app store ( http://apps.cytoscape.org/apps/cyrface).

  1. Self Cooling/Heating Devices

    NASA Technical Reports Server (NTRS)

    1992-01-01

    A NASA RTTC assisted International Thermal Packaging (ITP) in the identification of a NASA-developed synthetic polymer that can absorb 1,000 times its weight in water. The desiccant was used in the manufacture of a self-chilling can, a major innovation for packaging food and beverages. The refrigeration device is a small cylinder, fully incorporated into the can. When used in beverage containers, the top of the can is popped, the internal carbonization pressure is relieved, and the self-chilling reaction is induced. Company product line has been expanded to include two related products, and several licensing agreements have been signed.

  2. Design automation for complex CMOS/SOS LSI hybrid substrates

    NASA Technical Reports Server (NTRS)

    Ramondetta, P. W.; Smiley, J. W.

    1976-01-01

    A design automated approach used to develop thick-film hybrid packages is described. The hybrid packages produced combine thick-film and silicon on sapphire (SOS) laser surface interaction technologies to bring the on-chip performance level of SOS to the subsystem level. Packing densities are improved by a factor of eight over ceramic dual in-line packing; interchip wiring capacitance is low. Due to significant time savings, the design automated approach presented can be expected to yield a 3:1 reduction in cost over the use of manual methods for the initial design of a hybrid.

  3. Packaging - Materials review

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Herrmann, Matthias

    2014-06-16

    Nowadays, a large number of different electrochemical energy storage systems are known. In the last two decades the development was strongly driven by a continuously growing market of portable electronic devices (e.g. cellular phones, lap top computers, camcorders, cameras, tools). Current intensive efforts are under way to develop systems for automotive industry within the framework of electrically propelled mobility (e.g. hybrid electric vehicles, plug-in hybrid electric vehicles, full electric vehicles) and also for the energy storage market (e.g. electrical grid stability, renewable energies). Besides the different systems (cell chemistries), electrochemical cells and batteries were developed and are offered in manymore » shapes, sizes and designs, in order to meet performance and design requirements of the widespread applications. Proper packaging is thereby one important technological step for designing optimum, reliable and safe batteries for operation. In this contribution, current packaging approaches of cells and batteries together with the corresponding materials are discussed. The focus is laid on rechargeable systems for industrial applications (i.e. alkaline systems, lithium-ion, lead-acid). In principle, four different cell types (shapes) can be identified - button, cylindrical, prismatic and pouch. Cell size can be either in accordance with international (e.g. International Electrotechnical Commission, IEC) or other standards or can meet application-specific dimensions. Since cell housing or container, terminals and, if necessary, safety installations as inactive (non-reactive) materials reduce energy density of the battery, the development of low-weight packages is a challenging task. In addition to that, other requirements have to be fulfilled: mechanical stability and durability, sealing (e.g. high permeation barrier against humidity for lithium-ion technology), high packing efficiency, possible installation of safety devices (current interrupt device, valve, etc.), chemical inertness, cost issues, and others. Finally, proper cell design has to be considered for effective thermal management (i.e. cooling and heating) of battery packs.« less

  4. Ontological representation, integration, and analysis of LINCS cell line cells and their cellular responses.

    PubMed

    Ong, Edison; Xie, Jiangan; Ni, Zhaohui; Liu, Qingping; Sarntivijai, Sirarat; Lin, Yu; Cooper, Daniel; Terryn, Raymond; Stathias, Vasileios; Chung, Caty; Schürer, Stephan; He, Yongqun

    2017-12-21

    Aiming to understand cellular responses to different perturbations, the NIH Common Fund Library of Integrated Network-based Cellular Signatures (LINCS) program involves many institutes and laboratories working on over a thousand cell lines. The community-based Cell Line Ontology (CLO) is selected as the default ontology for LINCS cell line representation and integration. CLO has consistently represented all 1097 LINCS cell lines and included information extracted from the LINCS Data Portal and ChEMBL. Using MCF 10A cell line cells as an example, we demonstrated how to ontologically model LINCS cellular signatures such as their non-tumorigenic epithelial cell type, three-dimensional growth, latrunculin-A-induced actin depolymerization and apoptosis, and cell line transfection. A CLO subset view of LINCS cell lines, named LINCS-CLOview, was generated to support systematic LINCS cell line analysis and queries. In summary, LINCS cell lines are currently associated with 43 cell types, 131 tissues and organs, and 121 cancer types. The LINCS-CLO view information can be queried using SPARQL scripts. CLO was used to support ontological representation, integration, and analysis of over a thousand LINCS cell line cells and their cellular responses.

  5. Genotoxic potential of montmorillonite clay mineral and alteration in the expression of genes involved in toxicity mechanisms in the human hepatoma cell line HepG2.

    PubMed

    Maisanaba, Sara; Hercog, Klara; Filipic, Metka; Jos, Ángeles; Zegura, Bojana

    2016-03-05

    Montmorillonite, also known as Cloisite(®)Na(+) (CNa(+)), is a natural clay with a wide range of well-documented and novel applications, such as pharmaceutical products or food packaging. Although considered a low toxic product, the expected increased exposure to CNa(+) arises concern on the potential consequences on human and environmental health especially as its genotoxicity has scarcely been investigated so far. Thus, we investigated, for the first time, the influence of non-cytotoxic concentrations of CNa(+) (15.65, 31.25 and 62.5 μg/mL) on genomic instability of human hepatoma cell line (HepG2) by determining the formation of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) with the Cytokinesis block micronucleus cytome assay. Further on we studied the influence of CNa(+) on the expression of several genes involved in toxicity mechanisms using the real-time quantitative PCR. The results showed that CNa(+) increased the number of MNi, while the numbers of NBUDs and NPBs were not affected. In addition it deregulated genes in all the groups studied, mainly after longer time of exposure. These findings provide the evidence that CNa(+) is potentially genotoxic. Therefore further studies that will elucidate the molecular mechanisms involved in toxic activity of CNa(+) are needed for hazard identification and human safety assessment. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Spatial Normalization of Reverse Phase Protein Array Data

    PubMed Central

    Kaushik, Poorvi; Molinelli, Evan J.; Miller, Martin L.; Wang, Weiqing; Korkut, Anil; Liu, Wenbin; Ju, Zhenlin; Lu, Yiling; Mills, Gordon; Sander, Chris

    2014-01-01

    Reverse phase protein arrays (RPPA) are an efficient, high-throughput, cost-effective method for the quantification of specific proteins in complex biological samples. The quality of RPPA data may be affected by various sources of error. One of these, spatial variation, is caused by uneven exposure of different parts of an RPPA slide to the reagents used in protein detection. We present a method for the determination and correction of systematic spatial variation in RPPA slides using positive control spots printed on each slide. The method uses a simple bi-linear interpolation technique to obtain a surface representing the spatial variation occurring across the dimensions of a slide. This surface is used to calculate correction factors that can normalize the relative protein concentrations of the samples on each slide. The adoption of the method results in increased agreement between technical and biological replicates of various tumor and cell-line derived samples. Further, in data from a study of the melanoma cell-line SKMEL-133, several slides that had previously been rejected because they had a coefficient of variation (CV) greater than 15%, are rescued by reduction of CV below this threshold in each case. The method is implemented in the R statistical programing language. It is compatible with MicroVigene and SuperCurve, packages commonly used in RPPA data analysis. The method is made available, along with suggestions for implementation, at http://bitbucket.org/rppa_preprocess/rppa_preprocess/src. PMID:25501559

  7. Increased Serotonin Signaling Contributes to the Warburg Effect in Pancreatic Tumor Cells Under Metabolic Stress and Promotes Growth of Pancreatic Tumors in Mice.

    PubMed

    Jiang, Shu-Heng; Li, Jun; Dong, Fang-Yuan; Yang, Jian-Yu; Liu, De-Jun; Yang, Xiao-Mei; Wang, Ya-Hui; Yang, Min-Wei; Fu, Xue-Liang; Zhang, Xiao-Xin; Li, Qing; Pang, Xiu-Feng; Huo, Yan-Miao; Li, Jiao; Zhang, Jun-Feng; Lee, Ho-Young; Lee, Su-Jae; Qin, Wen-Xin; Gu, Jian-Ren; Sun, Yong-Wei; Zhang, Zhi-Gang

    2017-07-01

    Desmoplasia and poor vascularity cause severe metabolic stress in pancreatic ductal adenocarcinomas (PDACs). Serotonin (5-HT) is a neuromodulator with neurotransmitter and neuroendocrine functions that contributes to tumorigenesis. We investigated the role of 5-HT signaling in the growth of pancreatic tumors. We measured the levels of proteins that regulate 5-HT synthesis, packaging, and degradation in pancreata from Kras G12D/+ /Trp53 R172H/+ /Pdx1-Cre (KPC) mice, which develop pancreatic tumors, as well as in PDAC cell lines and a tissue microarray containing 81 human PDAC samples. We also analyzed expression levels of proteins involved in 5-HT synthesis and degradation by immunohistochemical analysis of a tissue microarray containing 311 PDAC specimens, and associated expression levels with patient survival times. 5-HT level in 14 matched PDAC tumor and non-tumor tissues were analyzed by ELISA. PDAC cell lines were incubated with 5-HT and cell survival and apoptosis were measured. We analyzed expression of the 5-HT receptor HTR2B in PDAC cells and effects of receptor agonists and antagonists, as well as HTR2B knockdown with small hairpin RNAs. We determined the effects of 5-HT stimulation on gene expression profiles of BxPC-3 cells. Regulation of glycolysis by 5-HT signaling via HTR2B was assessed by immunofluorescence and immunoprecipitation analyses, as well as by determination of the extracellular acid ratio, glucose consumption, and lactate production. Primary PDACs, with or without exposure to SB204741 (a selective antagonist of HTR2B), were grown as xenograft tumors in mice, and SB204741 was administered to tumor-bearing KPC mice; tumor growth and metabolism were measured by imaging analyses. In immunohistochemical analysis of a tissue microarray of PDAC specimens, increased levels of TPH1 and decreased level of MAOA, which regulate 5-HT synthesis and degradation, correlated with stage and size of PDACs and shorter patient survival time. We found levels of 5-HT to be increased in human PDAC tissues compared with non-tumor pancreatic tissues, and PDAC cell lines compared with non-transformed pancreatic cells. Incubation of PDAC cell lines with 5-HT increased proliferation and prevented apoptosis. Agonists of HTR2B, but not other 5-HT receptors, promoted proliferation and prevented apoptosis of PDAC cells. Knockdown of HTR2B in PDAC cells, or incubation of cells with HTR2B inhibitors, reduced their growth as xenograft tumors in mice. We observed a correlation between 5-HT and glycolytic flux in PDAC cells; levels of metabolic enzymes involved in glycolysis, the phosphate pentose pathway, and hexosamine biosynthesis pathway increased significantly in PDAC cells following 5-HT stimulation. 5-HT stimulation led to formation of the HTR2B-LYN-p85 complex, which increased PI3K-Akt-mTOR signaling and the Warburg effect by increasing protein levels of MYC and HIF1A. Administration of SB204741 to KPC mice slowed growth and metabolism of established pancreatic tumors and prolonged survival of the mice. Human PDACs have increased levels of 5-HT, and PDAC cells increase expression of its receptor, HTR2B. These increases allow for tumor glycolysis under metabolic stress and promote growth of pancreatic tumors and PDAC xenograft tumors in mice. Copyright © 2017 AGA Institute. Published by Elsevier Inc. All rights reserved.

  8. Incorrect strain information for mouse cell lines: sequential influence of misidentification on sublines.

    PubMed

    Uchio-Yamada, Kozue; Kasai, Fumio; Ozawa, Midori; Kohara, Arihiro

    2017-03-01

    Misidentification or cross-contamination of cell lines can cause serious issues. Human cell lines have been authenticated by short tandem repeat profiling; however, mouse cell lines have not been adequately assessed. In this study, mouse cell lines registered with the JCRB cell bank were examined by simple sequence length polymorphism (SSLP) analysis to identify their strains. Based on comparisons with 7 major inbred strains, our results revealed their strains in 80 of 90 cell lines. However, 12 of the 80 cell lines (15%) were found to differ from registered information. Of them, 4 cell lines originated from the same mouse, which had been generated through mating between two different inbred strains. The genotype of the mouse sample had not been examined after the backcross, leading to strain misidentification in those cell lines. Although 8 other cell lines had been established as sublines of a BALB/c cell line, their SSLP profiles are similar to a Swiss cell line. This affects differences in genotypes between inbred and outbred strains. Because the use of inbred samples and interbreeding between strains are not involved in human materials, our results suggest that the cause and influence of misidentification in mouse cell lines are different from those in human.

  9. Two-Photon Processor and SeNeCA: a freely available software package to process data from two-photon calcium imaging at speeds down to several milliseconds per frame.

    PubMed

    Tomek, Jakub; Novak, Ondrej; Syka, Josef

    2013-07-01

    Two-Photon Processor (TPP) is a versatile, ready-to-use, and freely available software package in MATLAB to process data from in vivo two-photon calcium imaging. TPP includes routines to search for cell bodies in full-frame (Search for Neural Cells Accelerated; SeNeCA) and line-scan acquisition, routines for calcium signal calculations, filtering, spike-mining, and routines to construct parametric fields. Searching for somata in artificial in vivo data, our algorithm achieved better performance than human annotators. SeNeCA copes well with uneven background brightness and in-plane motion artifacts, the major problems in simple segmentation methods. In the fast mode, artificial in vivo images with a resolution of 256 × 256 pixels containing ≈ 100 neurons can be processed at a rate up to 175 frames per second (tested on Intel i7, 8 threads, magnetic hard disk drive). This speed of a segmentation algorithm could bring new possibilities into the field of in vivo optophysiology. With such a short latency (down to 5-6 ms on an ordinary personal computer) and using some contemporary optogenetic tools, it will allow experiments in which a control program can continuously evaluate the occurrence of a particular spatial pattern of activity (a possible correlate of memory or cognition) and subsequently inhibit/stimulate the entire area of the circuit or inhibit/stimulate a different part of the neuronal system. TPP will be freely available on our public web site. Similar all-in-one and freely available software has not yet been published.

  10. 46 CFR 160.037-4 - Approval and production tests.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... signal plus packaging in a sealed plastic waterproof bag, the 24-hour water immersion conditioning will... discontinued production line. (2) Inspections and tests by the manufacturer. The manufacturer's quality control...: (i) Conditioning of test specimens—water resistance. Immerse specimen horizontally with uppermost...

  11. 46 CFR 160.037-4 - Approval and production tests.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... signal plus packaging in a sealed plastic waterproof bag, the 24-hour water immersion conditioning will... discontinued production line. (2) Inspections and tests by the manufacturer. The manufacturer's quality control...: (i) Conditioning of test specimens—water resistance. Immerse specimen horizontally with uppermost...

  12. 46 CFR 160.037-4 - Approval and production tests.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... signal plus packaging in a sealed plastic waterproof bag, the 24-hour water immersion conditioning will... discontinued production line. (2) Inspections and tests by the manufacturer. The manufacturer's quality control...: (i) Conditioning of test specimens—water resistance. Immerse specimen horizontally with uppermost...

  13. Ultraviolet observations of cool stars. VI - L alpha and Mg II emission line profiles /and a search for flux variability/ in Arcturus

    NASA Technical Reports Server (NTRS)

    Mcclintock, W.; Moos, H. W.; Henry, R. C.; Linsky, J. L.; Barker, E. S.

    1978-01-01

    High-precision, high-resolution profiles of the L alpha and Mg II k chromospheric emission lines from Arcturus (alpha Boo) obtained with the Princeton Experimental Package aboard the Copernicus satellite are presented. Asymmetries seen in the profiles of these lines are probably intrinsic to the star, rather than the result of interstellar absorption. In contrast to previous observations of the Ca II K emission line, no evidence is found during a three-year period for variability in the profiles or in the total fluxes from these lines on time scales ranging from hours to months. Also presented is a flux profile of the O I 1302 line and flux upper limits for L beta, O VI 1032, Si III 1206, and O V 1218.

  14. Establishment and characterization of three immortal bovine muscular epithelial cell lines.

    PubMed

    Jin, Xun; Lee, Joong-Seob; Kwak, Sungwook; Lee, Soo-Yeon; Jung, Ji-Eun; Kim, Tae-Kyung; Xu, Chenxiong; Hong, Zhongshan; Li, Zhehu; Kim, Sun-Myung; Pian, Xumin; Lee, Dong-Hee; Yoon, Jong-Taek; You, Seungkwon; Choi, Yun-Jaie; Kim, Huunggee

    2006-02-28

    We have established three immortal bovine muscular epithelial (BME) cell lines, one spontaneously immortalized (BMES), the second SV40LT-mediated (BMEV) and the third hTERT-mediated (BMET). The morphology of the three immortal cell lines was similar to that of early passage primary BME cells. Each of the immortal cell lines made cytokeratin, a typical epithelial marker. BMET grew faster than the other immortal lines and the BME cells, in 10% FBS-DMEM medium, whereas neither the primary cells nor the three immortal cell lines grew in 0.5% FBS-DMEM. The primary BME cells and the immortal cell lines, with the exception of BMES, made increasing amounts of p53 protein when treated with doxorubicin, a DNA damaging agent. On the other hand, almost half of the cells in populations of the three immortal cell lines may lack p16(INK4a) regulatory function, compared to primary BME cells that were growth arrested by enforced expression of p16(INK4a). In soft-agar assays, the primary cells and immortal cell lines proved to be less transformed in phenotype than HeLa cells. The three immortal epithelial-type cell lines reported here are the first cell lines established from muscle tissue of bovine or other species.

  15. Investigation of the expression of RIF1 gene on head and neck, pancreatic and brain cancer and cancer stem cells.

    PubMed

    GursesCila, Hacer E; Acar, Muradiye; Barut, Furkan B; Gunduz, Mehmet; Grenman, Reidar; Gunduz, Esra

    2016-12-01

    Recent studies have shown that cancer stem cells are resistant to chemotherapy. The aim of this study was to compare RIF1 gene expression in head and neck, pancreatic cancer and glioma cell lines and the cancer stem cells isolated from these cell lines. UT-SCC-74 from Turku University and UT-SCC-74B primary tumor metastasis and neck cancer cell lines, YKG-1 glioma cancer cell line from RIKEN, pancreatic cancer cell lines and ASPC-1 cells from ATCC were grown in cell culture. To isolate cancer stem cells, ALDH-1 for UT-SCC-74 and UT-SCC-74B cell line, CD-133 for YKG-1 cell line and CD-24 for ASPC-1 cell line, were used as markers of cancer stem cells. RNA isolation was performed for both cancer lines and cancer stem cells. RNAs were converted to cDNA. RIF1 gene expression was performed by qRT-PCR analysis. RIF1 gene expression was compared with cancer cell lines and cancer stem cells isolated from these cell lines. The possible effect of RIF1 gene was evaluated. In the pancreatic cells, RIF1 gene expression in the stem cell-positive cell line was 256 time that seen in the stem cell-negative cell line. Considering the importance of RIF1 in NHEJ and of NHEJ in pancreatic cancer, RIF1 may be one of the genes that plays an important role in the diagnoses and therapeutic treatment of pancreatic cancer. The results of head and neck and brain cancers are inconclusive and further studies are required to elucidate the connection between RIF1 gene and these other types of cancers.

  16. Highly efficient in vitro and in vivo delivery of functional RNAs using new versatile MS2-chimeric retrovirus-like particles

    PubMed Central

    Prel, Anne; Caval, Vincent; Gayon, Régis; Ravassard, Philippe; Duthoit, Christine; Payen, Emmanuel; Maouche-Chretien, Leila; Creneguy, Alison; Nguyen, Tuan Huy; Martin, Nicolas; Piver, Eric; Sevrain, Raphaël; Lamouroux, Lucille; Leboulch, Philippe; Deschaseaux, Frédéric; Bouillé, Pascale; Sensébé, Luc; Pagès, Jean-Christophe

    2015-01-01

    RNA delivery is an attractive strategy to achieve transient gene expression in research projects and in cell- or gene-based therapies. Despite significant efforts investigating vector-directed RNA transfer, there is still a requirement for better efficiency of delivery to primary cells and in vivo. Retroviral platforms drive RNA delivery, yet retrovirus RNA-packaging constraints limit gene transfer to two genome-molecules per viral particle. To improve retroviral transfer, we designed a dimerization-independent MS2-driven RNA packaging system using MS2-Coat-retrovirus chimeras. The engineered chimeric particles promoted effective packaging of several types of RNAs and enabled efficient transfer of biologically active RNAs in various cell types, including human CD34+ and iPS cells. Systemic injection of high-titer particles led to gene expression in mouse liver and transferring Cre-recombinase mRNA in muscle permitted widespread editing at the ROSA26 locus. We could further show that the VLPs were able to activate an osteoblast differentiation pathway by delivering RUNX2- or DLX5-mRNA into primary human bone-marrow mesenchymal-stem cells. Thus, the novel chimeric MS2-lentiviral particles are a versatile tool for a wide range of applications including cellular-programming or genome-editing. PMID:26528487

  17. Mix-ups and mycoplasma: the enemies within.

    PubMed

    Drexler, Hans G; Uphoff, Cord C; Dirks, Willy G; MacLeod, Roderick A F

    2002-04-01

    Human leukemia-lymphoma (LL) cell lines represent important tools for experimental research. Among the various problems associated with cell lines, the two most common concern contaminations: (1) cross-contamination with unrelated cells and (2) contamination with microorganisms, in particular mycoplasma. The bad news is that about one-third of the cell lines are either cross-contaminated or mycoplasma-infected or both. The good news is that there are means to recognize and overcome these problems. In cases where, during attempts to establish new LL cell lines, primary LL cultures are cross-contaminated with continuous cell lines, intended new cell lines simply cannot be established ("early" cross-contamination). In cases of "late" cross-contamination of existing LL cell lines where the intrusive cells have a growth advantage, the original ("uncontaminated") cell lines may still be available elsewhere. DNA fingerprinting and cytogenetic analysis appear to be the most suitable approaches to detect cross-contaminations and to authenticate LL cell lines. A different but related aspect of "false" LL cell lines is the frequent misclassification of cell lines whereby the actual cell type of the cell line does not correspond to the purported model character of the cell line. Mycoplasma infection can have a multitude of effects on the eukaryotic cells which, due to the variety of infecting mycoplasma species and many other contributing parameters, cannot be predicted, rendering resulting data questionable at best. Practical procedures for the detection and elimination of mycoplasma contamination have been developed. Diagnostic and preventive strategies in order to hem the alarming increase in "false" and mycoplasma-positive LL cell lines are recommended.

  18. [Experimental study of interleukin-12 gene vaccines in the treatment of low-load malignant lymphoma (EL4)].

    PubMed

    Jiang, Q; Da, W; Ou, Y

    2001-11-01

    Two kinds of murine interleukin-12 (mIL-12) fusion gene vaccines were used to treat the murine low-load malignant T cell lymphoma EL4 as minimal residual disease (MRD) model. C57BL/6 synergistical mice were subcutaneously inoculated with 1 x 10(6) wild-type (wt) EL4 tumor cells as low-load lymphoma model treated with two mIL-12 gene vaccines. Package cell line PA317/12 producing mIL-12 retrovirus (RV) was used as in vivo vaccine and EL4 tumor cells transferred with mIL-12 gene as ex vivo vaccine. In both mIL-12 gene vaccine-treated groups, there was no tumor growth in 50% mice 60 days after inoculation. Nine of these no tumor growth mice were re-challenged with 5 x 10(5) wt EL4 cells, and 5 of them survived without tumors in another 60 days. All control mice died with tumors within one month after inoculation. Among those developed tumors in both vaccine-treated groups, the development of tumors was delayed, the survival period prolonged (P < 0.01), and the tumors size at death smaller (P < 0.05) as compared with the controls. In the long-survived vaccine-treated mice, no residual tumor cells were found by morphological examination. Both IL-12 gene vaccines can efficiently eliminate wt EL4 MRD in C57BL/6 mice.

