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Sample records for hesc lines reveal

  1. Derivation, culture, and characterization of VUB hESC lines.

    PubMed

    Mateizel, Ileana; Spits, Claudia; De Rycke, Martine; Liebaers, Inge; Sermon, Karen

    2010-04-01

    In this report, we present the derivation and characterization of 15 hESC lines established at the Vrije Universiteit Brussel, Belgium in collaboration with the Universitair Ziekenhuis Brussel, Belgium, using surplus in vitro fertilization embryos and embryos carrying monogenic disorders donated for research. Four lines were derived from blastocyst-stage embryos presumed to be genetically normal, and 11 hESC lines were obtained from embryos shown to carry genetic mutations by preimplantation genetic diagnosis. All the lines express markers of pluripotency as determined by immunocytochemistry and RT-PCR, and formed teratomas when injected into SCID mice. All VUB hESC lines, except for VUB17, are reported in the European hESC registry and are available upon request after signing a Material Transfer Agreement from the VUB (contact person: Prof. Dr. Karen Sermon; Karen.Sermon@uzbrussel.be). PMID:20224973

  2. Effect of Chromatin Structure on the Extent and Distribution of DNA Double Strand Breaks Produced by Ionizing Radiation; Comparative Study of hESC and Differentiated Cells Lines

    PubMed Central

    Venkatesh, Priyanka; Panyutin, Irina V.; Remeeva, Evgenia; Neumann, Ronald D.; Panyutin, Igor G.

    2016-01-01

    Chromatin structure affects the extent of DNA damage and repair. Thus, it has been shown that heterochromatin is more protective against DNA double strand breaks (DSB) formation by ionizing radiation (IR); and that DNA DSB repair may proceed differently in hetero- and euchromatin regions. Human embryonic stem cells (hESC) have a more open chromatin structure than differentiated cells. Here, we study the effect of chromatin structure in hESC on initial DSB formation and subsequent DSB repair. DSB were scored by comet assay; and DSB repair was assessed by repair foci formation via 53BP1 antibody staining. We found that in hESC, heterochromatin is confined to distinct regions, while in differentiated cells it is distributed more evenly within the nuclei. The same dose of ionizing radiation produced considerably more DSB in hESC than in differentiated derivatives, normal human fibroblasts; and one cancer cell line. At the same time, the number of DNA repair foci were not statistically different among these cells. We showed that in hESC, DNA repair foci localized almost exclusively outside the heterochromatin regions. We also noticed that exposure to ionizing radiation resulted in an increase in heterochromatin marker H3K9me3 in cancer HT1080 cells, and to a lesser extent in IMR90 normal fibroblasts, but not in hESCs. These results demonstrate the importance of chromatin conformation for DNA protection and DNA damage repair; and indicate the difference of these processes in hESC. PMID:26729112

  3. A comparative transcriptomic study on the effects of valproic acid on two different hESCs lines in a neural teratogenicity test system.

    PubMed

    Colleoni, Silvia; Galli, Cesare; Gaspar, John Antony; Meganathan, Kesavan; Jagtap, Smita; Hescheler, Jurgen; Zagoura, Dimitra; Bremer, Susanne; Sachinidis, Agapios; Lazzari, Giovanna

    2014-11-18

    A number of in vitro toxicity assays based on human embryonic stem cells (hESCs) are under development in order to provide alternative methods for the screening of chemicals and drugs and to reduce the number of animals needed for developmental toxicity assessment. The major challenge is to demonstrate the reliability of these in vitro methods by correlating the in vitro produced results to the available in vivo data. In this context transcriptomic approaches associated to toxicogenomic database analysis give the possibility to screen, annotate and cluster high numbers of genes and to identify the molecular changes that univocally mark the toxicity induced processes or are indicative of the early initiating events that lead to cellular toxicity. In this retrospective study we compare microarray transcriptomic data derived from two different hESCs lines (HUES1 and H9) exposed to valproic acid (VA) while applying the same differentiation protocol. We present the results of this comparative analysis in light of the known teratogenic effects of VA. The results show molecular changes in the processes of neural development, neural crest migration, apoptosis and regulation of transcription, indicating a good correspondence with the available in vivo data. We also describe common toxicological signatures and provide an interpretation of the observed qualitative differences referring to known biological features of the two hESCs lines. PMID:25192806

  4. Generating hESCs with reduced immunogenicity by disrupting TAP1 or TAPBP.

    PubMed

    Cui, Di; Wang, Jinping; Zeng, Yelin; Rao, Lingjun; Chen, Haide; Li, Wenling; Li, Yang; Li, Hui; Cui, Chun; Xiao, Lei

    2016-08-01

    Human embryonic stem cells (hESCs) are thought to be a promising resource for cell therapy, while it has to face the major problem of graft immunological rejection. Major histocompatibility complex (MHC) class I expressed on the cell surface is the major cause of graft rejection. Transporter associated with antigen presentation 1 (TAP1) and TAP-associated glycoprotein (TAPBP) play important roles in regulating MHC class I expression. In this study, we generated TAP1- and TAPBP-deficient hESC lines, respectively, using transcription activator-like effector nucleases technique. These cells showed deficient expression of MHC class I on the cell surface and reduced immunogenicity compared with wild types, but maintained normal pluripotency, karyotypes, and differentiation ability. Thus, our findings are instrumental in developing a universal cell resource with both pluripotency and hypo-immunogenicity for transplantation therapy in the future. PMID:27068360

  5. Derivation of new human embryonic stem cell lines reveals rapid epigenetic progression in vitro that can be prevented by chemical modification of chromatin

    PubMed Central

    Diaz Perez, Silvia V.; Kim, Rachel; Li, Ziwei; Marquez, Victor E.; Patel, Sanjeet; Plath, Kathrin; Clark, Amander T.

    2012-01-01

    Human embryonic stem cells (hESCs) are pluripotent cell types derived from the inner cell mass of human blastocysts. Recent data indicate that the majority of established female XX hESC lines have undergone X chromosome inactivation (XCI) prior to differentiation, and XCI of hESCs can be either XIST-dependent (class II) or XIST-independent (class III). XCI of female hESCs precludes the use of XX hESCs as a cell-based model for examining mechanisms of XCI, and will be a challenge for studying X-linked diseases unless strategies are developed to reactivate the inactive X. In order to recover nuclei with two active X chromosomes (class I), we developed a reprogramming strategy by supplementing hESC media with the small molecules sodium butyrate and 3-deazaneplanocin A (DZNep). Our data demonstrate that successful reprogramming can occur from the XIST-dependent class II nuclear state but not class III nuclear state. To determine whether these small molecules prevent XCI, we derived six new hESC lines under normoxic conditions (UCLA1–UCLA6). We show that class I nuclei are present within the first 20 passages of hESC derivation prior to cryopreservation, and that supplementation with either sodium butyrate or DZNep preserve class I nuclei in the self-renewing state. Together, our data demonstrate that self-renewal and survival of class I nuclei are compatible with normoxic hESC derivation, and that chemical supplementation after derivation provides a strategy to prevent epigenetic progression and retain nuclei with two active X chromosomes in the self-renewing state. PMID:22058289

  6. Fibrochondrogenesis of hESCs: growth factor combinations and cocultures.

    PubMed

    Hoben, Gwendolyn M; Willard, Vincent P; Athanasiou, Kyriacos A

    2009-03-01

    The successful differentiation of human embryonic stem cells (hESCs) to fibrochondrocyte-like cells and characterization of these differentiated cells is a critical step toward tissue engineering of musculoskeletal fibrocartilages (e.g., knee meniscus, temporomandibular joint disc, and intervertebral disc). In this study, growth factors and primary cell cocultures were applied to hESC embryoid bodies (EBs) for 3 weeks and evaluated for their effect on the synthesis of critical fibrocartilage matrix components: glycosaminoglycans (GAG) and collagens (types I, II, and VI). Changes in surface markers (CD105, CD44, SSEA, PDGFR alpha) after the differentiation treatments were also analyzed. The study was conducted in three phases: (1) examination of growth factors (TGF-beta 3, BMP-2, BMP-4, BMP-6, PDGF-BB, sonic hedgehog protein); (2) comparison of two cocultures (primary chondrocytes or fibrochondrocytes); and (3) the combination of the most effective growth factor and coculture regimen. TGF-beta 3 with BMP-4 yielded EBs positive for collagens I, II, and VI, with up to 6.7- and 4.8-fold increases in GAG and collagen, respectively. Analysis of cell surface markers showed a significant increase in CD44 with the TGF-beta 3 + BMP-4 treatment compared to the controls. Coculture with fibrochondrocytes resulted in up to a 9.8-fold increase in collagen II production. The combination of the growth factors BMP-4 + TGF-beta 3 with the fibrochondrocyte coculture led to an increase in cell proliferation and GAG production compared to either treatment alone. This study determined two powerful treatments for inducing fibrocartilaginous differentiation of hESCs and provides a foundation for using flow cytometry to purify these differentiated cells. PMID:18454697

  7. Alginate microcapsule for propagation and directed differentiation of hESCs to definitive endoderm.

    PubMed

    Chayosumrit, Methichit; Tuch, Bernard; Sidhu, Kuldip

    2010-01-01

    Human embryonic stem cells (hESCs) are potential renewable sources of cells in replacement therapies for many diseases including type 1 diabetes. We have established a three dimensional (3D) model to culture and differentiate hESCs that are encapsulated in calcium alginate microcapsules. This system promotes cellular interactions that are essential for both maintaining pluripotency and differentiation. This 3D model also provides opportunity to separate out hESCs from fibroblasts used as feeder layer during culture. In this study, we compared the viability and proliferation of the encapsulated hESCs cultured in serum replacement (SR) medium, human fetal fibroblast-conditioned medium (hFF-CM), in the presence and absence of Y-27632, a ROCK inhibitor. Treatment of hESCs with Y-27632 promoted cell survival, cell cluster formation and proliferation rate in both SR medium and hFF-CM. These encapsulated hESC clusters were then directly differentiated to definitive endoderm cells that expressed mesendoderm (Brachyury 70-fold), definitive endoderm (SOX17>300-fold, FOXA2>800-fold, and CXCR4>100-fold) and primitive gut tube (HNF1beta>120-fold) as compared to the undifferentiated hESCs. These data show that microcapsules can be used for differentiation of hESCs into definitive endoderm in 3D and could have potential application for immune-isolation and prevention of teratomas formation of hESCs during transplantation. PMID:19833385