  19. Evaluation of the cytotoxicity of elastomeric ligatures after sterilisation with 0.25% peracetic acid.

    PubMed

    Pithon, Matheus Melo; dos Santos, Rogerio Lacerda; Judice, Renata Lima Pasini; de Assuncao, Paulo Sergio; Restle, Luciana

    2013-11-01

    Sterilisation using peracetic acid (PAA) has been advocated for orthodontic elastic bands. However, cane-loaded elastomeric ligatures can also become contaminated during processing, packaging, and manipulation before placement in the oral cavity and are therefore susceptible, and possible causes, of cross-contamination. To test the hypothesis that 0.25% peracetic acid (PAA), following the sterilisation of elastomers, influences the cytotoxicity of elastomeric ligatures on L929 cell lines. Four hundred and eighty silver elastomeric ligatures were divided into 4 groups of 120 ligatures to produce, Group TP (latex natural, bulk pack, TP Orthodontics), Group M1 (Polyurethane, bulk pack, Morelli), Group M2 (Polyurethane, cane-loaded, Morelli) and Group U (Polyurethane, cane-loaded, Uniden). Of the 120 ligatures in each group, 100 were sterilised in 0.25% PAA at time intervals (N = 20) of 1 hour, 2 hours, 3 hours, 4 hours and 5 hours. The 20 remaining elastomeric ligatures in each group were not sterilised and served as controls. Cytotoxicity was assessed using L929 cell lines and a dye-uptake method. Analysis of variance (ANOVA), followed by the Tukey post hoc test (p < 0.05) determined statistical relevance. There was a significant difference between TP, Morelli and Uniden elastomerics (p < 0.05), but no difference between the two types of Morelli elastomerics at the 1 hour time interval. In addition, there was a significant difference between Group CC and the other groups assessed, except between Groups CC and TP at the 1 hour time interval. The non-sterilised elastomeric ligatures showed similar cell viability to that observed after 1 hour of standard sterilisation. PAA did not significantly influence the cytotoxicity of elastomeric ligatures after a sterilisation time of 1 hour and is therefore recommended for clinical use.

  20. Robust identification of transcriptional regulatory networks using a Gibbs sampler on outlier sum statistic.

    PubMed

    Gu, Jinghua; Xuan, Jianhua; Riggins, Rebecca B; Chen, Li; Wang, Yue; Clarke, Robert

    2012-08-01

    Identification of transcriptional regulatory networks (TRNs) is of significant importance in computational biology for cancer research, providing a critical building block to unravel disease pathways. However, existing methods for TRN identification suffer from the inclusion of excessive 'noise' in microarray data and false-positives in binding data, especially when applied to human tumor-derived cell line studies. More robust methods that can counteract the imperfection of data sources are therefore needed for reliable identification of TRNs in this context. In this article, we propose to establish a link between the quality of one target gene to represent its regulator and the uncertainty of its expression to represent other target genes. Specifically, an outlier sum statistic was used to measure the aggregated evidence for regulation events between target genes and their corresponding transcription factors. A Gibbs sampling method was then developed to estimate the marginal distribution of the outlier sum statistic, hence, to uncover underlying regulatory relationships. To evaluate the effectiveness of our proposed method, we compared its performance with that of an existing sampling-based method using both simulation data and yeast cell cycle data. The experimental results show that our method consistently outperforms the competing method in different settings of signal-to-noise ratio and network topology, indicating its robustness for biological applications. Finally, we applied our method to breast cancer cell line data and demonstrated its ability to extract biologically meaningful regulatory modules related to estrogen signaling and action in breast cancer. The Gibbs sampler MATLAB package is freely available at http://www.cbil.ece.vt.edu/software.htm. xuan@vt.edu Supplementary data are available at Bioinformatics online.

  1. Robust identification of transcriptional regulatory networks using a Gibbs sampler on outlier sum statistic

    PubMed Central

    Gu, Jinghua; Xuan, Jianhua; Riggins, Rebecca B.; Chen, Li; Wang, Yue; Clarke, Robert

    2012-01-01

    Motivation: Identification of transcriptional regulatory networks (TRNs) is of significant importance in computational biology for cancer research, providing a critical building block to unravel disease pathways. However, existing methods for TRN identification suffer from the inclusion of excessive ‘noise’ in microarray data and false-positives in binding data, especially when applied to human tumor-derived cell line studies. More robust methods that can counteract the imperfection of data sources are therefore needed for reliable identification of TRNs in this context. Results: In this article, we propose to establish a link between the quality of one target gene to represent its regulator and the uncertainty of its expression to represent other target genes. Specifically, an outlier sum statistic was used to measure the aggregated evidence for regulation events between target genes and their corresponding transcription factors. A Gibbs sampling method was then developed to estimate the marginal distribution of the outlier sum statistic, hence, to uncover underlying regulatory relationships. To evaluate the effectiveness of our proposed method, we compared its performance with that of an existing sampling-based method using both simulation data and yeast cell cycle data. The experimental results show that our method consistently outperforms the competing method in different settings of signal-to-noise ratio and network topology, indicating its robustness for biological applications. Finally, we applied our method to breast cancer cell line data and demonstrated its ability to extract biologically meaningful regulatory modules related to estrogen signaling and action in breast cancer. Availability and implementation: The Gibbs sampler MATLAB package is freely available at http://www.cbil.ece.vt.edu/software.htm. Contact: xuan@vt.edu Supplementary information: Supplementary data are available at Bioinformatics online. PMID:22595208

  2. Stability Studies of a Freeze-Dried Recombinant Human Epidermal Growth Factor Formulation for Wound Healing.

    PubMed

    Santana, Héctor; García, Gerardo; Vega, Maribel; Beldarraín, Alejandro; Páez, Rolando

    2015-01-01

    We report on the stability assessment of a recombinant human epidermal growth factor (rhEGF) freeze-dried formulation for wound healing by intra-lesional injections. The suitability of packaging material for the light protection of finished dried powder was evaluated after stressed exposure conditions. Degradation kinetics of powder for injection was investigated at concentrations of 25-250 μg/vial and temperatures of 45, 60, and 70 °C. The long-term stability was evaluated after storage at 25 ± 2 °C/60 ± 5% relative humidity (6 months) and 2-8 °C (24 months) in the dark and analyzed at several time points. The stability after reconstitution with various diluents was also assessed after 24 h storage at 2-8 °C. The rhEGF samples were analyzed for structural integrity by reversed-phase high-performance liquid chromatography (RP-HPLC), size-exclusion HPLC, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Biological activity was investigated by measuring the cell proliferation in a murine fibroblast cell line. Results show that freeze-dried rhEGF in primary packaging only was photosensitive, as degradation by RP-HPLC that was completely suppressed by the secondary carton package was revealed. An increase in freeze-dried rhEGF stability was observed with the increase in protein concentration from 25 to 250 μg/vial. The long-term stability study showed no significant rhEGF degradation or physical change within the freeze-dried formulations containing 25 or 250 μg/vial of rhEGF. No physical, chemical or biological changes were observed for rhEGF after reconstitution in water for injection or 0.9% sodium chloride during the storage conditions studied. The stability of a recombinant human epidermal growth factor (rhEGF) freeze-dried formulation for wound healing by intra-lesional injections was assessed. The suitability of packaging material for the light protection of finished dried powder was evaluated after stressed exposure conditions. Degradation kinetics of powder for injection was investigated at concentrations of 25-250 μg/vial and temperatures of 45, 60, and 70 °C. The accelerated, long-term, and reconstitution stabilities were examined according to ICH guidelines for their utility time. The stability of rhEGF samples was analyzed by different chemical, physical, and biological activity assays. Results show that freeze-dried rhEGF in primary packaging only was photosensitive, as degradation by reversed-phase high performance liquid chromatography that was completely suppressed by the secondary carton package was revealed. An increase in freeze-dried rhEGF stability was observed with the increase in protein concentration. No significant rhEGF degradation or physical changes were observed within the freeze-dried formulations after 6 months storage at 25 ± 2 °C/60 ± 5% relative humidity or 24 months storage at 2-8 °C. No physical, chemical, or biological changes were observed for rhEGF after reconstitution in water for injection or 0.9% sodium chloride after 24 h storage at 2-8 °C. © PDA, Inc. 2015.

  3. Feeder-cell-independent culture of the pig-embryonic-stem-cell-derived exocrine pancreatic cell line, PICM-31

    USDA-ARS?s Scientific Manuscript database

    The adaptation to feeder-independent growth of a pig embryonic stem cell-derived pancreatic cell line is described. The parental PICM-31 cell line, previously characterized as an exocrine pancreas cell line, was colony-cloned two times in succession resulting in the subclonal cell line, PICM-31A1. P...

  4. DIELECTROPHORESIS-BASED MICROFLUIDIC SEPARATION AND DETECTION SYSTEMS

    PubMed Central

    Yang, Jun; Vykoukal, Jody; Noshari, Jamileh; Becker, Frederick; Gascoyne, Peter; Krulevitch, Peter; Fuller, Chris; Ackler, Harold; Hamilton, Julie; Boser, Bernhard; Eldredge, Adam; Hitchens, Duncan; Andrews, Craig

    2009-01-01

    Diagnosis and treatment of human diseases frequently requires isolation and detection of certain cell types from a complex mixture. Compared with traditional separation and detection techniques, microfluidic approaches promise to yield easy-to-use diagnostic instruments tolerant of a wide range of operating environments and capable of accomplishing automated analyses. These approaches will enable diagnostic advances to be disseminated from sophisticated clinical laboratories to the point-of-care. Applications will include the separation and differential analysis of blood cell subpopulations for host-based detection of blood cell changes caused by disease, infection, or exposure to toxins, and the separation and analysis of surface-sensitized, custom dielectric beads for chemical, biological, and biomolecular targets. Here we report a new particle separation and analysis microsystem that uses dielectrophoretic field-flow fractionation (DEP-FFF). The system consists of a microfluidic chip with integrated sample injector, a DEP-FFF separator, and an AC impedance sensor. We show the design of a miniaturized impedance sensor integrated circuit (IC) with improved sensitivity, a new packaging approach for micro-flumes that features a slide-together compression package and novel microfluidic interconnects, and the design, control, integration and packaging of a fieldable prototype. Illustrative applications will be shown, including the separation of different sized beads and different cell types, blood cell differential analysis, and impedance sensing results for beads, spores and cells. PMID:22025905

  5. First-Time Analysis of Completely Restored DTREM Instrument Data from Apollo 14 and 15

    NASA Technical Reports Server (NTRS)

    McBride, Marie J.; Williams, David R.; Hills, H. Kent; Turner, Niescja

    2013-01-01

    The Dust, Thermal and Radiation Engineering Measurement (DTREM) packages (figure 1) mounted on the central stations of the Apollo 11, 12, 14, and 15 ALSEPs (Apollo Lunar Surface Experiments Packages) measured the outputs of exposed solar cells and thermistors over time. The goal of the experiment, also commonly known as the dust detector, was to study the long-term effects of dust, radiation, and temperature at the lunar surface on solar cells. The monitors returned data for up to almost 8 years from the lunar surface.

  6. Submergible barge retrievable storage and permanent disposal system for radioactive waste

    DOEpatents

    Goldsberry, Fred L.; Cawley, William E.

    1981-01-01

    A submergible barge and process for submerging and storing radioactive waste material along a seabed. A submergible barge receives individual packages of radwaste within segregated cells. The cells are formed integrally within the barge, preferably surrounded by reinforced concrete. The cells are individually sealed by a concrete decking and by concrete hatch covers. Seawater may be vented into the cells for cooling, through an integral vent arrangement. The vent ducts may be attached to pumps when the barge is bouyant. The ducts are also arranged to promote passive ventilation of the cells when the barge is submerged. Packages of the radwaste are loaded into individual cells within the barge. The cells are then sealed and the barge is towed to the designated disposal-storage site. There, the individual cells are flooded and the barge will begin descent controlled by a powered submarine control device to the seabed storage site. The submerged barge will rest on the seabed permanently or until recovered by a submarine control device.

  7. Molecularly Targeted Dose-Enhancement Radiotherapy Using Gold and Luminescent Nanoparticles in an Orthotopic Human Prostate Cancer Rat Model

    DTIC Science & Technology

    2013-10-01

    cell lines, such as cervix cancer cell line (HeLa) and breast cancer cell line (MDA-MB-231), were also employed. The experiments with other cell lines...breast cancer cell line (MDA-MB- 231), and cervix cancer cell line (HeLa). Different from our hypothesis, prostate cancer cell lines did not present...Radiotherapy Using Gold and Luminescent Nanoparticles in an Orthotopic Human Prostate Cancer Rat Model PRINCIPAL INVESTIGATOR: Kwang Song

  8. Computational methods for evaluation of cell-based data assessment--Bioconductor.

    PubMed

    Le Meur, Nolwenn

    2013-02-01

    Recent advances in miniaturization and automation of technologies have enabled cell-based assay high-throughput screening, bringing along new challenges in data analysis. Automation, standardization, reproducibility have become requirements for qualitative research. The Bioconductor community has worked in that direction proposing several R packages to handle high-throughput data including flow cytometry (FCM) experiment. Altogether, these packages cover the main steps of a FCM analysis workflow, that is, data management, quality assessment, normalization, outlier detection, automated gating, cluster labeling, and feature extraction. Additionally, the open-source philosophy of R and Bioconductor, which offers room for new development, continuously drives research and improvement of theses analysis methods, especially in the field of clustering and data mining. This review presents the principal FCM packages currently available in R and Bioconductor, their advantages and their limits. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. [The factors involved in invasive ability of endometrial carcinoma cells].

    PubMed

    Mori, Y; Mizuuchi, H; Sato, K; Okamura, N; Kudo, R

    1994-06-01

    The in vitro invasive ability, the expression of cell adhesion molecule E-cadherin, activity of matrix metalloproteinase (MMP) and K-ras point mutation were investigated in eight human endometrial carcinoma cell lines. 1) In vitro invasive abilities of endometrial carcinoma cell lines depend on the degree of cell differentiation and the origin of cell lines. A poorly-differentiated carcinoma cell line (NUE-1) and a cell line derived from metastatic lymph node (SNG-M) were more invasive than moderately-(HEC-1A, HEC-1BE) and well-differentiated (HEC-6, Ishikawa) cell lines. 2) Immunohistochemically, less or non-invasive cell lines expressed E-cadherin strongly, whereas a highly invasive cell line (NUE-1) expressed E-cadherin weakly. 3) When cultured on Matrigel-coated dishes, the tumor cells derived from moderately- and well-differentiated carcinoma aggregated with each other and did not invade Matrigel in the invasion assay. The aggregated cells expressed E-cadherin more strongly when cultured on Matrigel. 4) 72-kD gelatinase (MMP-2) was secreted in serum-free conditioned medium of all cell lines. In an invasive cell line (NUE-1,SNG-M), the activity of MMP-2 was stronger than in other cell lines. And the activity of 92-kDa gelatinase (MMP-9) was detected in most invasive cell line (NUE-1). 5) Point mutation of K-ras codon 12 was detected in four of eight (50%) cell lines by the PCR-RFLP method. The changes in the DNA sequence were identified, but K-ras point mutation was not correlated with in vitro invasiveness of the tumor cells.

  10. In vitro Development of Chemotherapy and Targeted Therapy Drug-Resistant Cancer Cell Lines: A Practical Guide with Case Studies

    PubMed Central

    McDermott, Martina; Eustace, Alex J.; Busschots, Steven; Breen, Laura; Crown, John; Clynes, Martin; O’Donovan, Norma; Stordal, Britta

    2014-01-01

    The development of a drug-resistant cell line can take from 3 to 18 months. However, little is published on the methodology of this development process. This article will discuss key decisions to be made prior to starting resistant cell line development; the choice of parent cell line, dose of selecting agent, treatment interval, and optimizing the dose of drug for the parent cell line. Clinically relevant drug-resistant cell lines are developed by mimicking the conditions cancer patients experience during chemotherapy and cell lines display between two- and eight-fold resistance compared to their parental cell line. Doses of drug administered are low, and a pulsed treatment strategy is often used where the cells recover in drug-free media. High-level laboratory models are developed with the aim of understanding potential mechanisms of resistance to chemotherapy agents. Doses of drug are higher and escalated over time. It is common to have difficulty developing stable clinically relevant drug-resistant cell lines. A comparative selection strategy of multiple cell lines or multiple chemotherapeutic agents mitigates this risk and gives insight into which agents or type of cell line develops resistance easily. Successful selection strategies from our research are presented. Pulsed-selection produced platinum or taxane-resistant large cell lung cancer (H1299 and H460) and temozolomide-resistant melanoma (Malme-3M and HT144) cell lines. Continuous selection produced a lapatinib-resistant breast cancer cell line (HCC1954). Techniques for maintaining drug-resistant cell lines are outlined including; maintaining cells with chemotherapy, pulse treating with chemotherapy, or returning to master drug-resistant stocks. The heterogeneity of drug-resistant models produced from the same parent cell line with the same chemotherapy agent is explored with reference to P-glycoprotein. Heterogeneity in drug-resistant cell lines reflects the heterogeneity that can occur in clinical drug resistance. PMID:24639951

  11. Bonded polyimide fuel cell package

    DOEpatents

    Morse, Jeffrey D.; Jankowski, Alan; Graff, Robert T.; Bettencourt, Kerry

    2010-06-08

    Described herein are processes for fabricating microfluidic fuel cell systems with embedded components in which micron-scale features are formed by bonding layers of DuPont Kapton.TM. polyimide laminate. A microfluidic fuel cell system fabricated using this process is also described.

  12. Immortal, telomerase-negative cell lines derived from a Li-Fraumeni syndrome patient exhibit telomere length variability and chromosomal and minisatellite instabilities.

    PubMed

    Tsutsui, Takeki; Kumakura, Shin-Ichi; Tamura, Yukiko; Tsutsui, Takeo W; Sekiguchi, Mizuki; Higuchi, Tokihiro; Barrett, J Carl

    2003-05-01

    Five immortal cell lines derived from a Li-Fraumeni syndrome patient (MDAH 087) with a germline mutant p53 allele were characterized with respect to telomere length and genomic instability. The remaining wild-type p53 allele is lost in the cell lines. Telomerase activity was undetectable in all immortal cell lines. Five subclones of each cell line and five re-subclones of each of the subclones also showed undetectable telomerase activity. All five immortal cell lines exhibited variability in the mean length of terminal restriction fragments (TRFs). Subclones of each cell line, and re-subclones of the subclones also showed TRF variability, indicating that the variability is owing to clonal heterogeneity. Chromosome aberrations were observed at high frequencies in these cell lines including the subclones and re-subclones, and the principal types of aberrations were breaks, double minute chromosomes and dicentric chromosomes. In addition, minisatellite instability detected by DNA fingerprints was observed in the immortal cell lines. However, all of the cell lines were negative for microsatellite instability. As minisatellite sequences are considered recombinogenic in mammalian cells, these results suggest that recombination rates can be increased in these cell lines. Tumor-derived human cell lines, HT1080 cells and HeLa cells that also lack p53 function, exhibited little genomic instability involving chromosomal and minisatellite instabilities, indicating that chromosomal and minisatellite instabilities observed in the immortal cell lines lacking telomerase activity could not result from loss of p53 function.

  13. SIRB, sans iron oxide rhodamine B, a novel cross-linked dextran nanoparticle, labels human neuroprogenitor and SH-SY5Y neuroblastoma cells and serves as a USPIO cell labeling control.

    PubMed

    Shen, Wei-Bin; Vaccaro, Dennis E; Fishman, Paul S; Groman, Ernest V; Yarowsky, Paul

    2016-05-01

    This is the first report of the synthesis of a new nanoparticle, sans iron oxide rhodamine B (SIRB), an example of a new class of nanoparticles. SIRB is designed to provide all of the cell labeling properties of the ultrasmall superparamagnetic iron oxide (USPIO) nanoparticle Molday ION Rhodamine B (MIRB) without containing the iron oxide core. MIRB was developed to label cells and allow them to be tracked by MRI or to be manipulated by magnetic gradients. SIRB possesses a similar size, charge and cross-linked dextran coating as MIRB. Of great interest is understanding the biological and physiological changes in cells after they are labeled with a USPIO. Whether these effects are due to the iron oxide buried within the nanoparticle or to the surface coating surrounding the iron oxide core has not been considered previously. MIRB and SIRB represent an ideal pairing of nanoparticles to identify nanoparticle anatomy responsible for post-labeling cytotoxicity. Here we report the effects of SIRB labeling on the SH-SY5Y neuroblastoma cell line and primary human neuroprogenitor cells (hNPCs). These effects are contrasted with the effects of labeling SH-SY5Y cells and hNPCs with MIRB. We find that SIRB labeling, like MIRB labeling, (i) occurs without the use of transfection reagents, (ii) is packaged within lysosomes distributed within cell cytoplasm, (iii) is retained within cells with no loss of label after cell storage, and (iv) does not alter cellular viability or proliferation, and (v) SIRB labeled hNPCs differentiate normally into neurons or astrocytes. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  14. Miniature spectrally selective dosimeter

    NASA Technical Reports Server (NTRS)

    Adams, R. R.; Macconochie, I. O.; Poole, B. D., Jr. (Inventor)

    1980-01-01

    A miniature spectrally selective dosimeter capable of measuring selected bandwidths of radiation exposure on small mobile areas is described. This is achieved by the combination of photovoltaic detectors, electrochemical integrators (E-cells) and filters in a small compact case which can be easily attached in close proximity to and substantially parallel to the surface being measured. In one embodiment two photovoltaic detectors, two E-cells, and three filters are packaged in a small case with attaching means consisting of a safety pin. In another embodiment, two detectors, one E-cell, three filters are packaged in a small case with attaching means consisting of a clip to clip over a side piece of an eye glass frame.

  15. Community-based Ontology Development, Annotation and Discussion with MediaWiki extension Ontokiwi and Ontokiwi-based Ontobedia

    PubMed Central

    Ong, Edison; He, Yongqun

    2016-01-01

    Hundreds of biological and biomedical ontologies have been developed to support data standardization, integration and analysis. Although ontologies are typically developed for community usage, community efforts in ontology development are limited. To support ontology visualization, distribution, and community-based annotation and development, we have developed Ontokiwi, an ontology extension to the MediaWiki software. Ontokiwi displays hierarchical classes and ontological axioms. Ontology classes and axioms can be edited and added using Ontokiwi form or MediaWiki source editor. Ontokiwi also inherits MediaWiki features such as Wikitext editing and version control. Based on the Ontokiwi/MediaWiki software package, we have developed Ontobedia, which targets to support community-based development and annotations of biological and biomedical ontologies. As demonstrations, we have loaded the Ontology of Adverse Events (OAE) and the Cell Line Ontology (CLO) into Ontobedia. Our studies showed that Ontobedia was able to achieve expected Ontokiwi features. PMID:27570653

  16. Ephus: Multipurpose Data Acquisition Software for Neuroscience Experiments

    PubMed Central

    Suter, Benjamin A.; O'Connor, Timothy; Iyer, Vijay; Petreanu, Leopoldo T.; Hooks, Bryan M.; Kiritani, Taro; Svoboda, Karel; Shepherd, Gordon M. G.

    2010-01-01

    Physiological measurements in neuroscience experiments often involve complex stimulus paradigms and multiple data channels. Ephus (http://www.ephus.org) is an open-source software package designed for general-purpose data acquisition and instrument control. Ephus operates as a collection of modular programs, including an ephys program for standard whole-cell recording with single or multiple electrodes in typical electrophysiological experiments, and a mapper program for synaptic circuit mapping experiments involving laser scanning photostimulation based on glutamate uncaging or channelrhodopsin-2 excitation. Custom user functions allow user-extensibility at multiple levels, including on-line analysis and closed-loop experiments, where experimental parameters can be changed based on recently acquired data, such as during in vivo behavioral experiments. Ephus is compatible with a variety of data acquisition and imaging hardware. This paper describes the main features and modules of Ephus and their use in representative experimental applications. PMID:21960959

  17. Fungal-derived semiochemical 1-octen-3-ol disrupts dopamine packaging and causes neurodegeneration

    PubMed Central

    Inamdar, Arati A.; Hossain, Muhammad M.; Bernstein, Alison I.; Miller, Gary W.; Richardson, Jason R.; Bennett, Joan Wennstrom

    2013-01-01

    Parkinson disease (PD) is the most common movement disorder and, although the exact causes are unknown, recent epidemiological and experimental studies indicate that several environmental agents may be significant risk factors. To date, these suspected environmental risk factors have been man-made chemicals. In this report, we demonstrate via genetic, biochemical, and immunological studies that the common volatile fungal semiochemical 1-octen-3-ol reduces dopamine levels and causes dopamine neuron degeneration in Drosophila melanogaster. Overexpression of the vesicular monoamine transporter (VMAT) rescued the dopamine toxicity and neurodegeneration, whereas mutations decreasing VMAT and tyrosine hydroxylase exacerbated toxicity. Furthermore, 1-octen-3-ol also inhibited uptake of dopamine in human cell lines expressing the human plasma membrane dopamine transporter (DAT) and human VMAT ortholog, VMAT2. These data demonstrate that 1-octen-3-ol exerts toxicity via disruption of dopamine homeostasis and may represent a naturally occurring environmental agent involved in parkinsonism. PMID:24218591

  18. Development of apoptosis-resistant dihydrofolate reductase-deficient Chinese hamster ovary cell line.

    PubMed

    Lee, Suk Kyoo; Lee, Gyun Min

    2003-06-30

    Apoptosis-resistant dihydrofolate reductase-deficient CHO cell line (dhfr(-) CHO-bcl2) was developed by introduction of the bcl-2 gene into the dhfr(-) CHO cell line (DUKX-B11, ATCC CRL-9096) and subsequent selection of clones stably overexpressing Bcl-2 in the absence of selection pressure. When the dhfr(-) CHO-bcl2 cell line was used as a host cell line for development of a recombinant CHO (rCHO) cell line expressing a humanized antibody, it displayed stable expression of the bcl-2 gene during rCHO cell line development and no detrimental effect of Bcl-2 overexpression on specific antibody productivity. Taken together, the results obtained demonstrate that the use of an apoptosis-resistant dhfr(-) CHO cell line as the host cell line saves the effort of establishing an apoptosis-resistant rCHO cell line and expedites the development process of apoptosis-resistant rCHO cells producing therapeutic proteins. Copyright 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 872-876, 2003.

  19. Occasional Papers in Open and Distance Learning, Number 18.

    ERIC Educational Resources Information Center

    Donnan, Peter, Ed.

    Six papers examine innovations and trends in distance learning, frequently drawing upon empirical research or informal observations on distance learning students at Charles Sturt University (Australia). "On-Line Study Packages for Distance Education: Some Considerations of Conceptual Parameters" (Dirk M. R. Spennemann) discusses issues…

  20. QTL mapping of potato chip color and tuber traits within an autotetraploid family

    USDA-ARS?s Scientific Manuscript database

    Cultivated potato (Solanum tuberosum L.) is a highly heterozygous autotetraploid crop species, and this presents challenges for traditional line development and molecular breeding. Recent availability of a single nucleotide polymorphism (SNP) array with 8303 features and software packages for linkag...

  1. 7 CFR 2902.27 - Films.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 15 2010-01-01 2010-01-01 false Films. 2902.27 Section 2902.27 Agriculture... Films. (a) Definition. (1) Products that are used in packaging, wrappings, linings, and other similar applications. (2) Films for which preferred procurement applies are: (i) Semi-durable films. Films that are...

  2. 49 CFR 178.507 - Standards for plywood drums.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) SPECIFICATIONS FOR PACKAGINGS Non... effectiveness of the drum for the purpose intended. A material other than plywood, of at least equivalent..., lids must be lined with kraft paper or some other equivalent material which must be securely fastened...

  3. Virtual rough samples to test 3D nanometer-scale scanning electron microscopy stereo photogrammetry.

    PubMed

    Villarrubia, J S; Tondare, V N; Vladár, A E

    2016-01-01

    The combination of scanning electron microscopy for high spatial resolution, images from multiple angles to provide 3D information, and commercially available stereo photogrammetry software for 3D reconstruction offers promise for nanometer-scale dimensional metrology in 3D. A method is described to test 3D photogrammetry software by the use of virtual samples-mathematical samples from which simulated images are made for use as inputs to the software under test. The virtual sample is constructed by wrapping a rough skin with any desired power spectral density around a smooth near-trapezoidal line with rounded top corners. Reconstruction is performed with images simulated from different angular viewpoints. The software's reconstructed 3D model is then compared to the known geometry of the virtual sample. Three commercial photogrammetry software packages were tested. Two of them produced results for line height and width that were within close to 1 nm of the correct values. All of the packages exhibited some difficulty in reconstructing details of the surface roughness.

  4. Flagstaff, Arizona seen in Earth Resources Experiments package

    NASA Image and Video Library

    1974-02-01

    SL4-93-067 (16 Nov. 1973-8 Feb. 1974) --- A spectacular winter view of the Flagstaff, Arizona area is seen in this Skylab 4 Earth Resources Experiments package S190-B (five-inch earth terrain camera) infrared photograph taken from the Skylab space station in Earth orbit. Included in the scene are the San Francisco Mountains, Oak Creek Canyon, Painted Desert and Meteor Crater. The infrared picture depicts in red living vegetation, in white the snow, and in bright blue the water. Major features identified in this photograph are Humphrey's peak, top center, Flagstaff at foot of the peak, Sunset Crater volcanic field with numerous vents and craters right of Flagstaff and Meteor Crater (right center). Within the mountainous areas several clear areas generally rectangular are visible and represent the areas where lumbering has removed the forest. The thin white line extending from left corner to Sunset Crater fields is the power transmission line cleared area. Roads are subdued and are not easily visible. Photo credit: NASA

  5. ISAP: ISO Spectral Analysis Package

    NASA Astrophysics Data System (ADS)

    Ali, Babar; Bauer, Otto; Brauher, Jim; Buckley, Mark; Harwood, Andrew; Hur, Min; Khan, Iffat; Li, Jing; Lord, Steve; Lutz, Dieter; Mazzarella, Joe; Molinari, Sergio; Morris, Pat; Narron, Bob; Seidenschwang, Karla; Sidher, Sunil; Sturm, Eckhard; Swinyard, Bruce; Unger, Sarah; Verstraete, Laurent; Vivares, Florence; Wieprecht, Ecki

    2014-03-01

    ISAP, written in IDL, simplifies the process of visualizing, subsetting, shifting, rebinning, masking, combining scans with weighted means or medians, filtering, and smoothing Auto Analysis Results (AARs) from post-pipeline processing of the Infrared Space Observatory's (ISO) Short Wavelength Spectrometer (SWS) and Long Wavelength Spectrometer (LWS) data. It can also be applied to PHOT-S and CAM-CVF data, and data from practically any spectrometer. The result of a typical ISAP session is expected to be a "simple spectrum" (single-valued spectrum which may be resampled to a uniform wavelength separation if desired) that can be further analyzed and measured either with other ISAP functions, native IDL functions, or exported to other analysis package (e.g., IRAF, MIDAS) if desired. ISAP provides many tools for further analysis, line-fitting, and continuum measurements, such as routines for unit conversions, conversions from wavelength space to frequency space, line and continuum fitting, flux measurement, synthetic photometry and models such as a zodiacal light model to predict and subtract the dominant foreground at some wavelengths.

  6. Retrovirus-specific differences in matrix and nucleocapsid protein-nucleic acid interactions: implications for genomic RNA packaging.

    PubMed

    Sun, Meng; Grigsby, Iwen F; Gorelick, Robert J; Mansky, Louis M; Musier-Forsyth, Karin

    2014-01-01

    Retroviral RNA encapsidation involves a recognition event between genomic RNA (gRNA) and one or more domains in Gag. In HIV-1, the nucleocapsid (NC) domain is involved in gRNA packaging and displays robust nucleic acid (NA) binding and chaperone functions. In comparison, NC of human T-cell leukemia virus type 1 (HTLV-1), a deltaretrovirus, displays weaker NA binding and chaperone activity. Mutation of conserved charged residues in the deltaretrovirus bovine leukemia virus (BLV) matrix (MA) and NC domains affects virus replication and gRNA packaging efficiency. Based on these observations, we hypothesized that the MA domain may generally contribute to NA binding and genome encapsidation in deltaretroviruses. Here, we examined the interaction between HTLV-2 and HIV-1 MA proteins and various NAs in vitro. HTLV-2 MA displays higher NA binding affinity and better chaperone activity than HIV-1 MA. HTLV-2 MA also binds NAs with higher affinity than HTLV-2 NC and displays more robust chaperone function. Mutation of two basic residues in HTLV-2 MA α-helix II, previously implicated in BLV gRNA packaging, reduces NA binding affinity. HTLV-2 MA binds with high affinity and specificity to RNA derived from the putative packaging signal of HTLV-2 relative to nonspecific NA. Furthermore, an HIV-1 MA triple mutant designed to mimic the basic character of HTLV-2 MA α-helix II dramatically improves binding affinity and chaperone activity of HIV-1 MA in vitro and restores RNA packaging to a ΔNC HIV-1 variant in cell-based assays. Taken together, these results are consistent with a role for deltaretrovirus MA proteins in viral RNA packaging.

  7. Cigarette brand variant portfolio strategy and the use of colour in a darkening market.

    PubMed

    Greenland, Steven J

    2015-03-01

    To evaluate cigarette branding strategies used to segment a market with some of the toughest tobacco controls. To document brand variant and packaging portfolios and assess the role played by colour before plain packaging, as well as consider the threat that recently implemented legislation poses for tobacco manufacturers. Brand variant and packaging details were extracted from manufacturer ingredient reports, as well as a retail audit of Australian supermarkets. Details were also collected for other product categories to provide perspective on cigarette portfolios. Secondary and primary data sources were analysed to evaluate variant and packaging portfolio strategy. In Australia, 12 leading cigarette brands supported 120 brand variants. Of these 61 had names with a specific colour and a further 26 had names with colour connotation. There were 338 corresponding packaging configurations, with most variants available in the primary cigarette distribution channel in four pack size options. Tobacco companies microsegment Australian consumers with highly differentiated product offerings and a family branding strategy that helps ameliorate the effects of marketing restrictions. To date, tobacco controls have had little negative impact upon variant and packaging portfolios, which have continued to expand. Colour has become a key visual signifier differentiating one variant from the next, and colour names are used to extend brand lines. However, the role of colour, as a heuristic to simplify consumer decision-making processes, becomes largely redundant with plain packaging. Plain packaging's impact upon manufacturers' branding strategies is therefore likely to be significant. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  8. BoolFilter: an R package for estimation and identification of partially-observed Boolean dynamical systems.

    PubMed

    Mcclenny, Levi D; Imani, Mahdi; Braga-Neto, Ulisses M

    2017-11-25

    Gene regulatory networks govern the function of key cellular processes, such as control of the cell cycle, response to stress, DNA repair mechanisms, and more. Boolean networks have been used successfully in modeling gene regulatory networks. In the Boolean network model, the transcriptional state of each gene is represented by 0 (inactive) or 1 (active), and the relationship among genes is represented by logical gates updated at discrete time points. However, the Boolean gene states are never observed directly, but only indirectly and incompletely through noisy measurements based on expression technologies such as cDNA microarrays, RNA-Seq, and cell imaging-based assays. The Partially-Observed Boolean Dynamical System (POBDS) signal model is distinct from other deterministic and stochastic Boolean network models in removing the requirement of a directly observable Boolean state vector and allowing uncertainty in the measurement process, addressing the scenario encountered in practice in transcriptomic analysis. BoolFilter is an R package that implements the POBDS model and associated algorithms for state and parameter estimation. It allows the user to estimate the Boolean states, network topology, and measurement parameters from time series of transcriptomic data using exact and approximated (particle) filters, as well as simulate the transcriptomic data for a given Boolean network model. Some of its infrastructure, such as the network interface, is the same as in the previously published R package for Boolean Networks BoolNet, which enhances compatibility and user accessibility to the new package. We introduce the R package BoolFilter for Partially-Observed Boolean Dynamical Systems (POBDS). The BoolFilter package provides a useful toolbox for the bioinformatics community, with state-of-the-art algorithms for simulation of time series transcriptomic data as well as the inverse process of system identification from data obtained with various expression technologies such as cDNA microarrays, RNA-Seq, and cell imaging-based assays.

  9. Labeled line drawing of launch and entry suit identifies various components

    NASA Technical Reports Server (NTRS)

    1988-01-01

    Line drawings illustrate the front and back of the space shuttle launch and entry suit (LES) and labels identify various components. LES was designed for STS-26, the return to flight mission, and subsequent missions. Included in the crew escape system (CES) package are launch and entry helmet (LEH) with communications carrier (COMM CAP), parachute pack and harness, life preserver unit (LPU), life raft unit (LRU), LES gloves, suit oxygen manifold and valves, boots, and survival gear. Details of larger components are also identified.

  10. Applications of Multiconductor Transmission Line Theory to the Prediction of Cable Coupling. Volume 7. Digital Computer Programs for the Analysis of Multiconductor Transmission Lines

    DTIC Science & Technology

    1977-07-01

    on an IBM 370/165 computer at The University of Kentucky using the Fortran IV, G level compiler and should be easily implemented on other computers...order as the columns of T. 3.5.3 Subroutines NROOT and EIGEN Subroutines NROOT and EIGEN are a set of subroutines from the IBM Scientific Subroutine...November 1975). [10] System/360 Scientific Subroutine Package, Version III, Fifth Edition (August 1970), IBM Corporation, Technical Publications

  11. Platforms for Persistent communications Surveillance and Reconnaissance - II

    DTIC Science & Technology

    2009-11-01

    Lifter Airship with a comprehensive suite of sensors as indicated by the green line at the top of the chart with the red arrow. It had a value more than...package, but it was not as capable as the robust sensor suite on the red arrow option. Platforms for Persistent CSR II - 30...sensors remained the top-ranked option as indicated by the green line at the top of the chart with the red arrow. However, its value dropped from 0.253 on

  12. UV spectroscopy including ISM line absorption: of the exciting star of Abell 35

    NASA Astrophysics Data System (ADS)

    Ziegler, M.; Rauch, T.; Werner, K.; Kruk, J. W.

    Reliable spectral analysis that is based on high-resolution UV observations requires an adequate, simultaneous modeling of the interstellar line absorption and reddening. In the case of the central star of the planetary nebula Abell 35, BD-22 3467, we demonstrate our current standard spectral-analysis method that is based on the Tübingen NLTE Model-Atmosphere Package (TMAP). We present an on- going spectral analysis of FUSE and HST/STIS observations of BD-22 3467.

  13. Labeled line drawing of launch and entry suit identifies various components

    NASA Image and Video Library

    1988-09-22

    Line drawings illustrate the front and back of the space shuttle launch and entry suit (LES) and labels identify various components. LES was designed for STS-26, the return to flight mission, and subsequent missions. Included in the crew escape system (CES) package are launch and entry helmet (LEH) with communications carrier (COMM CAP), parachute pack and harness, life preserver unit (LPU), life raft unit (LRU), LES gloves, suit oxygen manifold and valves, boots, and survival gear. Details of larger components are also identified.

  14. Leukemia-lymphoma cell lines as model systems for hematopoietic research.

    PubMed

    Drexler, Hans G; MacLeod, Roderick A F

    2003-01-01

    Continuous human leukemia-lymphoma (LL) cell lines comprise a rich self-renewing resource of accessible and manipulable living cells which has illuminated the pathophysiology of hematopoietic tumors as well as basic cell biology. The major key advantages of continuous cell lines are the unlimited supply and worldwide availability of identical cell material and their cryopreservation. LL cell lines are characterized generally by monoclonal origin and differentiation arrest, sustained proliferation in vitro with preservation of most cellular features, and specific genetic alterations. The most practical classification of LL cell lines assigns them to one of the physiologically occurring cell lineages, based on their immunophenotype, genotype and functional features. Truly malignant cell lines may be distinguished from Epstein-Barr virus (EBV)-immortalized normal cells, using various operational and conceptual parameters. The characterization and publication of new LL cell lines provides important and informative core data which, by opening new avenues for investigation, have become ubiquitous powerful research tools that are available to every investigator by reference cell repositories. There is a need in the scientific community for clean and authenticated LL cell lines to which every scientist has access as offered by these institutionalized public cell line banks. A list of the most useful, robust and freely available reference cell lines is proposed in this review. Clearly, studies of LL cell lines have provided seminal insights into the biology of hematopoietic neoplasia.

  15. RIP-ET: A riparian evapotranspiration package for MODFLOW-2005

    USGS Publications Warehouse

    Maddock, Thomas; Baird, Kathryn J.; Hanson, R.T.; Schmid, Wolfgang; Ajami, Hoori

    2012-01-01

    A new evapotranspiration package for the U.S. Geological Survey's groundwater-flow model, MODFLOW, is documented. The Riparian Evapotranspiration Package (RIP-ET) provides flexibility in simulating riparian and wetland transpiration not provided by the Evapotranspiration (EVT) or Segmented Function Evapotranspiration (ETS1) Packages for MODFLOW 2005. This report describes how the RIP-ET package was conceptualized and provides input instructions, listings and explanations of the source code, and an example. Traditional approaches to modeling evapotranspiration (ET) processes assume a piecewise linear relationship between ET flux and hydraulic head. The RIP-ET replaces this traditional relationship with a segmented, nonlinear dimensionless curve that reflects the eco-physiology of riparian and wetland ecosystems. Evapotranspiration losses from these ecosystems are dependent not only on hydraulic head, but on the plant types present. User-defined plant functional groups (PFGs) are used to elucidate the interaction between plant transpiration and groundwater conditions. Five generalized plant functional groups based on transpiration rates, plant rooting depth, and water tolerance ranges are presented: obligate wetland, shallow-rooted riparian, deep-rooted riparian, transitional riparian and bare ground/open water. Plant functional groups can be further divided into subgroups (PFSGs) based on plant size, density or other characteristics. The RIP-ET allows for partial habitat coverage and mixtures of plant functional subgroups to be present in a single model cell. RIP-ET also distinguishes between plant transpiration and bare-ground evaporation. Habitat areas are designated by polygons; each polygon can contain a mixture of PFSGs and bare ground, and is assigned a surface elevation. This process requires a determination of fractional coverage for each of the plant functional subgroups present in a polygon to account for the mixture of coverage types and resulting transpiration. The fractional cover within a cell has two components: (1) the polygonal fraction of active habitat (excluding area of bare ground, dead trees, or brush) in a cell, and (2) fraction of plant type area or bare ground area in a polygon. RIP-ET determines the transpiration rate for each plant functional group and evaporation from bare ground/open water in a cell, the total ET in the cell, and the total ET rate over the region of simulation.

  16. 21 CFR 864.7825 - Sickle cell test.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Sickle cell test. 864.7825 Section 864.7825 Food... DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7825 Sickle cell test. (a) Identification. A sickle cell test is a device used to determine the sickle cell hemoglobin content of human...

  17. 21 CFR 864.7825 - Sickle cell test.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Sickle cell test. 864.7825 Section 864.7825 Food... DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7825 Sickle cell test. (a) Identification. A sickle cell test is a device used to determine the sickle cell hemoglobin content of human...

  18. 21 CFR 864.7825 - Sickle cell test.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Sickle cell test. 864.7825 Section 864.7825 Food... DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7825 Sickle cell test. (a) Identification. A sickle cell test is a device used to determine the sickle cell hemoglobin content of human...

  19. 21 CFR 864.7825 - Sickle cell test.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Sickle cell test. 864.7825 Section 864.7825 Food... DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7825 Sickle cell test. (a) Identification. A sickle cell test is a device used to determine the sickle cell hemoglobin content of human...

  20. 21 CFR 864.7825 - Sickle cell test.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Sickle cell test. 864.7825 Section 864.7825 Food... DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7825 Sickle cell test. (a) Identification. A sickle cell test is a device used to determine the sickle cell hemoglobin content of human...

  1. Transcriptome analysis of PCOS arrested 2-cell embryos.

    PubMed

    Lu, Cuiling; Chi, Hongbin; Wang, Yapeng; Feng, Xue; Wang, Lina; Huang, Shuo; Yan, Liying; Lin, Shengli; Liu, Ping; Qiao, Jie

    2018-06-18

    In an attempt to explore the early developmental arrest in embryos from polycystic ovarian syndrome (PCOS) patients, we sequenced the transcriptome profiles of PCOS arrested 2-cell embryos, non-PCOS arrested 2-cell embryos and non-arrested 2-cell embryos using single-cell RNA-Seq technique. Differential expression analysis was performed using the DEGSeq R package. Gene Ontology (GO) enrichment was analyzed using the GOseq R package. Data revealed 62 differentially expressed genes between non-PCOS arrested and PCOS arrested embryos and 2217 differentially expressed genes between PCOS arrested and non-arrested 2-cell embryos. A total of 49 differently expressed genes (DEGs) were annotated with GO terms in the up-regulated genes between PCOS arrested and non-PCOS arrested embryos after GO enrichment. A total of 29 DEGs were annotated with GO terms in the down-regulated genes between PCOS arrested and non-arrested 2-cell embryos after GO enrichment. These data can provide a reference for screening specific genes involved in the arrest of PCOS embryos.

  2. [Establishment of human embryonic stem cell lines and their therapeutic application].

    PubMed

    Suemori, Hirofumi

    2004-03-01

    Embryonic stem (ES) cell lines are pluripotent stem cell lines that can be propagated indefinitely in culture, retaining their potency to differentiate into every type of cell and tissue in the body. ES cell lines were first established from mouse blastocysts, and have been used for research in developmental biology. ES cells have been proven to be very valuable in the genetic modification of the mouse, especially in producing knockout mice. Since establishment of human ES cell lines was reported, their use in cell replacement therapies has been enthusiastically expected. There have been reports of the differentiation of several useful cell types from human ES cell lines, and clinical use of functional tissues and cells from human ES cells is anticipated. In Japan, there have also been many demands for the use of human ES cells in basic and pre-clinical research. We obtained governmental permission to establish human ES cell lines in April 2002 and started research using donated frozen embryos in January 2003. We successfully established three ES cell line from three blastocysts. These cell lines will be distributed at cost to researchers who have governmental permission to use human ES cells.

  3. Chandipura virus growth kinetics in vertebrate cell lines, insect cell lines & embryonated eggs.

    PubMed

    Jadi, R S; Sudeep, A B; Kumar, Satyendra; Arankalle, V A; Mishra, A C

    2010-08-01

    Since not much information on Chandipura virus is available, an attempt was made to study the growth kinetics of the virus in certain vertebrate, invertebrate cell lines and embryonated chicken eggs. Comparative study of Chandipura virus (CHPV) growth kinetics in three vertebrate cell lines [Vero E6, Rhabdo myosarcoma (RD), Porcine stable kidney (PS) cell lines], two insect cell lines [Aedes aegypti (AA) and Phlebotomus papatasi (PP-9) cell lines] and embryonated pathogen free chicken eggs was conducted, by tissue culture infective dose 50 per cent (TCID(50)) and indirect immunofluorescence assay (IFA). All the cell lines and embryonated egg supported the growth of CHPV and yielded high virus titre. The vertebrate cell lines showed distinct cytopathic effect (CPE) within 4-6 h post infection (PI), while no CPE was observed in insect cell lines. PP-9 cell line was the most sensitive system to CHPV as viral antigen could be detected at 1 h PI by IFA. Our results demonstrated that all the systems were susceptible to CHPV and achieved high yield of virus. However, the PP-9 cell line had an edge over the others due to its high sensitivity to the virus which might be useful for detection and isolation of the virus during epidemics.

  4. Fourteen Years of R/qtl: Just Barely Sustainable

    PubMed Central

    Broman, Karl W.

    2014-01-01

    R/qtl is an R package for mapping quantitative trait loci (genetic loci that contribute to variation in quantitative traits) in experimental crosses. Its development began in 2000. There have been 38 software releases since 2001. The latest release contains 35k lines of R code and 24k lines of C code, plus 15k lines of code for the documentation. Challenges in the development and maintenance of the software are discussed. A key to the success of R/qtl is that it remains a central tool for the chief developer's own research work, and so its maintenance is of selfish importance. PMID:25364504

  5. Advanced nickel-hydrogen cell configuration study

    NASA Technical Reports Server (NTRS)

    1983-01-01

    Long-term trends in the evolution of space power technology point toward increased payload power demand which in turn translates into both higher battery system charge storage capability and higher operating voltages. State of the art nickel-hydrogen cells of the 50 to 60 Wh size, packaged in individual pressure vessels, are capable of meeting the required cycle life for a wide range of anticipated operating conditions; however, they provided several drawbacks to battery system integrated efforts. Because of size, high voltage/high power systems require integrating hundreds of cells into the operating system. Packaging related weight and volume inefficiencies degrade the energy density and specific energy of individual cells currently at 30 Wh/cudm and 40 Wh/kg respectively. In addition, the increased parts count and associated handling significantly affect the overall battery related costs. Spacecraft battery systems designers within industry and Government realize that to reduce weight, volume, and cost requires increases in the capacity of nickel-hydrogen cells.

  6. esATAC: An Easy-to-use Systematic pipeline for ATAC-seq data analysis.