  8. AF9 promotes hESC neural differentiation through recruiting TET2 to neurodevelopmental gene loci for methylcytosine hydroxylation

    PubMed Central

    Qiao, Yunbo; Wang, Xiongjun; Wang, Ran; Li, Yuanyuan; Yu, Fang; Yang, Xianfa; Song, Lu; Xu, Guoliang; Chin, Y Eugene; Jing, Naihe

    2015-01-01

    AF9 mutations have been implicated in human neurodevelopmental diseases and murine Af9 mediates histone methylation during cortical neuron generation. However, AF9 function and related mechanisms in human neurodevelopment remain unknown. Here we show that AF9 is necessary and sufficient for human embryonic stem cell (hESC) neural differentiation and neurodevelopmental gene activation. The 5-methylcytosine (5mC) dioxygenase TET2, which was identified in an AF9-associated protein complex, physically interacted with AF9. Both AF9 and TET2 co-localized in 5-hydroxymethylcytosine (5hmC)-positive hESC-derived neurons and were required for appropriate hESC neural differentiation. Upon binding to AAC-containing motifs, AF9 recruited TET2 to occupy the common neurodevelopmental gene loci to direct 5mC-to-5hmC conversion, which was followed by sequential activation of neural target genes and hESC neural commitment. These findings define an AF9–TET2 regulatory complex for modulating human neural development and reveal a novel mechanism by which the AF9 recognition specificity and TET2 hydroxylation activity cooperate to control neurodevelopmental gene activation. PMID:27462416

  9. Molecular profiling reveals primary mesothelioma cell lines recapitulate human disease.

    PubMed

    Chernova, T; Sun, X M; Powley, I R; Galavotti, S; Grosso, S; Murphy, F A; Miles, G J; Cresswell, L; Antonov, A V; Bennett, J; Nakas, A; Dinsdale, D; Cain, K; Bushell, M; Willis, A E; MacFarlane, M

    2016-07-01

    Malignant mesothelioma (MM) is an aggressive, fatal tumor strongly associated with asbestos exposure. There is an urgent need to improve MM patient outcomes and this requires functionally validated pre-clinical models. Mesothelioma-derived cell lines provide an essential and relatively robust tool and remain among the most widely used systems for candidate drug evaluation. Although a number of cell lines are commercially available, a detailed comparison of these commercial lines with freshly derived primary tumor cells to validate their suitability as pre-clinical models is lacking. To address this, patient-derived primary mesothelioma cell lines were established and characterized using complementary multidisciplinary approaches and bioinformatic analysis. Clinical markers of mesothelioma, transcriptional and metabolic profiles, as well as the status of p53 and the tumor suppressor genes CDKN2A and NF2, were examined in primary cell lines and in two widely used commercial lines. Expression of MM-associated markers, as well as the status of CDKN2A, NF2, the 'gatekeeper' in MM development, and their products demonstrated that primary cell lines are more representative of the tumor close to its native state and show a degree of molecular diversity, thus capturing the disease heterogeneity in a patient cohort. Molecular profiling revealed a significantly different transcriptome and marked metabolic shift towards a greater glycolytic phenotype in commercial compared with primary cell lines. Our results highlight that multiple, appropriately characterised, patient-derived tumor cell lines are required to enable concurrent evaluation of molecular profiles versus drug response. Furthermore, application of this approach to other difficult-to-treat tumors would generate improved cellular models for pre-clinical evaluation of novel targeted therapies. PMID:26891694

  10. Molecular profiling reveals primary mesothelioma cell lines recapitulate human disease

    PubMed Central

    Chernova, T; Sun, X M; Powley, I R; Galavotti, S; Grosso, S; Murphy, F A; Miles, G J; Cresswell, L; Antonov, A V; Bennett, J; Nakas, A; Dinsdale, D; Cain, K; Bushell, M; Willis, A E; MacFarlane, M

    2016-01-01

    Malignant mesothelioma (MM) is an aggressive, fatal tumor strongly associated with asbestos exposure. There is an urgent need to improve MM patient outcomes and this requires functionally validated pre-clinical models. Mesothelioma-derived cell lines provide an essential and relatively robust tool and remain among the most widely used systems for candidate drug evaluation. Although a number of cell lines are commercially available, a detailed comparison of these commercial lines with freshly derived primary tumor cells to validate their suitability as pre-clinical models is lacking. To address this, patient-derived primary mesothelioma cell lines were established and characterized using complementary multidisciplinary approaches and bioinformatic analysis. Clinical markers of mesothelioma, transcriptional and metabolic profiles, as well as the status of p53 and the tumor suppressor genes CDKN2A and NF2, were examined in primary cell lines and in two widely used commercial lines. Expression of MM-associated markers, as well as the status of CDKN2A, NF2, the ‘gatekeeper' in MM development, and their products demonstrated that primary cell lines are more representative of the tumor close to its native state and show a degree of molecular diversity, thus capturing the disease heterogeneity in a patient cohort. Molecular profiling revealed a significantly different transcriptome and marked metabolic shift towards a greater glycolytic phenotype in commercial compared with primary cell lines. Our results highlight that multiple, appropriately characterised, patient-derived tumor cell lines are required to enable concurrent evaluation of molecular profiles versus drug response. Furthermore, application of this approach to other difficult-to-treat tumors would generate improved cellular models for pre-clinical evaluation of novel targeted therapies. PMID:26891694

  11. Genomic Analysis of hESC Pedigrees Identifies De Novo Mutations and Enables Determination of the Timing and Origin of Mutational Events

    PubMed Central

    Ben-Yosef, Dalit; Boscolo, Francesca S.; Amir, Hadar; Malcov, Mira; Amit, Ami; Laurent, Louise C.

    2013-01-01

    Summary Given the association between mutational load and cancer, the observation that genetic aberrations are frequently found in human pluripotent stem cells (hPSCs) is of concern. Prior studies in human induced pluripotent stem cells (hiPSCs) have shown that deletions and regions of loss of heterozygosity (LOH) tend to arise during reprogramming and early culture, whereas duplications more frequently occur during long-term culture. For the corresponding experiments in human embryonic stem cells (hESCs), we studied two sets of hESC lines: one including the corresponding parental DNA and the other generated from single blastomeres from four sibling embryos. Here, we show that genetic aberrations observed in hESCs can originate during preimplantation embryo development and/or early derivation. These early aberrations are mainly deletions and LOH, whereas aberrations arising during long-term culture of hESCs are more frequently duplications. Our results highlight the importance of close monitoring of genomic integrity and the development of improved methods for derivation and culture of hPSCs. PMID:24035391

  12. Genome-Wide Transcriptome and Binding Sites Analyses Identify Early FOX Expressions for Enhancing Cardiomyogenesis Efficiency of hESC Cultures

    PubMed Central

    Yeo, Hock Chuan; Ting, Sherwin; Brena, Romulo Martin; Koh, Geoffrey; Chen, Allen; Toh, Siew Qi; Lim, Yu Ming; Oh, Steve Kah Weng; Lee, Dong-Yup

    2016-01-01

    The differentiation efficiency of human embryonic stem cells (hESCs) into heart muscle cells (cardiomyocytes) is highly sensitive to culture conditions. To elucidate the regulatory mechanisms involved, we investigated hESCs grown on three distinct culture platforms: feeder-free Matrigel, mouse embryonic fibroblast feeders, and Matrigel replated on feeders. At the outset, we profiled and quantified their differentiation efficiency, transcriptome, transcription factor binding sites and DNA-methylation. Subsequent genome-wide analyses allowed us to reconstruct the relevant interactome, thereby forming the regulatory basis for implicating the contrasting differentiation efficiency of the culture conditions. We hypothesized that the parental expressions of FOXC1, FOXD1 and FOXQ1 transcription factors (TFs) are correlative with eventual cardiomyogenic outcome. Through WNT induction of the FOX TFs, we observed the co-activation of WNT3 and EOMES which are potent inducers of mesoderm differentiation. The result strengthened our hypothesis on the regulatory role of the FOX TFs in enhancing mesoderm differentiation capacity of hESCs. Importantly, the final proportions of cells expressing cardiac markers were directly correlated to the strength of FOX inductions within 72 hours after initiation of differentiation across different cell lines and protocols. Thus, we affirmed the relationship between early FOX TF expressions and cardiomyogenesis efficiency. PMID:27501774

  13. Genome-Wide Transcriptome and Binding Sites Analyses Identify Early FOX Expressions for Enhancing Cardiomyogenesis Efficiency of hESC Cultures.

    PubMed

    Yeo, Hock Chuan; Ting, Sherwin; Brena, Romulo Martin; Koh, Geoffrey; Chen, Allen; Toh, Siew Qi; Lim, Yu Ming; Oh, Steve Kah Weng; Lee, Dong-Yup

    2016-01-01

    The differentiation efficiency of human embryonic stem cells (hESCs) into heart muscle cells (cardiomyocytes) is highly sensitive to culture conditions. To elucidate the regulatory mechanisms involved, we investigated hESCs grown on three distinct culture platforms: feeder-free Matrigel, mouse embryonic fibroblast feeders, and Matrigel replated on feeders. At the outset, we profiled and quantified their differentiation efficiency, transcriptome, transcription factor binding sites and DNA-methylation. Subsequent genome-wide analyses allowed us to reconstruct the relevant interactome, thereby forming the regulatory basis for implicating the contrasting differentiation efficiency of the culture conditions. We hypothesized that the parental expressions of FOXC1, FOXD1 and FOXQ1 transcription factors (TFs) are correlative with eventual cardiomyogenic outcome. Through WNT induction of the FOX TFs, we observed the co-activation of WNT3 and EOMES which are potent inducers of mesoderm differentiation. The result strengthened our hypothesis on the regulatory role of the FOX TFs in enhancing mesoderm differentiation capacity of hESCs. Importantly, the final proportions of cells expressing cardiac markers were directly correlated to the strength of FOX inductions within 72 hours after initiation of differentiation across different cell lines and protocols. Thus, we affirmed the relationship between early FOX TF expressions and cardiomyogenesis efficiency. PMID:27501774

  14. Impact of transient down-regulation of DREAM in human embryonic stem cell pluripotency: The role of DREAM in the maintenance of hESCs.