    PubMed

    Wei, Zheng; Zhang, Wei; Fang, Huan; Li, Yanda; Wang, Xiaowo

    2018-03-07

    ATAC-seq is rapidly emerging as one of the major experimental approaches to probe chromatin accessibility genome-wide. Here, we present "esATAC", a highly integrated easy-to-use R/Bioconductor package, for systematic ATAC-seq data analysis. It covers essential steps for full analyzing procedure, including raw data processing, quality control and downstream statistical analysis such as peak calling, enrichment analysis and transcription factor footprinting. esATAC supports one command line execution for preset pipelines, and provides flexible interfaces for building customized pipelines. esATAC package is open source under the GPL-3.0 license. It is implemented in R and C ++. Source code and binaries for Linux, MAC OS X and Windows are available through Bioconductor https://www.bioconductor.org/packages/release/bioc/html/esATAC.html). xwwang@tsinghua.edu.cn. Supplementary data are available at Bioinformatics online.

  7. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Langer, Steven H.; Karlin, Ian; Marinak, Marty M.

    HYDRA is used to simulate a variety of experiments carried out at the National Ignition Facility (NIF) [4] and other high energy density physics facilities. HYDRA has packages to simulate radiation transfer, atomic physics, hydrodynamics, laser propagation, and a number of other physics effects. HYDRA has over one million lines of code and includes both MPI and thread-level (OpenMP and pthreads) parallelism. This paper measures the performance characteristics of HYDRA using hardware counters on an IBM BlueGene/Q system. We report key ratios such as bytes/instruction and memory bandwidth for several different physics packages. The total number of bytes read andmore » written per time step is also reported. We show that none of the packages which use significant time are memory bandwidth limited on a Blue Gene/Q. HYDRA currently issues very few SIMD instructions. The pressure on memory bandwidth will increase if high levels of SIMD instructions can be achieved.« less

  8. Room-temperature-deposited dielectrics and superconductors for integrated photonics.

    PubMed

    Shainline, Jeffrey M; Buckley, Sonia M; Nader, Nima; Gentry, Cale M; Cossel, Kevin C; Cleary, Justin W; Popović, Miloš; Newbury, Nathan R; Nam, Sae Woo; Mirin, Richard P

    2017-05-01

    We present an approach to fabrication and packaging of integrated photonic devices that utilizes waveguide and detector layers deposited at near-ambient temperature. All lithography is performed with a 365 nm i-line stepper, facilitating low cost and high scalability. We have shown low-loss SiN waveguides, high-Q ring resonators, critically coupled ring resonators, 50/50 beam splitters, Mach-Zehnder interferometers (MZIs) and a process-agnostic fiber packaging scheme. We have further explored the utility of this process for applications in nonlinear optics and quantum photonics. We demonstrate spectral tailoring and octave-spanning supercontinuum generation as well as the integration of superconducting nanowire single photon detectors with MZIs and channel-dropping filters. The packaging approach is suitable for operation up to 160 °C as well as below 1 K. The process is well suited for augmentation of existing foundry capabilities or as a stand-alone process.

  9. Malignant hematopoietic cell lines: in vitro models for the study of natural killer cell leukemia-lymphoma.

    PubMed

    Drexler, H G; Matsuo, Y

    2000-05-01

    Malignancies involving natural killer (NK) cells are rare disorders. The complexity of NK cell-involving disorders has only recently been appreciated. Modern classifications discern immature (precursor) from mature NK cell leukemias-lymphomas. Continuous NK leukemia-lymphoma cell lines represent important model systems to study these neoplasms. While there are a number of putative NK cell lines which are, however, either not characterized, not immortalized, non-malignant, non-NK, or plain false cell lines, six bona fide malignant NK cell lines have been established and are sufficiently well characterized: HANK1, KHYG-1, NK-92, NKL, NK-YS and YT. Except for YT which was derived from a not further defined acute lymphoblastic lymphoma, these cell lines were established from patients with various NK cell malignancies. Five of the six cell lines are constitutively interleukin-2-dependent. Their immunoprofile is remarkably similar: CD1-, CD2+, surface CD3 (but cytoplasmic CD3epsilon+), CD4-, CD5-, CD7+, CD8-, CD16-, CD56+, CD57-, TCRalphabeta-, TCRgammadelta-, negative for B cell and myelomonocytic markers. The immunoglobulin heavy chain and T cell receptor genes are all in germline configuration. All six lines show complex chromosomal alterations, with both numerical and structural aberrations, attesting to their malignant and monoclonal nature. Functionally, these cells which contain azurophilic granules in their cytoplasm are nearly universally positive in NK activity assays. Three of five cell lines are Epstein-Barr virus-positive (type II latency). The composite data on these six cell lines allow for the operational definition of a typical malignant NK cell line profile. NK leukemia-lymphoma cell lines will prove invaluable for studies of normal and malignant NK cell biology.

  10. Replication-guided nucleosome packing and nucleosome breathing expedite the formation of dense arrays

    PubMed Central

    Osberg, Brendan; Nuebler, Johannes; Korber, Philipp; Gerland, Ulrich

    2014-01-01

    The first level of genome packaging in eukaryotic cells involves the formation of dense nucleosome arrays, with DNA coverage near 90% in yeasts. How cells achieve such high coverage within a short time, e.g. after DNA replication, remains poorly understood. It is known that random sequential adsorption of impenetrable particles on a line reaches high density extremely slowly, due to a jamming phenomenon. The nucleosome-shifting action of remodeling enzymes has been proposed as a mechanism to resolve such jams. Here, we suggest two biophysical mechanisms which assist rapid filling of DNA with nucleosomes, and we quantitatively characterize these mechanisms within mathematical models. First, we show that the ‘softness’ of nucleosomes, due to nucleosome breathing and stepwise nucleosome assembly, significantly alters the filling behavior, speeding up the process relative to ‘hard’ particles with fixed, mutually exclusive DNA footprints. Second, we explore model scenarios in which the progression of the replication fork could eliminate nucleosome jamming, either by rapid filling in its wake or via memory of the parental nucleosome positions. Taken together, our results suggest that biophysical effects promote rapid nucleosome filling, making the reassembly of densely packed nucleosomes after DNA replication a simpler task for cells than was previously thought. PMID:25428353

  11. A Partial E3 Deletion in Replication-Defective Adenoviral Vectors Allows for Stable Expression of Potentially Toxic Transgene Products.

    PubMed

    Haut, Larissa H; Gill, Amanda L; Kurupati, Raj K; Bian, Ang; Li, Yan; Giles-Davis, Wynetta; Xiang, Zhiquan; Zhou, Xiang Yang; Ertl, Hildegund C J

    2016-10-01

    Adenovirus (Ad) is used extensively for construction of viral vectors, most commonly with deletion in its E1 and/or E3 genomic regions. Previously, our attempts to insert envelope proteins (Env) of HIV-1 into such vectors based on chimpanzee-derived Ad (AdC) viruses were thwarted. Here, we describe that genetic instability of an E1- and E3-deleted AdC vector of serotype C6 expressing Env of HIV-1 can be overcome by reinsertion of E3 sequences with anti-apoptotic activities. This partial E3 deletion presumably delays premature death of HEK-293 packaging cell lines due to Env-induced cell apoptosis. The same partial E3 deletion also allows for the generation of stable glycoprotein 140 (gp140)- and gp160-expressing Ad vectors based on AdC7, a distinct AdC serotype. Env-expressing AdC vectors containing the partial E3 deletion are genetically stable upon serial cell culture passaging, produce yields comparable to those of other AdC vectors, and induce transgene product-specific antibody responses in mice. A partial E3 deletion thereby allows expansion of the repertoire of transgenes that can be expressed by Ad vectors.

  12. A New Image Processing and GIS Package

    NASA Technical Reports Server (NTRS)

    Rickman, D.; Luvall, J. C.; Cheng, T.

    1998-01-01

    The image processing and GIS package ELAS was developed during the 1980's by NASA. It proved to be a popular, influential and powerful in the manipulation of digital imagery. Before the advent of PC's it was used by hundreds of institutions, mostly schools. It is the unquestioned, direct progenitor or two commercial GIS remote sensing packages, ERDAS and MapX and influenced others, such as PCI. Its power was demonstrated by its use for work far beyond its original purpose, having worked several different types of medical imagery, photomicrographs of rock, images of turtle flippers and numerous other esoteric imagery. Although development largely stopped in the early 1990's the package still offers as much or more power and flexibility than any other roughly comparable package, public or commercial. It is a huge body or code, representing more than a decade of work by full time, professional programmers. The current versions all have several deficiencies compared to current software standards and usage, notably its strictly command line interface. In order to support their research needs the authors are in the process of fundamentally changing ELAS, and in the process greatly increasing its power, utility, and ease of use. The new software is called ELAS II. This paper discusses the design of ELAS II.

  13. Analysis of the type II robotic mixed-model assembly line balancing problem

    NASA Astrophysics Data System (ADS)

    Çil, Zeynel Abidin; Mete, Süleyman; Ağpak, Kürşad

    2017-06-01

    In recent years, there has been an increasing trend towards using robots in production systems. Robots are used in different areas such as packaging, transportation, loading/unloading and especially assembly lines. One important step in taking advantage of robots on the assembly line is considering them while balancing the line. On the other hand, market conditions have increased the importance of mixed-model assembly lines. Therefore, in this article, the robotic mixed-model assembly line balancing problem is studied. The aim of this study is to develop a new efficient heuristic algorithm based on beam search in order to minimize the sum of cycle times over all models. In addition, mathematical models of the problem are presented for comparison. The proposed heuristic is tested on benchmark problems and compared with the optimal solutions. The results show that the algorithm is very competitive and is a promising tool for further research.

  14. Derivation and characterization of Chinese human embryonic stem cell line with high potential to differentiate into pancreatic and hepatic cells.

    PubMed

    Shi, Cheng; Shen, Huan; Jiang, Wei; Song, Zhi-Hua; Wang, Cheng-Yan; Wei, Li-Hui

    2011-04-01

    Human embryonic stem cells have prospective uses in regenerative medicine and drug screening. Every human embryonic stem cell line has its own genetic background, which determines its specific ability for differentiation as well as susceptibility to drugs. It is necessary to compile many human embryonic stem cell lines with various backgrounds for future clinical use, especially in China due to its large population. This study contributes to isolating new Chinese human embryonic stem cell lines with clarified directly differentiation ability. Donated embryos that exceeded clinical use in our in vitro fertilization-embryo transfer (IVF-ET) center were collected to establish human embryonic stem cells lines with informed consent. The classic growth factors of basic fibroblast growth factor (bFGF) and recombinant human leukaemia inhibitory factor (hLIF) for culturing embryonic stem cells were used to capture the stem cells from the plated embryos. Mechanical and enzymetic methods were used to propagate the newly established human embryonic stem cells line. The new cell line was checked for pluripotent characteristics with detecting the expression of stemness genes and observing spontaneous differentiation both in vitro and in vivo. Finally similar step-wise protocols from definitive endoderm to target specific cells were used to check the cell line's ability to directly differentiate into pancreatic and hepatic cells. We generated a new Chinese human embryonic stem cells line, CH1. This cell line showed the same characteristics as other reported Chinese human embryonic stem cells lines: normal morphology, karyotype and pluripotency in vitro and in vivo. The CH1 cells could be directly differentiated towards pancreatic and hepatic cells with equal efficiency compared to the H1 cell line. This newly established Chinese cell line, CH1, which is pluripotent and has high potential to differentiate into pancreatic and hepatic cells, will provide a useful tool for embryo development research, along with clinical treatments for diabetes and some hepatic diseases.

  15. Effect of specific silencing of EMMPRIN on the growth and cell cycle distribution of MCF-7 breast cancer cells.

    PubMed

    Yang, X Q; Yang, J; Wang, R; Zhang, S; Tan, Q W; Lv, Q; Meng, W T; Mo, X M; Li, H J

    2015-12-02

    The extracellular matrix metalloproteinase inducer (EMMPRIN, CD147) is a member of the immunoglobulin family and shows increased expression in tumor cells. We examined the effect of RNAi-mediated EMMPRIN gene silencing induced by lentiviral on the growth and cycle distribution of MCF-7 breast cancer cells. Lentiviral expressing EMMPRIN-short hairpin RNA were packaged to infect MCF-7 cells. The inhibition efficiency of EMMPRIN was validated by real-time fluorescent quantitation polymerase chain reaction and western blotting. The effect of EMMPRIN on cell proliferation ability was detected using the MTT assay and clone formation experiments. Changes in cell cycle were detected by flow cytometry. EMMPRIN-short hairpin RNA-packaged lentiviral significantly down-regulated EMMPRIN mRNA and protein expression, significantly inhibited cell proliferation and in vitro tumorigenicity, and induced cell cycle abnormalities. Cells in the G0/G1 and G2/M phases were increased, while cells in the S phase were decreased after infection of MCF-7 cells for 3 days. The EMMPRIN gene facilitates breast cancer cell malignant proliferation by regulating cell cycle distribution and may be a molecular target for breast cancer gene therapy.

  16. Mosquito cell-derived West Nile virus replicon particles mimic arbovirus inoculum and have reduced spread in mice.

    PubMed

    Boylan, Brendan T; Moreira, Fernando R; Carlson, Tim W; Bernard, Kristen A

    2017-02-01

    Half of the human population is at risk of infection by an arthropod-borne virus. Many of these arboviruses, such as West Nile, dengue, and Zika viruses, infect humans by way of a bite from an infected mosquito. This infectious inoculum is insect cell-derived giving the virus particles distinct qualities not present in secondary infectious virus particles produced by infected vertebrate host cells. The insect cell-derived particles differ in the glycosylation of virus structural proteins and the lipid content of the envelope, as well as their induction of cytokines. Thus, in order to accurately mimic the inoculum delivered by arthropods, arboviruses should be derived from arthropod cells. Previous studies have packaged replicon genome in mammalian cells to produce replicon particles, which undergo only one round of infection, but no studies exist packaging replicon particles in mosquito cells. Here we optimized the packaging of West Nile virus replicon genome in mosquito cells and produced replicon particles at high concentration, allowing us to mimic mosquito cell-derived viral inoculum. These particles were mature with similar genome equivalents-to-infectious units as full-length West Nile virus. We then compared the mosquito cell-derived particles to mammalian cell-derived particles in mice. Both replicon particles infected skin at the inoculation site and the draining lymph node by 3 hours post-inoculation. The mammalian cell-derived replicon particles spread from the site of inoculation to the spleen and contralateral lymph nodes significantly more than the particles derived from mosquito cells. This in vivo difference in spread of West Nile replicons in the inoculum demonstrates the importance of using arthropod cell-derived particles to model early events in arboviral infection and highlights the value of these novel arthropod cell-derived replicon particles for studying the earliest virus-host interactions for arboviruses.

  17. Inorganic Kernel-Reconstituted Lipoprotein Biomimetic Nanovehicles Enable Efficient Targeting “Trojan Horse” Delivery of STAT3-Decoy Oligonucleotide for Overcoming TRAIL Resistance

    PubMed Central

    Shi, Kai; Xue, Jianxiu; Fang, Yan; Bi, Hongshu; Gao, Shan; Yang, Dongjuan; Lu, Anqi; Li, Yuai; Chen, Yao; Ke, Liyuan

    2017-01-01

    Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can selectively induce apoptosis in a variety of tumor cells, but not most normal cells. Nevertheless, its therapeutic potential is limited due to the frequent occurrence of resistance in tumor cells, especially hepatocellular carcinoma cell lines. Therefore, we investigated the reversal effect of STAT3-decoy oligonucleotides (ODNs) on TRAIL resistance. Methods. Considering that the drawback of poor cellular permeability and rapid degradation in vivo limited ODNs' further clinical applications, we developed a biomimetic calcium phosphate-reconstituted low density lipoprotein nanovehicle (CaP@LDL) that would serve as a “Trojan horse” to carry STAT3-decoy ODNs into tumor cells and then regulate TRAIL-induced apoptosis. Results. In comparison with native ODNs, the reconstituted CaP@LDL packaged ODNs showed significantly increased serum stability, cellular transfection, in vitro synergistic cytotoxicity and apoptosis in hepatoma cells, while there was no cytotoxicity to normal cells. The improved TRAIL sensitization is attributed to blocking of STAT3 signaling and consequent expression of the downstream target antiapoptotic gene. Following systemic administration, CaP@LDL displayed LDL-mimicking pharmacokinetic behavior such as attenuated blood clearance as well as enhanced accumulation in tumor and hepatorenal sites. With the synergistic combination of decoyODN/CaP@LDL, TRAIL dramatically inhibited hepatic tumor growth in a xenograft model and induced significant tumor apoptosis in vivo. Conclusion. These results suggested that CaP@LDL-mediated STAT3-decoy ODN delivery might be a promising new strategy for reversing TRAIL resistance in hepatocellular carcinoma therapy. PMID:29158840

  18. Inorganic Kernel-Reconstituted Lipoprotein Biomimetic Nanovehicles Enable Efficient Targeting "Trojan Horse" Delivery of STAT3-Decoy Oligonucleotide for Overcoming TRAIL Resistance.

    PubMed

    Shi, Kai; Xue, Jianxiu; Fang, Yan; Bi, Hongshu; Gao, Shan; Yang, Dongjuan; Lu, Anqi; Li, Yuai; Chen, Yao; Ke, Liyuan

    2017-01-01

    Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can selectively induce apoptosis in a variety of tumor cells, but not most normal cells. Nevertheless, its therapeutic potential is limited due to the frequent occurrence of resistance in tumor cells, especially hepatocellular carcinoma cell lines. Therefore, we investigated the reversal effect of STAT3-decoy oligonucleotides (ODNs) on TRAIL resistance. Methods . Considering that the drawback of poor cellular permeability and rapid degradation in vivo limited ODNs' further clinical applications, we developed a biomimetic calcium phosphate-reconstituted low density lipoprotein nanovehicle (CaP@LDL) that would serve as a "Trojan horse" to carry STAT3-decoy ODNs into tumor cells and then regulate TRAIL-induced apoptosis. Results . In comparison with native ODNs, the reconstituted CaP@LDL packaged ODNs showed significantly increased serum stability, cellular transfection, in vitro synergistic cytotoxicity and apoptosis in hepatoma cells, while there was no cytotoxicity to normal cells. The improved TRAIL sensitization is attributed to blocking of STAT3 signaling and consequent expression of the downstream target antiapoptotic gene. Following systemic administration, CaP@LDL displayed LDL-mimicking pharmacokinetic behavior such as attenuated blood clearance as well as enhanced accumulation in tumor and hepatorenal sites. With the synergistic combination of decoyODN/CaP@LDL, TRAIL dramatically inhibited hepatic tumor growth in a xenograft model and induced significant tumor apoptosis in vivo . Conclusion. These results suggested that CaP@LDL-mediated STAT3-decoy ODN delivery might be a promising new strategy for reversing TRAIL resistance in hepatocellular carcinoma therapy.

  19. 77 FR 5489 - Identification of Human Cell Lines Project

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-03

    ...-01] Identification of Human Cell Lines Project AGENCY: National Institute of Standards and Technology... cell line samples as part of the Identification of Human Cell Lines Project. All data and corresponding... cell lines accepted on the NIST Applied Genetics Group Web site at http://www.nist.gov/mml/biochemical...

  20. Multiscale smeared finite element model for mass transport in biological tissue: From blood vessels to cells and cellular organelles.

    PubMed

    Kojic, M; Milosevic, M; Simic, V; Koay, E J; Kojic, N; Ziemys, A; Ferrari, M

    2018-05-21

    One of the basic and vital processes in living organisms is mass exchange, which occurs on several levels: it goes from blood vessels to cells and organelles within cells. On that path, molecules, as oxygen, metabolic products, drugs, etc. Traverse different macro and micro environments - blood, extracellular/intracellular space, and interior of organelles; and also biological barriers such as walls of blood vessels and membranes of cells and organelles. Many aspects of this mass transport remain unknown, particularly the biophysical mechanisms governing drug delivery. The main research approach relies on laboratory and clinical investigations. In parallel, considerable efforts have been directed to develop computational tools for additional insight into the intricate process of mass exchange and transport. Along these lines, we have recently formulated a composite smeared finite element (CSFE) which is composed of the smeared continuum pressure and concentration fields of the capillary and lymphatic system, and of these fields within tissue. The element offers an elegant and simple procedure which opens up new lines of inquiry and can be applied to large systems such as organs and tumors models. Here, we extend this concept to a multiscale scheme which concurrently couples domains that span from large blood vessels, capillaries and lymph, to cell cytosol and further to organelles of nanometer size. These spatial physical domains are coupled by the appropriate connectivity elements representing biological barriers. The composite finite element has "degrees of freedom" which include pressures and concentrations of all compartments of the vessels-tissue assemblage. The overall model uses the standard, measurable material properties of the continuum biological environments and biological barriers. It can be considered as a framework into which we can incorporate various additional effects (such as electrical or biochemical) for transport through membranes or within cells. This concept and the developed FE software within our package PAK offers a computational tool that can be applied to whole-organ systems, while also including specific domains such as tumors. The solved examples demonstrate the accuracy of this model and its applicability to large biological systems. Copyright © 2018. Published by Elsevier Ltd.

  1. Computational Modeling | Photovoltaic Research | NREL

    Science.gov Websites

    performance of single- and multijunction cells and modules. We anticipate the upcoming completion of our next software package for a simplified electronic design of single- and multicrystalline silicon solar cells

  2. Liposomal Packaging Generates Wnt Protein with In Vivo Biological Activity

    PubMed Central

    Zhao, Ludan; Kim, Jae-Beom; ten Berge, Derk; Ponnusamy, Karthik; Carre, A. Lyonel; Dudek, Henryk; Zachlederova, Marie; McElhaney, Michael; Brunton, Shirley; Gunzner, Janet; Callow, Marinella; Polakis, Paul; Costa, Mike; Zhang, Xiaoyan M.; Helms, Jill A.; Nusse, Roel

    2008-01-01

    Wnt signals exercise strong cell-biological and regenerative effects of considerable therapeutic value. There are, however, no specific Wnt agonists and no method for in vivo delivery of purified Wnt proteins. Wnts contain lipid adducts that are required for activity and we exploited this lipophilicity by packaging purified Wnt3a protein into lipid vesicles. Rather than being encapsulated, Wnts are tethered to the liposomal surface, where they enhance and sustain Wnt signaling in vitro. Molecules that effectively antagonize soluble Wnt3a protein but are ineffective against the Wnt3a signal presented by a cell in a paracrine or autocrine manner are also unable to block liposomal Wnt3a activity, suggesting that liposomal packaging mimics the biological state of active Wnts. When delivered subcutaneously, Wnt3a liposomes induce hair follicle neogenesis, demonstrating their robust biological activity in a regenerative context. PMID:18698373

  3. Liposomal packaging generates Wnt protein with in vivo biological activity.

    PubMed

    Morrell, Nathan T; Leucht, Philipp; Zhao, Ludan; Kim, Jae-Beom; ten Berge, Derk; Ponnusamy, Karthik; Carre, A Lyonel; Dudek, Henryk; Zachlederova, Marie; McElhaney, Michael; Brunton, Shirley; Gunzner, Janet; Callow, Marinella; Polakis, Paul; Costa, Mike; Zhang, Xiaoyan M; Helms, Jill A; Nusse, Roel

    2008-08-13

    Wnt signals exercise strong cell-biological and regenerative effects of considerable therapeutic value. There are, however, no specific Wnt agonists and no method for in vivo delivery of purified Wnt proteins. Wnts contain lipid adducts that are required for activity and we exploited this lipophilicity by packaging purified Wnt3a protein into lipid vesicles. Rather than being encapsulated, Wnts are tethered to the liposomal surface, where they enhance and sustain Wnt signaling in vitro. Molecules that effectively antagonize soluble Wnt3a protein but are ineffective against the Wnt3a signal presented by a cell in a paracrine or autocrine manner are also unable to block liposomal Wnt3a activity, suggesting that liposomal packaging mimics the biological state of active Wnts. When delivered subcutaneously, Wnt3a liposomes induce hair follicle neogenesis, demonstrating their robust biological activity in a regenerative context.

  4. Attachment of Asaia bogorensis Originating in Fruit-Flavored Water to Packaging Materials

    PubMed Central

    Otlewska, Anna; Antolak, Hubert

    2014-01-01

    The objective of this study was to investigate the adhesion of isolated spoilage bacteria to packaging materials used in the food industry. Microorganisms were isolated from commercial fruit-flavored mineral water in plastic bottles with flocks as a visual defect. The Gram-negative rods were identified using the molecular method through the amplification of a partial region of the 16S rRNA gene. Based on the sequence identity (99.6%) between the spoilage organism and a reference strain deposited in GenBank, the spoilage isolate was identified as Asaia bgorensis. Experiments on bacterial adhesion were conducted using plates made of glass and polystyrene (packaging materials commonly used in the beverage industry). Cell adhesion ability was determined using luminometry, plate count, and the microscopic method. The strain of A. bogorensis was characterized by strong adhesion properties which were dependent on the surface type, with the highest cell adhesion detected on polystyrene. PMID:25295262

  5. Bonded polyimide fuel cell package and method thereof

    DOEpatents

    Morse, Jeffrey D.; Jankowski, Alan; Graff, Robert T.; Bettencourt, Kerry

    2005-11-01

    Described herein are processes for fabricating microfluidic fuel cell systems with embedded components in which micron-scale features are formed by bonding layers of DuPont Kapton.TM. polyimide laminate. A microfluidic fuel cell system fabricated using this process is also described.

  6. Method of preparation of bonded polyimide fuel cell package

    DOEpatents

    Morse, Jeffrey D [Martinez, CA; Jankowski, Alan [Livermore, CA; Graff, Robert T [Modesto, CA; Bettencourt, Kerry [Dublin, CA

    2011-04-26

    Described herein are processes for fabricating microfluidic fuel cell systems with embedded components in which micron-scale features are formed by bonding layers of DuPont Kapton.TM. polyimide laminate. A microfluidic fuel cell system fabricated using this process is also described.

  7. Development and characterization of two new cell lines from milkfish (Chanos chanos) and grouper (Epinephelus coioides) for virus isolation.

    PubMed

    Parameswaran, V; Ishaq Ahmed, V P; Shukla, Ravi; Bhonde, R R; Sahul Hameed, A S

    2007-01-01

    Two new cell lines, SIMH and SIGE, were derived from the heart of milkfish (Chanos chanos), a euryhaline teleost, and from the eye of grouper (Epinephelus coioides), respectively. These cell lines were maintained in Leibovitz's L-15 supplemented with 20% fetal bovine serum (FBS). The SIMH cell line was subcultured more than 50 times over a period of 210 days and SIGE cell line has been subcultured 100 times over a period of 1 1/2 years. The SIMH cell line consists predominantly of fibroblastic-like cells. The SIGE cell line consists predominantly of epithelial cells. Both the cell lines were able to grow at temperatures between 25 and 32 degrees C with an optimum temperature of 28 degrees C. The growth rate of these cells increased as the proportion of FBS increased from 2% to 20% at 28 degrees C with optimum growth at the concentrations of 15% or 20% FBS. Seven marine fish viruses were tested to determine the susceptibility of these cell lines. The SIGE cell line was found to be susceptible to nodavirus, MABV NC-1 and Y6, and the infection was confirmed by cytopathic effect (CPE) and reverse transcriptase-polymerase chain reaction. When these cells were transfected with pEGFP-N1 vector DNA, significant fluorescent signals were observed, suggesting that these cell lines can be a useful tool for transgenic and genetic manipulation studies. Further, these cell lines are characterized by immunocytochemistry using confocal laser scanning microscopy (CFLSM).

  8. Packaging of MEMS/MOEMS and nanodevices: reliability, testing, and characterization aspects

    NASA Astrophysics Data System (ADS)

    Tekin, Tolga; Ngo, Ha-Duong; Wittler, Olaf; Bouhlal, Bouchaib; Lang, Klaus-Dieter

    2011-02-01

    The last decade witnessed an explosive growth in research and development efforts devoted to MEMS devices and packaging. The successfully developed MEMS devices are, for example inkjet, pressure sensors, silicon microphones, accelerometers, gyroscopes, MOEMS, micro fuel cells and emerging MEMS. For the next decade, MEMS/MOEMS and nanodevice based products will penetrate into IT, telecommunications, automotive, defense, life sciences, medical and implantable applications. Forecasts say the MEMS market to be $14 billion by 2012. The packaging cost of MEMS/MOEMS products in general is about 70 percent. Unlike today's electronics IC packaging, their packaging are custom-built and difficult due to the moving structural elements. In order for the moving elements of a MEMS device to move effectively in a well-controlled atmosphere, hermetic sealing of the MEMS device in a cap is necessary. For some MEMS devices, such as resonators and gyroscopes, vacuum packaging is required. Usually, the cap is processed at the wafer level, and thus MEMS packaging is truly a wafer level packaging. In terms of MEMS/MOEMS and nanodevice packaging, there are still many critical issues need to be addressed due to the increasing integration density supported by 3D heterogeneous integration of multi-physic components/layers consisting of photonics, electronics, rf, plasmonics, and wireless. The infrastructure of MEMS/MOEMS and nanodevices and their packaging is not well established yet. Generic packaging platform technologies are not available. Some of critical issues have been studied intensively in the last years. In this paper we will discuss about processes, reliability, testing and characterization of MEMS/MOEMS and nanodevice packaging.

  9. Authentication of M14 melanoma cell line proves misidentification of MDA‐MB‐435 breast cancer cell line

    PubMed Central

    Korch, Christopher; Hall, Erin M.; Dirks, Wilhelm G.; Ewing, Margaret; Faries, Mark; Varella‐Garcia, Marileila; Robinson, Steven; Storts, Douglas; Turner, Jacqueline A.; Wang, Ying; Burnett, Edward C.; Healy, Lyn; Kniss, Douglas; Neve, Richard M.; Nims, Raymond W.; Reid, Yvonne A.; Robinson, William A.

    2017-01-01

    A variety of analytical approaches have indicated that melanoma cell line UCLA‐SO‐M14 (M14) and breast carcinoma cell line MDA‐MB‐435 originate from a common donor. This indicates that at some point in the past, one of these cell lines became misidentified, meaning that it ceased to correspond to the reported donor and instead became falsely identified (through cross‐contamination or other means) as a cell line from a different donor. Initial studies concluded that MDA‐MB‐435 was the misidentified cell line and M14 was the authentic cell line, although contradictory evidence has been published, resulting in further confusion. To address this question, we obtained early samples of the melanoma cell line (M14), a lymphoblastoid cell line from the same donor (ML14), and donor serum preserved at the originator's institution. M14 samples were cryopreserved in December 1975, before MDA‐MB‐435 cells were established in culture. Through a series of molecular characterizations, including short tandem repeat (STR) profiling and cytogenetic analysis, we demonstrated that later samples of M14 and MDA‐MB‐435 correspond to samples of M14 frozen in 1975, to the lymphoblastoid cell line ML14, and to the melanoma donor's STR profile, sex and blood type. This work demonstrates conclusively that M14 is the authentic cell line and MDA‐MB‐435 is misidentified. With clear provenance information and authentication testing of early samples, it is possible to resolve debates regarding the origins of problematic cell lines that are widely used in cancer research. PMID:28940260

  10. Integration of safety engineering into a cost optimized development program.

    NASA Technical Reports Server (NTRS)

    Ball, L. W.

    1972-01-01

    A six-segment management model is presented, each segment of which represents a major area in a new product development program. The first segment of the model covers integration of specialist engineers into 'systems requirement definition' or the system engineering documentation process. The second covers preparation of five basic types of 'development program plans.' The third segment covers integration of system requirements, scheduling, and funding of specialist engineering activities into 'work breakdown structures,' 'cost accounts,' and 'work packages.' The fourth covers 'requirement communication' by line organizations. The fifth covers 'performance measurement' based on work package data. The sixth covers 'baseline requirements achievement tracking.'

  11. Development of remote sensing technology in New Zealand, part 1. Mapping land use and environmental studies in New Zealand, part 2. Indigenous forest assessment, part 3. Seismotectonic, structural, volcanologic and geomorphic study of New Zealand, part 4

    NASA Technical Reports Server (NTRS)

    Probine, M. C.; Suggate, R. P.; Stirling, I. F.; Mcgreevy, M. G. (Principal Investigator)

    1976-01-01

    The author has identified the following significant results. As part of the tape reformatting process, a simple coded picture output program was developed. This represents Pixel's radiance level by one of a 47 character set on a nonoverprinting line printer. It not only has aided in locating areas for the reformatting process, but has also formed the foundation for a supervised clustering package. This in turn has led to a simplistic but effective thematic mapping package.

  12. A Comprehensive Computer Package for Ambulatory Surgical Facilities

    PubMed Central

    Kessler, Robert R.

    1980-01-01

    Ambulatory surgical centers are a cost effective alternative to hospital surgery. Their increasing popularity has contributed to heavy case loads, an accumulation of vast amounts of medical and financial data and economic pressures to maintain a tight control over “cash flow”. Computerization is now a necessity to aid ambulatory surgical centers to maintain their competitive edge. An on-line system is especially necessary as it allows interactive scheduling of surgical cases, immediate access to financial data and rapid gathering of medical and statistical information. This paper describes the significant features of the computer package in use at the Salt Lake Surgical Center, which processes 500 cases per month.

  13. Rapid high-throughput characterisation, classification and selection of recombinant mammalian cell line phenotypes using intact cell MALDI-ToF mass spectrometry fingerprinting and PLS-DA modelling.

    PubMed

    Povey, Jane F; O'Malley, Christopher J; Root, Tracy; Martin, Elaine B; Montague, Gary A; Feary, Marc; Trim, Carol; Lang, Dietmar A; Alldread, Richard; Racher, Andrew J; Smales, C Mark

    2014-08-20

    Despite many advances in the generation of high producing recombinant mammalian cell lines over the last few decades, cell line selection and development is often slowed by the inability to predict a cell line's phenotypic characteristics (e.g. growth or recombinant protein productivity) at larger scale (large volume bioreactors) using data from early cell line construction at small culture scale. Here we describe the development of an intact cell MALDI-ToF mass spectrometry fingerprinting method for mammalian cells early in the cell line construction process whereby the resulting mass spectrometry data are used to predict the phenotype of mammalian cell lines at larger culture scale using a Partial Least Squares Discriminant Analysis (PLS-DA) model. Using MALDI-ToF mass spectrometry, a library of mass spectrometry fingerprints was generated for individual cell lines at the 96 deep well plate stage of cell line development. The growth and productivity of these cell lines were evaluated in a 10L bioreactor model of Lonza's large-scale (up to 20,000L) fed-batch cell culture processes. Using the mass spectrometry information at the 96 deep well plate stage and phenotype information at the 10L bioreactor scale a PLS-DA model was developed to predict the productivity of unknown cell lines at the 10L scale based upon their MALDI-ToF fingerprint at the 96 deep well plate scale. This approach provides the basis for the very early prediction of cell lines' performance in cGMP manufacturing-scale bioreactors and the foundation for methods and models for predicting other mammalian cell phenotypes from rapid, intact-cell mass spectrometry based measurements. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Casein gene expression in mouse mammary epithelial cell lines: Dependence upon extracellular matrix and cell type

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Medina, D.; Oborn, C.J.; Li, M.L.

    1987-09-01

    The COMMA-D mammary cell line exhibits mammary-specific functional differentiation under appropriate conditions in cell culture. The cytologically heterogeneous COMMA-D parental line and the clonal lines DB-1, TA-5, and FA-1 derived from the COMMA-D parent were examined for similar properties of functional differentiation. In monolayer cell culture, the cell lines DB-1, TA-5, FA-1, and MA-4 were examined for expression of mammary-specific and epithelial-specific proteins by an indirect immunofluorescence assay. The clonal cell lines were relatively homogeneous in their respective staining properties and seemed to represent three subpopulations found in the heterogeneous parental COMMA-D lines. None of the four clonal lines appearedmore » to represent myoepithelial cells. The cell lines were examined for expression of {beta}-casein mRNA in the presence or absence of prolactin. The inducibility of {beta}-casein in the COMMA-D cell line was further enhanced by a reconstituted basement membrane preparation enriched in laminin, collagen IV, and proteoglycans. These results support the hypothesis that the functional response of inducible mammary cell populations is a result of interaction among hormones, multiple extracellular matrix components, and specific cell types.« less

  15. Adventitious viruses in insect cell lines used for recombinant protein expression.

    PubMed

    Geisler, Christoph; Jarvis, Donald L

    2018-04-01

    Insect cells are widely used for recombinant protein expression, typically as hosts for recombinant baculovirus vectors, but also for plasmid-mediated transient transfection or stable genetic transformation. Insect cells are used to express proteins for research, as well as to manufacture biologicals for human and veterinary medicine. Recently, several insect cell lines used for recombinant protein expression were found to be persistently infected with adventitious viruses. This has raised questions about how these infections might affect research performed using those cell lines. Furthermore, these findings raised serious concerns about the safety of biologicals produced using those cell lines. In response, new insect cell lines lacking adventitious viruses have been isolated for use as improved research tools and safer biological manufacturing platforms. Here, we review the scientific and patent literature on adventitious viruses found in insect cell lines, affected cell lines, and new virus-free cell lines. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Effects of curcumin on stem-like cells in human esophageal squamous carcinoma cell lines.

    PubMed

    Almanaa, Taghreed N; Geusz, Michael E; Jamasbi, Roudabeh J

    2012-10-24

    Many cancers contain cell subpopulations that display characteristics of stem cells. Because these cancer stem cells (CSCs) appear to provide resistance to chemo-radiation therapy, development of therapeutic agents that target CSCs is essential. Curcumin is a phytochemical agent that is currently used in clinical trials to test its effectiveness against cancer. However, the effect of curcumin on CSCs is not well established. The current study evaluated curcumin-induced cell death in six cancer cell lines derived from human esophageal squamous cell carcinomas. Moreover, these cell lines and the ones established from cells that survived curcumin treatments were characterized. Cell loss was assayed after TE-1, TE-8, KY-5, KY-10, YES-1, and YES-2 cells were exposed to 20-80 μM curcumin for 30 hrs. Cell lines surviving 40 or 60 μM curcumin were established from these six original lines. The stem cell markers aldehyde dehydrogenase-1A1 (ALDH1A1) and CD44 as well as NF-κB were used to compare CSC-like subpopulations within and among the original lines as well as the curcumin-surviving lines. YES-2 was tested for tumorsphere-forming capabilities. Finally, the surviving lines were treated with 40 and 60 μM curcumin to determine whether their sensitivity was different from the original lines. The cell loss after curcumin treatment increased in a dose-dependent manner in all cell lines. The percentage of cells remaining after 60 μM curcumin treatment varied from 10.9% to 36.3% across the six lines. The cell lines were heterogeneous with respect to ALDH1A1, NF-κB and CD44 expression. KY-5 and YES-1 were the least sensitive and had the highest number of stem-like cells whereas TE-1 had the lowest. The curcumin-surviving lines showed a significant loss in the high staining ALDH1A1 and CD44 cell populations. Tumorspheres formed from YES-2 but were small and rare in the YES-2 surviving line. The curcumin-surviving lines showed a small but significant decrease in sensitivity to curcumin when compared with the original lines. Our results suggest that curcumin not only eliminates cancer cells but also targets CSCs. Therefore, curcumin may be an effective compound for treating esophageal and possibly other cancers in which CSCs can cause tumor recurrence.

  17. Cytoplasmic sequestration of the tumor suppressor p53 by a heat shock protein 70 family member, mortalin, in human colorectal adenocarcinoma cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gestl, Erin E., E-mail: egestl@wcupa.edu; Anne Boettger, S., E-mail: aboettger@wcupa.edu

    2012-06-29

    Highlights: Black-Right-Pointing-Pointer Eight human colorectal cell lines were evaluated for p53 and mortalin localization. Black-Right-Pointing-Pointer Six cell lines displayed cytoplasmic sequestration of the tumor suppressor p53. Black-Right-Pointing-Pointer Direct interaction between mortalin and p53 was shown in five cell lines. Black-Right-Pointing-Pointer Cell lines positive for p53 sequestration yielded elevated p53 expression levels. Black-Right-Pointing-Pointer This study yields the first evidence of cytoplasmic sequestration p53 by mortalin. -- Abstract: While it is known that cytoplasmic retention of p53 occurs in many solid tumors, the mechanisms responsible for this retention have not been positively identified. Since heatshock proteins like mortalin have been associated withmore » p53 inactivation in other tumors, the current study sought to characterize this potential interaction in never before examined colorectal adenocarcinoma cell lines. Six cell lines, one with 3 different fractions, were examined to determine expression of p53 and mortalin and characterize their cellular localization. Most of these cell lines displayed punctate p53 and mortalin localization in the cell cytoplasm with the exception of HCT-8 and HCT116 379.2 cells, where p53 was not detected. Nuclear p53 was only observed in HCT-116 40-16, LS123, and HT-29 cell lines. Mortalin was only localized in the cytoplasm in all cell lines. Co-immunoprecipitation and immunohistochemistry revealed that p53 and mortalin were bound and co-localized in the cytoplasmic fraction of four cell lines, HCT-116 (40-16 and 386; parental and heterozygous fractions respectively of the same cell line), HT-29, LS123 and LoVo, implying that p53 nuclear function is limited in those cell lines by being restricted to the cytoplasm. Mortalin gene expression levels were higher than gene expression levels of p53 in all cell lines. Cell lines with cytoplasmic sequestration of p53, however, also displayed elevated p53 gene expression levels compared to cell lines without p53 sequestration. Our data reveal the characteristic cytoplasmic sequestration of p53 by the heat shock protein mortalin in human colorectal adenocarcinoma cell lines, as is the case for other cancers, such as glioblastomas and hepatocellular carcinomas.« less

  18. Enhancing the cellular uptake of siRNA duplexes following noncovalent packaging with protein transduction domain peptides.

    PubMed

    Meade, Bryan R; Dowdy, Steven F

    2008-03-01

    The major limitation in utilizing information rich macromolecules for basic science and therapeutic applications is the inability of these large molecules to readily diffuse across the cellular membrane. While this restriction represents an efficient defense system against cellular penetration of unwanted foreign molecules and thus a crucial component of cell survival, overcoming this cellular characteristic for the intracellular delivery of macromolecules has been the focus of a large number of research groups worldwide. Recently, with the discovery of RNA interference, many of these groups have redirected their attention and have applied previously characterized cell delivery methodologies to synthetic short interfering RNA duplexes (siRNA). Protein transduction domain and cell penetrating peptides have been shown to enhance the delivery of multiple types of macromolecular cargo including peptides, proteins and antisense oligonucleotides and are now being utilized to enhance the cellular uptake of siRNA molecules. The dense cationic charge of these peptides that is critical for interaction with cell membrane components prior to internalization has also been shown to readily package siRNA molecules into stable nanoparticles that are capable of traversing the cell membrane. This review discusses the recent advances in noncovalent packaging of siRNA molecules with cationic peptides and the potential for the resulting complexes to successfully induce RNA interference within both in vitro and in vivo settings.

  19. Ebola Virus VP35-VP40 Interaction Is Sufficient for Packaging 3E-5E Minigenome RNA into Virus-Like Particles

    PubMed Central

    Johnson, Reed F.; McCarthy, Sarah E.; Godlewski, Peter J.; Harty, Ronald N.

    2006-01-01

    The packaging of viral genomic RNA into nucleocapsids and subsequently into virions is not completely understood. Phosphoprotein (P) and nucleoprotein (NP) interactions link NP-RNA complexes with P-L (polymerase) complexes to form viral nucleocapsids. The nucleocapsid then interacts with the viral matrix protein, leading to specific packaging of the nucleocapsid into the virion. A mammalian two-hybrid assay and confocal microscopy were used to demonstrate that Ebola virus VP35 and VP40 interact and colocalize in transfected cells. VP35 was packaged into budding virus-like particles (VLPs) as observed by protease protection assays. Moreover, VP40 and VP35 were sufficient for packaging an Ebola virus minignome RNA into VLPs. Results from immunoprecipitation-reverse transcriptase PCR experiments suggest that VP35 confers specificity of the nucleocapsid for viral genomic RNA by direct VP35-RNA interactions. PMID:16698994

  20. Development of a new canine osteosarcoma cell line.

    PubMed

    Séguin, B; Zwerdling, T; McCallan, J L; DeCock, H E V; Dewe, L L; Naydan, D K; Young, A E; Bannasch, D L; Foreman, O; Kent, M S

    2006-12-01

    Establishing a canine osteosarcoma (OSA) cell line can be useful to develop in vivo and in vitro models of OSA. The goal of this study was to develop, characterize and authenticate a new canine OSA cell line and a clone. A cell line and a clone were developed with standard cell culture techniques from a naturally occurring OSA in a dog. The clonal cell line induced a tumour after injection in RAG 1-deficient mouse. Histology was consistent with OSA. The original tumour from the dog and the tumour induced in the mouse were both reactive with vimentin and osteonectin (ON). The parent cell line and clonal cell line were reactive with ON, osteocalcin and alkaline phosphatase. Loss of heterozygosity was found in the same three microsatellite markers in the parent and clonal cell lines, and the tumour tissue grown in the mouse.

  1. Validation and calibration of a TDLAS oxygen sensor for in-line measurement on flow-packed products

    NASA Astrophysics Data System (ADS)

    Cocola, L.; Fedel, M.; Allermann, H.; Landa, S.; Tondello, G.; Bardenstein, A.; Poletto, L.

    2016-05-01

    A device based on Tunable Diode Laser Absorption Spectroscopy has been developed for non-invasive evaluation of gaseous oxygen concentration inside packed food containers. This work has been done in the context of the SAFETYPACK European project in order to enable full, automated product testing on a production line. The chosen samples at the end of the manufacturing process are modified atmosphere bags of processed mozzarella, in which the target oxygen concentration is required to be below 5%. The spectrometer allows in-line measurement of moving samples which are passing on a conveyor belt, with an optical layout optimized for bags made of a flexible scattering material, and works by sensing the gas phase in the headspace at the top of the package. A field applicable method for the calibration of this device has been identified and validated against traditional, industry standard, invasive measurement techniques. This allows some degrees of freedom for the end-user regarding packaging dimensions and shape. After deployment and setup of the instrument at the end-user manufacturing site, performance has been evaluated on a different range of samples in order to validate the choice of electro optical and geometrical parameters regarding sample handling and measurement timing at the actual measurement conditions.

  2. Reference Maps of Human ES and iPS Cell Variation Enable High-Throughput Characterization of Pluripotent Cell Lines

    PubMed Central

    Bock, Christoph; Kiskinis, Evangelos; Verstappen, Griet; Gu, Hongcang; Boulting, Gabriella; Smith, Zachary D.; Ziller, Michael; Croft, Gist F.; Amoroso, Mackenzie W.; Oakley, Derek H.; Gnirke, Andreas; Eggan, Kevin; Meissner, Alexander

    2011-01-01

    SUMMARY The developmental potential of human pluripotent stem cells suggests that they can produce disease-relevant cell types for biomedical research. However, substantial variation has been reported among pluripotent cell lines, which could affect their utility and clinical safety. Such cell-line-specific differences must be better understood before one can confidently use embryonic stem (ES) or induced pluripotent stem (iPS) cells in translational research. Toward this goal we have established genome-wide reference maps of DNA methylation and gene expression for 20 previously derived human ES lines and 12 human iPS cell lines, and we have measured the in vitro differentiation propensity of these cell lines. This resource enabled us to assess the epigenetic and transcriptional similarity of ES and iPS cells and to predict the differentiation efficiency of individual cell lines. The combination of assays yields a scorecard for quick and comprehensive characterization of pluripotent cell lines. PMID:21295703

  3. Documentation of a computer program to simulate transient leakage from confining units using the modular finite-difference, ground-water flow model

    USGS Publications Warehouse

    Leake, S.A.; Leahy, P.P.; Navoy, A.S.

    1994-01-01

    Transient leakage into or out of a compressible fine-grained confining unit results from ground- water storage changes within the unit. The computer program described in this report provides a new method of simulating transient leakage using the U.S. Geological Survey modular finite- difference ground-water flow model (MODFLOW). The new program is referred to as the Transient- Leakage Package. The Transient-Leakage Package solves integrodifferential equations that describe flow across the upper and lower boundaries of confining units. For each confining unit, vertical hydraulic conductivity, thickness, and specific storage are specified in input arrays. These properties can vary from cell to cell and the confining unit need not be present at all locations in the grid; however, the confining units must be bounded above and below by model layers in which head is calculated or specified. The package was used in an example problem to simulate drawdown around a pumping well in a system with two aquifers separated by a confining unit. For drawdown values in excess of 1 centimeter, the solution using the new package closely matched an exact analytical solution. The problem also was simulated without the new package by using a separate model layer to represent the confining unit. That simulation was refined by using two model layers to represent the confining unit. The simulation using the Transient-Leakage Package was faster and more accurate than either of the simulations using model layers to represent the confining unit.