    PubMed

    Fontán-Lozano, A; Capilla-Gonzalez, V; Aguilera, Y; Mellado, N; Carrión, A M; Soria, B; Hmadcha, A

    2016-05-01

    Little is known about the functions of downstream regulatory element antagonist modulator (DREAM) in embryonic stem cells (ESCs). However, DREAM interacts with cAMP response element-binding protein (CREB) in a Ca(2+)-dependent manner, preventing CREB binding protein (CBP) recruitment. Furthermore, CREB and CBP are involved in maintaining ESC self-renewal and pluripotency. However, a previous knockout study revealed the protective function of DREAM depletion in brain aging degeneration and that aging is accompanied by a progressive decline in stem cells (SCs) function. Interestingly, we found that DREAM is expressed in different cell types, including human ESCs (hESCs), human adipose-derived stromal cells (hASCs), human bone marrow-derived stromal cells (hBMSCs), and human newborn foreskin fibroblasts (hFFs), and that transitory inhibition of DREAM in hESCs reduces their pluripotency, increasing differentiation. We stipulate that these changes are partly mediated by increased CREB transcriptional activity. Overall, our data indicates that DREAM acts in the regulation of hESC pluripotency and could be a target to promote or prevent differentiation in embryonic cells. PMID:26999760

  15. Modeling Type 2 Diabetes GWAS Candidate Gene Function in hESCs.

    PubMed

    Rutter, Guy A

    2016-09-01

    Type 2 diabetes is a complex polygenic disorder that affects about 1 in 12 adults. In this issue of Cell Stem Cell, Zeng et al. (2016) elegantly combine CRISPR-based gene editing in hESCs with directed β cell differentiation to investigate the functions of genes highlighted by genome-wide association studies (GWAS) for this disease. PMID:27588741

  16. A human embryonic stem cell line adapted for high throughput screening.

    PubMed

    Caron, Nicolas J; Gage, Blair K; O'Connor, Michael D; Eaves, Connie J; Kieffer, Timothy J; Piret, James M

    2013-10-01

    Human embryonic stem cells (hESCs) can be differentiated into multiple cell types with great therapeutic potential. However, optimizing the often multi-week cultures to obtain sufficient differentiated cell yields has been in part limited by the high variability of even parallel hESC differentiation cultures. We describe the isolation and features of a subline of CA1 hESCs (CA1S) that display a very high 25% cloning efficiency while retaining many properties of the parental hESCs, including being karyotypically normal and their ability to generate teratomas containing all three germ layers. Although more detailed analysis revealed that CA1S cells have a 3.8 Mb genomic duplication on chromosome 20, they remain highly useful. In particular, CA1S cells are readily expanded at high yields in culture and possess greatly reduced well-to-well variation even when seeded at 100 cells/well. Thus, 10(8) CA1S cells can be generated within one week from 10(6) cells to seed 10(6) wells. We determined that CA1S cells have the capacity to follow established in vitro differentiation protocols to pancreatic progenitors and subsequent hormone-positive cell types and used CA1S cells to explore definitive endoderm induction in a high performance screen (Z-factor = 0.97). This system revealed that CA1S cells do not require WNT3A to efficiently form definitive endoderm, a finding that was confirmed with H1 hESCs, although H1 cells did show modest benefits of high WNT3A doses. Proliferative index measurements of CA1S cells were shown to rapidly reflect their differentiation status in a high throughput system. Though results obtained with CA1S cells will need to be confirmed using conventional hESC lines, these cells should ease the development of optimized hESC growth and differentiation protocols. In particular, they should limit the more arduous secondary screens using hESCs to a smaller number of variables and doses. PMID:23613129

  17. Label-free concentration of viable neurons, hESCs and cancer cells by means of acoustophoresis.

    PubMed

    Zalis, Marina C; Reyes, Juan F; Augustsson, Per; Holmqvist, Staffan; Roybon, Laurent; Laurell, Thomas; Deierborg, Tomas

    2016-03-14

    Concentration of viable cell populations in suspension is of interest for several clinical and pre-clinical applications. Here, we report that microfluidic acoustophoresis is an effective method to efficiently concentrate live and viable cells with high target purity without any need for protein fluorescent labeling using antibodies or over-expression. We explored the effect of the acoustic field acoustic energy density and systematically used different protocols to induce apoptosis or cell death and then determined the efficiency of live and dead cell separation. We used the breast cancer cell line MCF-7, the mouse neuroblastoma N2a as well as human embryonic stem cells (hESCs) to demonstrate that this method is gentle and can be applied to different cell populations. First, we induced cell death by means of high osmotic shock using a high concentration of PBS (10×), the protein kinase inhibitor staurosporine, high concentrations of dimethyl sulfoxide (DMSO, 10%), and finally, cell starvation. In all the methods employed, we successfully induced cell death and were able to purify and concentrate the remaining live cells using acoustophoresis. Importantly, the concentration of viable cells was not dependent on a specific cell type. Further, we demonstrate that different death inducing stimuli have different effects on the intrinsic cell properties and therefore affect the efficiency of the acoustophoretic separation. PMID:26915333

  18. A whole-mechanical method to establish human embryonic stem cell line HN4 from discarded embryos.

    PubMed

    Li, Bin; Xu, Lan; Lu, Wei-Ying; Xu, Wen; Wang, Mei-Hong; Yang, Ke; Dong, Juan; Ding, Xiao-Yan; Huang, Yuan-Hua

    2010-12-01

    Since the first human embryonic stem cell (hESC) line was generated by Thomson et al. (in Science 282:1145-1147, 1998), hundreds of hESC lines have been reported by different labs, providing resources for basic research and regenerative medicine as well. However it has been widely recognized that hESC lines varied on their properties, in terms of gene expression profile, epigenetic modify profile, and differentiation tendency. Generation of more hESC lines will largely enhance our knowledge of hESCs innate character. In this current work, we reported the generation of HN4, a hESC line derived from grade III IVF human embryo by using a mixture of human foreskin fibroblast (HFF) and mouse embryonic fibroblast (MEF) as feeder layers, and a whole-mechanical method in inner cell mass (ICM) isolation. HN4 satisfied the criteria of hESCs pluripotency, with high expression of hESC surface markers (SSEA-3, SSEA-4, TRA-1-60, TRA-1-81), transcription factors (OCT-4, NANOG, REX-1), and alkaline phosphatase. It is able to differentiate to three germ layer derivatives when cultured in vitro, or in teratoma formation. Moreover, it displayed promising potential in neural differentiation under a proper culture condition, suggesting the advantage of HN4 in further investigation. Additionally, the whole-mechanical protocol for ICM isolation facilitates hESC line generation for its ease to handle. PMID:20976554

  19. Highly efficient differentiation of hESCs to functional hepatic endoderm requires ActivinA and Wnt3a signaling

    PubMed Central

    Hay, David C.; Fletcher, Judy; Payne, Catherine; Terrace, John D.; Gallagher, Ronald C. J.; Snoeys, Jan; Black, James R.; Wojtacha, Davina; Samuel, Kay; Hannoun, Zara; Pryde, Anne; Filippi, Celine; Currie, Ian S.; Forbes, Stuart J.; Ross, James A.; Newsome, Philip N.; Iredale, John P.

    2008-01-01

    Human embryonic stem cells (hESCs) are a valuable source of pluripotential primary cells. To date, however, their homogeneous cellular differentiation to specific cell types in vitro has proven difficult. Wnt signaling has been shown to play important roles in coordinating development, and we demonstrate that Wnt3a is differentially expressed at critical stages of human liver development in vivo. The essential role of Wnt3a in hepatocyte differentiation from hESCs is paralleled by our in vitro model, demonstrating the importance of a physiologic approach to cellular differentiation. Our studies provide compelling evidence that Wnt3a signaling is important for coordinated hepatocellular function in vitro and in vivo. In addition, we demonstrate that Wnt3a facilitates clonal plating of hESCs exhibiting functional hepatic differentiation. These studies represent an important step toward the use of hESC-derived hepatocytes in high-throughput metabolic analysis of human liver function. PMID:18719101

  20. Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells.

    PubMed

    Unger, Christian; Felldin, Ulrika; Rodin, Sergey; Nordenskjöld, Agneta; Dilber, Sirac; Hovatta, Outi

    2016-01-01

    After the first derivations of human embryonic stem cell (hESC) lines on fetal mouse feeder cell layers, the idea of using human cells instead of mouse cells as feeder cells soon arose. Mouse cells bear a risk of microbial contamination, and nonhuman immunogenic proteins are absorbed from the feeders to hESCs. Human skin fibroblasts can be effectively used as feeder cells for hESCs. The same primary cell line, which can be safely used for up to 15 passages after stock preparations, can be expanded and used for large numbers of hESC derivations and cultures. These cells are relatively easy to handle and maintain. No animal facilities or animal work is needed. Here, we describe the derivation, culture, and cryopreservation procedures for research-grade human skin fibroblast lines. We also describe how to make feeder layers for hESCs using these fibroblasts. PMID:26840224

  1. Label-free separation of human embryonic stem cells (hESCs) and their cardiac derivatives using Raman spectroscopy

    SciTech Connect

    Chan, J W; Lieu, D K; Huser, T R; Li, R A

    2008-09-08

    Self-renewable, pluripotent human embryonic stem cells (hESCs) can be differentiated into cardiomyocytes (CMs), providing an unlimited source of cells for transplantation therapies. However, unlike certain cell lineages such as hematopoietic cells, CMs lack specific surface markers for convenient identification, physical separation, and enrichment. Identification by immunostaining of cardiac-specific proteins such as troponin requires permeabilization, which renders the cells unviable and non-recoverable. Ectopic expression of a reporter protein under the transcriptional control of a heart-specific promoter for identifying hESC-derived CMs (hESC-CMs) is useful for research but complicates potential clinical applications. The practical detection and removal of undifferentiated hESCs in a graft, which may lead to tumors, is also critical. Here, we demonstrate a non-destructive, label-free optical method based on Raman scattering to interrogate the intrinsic biochemical signatures of individual hESCs and their cardiac derivatives, allowing cells to be identified and classified. By combining the Raman spectroscopic data with multivariate statistical analysis, our results indicate that hESCs, human fetal left ventricular CMs, and hESC-CMs can be identified by their intrinsic biochemical characteristics with an accuracy of 96%, 98% and 66%, respectively. The present study lays the groundwork for developing a systematic and automated method for the non-invasive and label-free sorting of (i) high-quality hESCs for expansion, and (ii) ex vivo CMs (derived from embryonic or adult stem cells) for cell-based heart therapies.

  2. Generation of Human Embryonic Stem Cell Line Expressing zsGreen in Cholinergic Neurons Using CRISPR/Cas9 System.

    PubMed

    Zhou, Jing; Wang, Chencheng; Zhang, Kunshan; Wang, Yingying; Gong, Xi; Wang, Yanlu; Li, Siguang; Luo, Yuping

    2016-08-01

    Lineage specific human embryonic stem cell (hESC) reporter cell line is a versatile tool for biological studies on real time monitoring of differentiation, physiological and biochemical features of special cell types and pathological mechanism of disease. Here we report the generation of ChAT-zsGreen reporter hESC line that express zsGreen under the control of the choline acetyltransferase (ChAT) promoter using CRISPR (Clustered Regularly Interspersed Short Palindromic Repeats)/Cas9 system. We show that the ChAT-zsGreen hESC reporter cell lines retain the features of undifferentiated hESC. After cholinergic neuronal differentiation, cholinergic neurons were clearly labeled with green fluorescence protein (zsGreen). The ChAT-zsGreen reporter hESC lines are invaluable not only for the monitoring cholinergic neuronal differentiation but also for study physiological and biochemical hallmarks of cholinergic neurons. PMID:27113041

  3. Single exosome study reveals subpopulations distributed among cell lines with variability related to membrane content

    PubMed Central

    Smith, Zachary J.; Lee, Changwon; Rojalin, Tatu; Carney, Randy P.; Hazari, Sidhartha; Knudson, Alisha; Lam, Kit; Saari, Heikki; Ibañez, Elisa Lazaro; Viitala, Tapani; Laaksonen, Timo; Yliperttula, Marjo; Wachsmann-Hogiu, Sebastian

    2015-01-01

    Current analysis of exosomes focuses primarily on bulk analysis, where exosome-to-exosome variability cannot be assessed. In this study, we used Raman spectroscopy to study the chemical composition of single exosomes. We measured spectra of individual exosomes from 8 cell lines. Cell-line-averaged spectra varied considerably, reflecting the variation in total exosomal protein, lipid, genetic, and cytosolic content. Unexpectedly, single exosomes isolated from the same cell type also exhibited high spectral variability. Subsequent spectral analysis revealed clustering of single exosomes into 4 distinct groups that were not cell-line specific. Each group contained exosomes from multiple cell lines, and most cell lines had exosomes in multiple groups. The differences between these groups are related to chemical differences primarily due to differing membrane composition. Through a principal components analysis, we identified that the major sources of spectral variation among the exosomes were in cholesterol content, relative expression of phospholipids to cholesterol, and surface protein expression. For example, exosomes derived from cancerous versus non-cancerous cell lines can be largely separated based on their relative expression of cholesterol and phospholipids. We are the first to indicate that exosome subpopulations are shared among cell types, suggesting distributed exosome functionality. The origins of these differences are likely related to the specific role of extracellular vesicle subpopulations in both normal cell function and carcinogenesis, and they may provide diagnostic potential at the single exosome level. PMID:26649679

  4. Female Sex Bias in Human Embryonic Stem Cell Lines

    PubMed Central

    Ben-Yosef, Dalit; Amit, Ami; Malcov, Mira; Frumkin, Tsvia; Ben-Yehudah, Ahmi; Eldar, Ido; Mey-Raz, Nava; Azem, Foad; Altarescu, Gheona; Renbaum, Paul; Beeri, Rachel; Varshaver, Irit; Eldar-Geva, Talia; Epsztejn-Litman, Silvina; Levy-Lahad, Ephrat

    2012-01-01

    The factors limiting the rather inefficient derivation of human embryonic stem cells (HESCs) are not fully understood. The aim of this study was to analyze the sex ratio in our 42 preimplantation genetic diagnosis (PGD)-HESC lines, in an attempt to verify its affect on the establishment of HESC lines. The ratio between male and female PGD-derived cell lines was compared. We found a significant increase in female cell lines (76%). This finding was further confirmed by a meta-analysis for combining the results of all PGD-derived HESC lines published to date (148) and all normal karyotyped HESC lines derived from spare in vitro fertilization embryos worldwide (397). Further, gender determination of embryos demonstrated that this difference originates from the actual derivation process rather than from unequal representation of male and female embryos. It can therefore be concluded that the clear-cut tendency for female preponderance is attributed to suboptimal culture conditions rather than from a true gender imbalance in embryos used for derivation of HESC lines. We propose a mechanism in which aberrant X chromosome inactivation and/or overexpression of critical metabolic X-linked genes might explain this sex dimorphism. PMID:21585244

  5. Female sex bias in human embryonic stem cell lines.

    PubMed

    Ben-Yosef, Dalit; Amit, Ami; Malcov, Mira; Frumkin, Tsvia; Ben-Yehudah, Ahmi; Eldar, Ido; Mey-Raz, Nava; Azem, Foad; Altarescu, Gheona; Renbaum, Paul; Beeri, Rachel; Varshaver, Irit; Eldar-Geva, Talia; Epsztejn-Litman, Silvina; Levy-Lahad, Ephrat; Eiges, Rachel

    2012-02-10

    The factors limiting the rather inefficient derivation of human embryonic stem cells (HESCs) are not fully understood. The aim of this study was to analyze the sex ratio in our 42 preimplantation genetic diagnosis (PGD)-HESC lines, in an attempt to verify its affect on the establishment of HESC lines. The ratio between male and female PGD-derived cell lines was compared. We found a significant increase in female cell lines (76%). This finding was further confirmed by a meta-analysis for combining the results of all PGD-derived HESC lines published to date (148) and all normal karyotyped HESC lines derived from spare in vitro fertilization embryos worldwide (397). Further, gender determination of embryos demonstrated that this difference originates from the actual derivation process rather than from unequal representation of male and female embryos. It can therefore be concluded that the clear-cut tendency for female preponderance is attributed to suboptimal culture conditions rather than from a true gender imbalance in embryos used for derivation of HESC lines. We propose a mechanism in which aberrant X chromosome inactivation and/or overexpression of critical metabolic X-linked genes might explain this sex dimorphism. PMID:21585244

  6. Myoblasts Derived From Normal hESCs and Dystrophic hiPSCs Efficiently Fuse With Existing Muscle Fibers Following Transplantation

    PubMed Central

    Goudenege, Sébastien; Lebel, Carl; Huot, Nicolas B; Dufour, Christine; Fujii, Isao; Gekas, Jean; Rousseau, Joël; Tremblay, Jacques P

    2012-01-01

    Human embryonic stem cells (hESCs) and human-induced pluripotent stem cells (hiPSCs) have an endless self-renewal capacity and can theoretically differentiate into all types of lineages. They thus represent an unlimited source of cells for therapies of regenerative diseases, such as Duchenne muscular dystrophy (DMD), and for tissue repair in specific medical fields. However, at the moment, the low number of efficient specific lineage differentiation protocols compromises their use in regenerative medicine. We developed a two-step procedure to differentiate hESCs and dystrophic hiPSCs in myogenic cells. The first step was a culture in a myogenic medium and the second step an infection with an adenovirus expressing the myogenic master gene MyoD. Following infection, the cells expressed several myogenic markers and formed abundant multinucleated myotubes in vitro. When transplanted in the muscle of Rag/mdx mice, these cells participated in muscle regeneration by fusing very well with existing muscle fibers. Our findings provide an effective method that will permit to use hESCs or hiPSCs for preclinical studies in muscle repair. PMID:22990676

  7. A negative genetic interaction map in isogenic cancer cell lines reveals cancer cell vulnerabilities

    PubMed Central

    Vizeacoumar, Franco J; Arnold, Roland; Vizeacoumar, Frederick S; Chandrashekhar, Megha; Buzina, Alla; Young, Jordan T F; Kwan, Julian H M; Sayad, Azin; Mero, Patricia; Lawo, Steffen; Tanaka, Hiromasa; Brown, Kevin R; Baryshnikova, Anastasia; Mak, Anthony B; Fedyshyn, Yaroslav; Wang, Yadong; Brito, Glauber C; Kasimer, Dahlia; Makhnevych, Taras; Ketela, Troy; Datti, Alessandro; Babu, Mohan; Emili, Andrew; Pelletier, Laurence; Wrana, Jeff; Wainberg, Zev; Kim, Philip M; Rottapel, Robert; O'Brien, Catherine A; Andrews, Brenda; Boone, Charles; Moffat, Jason

    2013-01-01

    Improved efforts are necessary to define the functional product of cancer mutations currently being revealed through large-scale sequencing efforts. Using genome-scale pooled shRNA screening technology, we mapped negative genetic interactions across a set of isogenic cancer cell lines and confirmed hundreds of these interactions in orthogonal co-culture competition assays to generate a high-confidence genetic interaction network of differentially essential or differential essentiality (DiE) genes. The network uncovered examples of conserved genetic interactions, densely connected functional modules derived from comparative genomics with model systems data, functions for uncharacterized genes in the human genome and targetable vulnerabilities. Finally, we demonstrate a general applicability of DiE gene signatures in determining genetic dependencies of other non-isogenic cancer cell lines. For example, the PTEN−/− DiE genes reveal a signature that can preferentially classify PTEN-dependent genotypes across a series of non-isogenic cell lines derived from the breast, pancreas and ovarian cancers. Our reference network suggests that many cancer vulnerabilities remain to be discovered through systematic derivation of a network of differentially essential genes in an isogenic cancer cell model. PMID:24104479

  8. Power-Laws and the Use of Pluripotent Stem Cell Lines

    PubMed Central

    Lenz, Michael; Kobold, Sabine; MacArthur, Ben D.; Schuppert, Andreas; Löser, Peter; Müller, Franz-Josef

    2013-01-01

    It is widely accepted that the (now reversed) Bush administration’s decision to restrict federal funding for human embryonic stem cell (hESC) research to a few “eligible” hESC lines is responsible for the sustained preferential use of a small subset of hESC lines (principally the H1 and H9 lines) in basic and preclinical research. Yet, international hESC usage patterns, in both permissive and restrictive political environments, do not correlate with a specific type of stem cell policy. Here we conducted a descriptive analysis of hESC line usage and compared the ability of policy-driven processes and collaborative processes inherent to biomedical research to recapitulate global hESC usage patterns. We find that current global hESC usage can be modelled as a cumulative advantage process, independent of restrictive or permissive policy influence, suggesting a primarily innovation-driven (rather than policy-driven) mechanism underlying human pluripotent stem cell usage in preclinical research. PMID:23300961