  4. Genetics Home Reference: cyclic vomiting syndrome

    MedlinePlus

    ... childhood, may be related to changes in mitochondrial DNA . Mitochondria are structures within cells that convert the ... a form that cells can use. Although most DNA is packaged in chromosomes within the nucleus, mitochondria ...

  5. CellNOptR: a flexible toolkit to train protein signaling networks to data using multiple logic formalisms.

    PubMed

    Terfve, Camille; Cokelaer, Thomas; Henriques, David; MacNamara, Aidan; Goncalves, Emanuel; Morris, Melody K; van Iersel, Martijn; Lauffenburger, Douglas A; Saez-Rodriguez, Julio

    2012-10-18

    Cells process signals using complex and dynamic networks. Studying how this is performed in a context and cell type specific way is essential to understand signaling both in physiological and diseased situations. Context-specific medium/high throughput proteomic data measured upon perturbation is now relatively easy to obtain but formalisms that can take advantage of these features to build models of signaling are still comparatively scarce. Here we present CellNOptR, an open-source R software package for building predictive logic models of signaling networks by training networks derived from prior knowledge to signaling (typically phosphoproteomic) data. CellNOptR features different logic formalisms, from Boolean models to differential equations, in a common framework. These different logic model representations accommodate state and time values with increasing levels of detail. We provide in addition an interface via Cytoscape (CytoCopteR) to facilitate use and integration with Cytoscape network-based capabilities. Models generated with this pipeline have two key features. First, they are constrained by prior knowledge about the network but trained to data. They are therefore context and cell line specific, which results in enhanced predictive and mechanistic insights. Second, they can be built using different logic formalisms depending on the richness of the available data. Models built with CellNOptR are useful tools to understand how signals are processed by cells and how this is altered in disease. They can be used to predict the effect of perturbations (individual or in combinations), and potentially to engineer therapies that have differential effects/side effects depending on the cell type or context.

  6. CellNOptR: a flexible toolkit to train protein signaling networks to data using multiple logic formalisms

    PubMed Central

    2012-01-01

    Background Cells process signals using complex and dynamic networks. Studying how this is performed in a context and cell type specific way is essential to understand signaling both in physiological and diseased situations. Context-specific medium/high throughput proteomic data measured upon perturbation is now relatively easy to obtain but formalisms that can take advantage of these features to build models of signaling are still comparatively scarce. Results Here we present CellNOptR, an open-source R software package for building predictive logic models of signaling networks by training networks derived from prior knowledge to signaling (typically phosphoproteomic) data. CellNOptR features different logic formalisms, from Boolean models to differential equations, in a common framework. These different logic model representations accommodate state and time values with increasing levels of detail. We provide in addition an interface via Cytoscape (CytoCopteR) to facilitate use and integration with Cytoscape network-based capabilities. Conclusions Models generated with this pipeline have two key features. First, they are constrained by prior knowledge about the network but trained to data. They are therefore context and cell line specific, which results in enhanced predictive and mechanistic insights. Second, they can be built using different logic formalisms depending on the richness of the available data. Models built with CellNOptR are useful tools to understand how signals are processed by cells and how this is altered in disease. They can be used to predict the effect of perturbations (individual or in combinations), and potentially to engineer therapies that have differential effects/side effects depending on the cell type or context. PMID:23079107

  7. The Encapsidated Genome of Microplitis demolitor Bracovirus Integrates into the Host Pseudoplusia includens ▿ ‡

    PubMed Central

    Beck, Markus H.; Zhang, Shu; Bitra, Kavita; Burke, Gaelen R.; Strand, Michael R.

    2011-01-01

    Polydnaviruses (PDVs) are symbionts of parasitoid wasps that function as gene delivery vehicles in the insects (hosts) that the wasps parasitize. PDVs persist in wasps as integrated proviruses but are packaged as circularized and segmented double-stranded DNAs into the virions that wasps inject into hosts. In contrast, little is known about how PDV genomic DNAs persist in host cells. Microplitis demolitor carries Microplitis demolitor bracovirus (MdBV) and parasitizes the host Pseudoplusia includens. MdBV infects primarily host hemocytes and also infects a hemocyte-derived cell line from P. includens called CiE1 cells. Here we report that all 15 genomic segments of the MdBV encapsidated genome exhibited long-term persistence in CiE1 cells. Most MdBV genes expressed in hemocytes were persistently expressed in CiE1 cells, including members of the glc gene family whose products transformed CiE1 cells into a suspension culture. PCR-based integration assays combined with cloning and sequencing of host-virus junctions confirmed that genomic segments J and C persisted in CiE1 cells by integration. These genomic DNAs also rapidly integrated into parasitized P. includens. Sequence analysis of wasp-viral junction clones showed that the integration of proviral segments in M. demolitor was associated with a wasp excision/integration motif (WIM) known from other bracoviruses. However, integration into host cells occurred in association with a previously unknown domain that we named the host integration motif (HIM). The presence of HIMs in most MdBV genomic DNAs suggests that the integration of each genomic segment into host cells occurs through a shared mechanism. PMID:21880747

  8. Stakeholder Views of Nanosilver Linings: Macroethics Education and Automated Text Analysis Through Participatory Governance Role Play in a Workshop Format.

    PubMed

    Dempsey, Joshua; Stamets, Justin; Eggleson, Kathleen

    2017-06-01

    The Nanosilver Linings role play case offers participants first-person experience with interpersonal interaction in the context of the wicked problems of emerging technology macroethics. In the fictional scenario, diverse societal stakeholders convene at a town hall meeting to consider whether a nanotechnology-enabled food packaging industry should be offered incentives to establish an operation in their economically struggling Midwestern city. This original creative work was built with a combination of elements, selected for their established pedagogical efficacy (e.g. active learning, case-based learning) and as topical dimensions of the realistic scenario (e.g. nanosilver in food packaging, occupational safety and health). The product life cycle is used as a framework for integrated consideration of scientific, societal, and ethical issues. The Nanosilver Linings hypothetical case was delivered through the format of the 3-hour workshop Ethics when Biocomplexity meets Human Complexity, providing an immersive, holistic ethics learning experience for STEM graduate students. Through their participation in the Nanosilver Linings case and Ethics when Biocomplexity meets Human Complexity workshop, four cohorts of science and engineering doctoral students reported the achievement of specific learning objectives pertaining to a range of macroethics concepts and professional practices, including stakeholder perspectives, communication, human values, and ethical frameworks. Automated text analysis of workshop transcripts revealed differences in sentiment and in ethical framework (consequentialism/deontology) preference between societal stakeholder roles. These resources have been recognized as ethics education exemplars by the U.S. National Academy of Engineering .

  9. Cancer stem cell-like cells from a single cell of oral squamous carcinoma cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Felthaus, O.; Department of Oral and Maxillofacial Surgery, University of Regensburg; Ettl, T.

    2011-04-01

    Research highlights: {yields} Four oral squamous cancer cell lines (OSCCL) were analyzed for cancer stem cells (CSCs). {yields} Single cell derived colonies of OSCCL express CSC-marker CD133 differentially. {yields} Monoclonal cell lines showed reduced sensitivity for Paclitaxel. {yields} In situ CD133{sup +} cells are slow cycling (Ki67-) indicating a reduced drug sensitivity. {yields} CD133{sup +} and CSC-like cells can be obtained from single colony forming cells of OSCCL. -- Abstract: Resistance of oral squamous cell carcinomas (OSCC) to conventional chemotherapy or radiation therapy might be due to cancer stem cells (CSCs). The development of novel anticancer drugs requires a simplemore » method for the enrichment of CSCs. CSCs can be enriched from OSCC cell lines, for example, after cultivation in serum-free cell culture medium (SFM). In our study, we analyzed four OSCC cell lines for the presence of CSCs. CSC-like cells could not be enriched with SFM. However, cell lines obtained from holoclone colonies showed CSC-like properties such as a reduced rate of cell proliferation and a reduced sensitivity to Paclitaxel in comparison to cells from the parental lineage. Moreover, these cell lines differentially expressed the CSC-marker CD133, which is also upregulated in OSCC tissues. Interestingly, CD133{sup +} cells in OSCC tissues expressed little to no Ki67, the cell proliferation marker that also indicates reduced drug sensitivity. Our study shows a method for the isolation of CSC-like cell lines from OSCC cell lines. These CSC-like cell lines could be new targets for the development of anticancer drugs under in vitro conditions.« less

  10. Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

    PubMed Central

    Gardell, Alison M.; Qin, Qin; Rice, Robert H.; Li, Johnathan; Kültz, Dietmar

    2014-01-01

    Fish cell cultures are becoming more widely used models for investigating molecular mechanisms of physiological response to environmental challenge. In this study, we derived two immortalized Mozambique tilapia (Oreochromis mossambicus) cell lines from brain (OmB) and lip epithelium (OmL), and compared them to a previously immortalized bulbus arteriosus (TmB) cell line. The OmB and OmL cell lines were generated without or with Rho-associated kinase (ROCK) inhibitor/3T3 feeder layer supplementation. Although both approaches were successful, ROCK inhibitor/feeder layer supplementation was found to offer the advantages of selecting for epithelial-like cell type and decreasing time to immortalization. After immortalization (≥ passage 5), we characterized the proteomes of the newly derived cell lines (OmB and OmL) using LCMS and identified several unique cell markers for each line. Subsequently, osmotolerance for each of the three cell lines following acute exposure to elevated sodium chloride was evaluated. The acute maximum osmotolerance of these tilapia cell lines (>700 mOsm/kg) was markedly higher than that of any other known vertebrate cell line, but was significantly higher in the epithelial-like OmL cell line. To validate the physiological relevance of these tilapia cell lines, we quantified the effects of acute hyperosmotic challenge (450 mOsm/kg and 700 mOsm/kg) on the transcriptional regulation of two enzymes involved in biosynthesis of the compatible organic osmolyte, myo-inositol. Both enzymes were found to be robustly upregulated in all three tilapia cell lines. Therefore, the newly established tilapia cells lines represent valuable tools for studying molecular mechanisms involved in the osmotic stress response of euryhaline fish. PMID:24797371

  11. HIV integration sites in latently infected cell lines: evidence of ongoing replication.

    PubMed

    Symons, Jori; Chopra, Abha; Malatinkova, Eva; De Spiegelaere, Ward; Leary, Shay; Cooper, Don; Abana, Chike O; Rhodes, Ajantha; Rezaei, Simin D; Vandekerckhove, Linos; Mallal, Simon; Lewin, Sharon R; Cameron, Paul U

    2017-01-13

    Assessing the location and frequency of HIV integration sites in latently infected cells can potentially inform our understanding of how HIV persists during combination antiretroviral therapy. We developed a novel high throughput sequencing method to evaluate HIV integration sites in latently infected cell lines to determine whether there was virus replication or clonal expansion in these cell lines observed as multiple integration events at the same position. We modified a previously reported method using random DNA shearing and PCR to allow for high throughput robotic processing to identify the site and frequency of HIV integration in latently infected cell lines. Latently infected cell lines infected with intact virus demonstrated multiple distinct HIV integration sites (28 different sites in U1, 110 in ACH-2 and 117 in J1.1 per 150,000 cells). In contrast, cell lines infected with replication-incompetent viruses (J-Lat cells) demonstrated single integration sites. Following in vitro passaging of the ACH-2 cell line, we observed a significant increase in the frequency of unique HIV integration sites and there were multiple mutations and large deletions in the proviral DNA. When the ACH-2 cell line was cultured with the integrase inhibitor raltegravir, there was a significant decrease in the number of unique HIV integration sites and a transient increase in the frequency of 2-LTR circles consistent with virus replication in these cells. Cell lines latently infected with intact HIV demonstrated multiple unique HIV integration sites indicating that these cell lines are not clonal and in the ACH-2 cell line there was evidence of low level virus replication. These findings have implications for the use of latently infected cell lines as models of HIV latency and for the use of these cells as standards.

  12. User's Guide for MapIMG 2: Map Image Re-projection Software Package

    USGS Publications Warehouse

    Finn, Michael P.; Trent, Jason R.; Buehler, Robert A.

    2006-01-01

    BACKGROUND Scientists routinely accomplish small-scale geospatial modeling in the raster domain, using high-resolution datasets for large parts of continents and low-resolution to high-resolution datasets for the entire globe. Direct implementation of point-to-point transformation with appropriate functions yields the variety of projections available in commercial software packages, but implementation with data other than points requires specific adaptation of the transformation equations or prior preparation of the data to allow the transformation to succeed. It seems that some of these packages use the U.S. Geological Survey's (USGS) General Cartographic Transformation Package (GCTP) or similar point transformations without adaptation to the specific characteristics of raster data (Usery and others, 2003a). Usery and others (2003b) compiled and tabulated the accuracy of categorical areas in projected raster datasets of global extent. Based on the shortcomings identified in these studies, geographers and applications programmers at the USGS expanded and evolved a USGS software package, MapIMG, for raster map projection transformation (Finn and Trent, 2004). Daniel R. Steinwand of Science Applications International Corporation, National Center for Earth Resources Observation and Science, originally developed MapIMG for the USGS, basing it on GCTP. Through previous and continuing efforts at the USGS' National Geospatial Technical Operations Center, this program has been transformed from an application based on command line input into a software package based on a graphical user interface for Windows, Linux, and other UNIX machines.

  13. Case study: Is the 'catch-all-plastics bin' useful in unlocking the hidden resource potential in the residual waste collection system?

    PubMed

    Kranzinger, Lukas; Schopf, Kerstin; Pomberger, Roland; Punesch, Elisabeth

    2017-02-01

    Austria's performance in the collection of separated waste is adequate. However, the residual waste still contains substantial amounts of recyclable materials - for example, plastics, paper and board, glass and composite packaging. Plastics (lightweight packaging and similar non-packaging materials) are detected at an average mass content of 13% in residual waste. Despite this huge potential, only 3% of the total amount of residual waste (1,687,000 t y -1 ) is recycled. This implies that most of the recyclable materials contained in the residual waste are destined for thermal recovery and are lost for recycling. This pilot project, commissioned by the Land of Lower Austria, applied a holistic approach, unique in Europe, to the Lower Austrian waste management system. It aims to transfer excess quantities of plastic packaging and non-packaging recyclables from the residual waste system to the separately collected waste system by introducing a so-called 'catch-all-plastics bin'. A quantity flow model was constructed and the results showed a realistic increase in the amount of plastics collected of 33.9 wt%. This equals a calculated excess quantity of 19,638 t y -1 . The increased plastics collection resulted in a positive impact on the climate footprint (CO 2 equivalent) in line with the targets of EU Directive 94/62/EG (Circular Economy Package) and its Amendments. The new collection system involves only moderate additional costs.

  14. Cell lines authentication and mycoplasma detection as minimun quality control of cell lines in biobanking.

    PubMed

    Corral-Vázquez, C; Aguilar-Quesada, R; Catalina, P; Lucena-Aguilar, G; Ligero, G; Miranda, B; Carrillo-Ávila, J A

    2017-06-01

    Establishment of continuous cell lines from human normal and tumor tissues is an extended and useful methodology for molecular characterization of cancer pathophysiology and drug development in research laboratories. The exchange of these cell lines between different labs is a common practice that can compromise assays reliability due to contamination with microorganism such as mycoplasma or cells from different flasks that compromise experiment reproducibility and reliability. Great proportions of cell lines are contaminated with mycoplasma and/or are replaced by cells derived for a different origin during processing or distribution process. The scientific community has underestimated this problem and thousand of research experiment has been done with cell lines that are incorrectly identified and wrong scientific conclusions have been published. Regular contamination and authentication tests are necessary in order to avoid negative consequences of widespread misidentified and contaminated cell lines. Cell banks generate, store and distribute cell lines for research, being mandatory a consistent and continuous quality program. Methods implementation for guaranteeing both, the absence of mycoplasma and authentication in the supplied cell lines, has been performed in the Andalusian Health System Biobank. Specifically, precise results were obtained using real time PCR detection for mycoplasma and 10 STRs identification by capillary electrophoresis for cell line authentication. Advantages and disadvantages of these protocols are discussed.

  15. The BRL-CAD Package: An Overview

    DTIC Science & Technology

    2013-04-01

    many different display devices to be supported. The types of primatives supported include: arbitrary boxes of up to eight verticies, ellipsoids...file size. Many algorithms simply run until all of the data is gone, and some don’t even care about scan lines at aiL 5.2. Format Conversion Several

  16. Back-illuminated CCD imager adapted for contrast transfer function measurements thereon

    NASA Technical Reports Server (NTRS)

    Levine, Peter A. (Inventor)

    1987-01-01

    Stripe patterns of varying spatial frequency, formed in the top-metalization of a back-illuminated solid-state imager, facilitate on-line measurement of contrast transfer function during wafer-probe testing. The imager may be packaged to allow front-illumination during in-the-field testing after its manufacture.

  17. Multiple Learning Strategies Project. Small Engine Repair. Visually Impaired.

    ERIC Educational Resources Information Center

    Foster, Don; And Others

    This instructional package designed for visually impaired students, focuses on the vocational area of small engine repair. Contained in this document are forty learning modules organized into fourteen units: engine block; starters; fuel tank, lines, filters and pumps; carburetors; electrical; test equipment; motorcycle; machining; tune-ups; short…

  18. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Doug Blankenship

    ArcGIS Map Package with MT Station Locations, 2D Seismic Lines, Well data, Known Regional Hydrothermal Systems, Regional Historic Earthquake Seismicity, Regional Temperature Gradient Data, Regional Heat Flow Data, Regional Radiogenic Heat Production, Local Geology, Land Status, Cultural Data, 2m Temperature Probe Data, and Gravity Data. Also a detailed down-hole lithology notes are provided.

  19. Uveal Melanoma Cell Lines: Where do they come from? (An American Ophthalmological Society Thesis).

    PubMed

    Jager, Martine J; Magner, J Antonio Bermudez; Ksander, Bruce R; Dubovy, Sander R

    2016-08-01

    To determine whether some of the most often used uveal melanoma cell lines resemble their original tumor. Analysis of the literature, patient charts, histopathology, mutations, chromosome status, HLA type, and expression of melanocyte markers on cell lines and their primary tumors. We examined five cell lines and the primary tumors from which they were derived. Four of the five examined primary tumors were unusual: one occupied the orbit, two were recurrences after prior irradiation, and one developed in an eye with a nevus of Ota. One cell line did not contain the GNA11 mutation, but it was present in the primary tumor. Three of the primary tumors had monosomy 3 (two of these lacked BAP1 expression); however, all five cell lines showed disomy 3 and BAP1 expression. All of the cell lines had gain of 8q. Two cell lines lacked expression of melanocyte markers, although these were present in the corresponding primary tumor. All cell lines could be traced back to their original uveal melanoma. Four of the five primary tumors were unusual. Cell lines often differed from their primary tumor in chromosome status and melanocyte markers. However, their specific chromosome aberrations and capacity to continue proliferation characterize them as uveal melanoma cell lines.

  20. Uveal Melanoma Cell Lines: Where do they come from? (An American Ophthalmological Society Thesis)

    PubMed Central

    Jager, Martine J.; Magner, J. Antonio Bermudez; Ksander, Bruce R.; Dubovy, Sander R.

    2016-01-01

    Purpose To determine whether some of the most often used uveal melanoma cell lines resemble their original tumor. Methods Analysis of the literature, patient charts, histopathology, mutations, chromosome status, HLA type, and expression of melanocyte markers on cell lines and their primary tumors. We examined five cell lines and the primary tumors from which they were derived. Results Four of the five examined primary tumors were unusual: one occupied the orbit, two were recurrences after prior irradiation, and one developed in an eye with a nevus of Ota. One cell line did not contain the GNA11 mutation, but it was present in the primary tumor. Three of the primary tumors had monosomy 3 (two of these lacked BAP1 expression); however, all five cell lines showed disomy 3 and BAP1 expression. All of the cell lines had gain of 8q. Two cell lines lacked expression of melanocyte markers, although these were present in the corresponding primary tumor. Conclusions All cell lines could be traced back to their original uveal melanoma. Four of the five primary tumors were unusual. Cell lines often differed from their primary tumor in chromosome status and melanocyte markers. However, their specific chromosome aberrations and capacity to continue proliferation characterize them as uveal melanoma cell lines. PMID:28018010

  1. Different toxic effects of YTX in tumor K-562 and lymphoblastoid cell lines

    PubMed Central

    Fernández-Araujo, Andrea; Sánchez, Jon A.; Alfonso, Amparo; Vieytes, Mercedes R.; Botana, Luis M.

    2015-01-01

    Yessotoxin (YTX) modulates cellular phosphodiesterases (PDEs). In this regard, opposite effects had been described in the tumor model K-562 cell line and fresh human lymphocytes in terms of cell viability, cyclic adenosine 3',5'-cyclic monophosphate (cAMP) production and protein expression after YTX treatment. Studies in depth of the pathways activated by YTX in K-562 cell line, have demonstrated the activation of two different cell death types, apoptosis, and autophagy after 24 and 48 h of treatment, respectively. Furthermore, the key role of type 4A PDE (PDE4A) in both pathways activated by YTX was demonstrated. Therefore, taking into account the differences between cellular lines and fresh cells, a study of cell death pathways activated by YTX in a non-tumor cell line with mitotic activity, was performed. The cellular model used was the lymphoblastoid cell line that represents a non-tumor model with normal apoptotic and mitotic machinery. In this context, cell viability and cell proliferation, expression of proteins involved in cell death activated by YTX and mitochondrial mass, were studied after the incubation with the toxin. Opposite to the tumor model, no cell death activation was observed in lymphoblastoid cell line in the presence of YTX. In this sense, variations in apoptosis hallmarks were not detected in the lymphoblastoid cell line after YTX incubation, whereas this type I of programmed cell death was observed in K-562 cells. On the other hand, autophagy cell death was triggered in this cellular line, while other autophagic process is suggested in lymphoblastoid cells. These YTX effects are related to PDE4A in both cellular lines. In addition, while cell death is triggered in K-562 cells after YTX treatment, in lymphoblastoid cells the toxin stops cellular proliferation. These results point to YTX as a specific toxic compound of tumor cells, since in the non-tumor lymphoblastoid cell line, no cell death hallmarks are observed. PMID:26136685

  2. Lung Cancer Cell Lines as Tools for Biomedical Discovery and Research

    PubMed Central

    Girard, Luc; Lockwood, William W.; Lam, Wan L.; Minna, John D.

    2010-01-01

    Lung cancer cell lines have made a substantial contribution to lung cancer translational research and biomedical discovery. A systematic approach to initiating and characterizing cell lines from small cell and non–small cell lung carcinomas has led to the current collection of more than 200 lung cancer cell lines, a number that exceeds those for other common epithelial cancers combined. The ready availability and widespread dissemination of the lines to investigators worldwide have resulted in more than 9000 citations, including multiple examples of important biomedical discoveries. The high (but not perfect) genomic similarities between lung cancer cell lines and the lung tumor type from which they were derived provide evidence of the relevance of their use. However, major problems including misidentification or cell line contamination remain. Ongoing studies and new approaches are expected to reveal the full potential of the lung cancer cell line panel. PMID:20679594

  3. Cell lines derived from feline fibrosarcoma display unstable chromosomal aneuploidy and additionally centrosome number aberrations.

    PubMed

    von Erichsen, J; Hecht, W; Löhberg-Gruene, C; Reinacher, M

    2012-07-01

    The purpose of the study was to evaluate clonality and presence of numerical chromosomal and centrosomal aberrations in 5 established feline fibrosarcoma cell lines and in a fetal dermal fibroblast cell line as a control. The clonality of all cell lines was examined using limited-dilution cloning. The number of chromosomes was counted in metaphase spreads. The immunocytochemical analysis of centrosome numbers was performed by indirect immunofluorescence using a monoclonal antibody that targets γ-tubulin, a well-characterized component of centrosomes. Monoclonal cell populations could be established from all cell lines. In all feline fibrosarcoma cell lines, the number of chromosomes deviated abnormally from the normal feline chromosome number of 2n = 38, ranging from 19 to 155 chromosomes per cell. Centrosome hyperamplification was observed in all 5 feline fibrosarcoma cell lines with a proportion of cells (5.7 to 15.2%) having more than 2 centrosomes. In the control cell line, only 0.6% of the cells had more than 2 centrosomes. In conclusion, the examinations revealed that centrosome hyperamplification occurs in feline fibrosarcoma cell lines. The feline fibrosarcoma cell lines possessed 10 to 25 times as many cells with centrosome hyperamplification as the control cell line. These observations suggest an association of numerical centrosome aberrations with karyotype instability by increasing the frequency of chromosome missegregation. The results of this study may be helpful for further characterization of feline fibrosarcomas and may contribute to the knowledge of cytogenetic factors that may be important for the pathogenesis of feline fibrosarcomas.

  4. Attempt to develop taste bud models in three-dimensional culture.

    PubMed

    Nishiyama, Miyako; Yuki, Saori; Fukano, Chiharu; Sako, Hideyuki; Miyamoto, Takenori; Tomooka, Yasuhiro

    2011-09-01

    Taste buds are the end organs of taste located in the gustatory papillae, which occur on the surface of the oral cavity. The goal of the present study was to establish a culture model mimicking the lingual taste bud of the mouse. To this end, three cell lines were employed: taste bud-derived cell lines (TBD cell lines), a lingual epithelial cell-derived cell line (20A cell line), and a mesenchymal cell-derived cell line (TMD cell line). TBD cells embedded in collagen gel formed three-dimensional clusters, which had an internal cavity equipped with a tight junction-like structure, a microvilluslike structure, and a laminin-positive layer surrounding the cluster. The cells with this epitheliumlike morphology expressed marker proteins of taste cells: gustducin and NCAM. TBD cells formed a monolayer on collagen gel when they were co-cultured with TMD cells. TBD, 20A, and TMD cell lines were maintained in a triple cell co-culture, in which TBD cells were pre-seeded as aggregates or in suspension on the collagen gel containing TMD cells, and 20A cells were laid over the TBD cells. TBD cells in the triple cell co-culture expressed NCAM. This result suggests that co-cultured TBD cells exhibited a characteristic of Type III taste cells. The culture model would be useful to study morphogenesis and functions of the gustatory organ.

  5. 16 CFR 503.4 - Net quantity of contents, numerical count.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... clearly expresses the fact that only one unit is contained in the package. Thus the unit synthetic sponge... sponge,” “one light bulb,” or “one dry cell battery.” However, there still exists the necessity to.... For example, the synthetic sponge which is packaged, requires dimensions such as “5 in. × 3 in. × 1 in...

  6. Enhancement of Radiation Response in Osteosarcoma and Rhabomyosarcoma Cell Lines by Histone Deacetylase Inhibition

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Blattmann, Claudia, E-mail: claudia.blattmann@med.uni-heidelberg.d; Oertel, Susanne; Ehemann, Volker

    2010-09-01

    Purpose: Histone deacetylase inhibitors (HDACIs) can enhance the sensitivity of cells to photon radiation treatment (XRT) by altering numerous molecular pathways. We investigated the effect of pan-HDACIs such as suberoylanilide hydroxamic acid (SAHA) on radiation response in two osteosarcoma (OS) and two rhabdomyosarcoma (RMS) cell lines. Methods and Materials: Clonogenic survival, cell cycle analysis, and apoptosis were examined in OS (KHOS-24OS, SAOS2) and RMS (A-204, RD) cell lines treated with HDACI and HDACI plus XRT, respectively. Protein expression was investigated via immunoblot analysis, and cell cycle analysis and measurement of apoptosis were performed using flow cytometry. Results: SAHA induced anmore » inhibition of cell proliferation and clonogenic survival in OS and RMS cell lines and led to a significant radiosensitization of all tumor cell lines. Other HDACI such as M344 and valproate showed similar effects as investigated in one OS cell line. Furthermore, SAHA significantly increased radiation-induced apoptosis in the OS cell lines, whereas in the RMS cell lines radiation-induced apoptosis was insignificant with and without SAHA. In all investigated sarcoma cell lines, SAHA attenuated radiation-induced DNA repair protein expression (Rad51, Ku80). Conclusion: Our results show that HDACIs enhance radiation action in OS and RMS cell lines. Inhibition of DNA repair, as well as increased apoptosis induction after exposure to HDACIs, can be mechanisms of radiosensitization by HDACIs.« less

  7. Generation of genome-modified Drosophila cell lines using SwAP.

    PubMed

    Franz, Alexandra; Brunner, Erich; Basler, Konrad

    2017-10-02

    The ease of generating genetically modified animals and cell lines has been markedly increased by the recent development of the versatile CRISPR/Cas9 tool. However, while the isolation of isogenic cell populations is usually straightforward for mammalian cell lines, the generation of clonal Drosophila cell lines has remained a longstanding challenge, hampered by the difficulty of getting Drosophila cells to grow at low densities. Here, we describe a highly efficient workflow to generate clonal Cas9-engineered Drosophila cell lines using a combination of cell pools, limiting dilution in conditioned medium and PCR with allele-specific primers, enabling the efficient selection of a clonal cell line with a suitable mutation profile. We validate the protocol by documenting the isolation, selection and verification of eight independently Cas9-edited armadillo mutant Drosophila cell lines. Our method provides a powerful and simple workflow that improves the utility of Drosophila cells for genetic studies with CRISPR/Cas9.

  8. Fluoroorotic acid-selected Nicotiana plumbaginifolia cell lines with a stable thymine starvation phenotype have lost the thymine-regulated transcriptional program.

    PubMed

    Santoso, D; Thornburg, R

    2000-08-01

    We have selected 143 independent Nicotiana plumbaginifolia cell lines that survive in the presence of 5-fluoroorotic acid. These lines show several diverse phenotypes. The majority of these cell lines showed reduced levels of UMP synthase. However, one particular phenotype, which represents 14% of the total independent lines (20 cell lines), showed an unexpected, high level of UMP synthase and was therefore analyzed in detail. The selected cell lines showed no differences with wild-type cells with respect to uptake of orotic acid, affinity of UMP synthase for its substrates, or UMP synthase gene-copy number. Alternative detoxification mechanisms were also excluded. The elevated enzyme activity was correlated with elevated UMP synthase protein levels as well as elevated UMP synthase mRNA levels. In contrast to wild-type cell lines, the fluoroorotic acid-selected cell lines did not respond to thymine or to other biochemicals that affect thymine levels. In addition, there was also a concomitant up-regulation of aspartate transcarbamoylase, however, dihydroorotase and dihydroorotate dehydrogenase are not up-regulated in these cell lines.

  9. Fluoroorotic Acid-Selected Nicotiana plumbaginifolia Cell Lines with a Stable Thymine Starvation Phenotype Have Lost the Thymine-Regulated Transcriptional Program1

    PubMed Central

    Santoso, Djoko; Thornburg, Robert

    2000-01-01

    We have selected 143 independent Nicotiana plumbaginifolia cell lines that survive in the presence of 5-fluoroorotic acid. These lines show several diverse phenotypes. The majority of these cell lines showed reduced levels of UMP synthase. However, one particular phenotype, which represents 14% of the total independent lines (20 cell lines), showed an unexpected, high level of UMP synthase and was therefore analyzed in detail. The selected cell lines showed no differences with wild-type cells with respect to uptake of orotic acid, affinity of UMP synthase for its substrates, or UMP synthase gene-copy number. Alternative detoxification mechanisms were also excluded. The elevated enzyme activity was correlated with elevated UMP synthase protein levels as well as elevated UMP synthase mRNA levels. In contrast to wild-type cell lines, the fluoroorotic acid-selected cell lines did not respond to thymine or to other biochemicals that affect thymine levels. In addition, there was also a concomitant up-regulation of aspartate transcarbamoylase, however, dihydroorotase and dihydroorotate dehydrogenase are not up-regulated in these cell lines. PMID:10938367

  10. The T47D cell line is an ideal experimental model to elucidate the progesterone-specific effects of a luminal A subtype of breast cancer.

    PubMed

    Yu, Sungryul; Kim, Taemook; Yoo, Kyung Hyun; Kang, Keunsoo

    2017-05-06

    Cell lines are often used as in vitro tools to mimic certain types of in vivo system; several cell lines, including MCF-7 and T47D, have been widely used in breast cancer studies without investigating the cell lines' characteristics. In this study, we compared the genome-wide binding profiles of ERα, PR, and P300, and the gene expression changes between MCF-7 and T47D cell lines that represent the luminal A subtype of breast cancer. Surprisingly, several thousand genes were differentially expressed under estrogenic condition. In addition, ERα, PR, and P300 binding to regulatory elements showed distinct genomic landscapes between MCF-7 and T47D cell lines in the same hormonal states. In particular, the T47D cell line was markedly susceptible to progesterone, whereas the MCF-7 cell line did not respond to progesterone in the presence of estrogen. Consistently, changes in the expression level of the PR-target gene, STAT5A, were only observed in the T47D cell line, not the MCF-7 cell line, when treated with progesterone. Overall, the results highlight the importance of selecting appropriate cell lines for breast cancer studies and suggest that T47D cell lines can be an ideal experimental model to elucidate the progesterone-specific effects of a luminal A subtype of breast cancer. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Discovery of HeLa Cell Contamination in HES Cells: Call for Cell Line Authentication in Reproductive Biology Research.

    PubMed

    Kniss, Douglas A; Summerfield, Taryn L

    2014-08-01

    Continuous cell lines are used frequently in reproductive biology research to study problems in early pregnancy events and parturition. It has been recognized for 50 years that many mammalian cell lines contain inter- or intraspecies contaminations with other cells. However, most investigators do not routinely test their culture systems for cross-contamination. The most frequent contributor to cross-contamination of cell lines is the HeLa cell isolated from an aggressive cervical adenocarcinoma. We report on the discovery of HeLa cell contamination of the human endometrial epithelial cell line HES isolated in our laboratory. Short tandem repeat analysis of 9 unique genetic loci demonstrated molecular identity between HES and HeLa cells. In addition, we verified that WISH cells, isolated originally from human amnion epithelium, were also contaminated with HeLa cells. Inasmuch as our laboratory did not culture HeLa cells at the time of HES cell derivations, the source of contamination was the WISH cell line. These data highlight the need for continued diligence in authenticating cell lines used in reproductive biology research. © The Author(s) 2014.

  12. Fatty acid binding proteins (FABPs) in prostate, bladder and kidney cancer cell lines and the use of IL-FABP as survival predictor in patients with renal cell carcinoma

    PubMed Central

    2011-01-01

    Background Fatty acid binding proteins (FABP) play an important role in carcinogenesis. Modified FABP expression patterns were described for prostate, bladder and for renal cell carcinoma. Studies on metabolic relationships and interactions in permanent cell lines allow a deeper insight into molecular processes. The aim of this study is therefore a systematic overview on mRNA and protein expressions of seven FABPs in frequently used urological cell lines. Methods Nine cell lines of renal carcinomas, seven of urinary bladder carcinomas, and five of prostate carcinomas were investigated. Quantitative RT-qPCR and western blotting were used to determine different FABPs. In addition, 46 paired cancerous and noncancerous tissue samples from nephrectomy specimen with renal cell carcinomas were investigated regarding the ileum FABP mRNA expression level and associated with survival outcome. Results General characteristics of all urological carcinoma cell lines were the expression of E-and IL-FABP on mRNA and protein level, while the expressions differed between the cell lines. The protein expression was not always congruent with the mRNA expression. Renal cell carcinoma cell lines showed expressions of L-, H- and B-FABP mRNA in addition to the general FABP expression in five out of the eight investigated cell lines. In bladder cancer cell lines, we additionally found the expression of A-FABP mRNA in six cell lines, while H-FABP was present only in three cell lines. In prostate cancer cell lines, a strong reduction of A- and E- FABP mRNA was observed. The expression of B-FABP mRNA and protein was observed only in the 22 RV-1 cells. IL-FABP mRNA was over-expressed in renal tumour tissue. The IL-FABP ratio was identified as an independent indicator of survival outcome. Conclusions Distinctly different FABP expression patterns were observed not only between the cell lines derived from the three cancer types, but also between the cell lines from the same cancer. The FABP patterns in the cell lines do not always reflect the real situation in the tumours. These facts have to be considered in functional studies concerning the different FABPs. PMID:21767383

  13. Utility of human embryonic kidney cell line HEK-293 for rapid isolation of fixed and street rabies viruses: comparison with Neuro-2a and BHK-21 cell lines.

    PubMed

    Madhusudana, Shampur Narayan; Sundaramoorthy, Subha; Ullas, Padinjaremattatthil Thankappan

    2010-12-01

    A confirmatory rabies diagnosis can be achieved by rapid virus isolation in cell culture using brain tissue from the suspect animal. Several cell lines have been used for this purpose and the murine neuroblastoma cell line Neuro-2a has been found to be the most sensitive. The human embryonic kidney cell line HEK-293 is known to express several neuronal proteins and is believed to be of neuronal origin. We hypothesized that this cell line could be susceptible to rabies virus, which is highly neurotropic. First we tested the sensitivity of HEK-293 cells to the laboratory strain, challenge virus standard (CVS). We then tested 120 brain samples from different animals and humans suspected to have died of rabies by fluorescent antibody test (FAT). Both FAT-positive and FAT-negative brains were tested for virus isolation using Neuro-2a, BHK-21, and HEK-293 cell lines and also by mouse inoculation. There was 100% correlation between FAT, virus isolation in Neuro-2a and HEK-293 cells, and mouse inoculation. However, the rate of virus isolation in the BHK-21 cell line was only 28% when compared to the other cell lines. The sensitivity of HEK-293 to CVS strain of virus was similar to that of Neuro-2a. We conclude that the HEK-293 cell line is as sensitive as the Neuro-2a cell line for the rapid isolation of rabies virus and may serve as an alternative cell line for rabies diagnosis and future research. Copyright © 2010 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  14. Establishment and characterization of fetal fibroblast cell lines for generating human lysozyme transgenic goats by somatic cell nuclear transfer.

    PubMed

    Liu, Jun; Luo, Yan; Zheng, Liming; Liu, Qingqing; Yang, Zhongcai; Wang, Yongsheng; Su, Jianmin; Quan, Fusheng; Zhang, Yong

    2013-10-01

    This study was performed to qualify goat fetal fibroblast (GFF) cell lines for genetic modification and somatic cell nuclear transfer (SCNT) to produce human lysozyme (hLYZ) transgenic goats. Nine GFF cell lines were established from different fetuses, and the proliferative lifespan and chromosomal stability were analyzed. The results suggested that cell lines with a longer lifespan had stable chromosomes compared with those of cells lines with a shorter lifespan. According to the proliferative lifespan, we divided GFF cell lines into two groups: cell lines with a long lifespan (GFF1/2/7/8/9; group L) and cell lines with a short lifespan (GFF3/4/5/6; group S). Next, a hLYZ expression vector was introduced into these cell lines by electroporation. The efficiencies of colony formation, expansion in culture, and the quality of transgenic clonal cell lines were significant higher in group L than those in group S. The mean fusion rate and blastocyst rate in group L were higher than those in group S (80.3 ± 1.7 vs. 65.1 ± 4.2 % and 19.5 ± 0.6 vs. 15.1 ± 1.1 %, respectively, P < 0.05). After transferring cloned embryos into the oviducts of recipient goats, three live kids were born. PCR and Southern blot analyses confirmed integration of the transgene in cloned goats. In conclusion, the lifespan of GFF cell lines has a major effect on the efficiency to produce transgenic cloned goats. Therefore, the proliferative lifespan of primary cells may be used as a criterion to characterize the quality of cell lines for genetic modification and SCNT.

  15. [Establishment of Z-HL16C cell line.].

    PubMed

    Chen, J P; Li, J; Zhao, S L; Tian, J Y; Ye, F

    2006-09-01

    To establish and study the nature and the application of Z-HL16C cell line. The cell line was continuously passed, frozen stored and recovered. Its application was expanded and the cell type was identified. The cell line had an epithelial-cell-like shape, the size appeared uniform, the cell boundary was distinct. It has been continuously passed, frozen stored and recovered for ten years. Its recovery rate was about 90%. It has been proved to be sensitive to the tested viruses which were enteroviruses (Polio, Cox, Echo), influenza viruses, parainfluenzaviruses, adenoviruses, measles virus. This cell line has been identified as a cancerization cell. The cell line Z-HL16C has been stably established, it has a broad spectrum in sensitivity for culturing viruses.

  16. DNA Packaging Specificity of Bacteriophage N15 with an Excursion into the Genetics of a Cohesive End Mismatch

    PubMed Central

    Feiss, Michael; Young Min, Jea; Sultana, Sawsan; Patel, Priyal; Sippy, Jean

    2015-01-01

    During DNA replication by the λ-like bacteriophages, immature concatemeric DNA is produced by rolling circle replication. The concatemers are processed into mature chromosomes with cohesive ends, and packaged into prohead shells, during virion assembly. Cohesive ends are generated by the viral enzyme terminase, which introduces staggered nicks at cos, an approx. 200 bp-long sequence containing subsites cosQ, cosN and cosB. Interactions of cos subsites of immature concatemeric DNA with terminase orchestrate DNA processing and packaging. To initiate DNA packaging, terminase interacts with cosB and nicks cosN. The cohesive ends of N15 DNA differ from those of λ at 2/12 positions. Genetic experiments show that phages with chromosomes containing mismatched cohesive ends are functional. In at least some infections, the cohesive end mismatch persists through cyclization and replication, so that progeny phages of both allelic types are produced in the infected cell. N15 possesses an asymmetric packaging specificity: N15 DNA is not packaged by phages λ or 21, but surprisingly, N15-specific terminase packages λ DNA. Implications for genetic interactions among λ-like bacteriophages are discussed. PMID:26633301

  17. 21 CFR 660.36 - Samples and protocols.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... ADDITIONAL STANDARDS FOR DIAGNOSTIC SUBSTANCES FOR LABORATORY TESTS Reagent Red Blood Cells § 660.36 Samples... a cell panel intended for identification of unexpected antibodies. The sample shall be packaged as... distribution of each lot of Reagent Red Blood Cells for detection or identification of unexpected antibodies...

  18. Development and characterization of a cell line WAF from freshwater shark Wallago attu.

    PubMed

    Dubey, Akhilesh; Goswami, Mukunda; Yadav, Kamalendra; Sharma, Bhagwati S

    2014-02-01

    A new epithelial cell line, WAF was developed from caudal fin of freshwater shark, Wallago attu. The cell line was optimally maintained at 28 °C in Leibovitz-15 (L-15) medium supplemented with 20 % fetal bovine serum. The cell line was characterized by various cytogenetic and molecular markers. The cytogenetic analysis revealed a diploid count of 86 chromosomes at different passages. The origin of the cell lines was confirmed by the amplification of 547 and 654 bp sequences of 16S rRNA and cytochrome oxidase subunit I genes of mitochondrial DNA, respectively. WAF cells were characterized for their growth characteristics at different temperature and serum concentration. Epithelial morphology of the cell line was confirmed using immunocytochemistry. Further cell plating efficiency, transfection efficiency and viability of cryopreserved WAF cells was also determined. Cytotoxicity and genotoxicity assessment of cadmium salts on WAF cells by MTT, NR and comet assay illustrated the utility of this cell line as an in vitro model for aquatic toxicological studies. The cell line will be further useful for studying oxidative stress markers against aquatic pollutants.

  19. Characterization of a Merkel Cell Polyomavirus-Positive Merkel Cell Carcinoma Cell Line CVG-1.

    PubMed

    Velásquez, Celestino; Amako, Yutaka; Harold, Alexis; Toptan, Tuna; Chang, Yuan; Shuda, Masahiro

    2018-01-01

    Merkel cell polyomavirus (MCV) plays a causal role in ∼80% of Merkel cell carcinomas (MCC). MCV is clonally integrated into the MCC tumor genome, which results in persistent expression of large T (LT) and small T (sT) antigen oncoproteins encoded by the early locus. In MCV-positive MCC tumors, LT is truncated by premature stop codons or deletions that lead to loss of the C-terminal origin binding (OBD) and helicase domains important for replication. The N-terminal Rb binding domain remains intact. MCV-positive cell lines derived from MCC explants have been valuable tools to study the molecular mechanism of MCV-induced Merkel cell carcinogenesis. Although all cell lines have integrated MCV and express truncated LT antigens, the molecular sizes of the LT proteins differ between cell lines. The copy number of integrated viral genome also varies across cell lines, leading to significantly different levels of viral protein expression. Nevertheless, these cell lines share phenotypic similarities in cell morphology, growth characteristics, and neuroendocrine marker expression. Several low-passage MCV-positive MCC cell lines have been established since the identification of MCV. We describe a new MCV-positive MCV cell line, CVG-1, with features distinct from previously reported cell lines. CVG-1 tumor cells grow in more discohesive clusters in loose round cell suspension, and individual cells show dramatic size heterogeneity. It is the first cell line to encode an MCV sT polymorphism resulting in a unique leucine (L) to proline (P) substitution mutation at amino acid 144. CVG-1 possesses a LT truncation pattern near identical to that of MKL-1 cells differing by the last two C-terminal amino acids and also shows an LT protein expression level similar to MKL-1. Viral T antigen knockdown reveals that, like other MCV-positive MCC cell lines, CVG-1 requires T antigen expression for cell proliferation.

  20. Analysis of differential protein expression in normal and neoplastic human breast epithelial cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Williams, K.; Chubb, C.; Huberman, E.

    High resolution two dimensional get electrophoresis (2DE) and database analysis was used to establish protein expression patterns for cultured normal human mammary epithelial cells and thirteen breast cancer cell lines. The Human Breast Epithelial Cell database contains the 2DE protein patterns, including relative protein abundances, for each cell line, plus a composite pattern that contains all the common and specifically expressed proteins from all the cell lines. Significant differences in protein expression, both qualitative and quantitative, were observed not only between normal cells and tumor cells, but also among the tumor cell lines. Eight percent of the consistently detected proteinsmore » were found in significantly (P < 0.001) variable levels among the cell lines. Using a combination of immunostaining, comigration with purified protein, subcellular fractionation, and amino-terminal protein sequencing, we identified a subset of the differentially expressed proteins. These identified proteins include the cytoskeletal proteins actin, tubulin, vimentin, and cytokeratins. The cell lines can be classified into four distinct groups based on their intermediate filament protein profile. We also identified heat shock proteins; hsp27, hsp60, and hsp70 varied in abundance and in some cases in the relative phosphorylation levels among the cell lines. Finally, we identified IMP dehydrogenase in each of the cell lines, and found the levels of this enzyme in the tumor cell lines elevated 2- to 20-fold relative to the levels in normal cells.« less

  1. Derivation and characterization of a pig embryonic stem cell-derived exocrine pancreatic cell line

    USDA-ARS?s Scientific Manuscript database

    The establishment and initial characterization of a pig embryonic stem cell-derived pancreatic cell line, PICM-31, and a colony-cloned derivative cell line, PICM-31A, is described. The cell lines were propagated for several months at split ratios of 1:3 or 1:5 at each passage on STO feeder cells af...