  9. Functional Coding Variation in Recombinant Inbred Mouse Lines Reveals Novel Serotonin Transporter-Associated Phenotypes

    SciTech Connect

    Carneiro, Ana; Airey, David; Thompson, Brent; Zhu, C; Rinchik, Eugene M; Lu, Lu; Chesler, Elissa J; Erikson, Keith; Blakely, Randy

    2009-01-01

    The human serotonin (5-hydroxytryptamine, 5-HT) transporter (hSERT, SLC6A4) figures prominently in the etiology or treatment of many prevalent neurobehavioral disorders including anxiety, alcoholism, depression, autism and obsessive-compulsive disorder (OCD). Here we utilize naturally occurring polymorphisms in recombinant inbred (RI) lines to identify novel phenotypes associated with altered SERT function. The widely used mouse strain C57BL/6J, harbors a SERT haplotype defined by two nonsynonymous coding variants (Gly39 and Lys152 (GK)). At these positions, many other mouse lines, including DBA/2J, encode Glu39 and Arg152 (ER haplotype), assignments found also in hSERT. Synaptosomal 5-HT transport studies revealed reduced uptake associated with the GK variant. Heterologous expression studies confirmed a reduced SERT turnover rate for the GK variant. Experimental and in silico approaches using RI lines (C57Bl/6J X DBA/2J=BXD) identifies multiple anatomical, biochemical and behavioral phenotypes specifically impacted by GK/ER variation. Among our findings are multiple traits associated with anxiety and alcohol consumption, as well as of the control of dopamine (DA) signaling. Further bioinformatic analysis of BXD phenotypes, combined with biochemical evaluation of SERT knockout mice, nominates SERT-dependent 5-HT signaling as a major determinant of midbrain iron homeostasis that, in turn, dictates ironregulated DA phenotypes. Our studies provide a novel example of the power of coordinated in vitro, in vivo and in silico approaches using murine RI lines to elucidate and quantify the system-level impact of gene variation.

  10. Transcript profiling reveals expression differences in wild-type and glabrous soybean lines

    PubMed Central

    2011-01-01

    Background Trichome hairs affect diverse agronomic characters such as seed weight and yield, prevent insect damage and reduce loss of water but their molecular control has not been extensively studied in soybean. Several detailed models for trichome development have been proposed for Arabidopsis thaliana, but their applicability to important crops such as cotton and soybean is not fully known. Results Two high throughput transcript sequencing methods, Digital Gene Expression (DGE) Tag Profiling and RNA-Seq, were used to compare the transcriptional profiles in wild-type (cv. Clark standard, CS) and a mutant (cv. Clark glabrous, i.e., trichomeless or hairless, CG) soybean isoline that carries the dominant P1 allele. DGE data and RNA-Seq data were mapped to the cDNAs (Glyma models) predicted from the reference soybean genome, Williams 82. Extending the model length by 250 bp at both ends resulted in significantly more matches of authentic DGE tags indicating that many of the predicted gene models are prematurely truncated at the 5' and 3' UTRs. The genome-wide comparative study of the transcript profiles of the wild-type versus mutant line revealed a number of differentially expressed genes. One highly-expressed gene, Glyma04g35130, in wild-type soybean was of interest as it has high homology to the cotton gene GhRDL1 gene that has been identified as being involved in cotton fiber initiation and is a member of the BURP protein family. Sequence comparison of Glyma04g35130 among Williams 82 with our sequences derived from CS and CG isolines revealed various SNPs and indels including addition of one nucleotide C in the CG and insertion of ~60 bp in the third exon of CS that causes a frameshift mutation and premature truncation of peptides in both lines as compared to Williams 82. Conclusion Although not a candidate for the P1 locus, a BURP family member (Glyma04g35130) from soybean has been shown to be abundantly expressed in the CS line and very weakly expressed in the

  11. The use of human amniotic fluid mesenchymal stem cells as the feeder layer to establish human embryonic stem cell lines.

    PubMed

    Soong, Yung-Kwei; Huang, Shang-Yu; Yeh, Chiu-Hsiang; Wang, Tzu-Hao; Chang, Kuo-Hsuan; Cheng, Po-Jen; Shaw, S W Steven

    2015-12-01

    Human embryonic stem cells (hESCs) are pluripotent cells that have the potential to differentiate into the three germ layers and possibly all tissues of the human body. To fulfil the clinical potentials for cell-based therapy, banks of hESC lines that express different combinations of the major histocompatibility genes should be established, preferably without exposing such cells to animal cells and proteins. In this study, we tested human amniotic fluid mesenchymal stem cells (AFMSCs) as feeder cells to support the growth of hESCs. Our results indicated that mitomycin-treated AFMSCs were able to support the newly established hESC lines CGLK-1 and CGLK-2. The hESC colonies cultured on AFMSCs expressed alkaline phosphatase (ALK-P), SSEA-4, TRA-1-60, TRA-1-81, Oct-4, Nanog and Sox-2, which are markers for undifferentiated hESCs. Chromosomal analyses of both hESC lines, CGLK-1 and CGLK-2, which were cultured on AFMSC feeders for 22 and 14 passages, respectively, were confirmed to be normal karyotypes (46, XX). The ability of AFMSCs as feeder cells to maintain the undifferentiated growth and pluripotency of hESCs was confirmed by in vivo formation of teratomas derived on AFMSC hESCs in severe combined immune-compromised mice. The use of AFMSCs for feeder cells to culture hESCs has several advantages, in that AFMSCs are not tumourigenic and can be expanded extensively with a short doubling time. PMID:23460275

  12. Transcriptome Profiling Reveals Degree of Variability in Induced Pluripotent Stem Cell Lines: Impact for Human Disease Modeling.

    PubMed

    Schuster, Jens; Halvardson, Jonatan; Pilar Lorenzo, Laureanne; Ameur, Adam; Sobol, Maria; Raykova, Doroteya; Annerén, Göran; Feuk, Lars; Dahl, Niklas

    2015-10-01

    Induced pluripotent stem cell (iPSC) technology has become an important tool for disease modeling. Insufficient data on the variability among iPSC lines derived from a single somatic parental cell line have in practice led to generation and analysis of several, usually three, iPSC sister lines from each parental cell line. We established iPSC lines from a human fibroblast line (HDF-K1) and used transcriptome sequencing to investigate the variation among three sister lines (iPSC-K1A, B, and C). For comparison, we analyzed the transcriptome of an iPSC line (iPSC-K5B) derived from a different fibroblast line (HDF-K5), a human embryonic stem cell (ESC) line (ESC-HS181), as well as the two parental fibroblast lines. All iPSC lines fulfilled stringent criteria for pluripotency. In an unbiased cluster analysis, all stem cell lines (four iPSCs and one ESC) clustered together as opposed to the parental fibroblasts. The transcriptome profiles of the three iPSC sister lines were indistinguishable from each other, and functional pathway analysis did not reveal any significant hits. In contrast, the expression profiles of the ESC line and the iPSC-K5B line were distinct from that of the sister lines iPSC-K1A, B, and C. Differentiation to embryoid bodies and subsequent analysis of germ layer markers in the five stem cell clones confirmed that the distribution of their expression profiles was retained. Taken together, our observations stress the importance of using iPSCs of different parental origin rather than several sister iPSC lines to distinguish disease-associated mechanisms from genetic background effects in disease modeling. PMID:26348590

  13. Comparative study of hematopoietic differentiation between human embryonic stem cell lines.

    PubMed

    Melichar, Heather; Li, Ou; Ross, Jenny; Haber, Hilary; Cado, Dragana; Nolla, Hector; Robey, Ellen A; Winoto, Astar

    2011-01-01

    Directed differentiation of human embryonic stem cells (hESCs) into any desired cell type has been hailed as a therapeutic promise to cure many human diseases. However, substantial roadblocks still exist for in vitro differentiation of hESCs into distinct cell types, including T lymphocytes. Here we examined the hematopoietic differentiation potential of six different hESC lines. We compare their ability to develop into CD34(+) or CD34(+)CD45(+) hematopoietic precursor populations under several differentiation conditions. Comparison of lymphoid potential of hESC derived- and fetal tissue derived-hematopoietic precursors was also made. We found diverse hematopoietic potential between hESC lines depending on the culture or passage conditions. In contrast to fetal-derived hematopoietic precursors, none of the CD34(+) precursors differentiated from hESCs were able to develop further into T cells. These data underscore the difficulties in the current strategy of hESC forward differentiation and highlight distinct differences between CD34(+) hematopoietic precursors generated in vitro versus in vivo. PMID:21603627

  14. Generation of Sheffield (Shef) human embryonic stem cell lines using a microdrop culture system.

    PubMed

    Aflatoonian, Behrouz; Ruban, Ludmila; Shamsuddin, Shamsul; Baker, Duncan; Andrews, Peter; Moore, Harry

    2010-04-01

    The conventional method for the derivation of human embryonic stem cells (hESCs) involves inner cell mass (ICM) co-culture with a feeder layer of inactivated mouse or human embryonic fibroblasts in an in vitro fertilisation culture dish. Growth factors potentially involved in primary derivation of hESCs may be lost or diluted in such a system. We established a microdrop method which maintained feeder cells and efficiently generated hESCs. Embryos were donated for stem cell research after fully informed patient consent. A feeder cell layer was made by incubating inactivated mouse embryonic fibroblasts (MEFs) feeder cells in a 50 microl drop of medium (DMEM/10% foetal calf serum) under mineral oil in a small tissue culture dish. MEFs formed a confluent layer and medium was replaced with human embryonic stem medium supplemented with 10% Plasmanate (Bayer) and incubated overnight. Cryopreserved embryos were thawed and cultured until the blastocyst stage and the zona pellucida removed with pronase (2 mg/ml; Calbiochem). A zona-free intact blastocyst was placed in the feeder microdrop and monitored for ES derivation with medium changed every 2-3 d. Proliferating hESCs were passaged into other feeder drops and standard feeder preparation by manual dissection until a stable cell line was established. Six hESC lines (Shef 3-8) were derived. From a total of 46 blastocysts (early to expanded), five hESC lines were generated (Shef 3-7). Shef 3-6 were generated on MEFs from 25 blastocysts. Shef7 was generated on human foetal gonadal embryonic fibroblasts from a further 21 blastocysts. From our experience, microdrop technique is more efficient than conventional method for derivation of hESCs and it is much easier to monitor early hESC derivation. The microdrop method lends itself to good manufacturing practice derivation of hESCs. PMID:20224972

  15. The Cellular Architecture of the Larval Zebrafish Tectum, as Revealed by Gal4 Enhancer Trap Lines

    PubMed Central

    Scott, Ethan K.; Baier, Herwig

    2009-01-01

    We have carried out a Gal4 enhancer trap screen in zebrafish, and have generated 184 stable transgenic lines with interesting expression patterns throughout the nervous system. Of these, three display clear expression in the tectum, each with a distinguishable and stereotyped distribution of Gal4 expressing cells. Detailed morphological analysis of single cells, using a genetic “Golgi-like” labelling method, revealed four common cell types (superficial, periventricular, shallow periventricular, and radial glial), along with a range of other less common neurons. The shallow periventricular (PV) and a subset of the PV neurons are tectal efferent neurons that target various parts of the reticular formation. We find that it is specifically PV neurons with dendrites in the deep tectal neuropil that target the reticular formation. This indicates that these neurons receive the tectum's highly processed visual information (which is fed from the superficial retinorecipient layers), and relay it to premotor regions. Our results show that the larval tectum, both broadly and at the single cell level, strongly resembles a miniature version of its adult counterpart, and that it has all of the necessary anatomical characteristics to inform motor responses based on sensory input. We also demonstrate that mosaic expression of GFP in Gal4 enhancer trap lines can be used to describe the types and abundance of cells in an expression pattern, including the architectures of individual neurons. Such detailed anatomical descriptions will be an important part of future efforts to describe the functions of discrete tectal circuits in the generation of behavior. PMID:19862330

  16. Chromatic line-profile tomography to reveal exoplanetary atmospheres: application to HD 189733b

    NASA Astrophysics Data System (ADS)

    Borsa, F.; Rainer, M.; Poretti, E.

    2016-05-01

    Context. Transmission spectroscopy can be used to constrain the properties of exoplanetary atmospheres. During a transit, the light blocked from the atmosphere of the planet leaves an imprint in the light coming from the star. This has been shown for many exoplanets with both photometry and spectroscopy, using different analysis methods. Aims: We test chromatic line-profile tomography as a new tool to investigate exoplanetary atmospheres. The signal imprinted on the cross-correlation function (CCF) by a planet transiting its star is dependent on the planet-to-star radius ratio. We want to verify whether the precision reachable on the CCF obtained from a subset of the spectral orders of the HARPS spectrograph is high enough to determine the radius of a planet at different wavelengths. Methods: We analyze HARPS archival data of three transits of HD 189733b. We divide the HARPS spectral range into seven broadbands, calculating for each band the ratio between the area of the out-of-transit CCF and the area of the signal imprinted by the planet on it during the full part of the transit. We take into account the effect of the limb darkening using the theoretical coefficients of a linear law. Averaging the results of three different transits allows us to obtain a good-quality broadband transmission spectrum of HD 189733b with a greater precision than that of the chromatic Rossiter McLaughlin effect. Results: We proved that chromatic line-profile tomography is an interesting way to reveal broadband transmission spectra of exoplanets: our analysis of the atmosphere of HD 189733b is in agreement with other ground- and space-based observations. The independent analysis of different transits emphasizes the probability that stellar activity plays a role in the extracted transmission spectrum. Therefore, care should be taken when claiming that Rayleigh scattering is present in the atmosphere of exoplanets orbiting active stars using only one transit.

  17. Inhibition of Lysine-Specific Demethylase-1 (LSD1/KDM1A) Promotes the Adipogenic Differentiation of hESCs Through H3K4 Methylation.

    PubMed

    Xiong, Yujing; Wang, Enyin; Huang, Yan; Guo, Xiaoyi; Yu, Yiping; Du, Qingyun; Ding, Xiaoyan; Sun, Yingpu

    2016-06-01

    Given their totipotency, human embryonic stem cells (hESCs) can differentiate into all types of cells, including adipocytes, and provide an excellent research model for studying diseases associated with the metabolism of adipocytes, such as obesity and diabetes mellitus. Epigenetic regulation, including DNA methylation and histone modification, plays an essential role in the development and differentiation of hESCs. Lysine-specific demethylase 1 (LSD1), a well-characterized histone-modifying enzyme, demethylates dimethylated histone H3 lysine 4 (H3K4) through a flavin adenine dinucleotide (FAD)-dependent oxidative reaction. LSD1 affects the growth and differentiation of human and mouse ES cells, and the deletion of this gene in mice leads to embryonic lethality. Here, we investigated the functional role of LSD1 during the adipogenic differentiation of hESCs involving the demethylation of H3K4. We also found that treating hESCs with the LSD1 inhibitor CBB1007 promotes the adipogenic differentiation of hESCs. PMID:27059868

  18. HDE 245059: A WEAK-LINED T TAURI BINARY REVEALED BY CHANDRA AND KECK

    SciTech Connect

    Baldovin-Saavedra, C.; Audard, M.; Duchene, G.; Guedel, M.; Skinner, S.L.; Paerels, F. B. S.; Ghez, A.; McCabe, C.

    2009-05-20

    We present the Chandra High Energy Transmission Grating Spectrometer and Keck observations of HDE 245059, a young weak-lined T Tauri star (WTTS), member of the pre-main-sequence group in the {lambda} Orionis Cluster. Our high spatial resolution, near-infrared observations with Keck reveal that HDE 245059 is in fact a binary separated by 0.''87, probably composed of two WTTS based on their color indices. Based on this new information we have obtained an estimate of the masses of the binary components; {approx}3 M {sub sun} and {approx}2.5 M {sub sun} for the north and south components, respectively. We have also estimated the age of the system to be {approx}2-3 Myr. We detect both components of the binary in the zeroth-order Chandra image and in the grating spectra. The light curves show X-ray variability of both sources and in particular a flaring event in the weaker southern component. The spectra of both stars show similar features: a combination of cool and hot plasma as demonstrated by several iron lines from Fe XVII to Fe XXV and a strong bremsstrahlung continuum at short wavelengths. We have fitted the combined grating and zeroth-order spectrum (considering the contribution of both stars) in XSPEC. The coronal abundances and emission measure distribution for the binary have been obtained using different methods, including a continuous emission measure distribution and a multi-temperature approximation. In all cases we have found that the emission is dominated by plasma between {approx}8 and {approx}15 MK a soft component at {approx}4 MK and a hard component at {approx}50 MK are also detected. The value of the hydrogen column density was low, N {sub H} {approx} 8 x 10{sup 19} cm{sup -2}, likely due to the clearing of the inner region of the {lambda} Orionis cloud, where HDE 245059 is located. The abundance pattern shows an inverse first ionization potential effect for all elements from O to Fe, the only exception being Ca. To obtain the properties of the

  19. Mapping of fiber quality QTLs reveals useful variation and footprints of cotton domestication using introgression lines.

    PubMed

    Zhang, Shu-Wen; Zhu, Xie-Fei; Feng, Liu-Chun; Gao, Xiang; Yang, Biao; Zhang, Tian-Zhen; Zhou, Bao-Liang

    2016-01-01

    Fiber quality improvement is a driving force for further cotton domestication and breeding. Here, QTLs for fiber quality were mapped in 115 introgression lines (ILs) first developed from two intraspecific populations of cultivated and feral cotton landraces. A total of 60 QTLs were found, which explained 2.03-16.85% of the phenotypic variance found in fiber quality traits. A total of 36 markers were associated with five fiber traits, 33 of which were found to be associated with QTLs in multiple environments. In addition, nine pairs of common QTLs were identified; namely, one pair of QTLs for fiber elongation, three pairs for fiber length, three pairs for fiber strength and two pairs for micronaire (qMICs). All common QTLs had additive effects in the same direction in both IL populations. We also found five QTL clusters, allowing cotton breeders to focus their efforts on regions of QTLs with the highest percentages of phenotypic variance. Our results also reveal footprints of domestication; for example, fourteen QTLs with positive effects were found to have remained in modern cultivars during domestication, and two negative qMICs that had never been reported before were found, suggesting that the qMICs regions may be eliminated during artificial selection. PMID:27549323

  20. Mapping of fiber quality QTLs reveals useful variation and footprints of cotton domestication using introgression lines

    PubMed Central

    Zhang, Shu-Wen; Zhu, Xie-Fei; Feng, Liu-Chun; Gao, Xiang; Yang, Biao; Zhang, Tian-Zhen; Zhou, Bao-Liang

    2016-01-01

    Fiber quality improvement is a driving force for further cotton domestication and breeding. Here, QTLs for fiber quality were mapped in 115 introgression lines (ILs) first developed from two intraspecific populations of cultivated and feral cotton landraces. A total of 60 QTLs were found, which explained 2.03–16.85% of the phenotypic variance found in fiber quality traits. A total of 36 markers were associated with five fiber traits, 33 of which were found to be associated with QTLs in multiple environments. In addition, nine pairs of common QTLs were identified; namely, one pair of QTLs for fiber elongation, three pairs for fiber length, three pairs for fiber strength and two pairs for micronaire (qMICs). All common QTLs had additive effects in the same direction in both IL populations. We also found five QTL clusters, allowing cotton breeders to focus their efforts on regions of QTLs with the highest percentages of phenotypic variance. Our results also reveal footprints of domestication; for example, fourteen QTLs with positive effects were found to have remained in modern cultivars during domestication, and two negative qMICs that had never been reported before were found, suggesting that the qMICs regions may be eliminated during artificial selection. PMID:27549323

  1. Human Embryonic Stem Cell Lines with Lesions in FOXP3 and NF1

    PubMed Central

    Zhu, Hui; Behr, Barry; Reddy, Vikrant V.; Hughes, Mark; Pan, Yuqiong; Baker, Julie

    2016-01-01

    Human embryonic stem cells (hESCs) are derived from the inner cell mass (ICM) of blastocyst staged embryos. Spare blastocyst staged embryos were obtained by in vitro fertilization (IVF) and donated for research purposes. hESCs carrying specific mutations can be used as a powerful cell system in modeling human genetic disorders. We obtained preimplantation genetic diagnosed (PGD) blastocyst staged embryos with genetic mutations that cause human disorders and derived hESCs from these embryos. We applied laser assisted micromanipulation to isolate the inner cell mass from the blastocysts and plated the ICM onto the mouse embryonic fibroblast cells. Two hESC lines with lesions in FOXP3 and NF1 were established. Both lines maintain a typical undifferentiated hESCs phenotype and present a normal karyotype. The two lines express a panel of pluripotency markers and have the potential to differentiate to the three germ layers in vitro and in vivo. The hESC lines with lesions in FOXP3 and NF1 are available for the scientific community and may serve as an important resource for research into these disease states. PMID:26990425

  2. A novel stem cell associated marker identified by monoclonal antibody HESC5:3 differentiates between neoplastic lesions in follicular thyroid neoplasms.