  2. The R package 'Luminescence': a history of unexpected complexity and concepts to deal with it

    NASA Astrophysics Data System (ADS)

    Kreutzer, Sebastian; Burow, Christoph; Dietze, Michael; Fuchs, Margret C.; Friedrich, Johannes; Fischer, Manfred; Schmidt, Christoph

    2017-04-01

    Overcoming limitations in the so far used standard software, developing an efficient solution of low weight for a very specific task or creating graphs of high quality: the reasons that may had initially lead a scientist to work with R are manifold. And as long as developed solutions, e.g., R scripts, are needed for personal use only, code can remain unstructured and a documentation is not compulsory. However, this changes with the first friendly request for help after the code has been reused by others. In contrast to single scripts, written without intention to ever get published, for R packages the CRAN policy demands a more structured and elaborated approach including a minimum of documentation. Nevertheless, growing projects with thousands of lines of code that need to be maintained can become overwhelming, in particular as researchers are not by definition experts on managing software projects. The R package 'Luminescence' (Kreutzer et al., 2017), a collection of tools dealing with the analysis of luminescence data in a geoscientific, geochronological context, started as one single R script, but quickly evolved into a comprehensive solution connected with various other R packages. We present (1) a very brief development history of the package 'Luminescence', before we (2) sketch technical challenges encountered over time and solutions that have been found to deal with it by using various open source tools. Our presentation is considered as a collection of concepts and approaches to set up R projects in geosciences. References. Kreutzer, S., Dietze, M., Burow, C., Fuchs, M. C., Schmidt, C., Fischer, M., Friedrich, J., 2017. Luminescence: Comprehensive Luminescence Dating Data Analysis. R package version 0.6.4. https://CRAN.R-project.org/package=Luminescence

  3. MODFLOW-2000 : the U.S. Geological Survey modular ground-water model--documentation of the Advective-Transport Observation (ADV2) Package

    USGS Publications Warehouse

    Anderman, Evan R.; Hill, Mary Catherine

    2001-01-01

    Observations of the advective component of contaminant transport in steady-state flow fields can provide important information for the calibration of ground-water flow models. This report documents the Advective-Transport Observation (ADV2) Package, version 2, which allows advective-transport observations to be used in the three-dimensional ground-water flow parameter-estimation model MODFLOW-2000. The ADV2 Package is compatible with some of the features in the Layer-Property Flow and Hydrogeologic-Unit Flow Packages, but is not compatible with the Block-Centered Flow or Generalized Finite-Difference Packages. The particle-tracking routine used in the ADV2 Package duplicates the semi-analytical method of MODPATH, as shown in a sample problem. Particles can be tracked in a forward or backward direction, and effects such as retardation can be simulated through manipulation of the effective-porosity value used to calculate velocity. Particles can be discharged at cells that are considered to be weak sinks, in which the sink applied does not capture all the water flowing into the cell, using one of two criteria: (1) if there is any outflow to a boundary condition such as a well or surface-water feature, or (2) if the outflow exceeds a user specified fraction of the cell budget. Although effective porosity could be included as a parameter in the regression, this capability is not included in this package. The weighted sum-of-squares objective function, which is minimized in the Parameter-Estimation Process, was augmented to include the square of the weighted x-, y-, and z-components of the differences between the simulated and observed advective-front locations at defined times, thereby including the direction of travel as well as the overall travel distance in the calibration process. The sensitivities of the particle movement to the parameters needed to minimize the objective function are calculated for any particle location using the exact sensitivity-equation approach; the equations are derived by taking the partial derivatives of the semi-analytical particle-tracking equation with respect to the parameters. The ADV2 Package is verified by showing that parameter estimation using advective-transport observations produces the true parameter values in a small but complicated test case when exact observations are used. To demonstrate how the ADV2 Package can be used in practice, a field application is presented. In this application, the ADV2 Package is used first in the Sensitivity-Analysis mode of MODFLOW-2000 to calculate measures of the importance of advective-transport observations relative to head-dependent flow observations when either or both are used in conjunction with hydraulic-head observations in a simulation of the sewage-discharge plume at Cape Cod, Massachusetts. The ADV2 Package is then used in the Parameter-Estimation mode of MODFLOW-2000 to determine best-fit parameter values. It is concluded that, for this problem, advective-transport observations improved the calibration of the model and the estimation of ground-water flow parameters, and the use of formal parameter-estimation methods and related techniques produced significant insight into the physical system.

  4. Inactivation of Bacillus cereus by Na-chlorophyllin-based photosensitization on the surface of packaging.

    PubMed

    Luksiene, Z; Buchovec, I; Paskeviciute, E

    2010-11-01

    This study was focused on the possibility to inactivate food-borne pathogen Bacillus cereus by Na-chlorophyllin (Na-Chl)-based photosensitization in vitro and after attachment to the surface of packaging material. Bacillus cereus in vitro or attached to the packaging was incubated with Na-Chl (7·5×10(-8) to 7·5×10(-5) mol l(-1) ) for 2-60min in phosphate buffer saline. Photosensitization was performed by illuminating cells under a light with a λ of 400nm and an energy density of 20mW cm(-2) . The illumination time varied 0-5min and subsequently the total energy dose was 0-6J cm(-2) . The results show that B. cereus vegetative cells in vitro or attached to the surface of packaging after incubation with 7·5×10(-7) mol l(-1) Na-Chl and following illumination were inactivated by 7log. The photoinactivation of B. cereus spores in vitro by 4log required higher (7·5×10(-6) mol l(-1) ) Na-Chl concentration. Decontamination of packaging material from attached spores by photosensitization reached 5log at 7·5×10(-5) mol l(-1) Na-Chl concentration. Comparative analysis of different packaging decontamination treatments indicates that washing with water can diminish pathogen population on the surface by <1log, 100ppm Na-hypochlorite reduces the pathogens about 1·7log and 200ppm Na-hypochlorite by 2·2log. Meanwhile, Na-Chl-based photosensitization reduces bacteria on the surface by 4·2 orders of magnitude. Food-borne pathogen B. cereus could be effectively inactivated (7log) by Na-Chl-based photosensitization in vitro and on the surface of packaging material. Spores are more resistant than vegetative cells to photosensitization-based inactivation. Comparison of different surface decontamination treatments indicates that Na-Chl-based photosensitization is much more effective antibacterial tool than washing with water or 200ppm Na-hypochlorite. Our data support the idea that Na-Chl-based photosensitization has great potential for future application as an environment-friendly, nonthermal surface decontamination technique. © 2010 The Authors. Journal of Applied Microbiology © 2010 The Society for Applied Microbiology.

  5. Evaluation of the drug sensitivity and expression of 16 drug resistance-related genes in canine histiocytic sarcoma cell lines

    PubMed Central

    ASADA, Hajime; TOMIYASU, Hirotaka; GOTO-KOSHINO, Yuko; FUJINO, Yasuhito; OHNO, Koichi; TSUJIMOTO, Hajime

    2015-01-01

    Canine histiocytic sarcoma (HS) is an aggressive tumor type originating from histiocytic cell lineages. This disease is characterized by poor response to chemotherapy and short survival time. Therefore, it is of critical importance to identify and develop effective antitumor drugs against HS. The objectives of this study were to examine the drug sensitivities of 10 antitumor drugs. Using a real-time RT-PCR system, the mRNA expression levels of 16 genes related to drug resistance in 4 canine HS cell lines established from dogs with disseminated HS were determined and compared to 2 canine lymphoma cell lines (B-cell and T-cell). These 4 canine HS cell lines showed sensitivities toward microtubule inhibitors (vincristine, vinblastine and paclitaxel), comparable to those in the canine B-cell lymphoma cell line. Moreover, it was shown that P-gp in the HS cell lines used in this study did not have enough function to efflux its substrate. Sensitivities to melphalan, nimustine, methotrexate, cytarabine, doxorubicin and etoposide were lower in the 4 HS cell lines than in the 2 canine lymphoma cell lines. The data obtained in this study using cultured cell lines could prove helpful in the developing of advanced and effective chemotherapies for treating dogs that are suffering from HS. PMID:25715778

  6. Evaluation of the drug sensitivity and expression of 16 drug resistance-related genes in canine histiocytic sarcoma cell lines.

    PubMed

    Asada, Hajime; Tomiyasu, Hirotaka; Goto-Koshino, Yuko; Fujino, Yasuhito; Ohno, Koichi; Tsujimoto, Hajime

    2015-06-01

    Canine histiocytic sarcoma (HS) is an aggressive tumor type originating from histiocytic cell lineages. This disease is characterized by poor response to chemotherapy and short survival time. Therefore, it is of critical importance to identify and develop effective antitumor drugs against HS. The objectives of this study were to examine the drug sensitivities of 10 antitumor drugs. Using a real-time RT-PCR system, the mRNA expression levels of 16 genes related to drug resistance in 4 canine HS cell lines established from dogs with disseminated HS were determined and compared to 2 canine lymphoma cell lines (B-cell and T-cell). These 4 canine HS cell lines showed sensitivities toward microtubule inhibitors (vincristine, vinblastine and paclitaxel), comparable to those in the canine B-cell lymphoma cell line. Moreover, it was shown that P-gp in the HS cell lines used in this study did not have enough function to efflux its substrate. Sensitivities to melphalan, nimustine, methotrexate, cytarabine, doxorubicin and etoposide were lower in the 4 HS cell lines than in the 2 canine lymphoma cell lines. The data obtained in this study using cultured cell lines could prove helpful in the developing of advanced and effective chemotherapies for treating dogs that are suffering from HS.

  7. Interactive Spectral Analysis and Computation (ISAAC)

    NASA Technical Reports Server (NTRS)

    Lytle, D. M.

    1992-01-01

    Isaac is a task in the NSO external package for IRAF. A descendant of a FORTRAN program written to analyze data from a Fourier transform spectrometer, the current implementation has been generalized sufficiently to make it useful for general spectral analysis and other one dimensional data analysis tasks. The user interface for Isaac is implemented as an interpreted mini-language containing a powerful, programmable vector calculator. Built-in commands provide much of the functionality needed to produce accurate line lists from input spectra. These built-in functions include automated spectral line finding, least squares fitting of Voigt profiles to spectral lines including equality constraints, various filters including an optimal filter construction tool, continuum fitting, and various I/O functions.

  8. UniPOPS: Unified data reduction suite

    NASA Astrophysics Data System (ADS)

    Maddalena, Ronald J.; Garwood, Robert W.; Salter, Christopher J.; Stobie, Elizabeth B.; Cram, Thomas R.; Morgan, Lorrie; Vance, Bob; Hudson, Jerome

    2015-03-01

    UniPOPS, a suite of programs and utilities developed at the National Radio Astronomy Observatory (NRAO), reduced data from the observatory's single-dish telescopes: the Tucson 12-m, the Green Bank 140-ft, and archived data from the Green Bank 300-ft. The primary reduction programs, 'line' (for spectral-line reduction) and 'condar' (for continuum reduction), used the People-Oriented Parsing Service (POPS) as the command line interpreter. UniPOPS unified previous analysis packages and provided new capabilities; development of UniPOPS continued within the NRAO until 2004 when the 12-m was turned over to the Arizona Radio Observatory (ARO). The submitted code is version 3.5 from 2004, the last supported by the NRAO.

  9. Genetic Profiling Reveals Cross-Contamination and Misidentification of 6 Adenoid Cystic Carcinoma Cell Lines: ACC2, ACC3, ACCM, ACCNS, ACCS and CAC2

    PubMed Central

    Phuchareon, Janyaporn; Ohta, Yoshihito; Woo, Jonathan M.; Eisele, David W.; Tetsu, Osamu

    2009-01-01

    Adenoid cystic carcinoma (ACC) is the second most common malignant neoplasm of the salivary glands. Most patients survive more than 5 years after surgery and postoperative radiation therapy. The 10 year survival rate, however, drops to 40%, due to locoregional recurrences and distant metastases. Improving long-term survival in ACC requires the development of more effective systemic therapies based on a better understanding of the biologic behavior of ACC. Much preclinical research in this field involves the use of cultured cells and, to date, several ACC cell lines have been established. Authentication of these cell lines, however, has not been reported. We performed DNA fingerprint analysis on six ACC cell lines using short tandem repeat (STR) examinations and found that all six cell lines had been contaminated with other cells. ACC2, ACC3, and ACCM were determined to be cervical cancer cells (HeLa cells), whereas the ACCS cell line was composed of T24 urinary bladder cancer cells. ACCNS and CAC2 cells were contaminated with cells derived from non-human mammalian species: the cells labeled ACCNS were mouse cells and the CAC2 cells were rat cells. These observations suggest that future studies using ACC cell lines should include cell line authentication to avoid the use of contaminated or non-human cells. PMID:19557180

  10. Molecular docking and dynamics simulations of A.niger RNase from Aspergillus niger ATCC26550: for potential prevention of human cancer.

    PubMed

    Kumar, Gundampati Ravi; Chikati, Rajasekhar; Pandrangi, Santhi Latha; Kandapal, Manoj; Sonkar, Kirti; Gupta, Neeraj; Mulakayala, Chaitanya; Jagannadham, Medicherla V; Kumar, Chitta Suresh; Saxena, Sunita; Das, Mira Debnath

    2013-02-01

    The aim of the present research was to study the anticancer effects of Aspergillus niger (A.niger) RNase. We found that RNase (A.niger RNase) significantly and dose dependently inhibited invasiveness of breast cancer cell line MDA MB 231 by 55 % (P<0.01) at 1 μM concentration. At a concentration of 2 μM, the anti invasive effect of the enzyme increased to 90 % (P<0.002). Keeping the aim to determine molecular level interactions (molecular simulations and protein docking) of human actin with A.niger RNase we extended our work in-vitro to in-silico studies. To gain better relaxation and accurate arrangement of atoms, refinement was done on the human actin and A.niger RNase by energy minimization (EM) and molecular dynamics (MD) simulations using 43A(2) force field of Gromacs96 implemented in the Gromacs 4.0.5 package, finally the interaction energies were calculated by protein-protein docking using the HEX. These in vitro and in-silico structural studies prove the effective inhibition of actin activity by A.niger RNase in neoplastic cells and thereby provide new insights for the development of novel anti cancer drugs.

  11. microMS: A Python Platform for Image-Guided Mass Spectrometry Profiling

    NASA Astrophysics Data System (ADS)

    Comi, Troy J.; Neumann, Elizabeth K.; Do, Thanh D.; Sweedler, Jonathan V.

    2017-09-01

    Image-guided mass spectrometry (MS) profiling provides a facile framework for analyzing samples ranging from single cells to tissue sections. The fundamental workflow utilizes a whole-slide microscopy image to select targets of interest, determine their spatial locations, and subsequently perform MS analysis at those locations. Improving upon prior reported methodology, a software package was developed for working with microscopy images. microMS, for microscopy-guided mass spectrometry, allows the user to select and profile diverse samples using a variety of target patterns and mass analyzers. Written in Python, the program provides an intuitive graphical user interface to simplify image-guided MS for novice users. The class hierarchy of instrument interactions permits integration of new MS systems while retaining the feature-rich image analysis framework. microMS is a versatile platform for performing targeted profiling experiments using a series of mass spectrometers. The flexibility in mass analyzers greatly simplifies serial analyses of the same targets by different instruments. The current capabilities of microMS are presented, and its application for off-line analysis of single cells on three distinct instruments is demonstrated. The software has been made freely available for research purposes. [Figure not available: see fulltext.

  12. microMS: A Python Platform for Image-Guided Mass Spectrometry Profiling.

    PubMed

    Comi, Troy J; Neumann, Elizabeth K; Do, Thanh D; Sweedler, Jonathan V

    2017-09-01

    Image-guided mass spectrometry (MS) profiling provides a facile framework for analyzing samples ranging from single cells to tissue sections. The fundamental workflow utilizes a whole-slide microscopy image to select targets of interest, determine their spatial locations, and subsequently perform MS analysis at those locations. Improving upon prior reported methodology, a software package was developed for working with microscopy images. microMS, for microscopy-guided mass spectrometry, allows the user to select and profile diverse samples using a variety of target patterns and mass analyzers. Written in Python, the program provides an intuitive graphical user interface to simplify image-guided MS for novice users. The class hierarchy of instrument interactions permits integration of new MS systems while retaining the feature-rich image analysis framework. microMS is a versatile platform for performing targeted profiling experiments using a series of mass spectrometers. The flexibility in mass analyzers greatly simplifies serial analyses of the same targets by different instruments. The current capabilities of microMS are presented, and its application for off-line analysis of single cells on three distinct instruments is demonstrated. The software has been made freely available for research purposes. Graphical Abstract ᅟ.

  13. Continuous human cell lines and method of making same

    DOEpatents

    Stampfer, Martha R.

    1989-01-01

    Substantially genetically stable continuous human cell lines derived from normal human mammary epithelial cells (HMEC) and processes for making and using the same. In a preferred embodiment, the cell lines are derived by treating normal human mammary epithelial tissue with a chemical carcinogen such as benzo[a]pyrene. The novel cell lines serve as useful substrates for elucidating the potential effects of a number of toxins, carcinogens and mutagens as well as of the addition of exogenous genetic material. The autogenic parent cells from which the cell lines are derived serve as convenient control samples for testing. The cell lines are not neoplastically transformed, although they have acquired several properties which distinguish them from their normal progenitors.

  14. Establishment of dermal sheath cell line from Cashmere goat and characterizing cytokeratin 13 as its novel biomarker.

    PubMed

    Zhu, Bing; Guo, Zhili; Jin, Muzi; Bai, Yujuan; Yang, Wenliang; Hui, Lihua

    2018-05-01

    To establish a dermal sheath cell line, a dermal papilla cell line and a outer root sheath cell line from Cashmere goat and clarify the similarities and differences among them. We established a dermal sheath cell line, a dermal papilla cell line and a outer root sheath cell line from the pelage skin hair follicles of Cashmere goat. The growth rate of dermal sheath cells was intermediate between that of dermal papilla cells and outer root sheath cells. Immunofluorescence experiments and reverse transcription-polymerase chain reaction analysis showed that at both the transcriptional and translational levels, the dermal sheath cells were alpha-smooth muscle actin (α-SMA) + /cytokeratin 13 + , while the dermal papilla cells were α-SMA + /cytokeratin 13 - and the outer root sheath cells were α-SMA - /cytokeratin 13 + . Patterns of cytokeratin 13 expression could distinguish the dermal sheath cells from the dermal papilla cells. These results suggest that cytokeratin 13 could serve as a novel biomarker for dermal sheath cells of Cashmere goat, and should prove useful for researchers investigating dermal stem cells or interaction of different types of cells during hair cycle.

  15. Cell lines, Md108 and Md66, from the hemocytes of Malacosoma disstria (Lepidoptera) display aspects of plasma-free innate non-self activities.

    PubMed

    Lapointe, Jason F; Dunphy, Gary B; Giannoulis, Paschalis; Mandato, Craig A; Nardi, James B; Gharib, Osama H; Niven, Donald F

    2011-11-01

    The innate non-self response systems of the deciduous tree pest, the forest tent caterpillar, Malacosoma disstria has been documented by us in terms of in vitro and in vivo reactions towards the Gram-positive nonpathogenic bacterium, Bacillus subtilis and Gram-negative pathogenic microbe, Xenorhabdus nematophila and their respective surface antigens, lipopoteichoic acids (LTA) and lipopolysaccharides (LPS). These studies, often conducted in whole and diluted hemolymph, preclude examination of plasma-free cellular (hemocyte) responses. Plasma-free hemocytes as primary cultures are difficult to obtain. The floating cell line Md66 and attached cell line Md108 from M. disstria hemocytes were examined as a model for plasma-free M. disstria hemocyte non-self responses. Herein, it was established that although both lines differed from each other and from the primary hemocyte cultures of M. disstria in growth parameters, cell composition and sizes both cell lines displayed granular cell-like (GL) cells and plasmatocyte-like (PL) cells according to morphological criteria and to some extent antigenic similarities based on labeling with anti-Chrysodeixis includens hemocyte monoclonal antibodies. Hemocyte-specific neuroglian-like protein was detected on cells of both cell lines and in the primary hemocyte cultures albeit with staining patterns differing according to culture and cell types, confluency levels and cell-cell adhesion. Both cell lines bound B. subtilis and X. nematophila, the reaction extent varying with the cell line and its cell types. LPS damaged both cell types in the two cell lines whereas LTA enhanced the adhesion of Md66 GL cells to flask surfaces followed by PL cell adhesion. PL cells of both lines, like the primary cultures, phagocytosed FITC-labeled B. subtilis; only Md108 GL cells phagocytosed B. subtilis. In either case phagocytosis was always less in frequency and intensity than the primary cultures. Proteins released from the cell lines differed in pattern and magnitude but contained bacterial binding proteins that enhanced differential bacterial adhesion to both cell types in both cell lines: the GL cells both cultures, and those of granular cells in primary cultures, were more involved than the primary plasmatocytes and PL cells. Only Md66 cells possessed lysozyme and both cell types of both lines contained phenoloxidase. Neither enzyme type was released during early phase reaction with the bacteria. LPS inhibited phenoloxidase activity. The similarities and differences between the lines and primary cultures make Md66 and Md108 useful for the systematic examination of plasma-free cellular non-self reactions. Copyright © 2011 Elsevier Inc. All rights reserved.

  16. Differential sensitivity of melanoma cell lines with BRAFV600E mutation to the specific Raf inhibitor PLX4032

    PubMed Central

    2010-01-01

    Blocking oncogenic signaling induced by the BRAFV600E mutation is a promising approach for melanoma treatment. We tested the anti-tumor effects of a specific inhibitor of Raf protein kinases, PLX4032/RG7204, in melanoma cell lines. PLX4032 decreased signaling through the MAPK pathway only in cell lines with the BRAFV600E mutation. Seven out of 10 BRAFV600E mutant cell lines displayed sensitivity based on cell viability assays and three were resistant at concentrations up to 10 μM. Among the sensitive cell lines, four were highly sensitive with IC50 values below 1 μM, and three were moderately sensitive with IC50 values between 1 and 10 μM. There was evidence of MAPK pathway inhibition and cell cycle arrest in both sensitive and resistant cell lines. Genomic analysis by sequencing, genotyping of close to 400 oncogeninc mutations by mass spectrometry, and SNP arrays demonstrated no major differences in BRAF locus amplification or in other oncogenic events between sensitive and resistant cell lines. However, metabolic tracer uptake studies demonstrated that sensitive cell lines had a more profound inhibition of FDG uptake upon exposure to PLX4032 than resistant cell lines. In conclusion, BRAFV600E mutant melanoma cell lines displayed a range of sensitivities to PLX4032 and metabolic imaging using PET probes can be used to assess sensitivity. PMID:20406486

  17. Molecular characterization of breast cancer cell lines through multiple omic approaches.

    PubMed

    Smith, Shari E; Mellor, Paul; Ward, Alison K; Kendall, Stephanie; McDonald, Megan; Vizeacoumar, Frederick S; Vizeacoumar, Franco J; Napper, Scott; Anderson, Deborah H

    2017-06-05

    Breast cancer cell lines are frequently used as model systems to study the cellular properties and biology of breast cancer. Our objective was to characterize a large, commonly employed panel of breast cancer cell lines obtained from the American Type Culture Collection (ATCC 30-4500 K) to enable researchers to make more informed decisions in selecting cell lines for specific studies. Information about these cell lines was obtained from a wide variety of sources. In addition, new information about cellular pathways that are activated within each cell line was generated. We determined key protein expression data using immunoblot analyses. In addition, two analyses on serum-starved cells were carried out to identify cellular proteins and pathways that are activated in these cells. These analyses were performed using a commercial PathScan array and a novel and more extensive phosphopeptide-based kinome analysis that queries 1290 phosphorylation events in major signaling pathways. Data about this panel of breast cancer cell lines was also accessed from several online sources, compiled and summarized for the following areas: molecular classification, mRNA expression, mutational status of key proteins and other possible cancer-associated mutations, and the tumorigenic and metastatic capacity in mouse xenograft models of breast cancer. The cell lines that were characterized included 10 estrogen receptor (ER)-positive, 12 human epidermal growth factor receptor 2 (HER2)-amplified and 18 triple negative breast cancer cell lines, in addition to 4 non-tumorigenic breast cell lines. Within each subtype, there was significant genetic heterogeneity that could impact both the selection of model cell lines and the interpretation of the results obtained. To capture the net activation of key signaling pathways as a result of these mutational combinations, profiled pathway activation status was examined. This provided further clarity for which cell lines were particularly deregulated in common or unique ways. These two new kinase or "Kin-OMIC" analyses add another dimension of important data about these frequently used breast cancer cell lines. This will assist researchers in selecting the most appropriate cell lines to use for breast cancer studies and provide context for the interpretation of the emerging results.

  18. 49 CFR 171.24 - Additional requirements for the use of the ICAO Technical Instructions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    .... (ii) Lithium metal cells and batteries. Lithium metal cells and batteries (UN3090) are forbidden for transport aboard passenger-carrying aircraft. The outside of each package that contains lithium metal cells or lithium metal batteries (UN3090) transported in accordance with Packing Instruction 968, Section...

  19. Authorized Course of Instruction for the Quinmester Program. Science: Cell Biology, Introduction to Life Science.

    ERIC Educational Resources Information Center

    Dade County Public Schools, Miami, FL.

    This instructional package contains two biological units developed for the Dade County Florida Quinmester Program. "Introduction to Life Sciences" develops student understandings of cell structure and function, and compares different levels of cellular organization. "Cell Biology" investigates the origin of modern cellular…

  20. Development and characterization of two cell lines PDF and PDH from Puntius denisonii (Day 1865).

    PubMed

    Lakra, Wazir S; Goswami, M; Yadav, Kamalendra; Gopalakrishnan, A; Patiyal, R S; Singh, M

    2011-02-01

    The Puntius denisonii colloquially and more popularly referred to as Miss Kerala is a subtropical fish belonging to the genus Puntius (Barb) and family Cyprinidae. Two cell lines PDF and PDH were developed from the caudal fin and heart of P. denisonii, respectively. The cell lines were optimally maintained at 26°C in Leibovitz-15 medium supplemented with 10% fetal bovine serum. A diploid count of 50 chromosomes at passage 50 was observed in both the cell lines. The high growth potential of the cell lines was reflected from the cell doubling time of 28 and 30 h of PDF and PDH cell lines, respectively. The viability of the PDF and PDH cell lines was 70% and 76%, respectively, after 4 mo of storage in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 653 bp fragments of cytochrome oxidase subunit I of mitochondrial DNA genes.

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