    PubMed

    Heikkilä, Annukka; Fermér, Christian; Hagström, Jaana; Louhimo, Johanna; Mäenpää, Hanna; Siironen, Päivi; Heiskanen, Ilkka; Nilsson, Olle; Arola, Johanna; Haglund, Caj

    2015-07-01

    Follicular thyroid lesions are the bane of cytopathology. Differentiation between adenoma and carcinoma is impossible, and often these neoplasms are indistinguishable even from uninodular goitre. In other cancers as well, a theory of stem cells as the origin of cancer has been discussed in thyroid carcinogenesis. We aimed to examine a novel stem cell associated marker identified by monoclonal antibody HESC5:3 in follicular lesions in an attempt to find a marker for differential diagnosis in thyroid cytopathology. HESC5:3 was raised against and is specific for undifferentiated human embryonic stem cells. The epitope of this novel antibody is to be defined. Immunohistochemical expression of HESC5:3 was examined in clinical material comprised of follicular neoplasms (83 adenomas, 43 carcinomas) and non-neoplastic lesions (41 goitrous, 22 hyperplastic, 23 normal tissue specimens). Staining differed significantly between neoplastic and non-neoplastic lesions. Nuclear staining was increased in non-neoplastic cells, whereas in neoplastic cells expression was mainly cytoplasmic. There was no difference between benign and malignant lesions, suggesting a role in early tumourigenesis. In conclusion, the HESC5:3 epitope may be of benefit as a neoplasia marker in distinguishing between uninodular goitre and neoplasia. Characterization of the epitope would increase the interest in this promising new stem cell associated marker. PMID:25960045

  3. The precision of experienced action video-game players: line bisection reveals reduced leftward response bias.

    PubMed

    Latham, Andrew J; Patston, Lucy L M; Tippett, Lynette J

    2014-11-01

    Twenty-two experienced action video-game players (AVGPs) and 18 non-VGPs were tested on a pen-and-paper line bisection task that was untimed. Typically, right-handers bisect lines 2 % to the left of true centre, a bias thought to reflect the dominance of the right-hemisphere for visuospatial attention. Expertise may affect this bias, with expert musicians showing no bias in line bisection performance. Our results show that experienced-AVGPs also bisect lines with no bias with their right hand and a significantly reduced bias with their left hand compared to non-AVGPs. Bisections by experienced-AVGPs were also more precise than those of non-AVGPs. These findings show the cognitive proficiencies of experienced-AVGPs can generalize beyond computer based tasks, which resemble their training environment. PMID:25341651

  4. Hemispheric asymmetries in perceived depth revealed through a radial line bisection task.

    PubMed

    Szpak, Ancrêt; Thomas, Nicole A; Nicholls, Michael E R

    2016-03-01

    Research suggests that the left cerebral hemisphere is predisposed for processing stimuli in 'near' space, whereas the right hemisphere is specialised for processing stimuli in 'far' space. This hypothesis was tested directly by asking 25 undergraduates to carry out a landmark radial line bisection task. To test the effect of hemispheric differences in processing, the lines were placed to the left, right or centre within the transverse plane. Consistent with predictions, lines in all three conditions were bisected distal to the true centre. More importantly, there was an asymmetry whereby the distal bias was stronger for lines presented in the left hemispace compared to the right hemispace. The results demonstrate that the perception of depth is affected by left/right placement along the lateral axis and highlight the cognitive/neural interplay between the radial and lateral axes. PMID:26645309

  5. Genetic Rearrangements of Six Wheat–Agropyron cristatum 6P Addition Lines Revealed by Molecular Markers

    PubMed Central

    Su, Junji; Zhang, Jinpeng; Song, Liqiang; Gao, Ainong; Yang, Xinming; Li, Xiuquan; Liu, Weihua; Li, Lihui

    2014-01-01

    Agropyron cristatum (L.) Gaertn. (2n = 4x = 28, PPPP) not only is cultivated as pasture fodder but also could provide many desirable genes for wheat improvement. It is critical to obtain common wheat–A. cristatum alien disomic addition lines to locate the desired genes on the P genome chromosomes. Comparative analysis of the homoeologous relationships between the P genome chromosome and wheat genome chromosomes is a key step in transferring different desirable genes into common wheat and producing the desired alien translocation line while compensating for the loss of wheat chromatin. In this study, six common wheat–A. cristatum disomic addition lines were produced and analyzed by phenotypic examination, genomic in situ hybridization (GISH), SSR markers from the ABD genomes and STS markers from the P genome. Comparative maps, six in total, were generated and demonstrated that all six addition lines belonged to homoeologous group 6. However, chromosome 6P had undergone obvious rearrangements in different addition lines compared with the wheat chromosome, indicating that to obtain a genetic compensating alien translocation line, one should recombine alien chromosomal regions with homoeologous wheat chromosomes. Indeed, these addition lines were classified into four types based on the comparative mapping: 6PI, 6PII, 6PIII, and 6PIV. The different types of chromosome 6P possessed different desirable genes. For example, the 6PI type, containing three addition lines, carried genes conferring high numbers of kernels per spike and resistance to powdery mildew, important traits for wheat improvement. These results may prove valuable for promoting the development of conventional chromosome engineering techniques toward molecular chromosome engineering. PMID:24595330

  6. Microarray reveals complement components are regulated in the serum-deprived rat retinal ganglion cell line

    PubMed Central

    Khalyfa, Abdelnaby; Chlon, Timothy; Qiang, He; Agarwal, Neeraj

    2007-01-01

    Purpose Glaucoma is a progressive eye disease that leads to blindness due to loss of retinal ganglion cells (RGCs). There are difficulties in using primary cultures of purified RGC to study this pathophysiology. RGC-5, a transformed not RGC line, expresses several markers characteristic of the RGCs. The aim of this study was to generate a genome-wide gene expression of RGC-5 following serum deprivation and to identify candidate genes that may be involved in the signal transduction pathways. Methods Apoptosis in the transformed rat RGC-5 was induced by serum deprivation for 0, 8, 24, 48, and 96 h. Briefly, 400 ng of RNA from each sample was reverse transcribed and labeled with Cy3 dye. Fragmented fluorescent cRNA was mixed with hybridization buffer and incubated at 60 °C for 16 h. Labeled cRNA was hybridized to Rat Genome Oligonucleotide Arrays. These arrays contain 22,775 transcripts with one oligonucleotide per transcript (60-mer). Gene expression from scanned images was quantified and analyzed using ArrayVision software. Reproducibility among triplicate arrays was determined by ANOVA statistical analysis. Significant differences in gene expression between apoptotic and nonapoptotic cells were determined based on p-values. Results Of the 22,775 transcripts present on the arrays (Agilent rat genome, 60-mer), 713 (8 h), 1,967 (24 h), 1,011 (48 h), and 1,161 (96 h) were differentially expressed relative to the 0 h time point (p-values <0.05). Twenty-three transcripts were common to 8, 24, 48, and 96 h and 130 transcripts were common to the 24, 48, and 96 h time points. The two most highly upregulated genes were Fdft1 and Lgals3 (8 h), C3 and Fcgrt (24 h), C and Lcn2 (48 h), and Mgp and C3 (96 h). A subset of the differentially expressed genes identified in microarray data (Ftl1, C3, C1s, Neu1, Polr2g, Acadm, Nupr1, Gch, Dia1, DNase1, Tgfb2, and Cyr61) were validated using quantitative real time polymerase chain reaction (QRT-PCR). Here we show that complement factor

  7. Integrative proteomic profiling of ovarian cancer cell lines reveals precursor cell associated proteins and functional status

    PubMed Central

    Coscia, F.; Watters, K. M.; Curtis, M.; Eckert, M. A.; Chiang, C. Y.; Tyanova, S.; Montag, A.; Lastra, R. R.; Lengyel, E.; Mann, M.

    2016-01-01

    A cell line representative of human high-grade serous ovarian cancer (HGSOC) should not only resemble its tumour of origin at the molecular level, but also demonstrate functional utility in pre-clinical investigations. Here, we report the integrated proteomic analysis of 26 ovarian cancer cell lines, HGSOC tumours, immortalized ovarian surface epithelial cells and fallopian tube epithelial cells via a single-run mass spectrometric workflow. The in-depth quantification of >10,000 proteins results in three distinct cell line categories: epithelial (group I), clear cell (group II) and mesenchymal (group III). We identify a 67-protein cell line signature, which separates our entire proteomic data set, as well as a confirmatory publicly available CPTAC/TCGA tumour proteome data set, into a predominantly epithelial and mesenchymal HGSOC tumour cluster. This proteomics-based epithelial/mesenchymal stratification of cell lines and human tumours indicates a possible origin of HGSOC either from the fallopian tube or from the ovarian surface epithelium. PMID:27561551

  8. Dense-gas properties in Arp 220 revealed by isotopologue lines

    NASA Astrophysics Data System (ADS)

    Wang, Junzhi; Zhang, Zhi-Yu; Zhang, Jiangshui; Shi, Yong; Fang, Min

    2016-02-01

    We present observations of isotopologue lines of dense-gas tracers at 3 mm and 1 mm towards the nearest ultra-luminous infrared galaxy Arp 220. The 3-mm and 1-mm observations were performed with the Institut de Radioastronomie Millimétrique 30-m telescope and the Atacama Pathfinder Experiment 12-m telescope, respectively. We detected H13CN and HN13C in 1-0 and 3-2, and HC15N 1-0, among which HC15N 1-0 and HN13C 1-0 are detected in Arp 220 for the first time. The H13CO+ 1-0 and 3-2 lines are unlikely to be detected because of the confusion of SiO lines. We find that the ratio of the line brightness temperatures of HN13C 3-2/1-0 is 2.4, which is significantly higher than that of H13CN 3-2/1-0 (0.73). This indicates that HN13C and HNC molecules are in denser regions than H13CN and HCN molecules. With the line ratio of H13CN 1-0 and HC15N 1-0, the 14N/15N ratio was estimated to be 440^{+140}_{-82}, which is larger than that of the local interstellar medium.

  9. Clumpy tori around type II active galactic nuclei as revealed by X-ray fluorescent lines

    NASA Astrophysics Data System (ADS)

    Liu, Jiren; Liu, Yuan; Li, Xiaobo; Xu, Weiwei; Gou, Lijun; Cheng, Cheng

    2016-06-01

    The reflection spectrum of a torus around an active galactic nucleus (AGN) is characterized by X-ray fluorescent lines, which are most prominent for type II AGNs. A clumpy torus allows photons reflected from the back-side of the torus to leak through the front regions that are free of obscuration. The observed X-ray fluorescent lines are therefore sensitive to the clumpiness of the torus. We analysed a sample of type II AGNs observed with the Chandra High Energy Transmission Grating Spectrometer (HETGS), and measured the fluxes for the Si Kα and Fe Kα lines. The measured Fe Kα/Si Kα ratios, spanning a range between 5 and 60, are far smaller than the ratios predicted from simulations of smooth tori, indicating that the tori of the studied sources have clumpy distributions rather than smooth ones. We compared the measured Fe Kα/Si Kα ratios with simulation results of clumpy tori. The Circinus galaxy has a Fe Kα/Si Kα ratio of ˜60, which is close to the simulation results for N = 5, where N is the average number of clumps along the line of sight. The Fe Kα/Si Kα ratios of the other sources are all below the simulation results for N = 2. Overall, this shows that the non-Fe fluorescent lines in the soft X-ray band are a potentially powerful probe of the clumpiness of tori around AGNs.

  10. Expression Profiling of Glucosinolate Biosynthetic Genes in Brassica oleracea L. var. capitata Inbred Lines Reveals Their Association with Glucosinolate Content.

    PubMed

    Robin, Arif Hasan Khan; Yi, Go-Eun; Laila, Rawnak; Yang, Kiwoung; Park, Jong-In; Kim, Hye Ran; Nou, Ill-Sup

    2016-01-01

    Glucosinolates are the biochemical compounds that provide defense to plants against pathogens and herbivores. In this study, the relative expression level of 48 glucosinolate biosynthesis genes was explored in four morphologically-different cabbage inbred lines by qPCR analysis. The content of aliphatic and indolic glucosinolate molecules present in those cabbage lines was also estimated by HPLC analysis. The possible association between glucosinolate accumulation and related gene expression level was explored by principal component analysis (PCA). The genotype-dependent variation in the relative expression level of different aliphatic and indolic glucosinolate biosynthesis genes is the novel result of this study. A total of eight different types of glucosinolates, including five aliphatic and three indolic glucosinolates, was detected in four cabbage lines. Three inbred lines BN3383, BN4059 and BN4072 had no glucoraphanin, sinigrin and gluconapin detected, but the inbred line BN3273 had these three aliphatic glucosinolate compounds. PCA revealed that a higher expression level of ST5b genes and lower expression of GSL-OH was associated with the accumulation of these three aliphatic glucosinolate compounds. PCA further revealed that comparatively higher accumulation of neoglucobrassicin in the inbred line, BN4072, was associated with a high level of expression of MYB34 (Bol017062) and CYP81F1 genes. The Dof1 and IQD1 genes probably trans-activated the genes related to biosynthesis of glucoerucin and methoxyglucobrassicin for their comparatively higher accumulation in the BN4059 and BN4072 lines compared to the other two lines, BN3273 and BN3383. A comparatively higher progoitrin level in BN3273 was probably associated with the higher expression level of the GSL-OH gene. The cabbage inbred line BN3383 accounted for the significantly higher relative expression level for the 12 genes out of 48, but this line had comparatively lower total glucosinolates detected

  11. Cell surface and secreted protein profiles of human thyroid cancer cell lines reveal distinct glycoprotein patterns.

    PubMed

    Arcinas, Arthur; Yen, Ten-Yang; Kebebew, Electron; Macher, Bruce A

    2009-08-01

    Cell surface proteins have been shown to be effective therapeutic targets. In addition, shed forms of these proteins and secreted proteins can serve as biomarkers for diseases, including cancer. Thus, identification of cell surface and secreted proteins has been a prime area of interest in the proteomics field. Most cell surface and secreted proteins are known to be glycosylated, and therefore, a proteomics strategy targeting these proteins was applied to obtain proteomic profiles from various thyroid cancer cell lines that represent the range of thyroid cancers of follicular cell origin. In this study, we oxidized the carbohydrates of secreted proteins and those on the cell surface with periodate and isolated them via covalent coupling to hydrazide resin. The glycoproteins obtained were identified from tryptic peptides and N-linked glycopeptides released from the hydrazide resin using two-dimensional liquid chromatography-tandem mass spectrometry in combination with the gas phase fractionation. Thyroid cancer cell lines derived from papillary thyroid cancer (TPC-1), follicular thyroid cancer (FTC-133), Hurthle cell carcinoma (XTC-1), and anaplastic thyroid cancer (ARO and DRO-1) were evaluated. An average of 150 glycoproteins were identified per cell line, of which more than 57% are known cell surface or secreted glycoproteins. The usefulness of the approach for identifying thyroid cancer associated biomarkers was validated by the identification of glycoproteins (e.g., CD44, galectin 3 and metalloproteinase inhibitor 1) that have been found to be useful markers for thyroid cancer. In addition to glycoproteins that are commonly expressed by all of the cell lines, we identified others that are only expressed in the more well-differentiated thyroid cancer cell lines (follicular, Hurthle cell and papillary), or by cell lines derived from undifferentiated tumors that are uniformly fatal forms of thyroid cancer (i.e., anaplastic). On the basis of the results obtained, a

  12. Ti-44 Gamma-Ray Emission Lines from SN1987A Reveal an Asymmetric Explosion

    NASA Technical Reports Server (NTRS)

    Boggs, S. E.; Harrison, F. A.; Miyasaka, H.; Grefenstette, B. W.; Zoglauer, A.; Fryer, C. L.; Reynolds, S. P.; Alexander, D. M.; An, H.; Barret, D.; Christensen, F. E.; Craig, W. W.; Forster, K.; Giommi, P.; Hailey, C. J.; Hornstrup, A.; Kitaguchi, T.; Koglin, J. E.; Madsen, K. K.; Zhang, W. W.

    2015-01-01

    In core-collapse supernovae, titanium-44 (Ti-44) is produced in the innermost ejecta, in the layer of material directly on top of the newly formed compact object. As such, it provides a direct probe of the supernova engine. Observations of supernova 1987A (SN1987A) have resolved the 67.87- and 78.32-kilo-electron volt emission lines from decay of Ti-44 produced in the supernova explosion. These lines are narrow and redshifted with a Doppler velocity of 700 kilometers per second, direct evidence of large-scale asymmetry in the explosion.

  13. The most powerful quasar outflows as revealed by the Civ λ1549 resonance line

    NASA Astrophysics Data System (ADS)

    Marziani, P.; Martínez Carballo, M. A.; Sulentic, J. W.; Del Olmo, A.; Stirpe, G. M.; Dultzin, D.

    2016-01-01

    Outflows from quasars may be almost ubiquitous, but there are significant differences on a source- by-source basis. These differences can be organized along the 4D Eigenvector 1 sequence: at low z, only the Population A sources radiating at relatively high Eddington ratio show evidences of prominent high- velocity outflows from the Civλ1549 line profiles. Here we discuss, starting from recent observations of high-luminosity sample of Hamburg-ESO quasars, the Civλ1549 emission line profiles and how they are affected by outflow motion as a function of the quasar luminosity. Our high-luminosity sample has the notable advantage that the rest frame has been set by previous Hβ observations in the J, H, and K band, therefore making measurements of inter-line shift accurate and free of systemic biases. As the redshift increases and the luminosity of the brightest quasars grows, powerful, high-velocity outflows may become more frequent. We then discuss the outflow contextualisation following the 4DE1 approach as a tool for unveiling the nature of the so-called Weak Lined Quasars (WLQs) that have emerged in recent years as a new, poorly understood class of quasars. We estimate the kinetic power associated with the Civλ1549 emitting gas in outflow, and we suggest that the host galaxies of the most luminous sources may experience a significant feedback effect.

  14. Resolved atomic lines reveal outflows in two ultraluminous X-ray sources

    NASA Astrophysics Data System (ADS)

    Pinto, Ciro; Middleton, Matthew J.; Fabian, Andrew C.

    2016-05-01

    Ultraluminous X-ray sources are extragalactic, off-nucleus, point sources in galaxies, and have X-ray luminosities in excess of 3 × 1039 ergs per second. They are thought to be powered by accretion onto a compact object. Possible explanations include accretion onto neutron stars with strong magnetic fields, onto stellar-mass black holes (of up to 20 solar masses) at or in excess of the classical Eddington limit, or onto intermediate-mass black holes (103-105 solar masses). The lack of sufficient energy resolution in previous analyses has prevented an unambiguous identification of any emission or absorption lines in the X-ray band, thereby precluding a detailed analysis of the accretion flow. Here we report the presence of X-ray emission lines arising from highly ionized iron, oxygen and neon with a cumulative significance in excess of five standard deviations, together with blueshifted (about 0.2 times light velocity) absorption lines of similar significance, in the high-resolution X-ray spectra of the ultraluminous X-ray sources NGC 1313 X-1 and NGC 5408 X-1. The blueshifted absorption lines must occur in a fast-outflowing gas, whereas the emission lines originate in slow-moving gas around the source. We conclude that the compact object in each source is surrounded by powerful winds with an outflow velocity of about 0.2 times that of light, as predicted by models of accreting supermassive black holes and hyper-accreting stellar-mass black holes.

  15. Resolved atomic lines reveal outflows in two ultraluminous X-ray sources.

    PubMed

    Pinto, Ciro; Middleton, Matthew J; Fabian, Andrew C

    2016-05-01

    Ultraluminous X-ray sources are extragalactic, off-nucleus, point sources in galaxies, and have X-ray luminosities in excess of 3 × 10(39) ergs per second. They are thought to be powered by accretion onto a compact object. Possible explanations include accretion onto neutron stars with strong magnetic fields, onto stellar-mass black holes (of up to 20 solar masses) at or in excess of the classical Eddington limit, or onto intermediate-mass black holes (10(3)-10(5) solar masses). The lack of sufficient energy resolution in previous analyses has prevented an unambiguous identification of any emission or absorption lines in the X-ray band, thereby precluding a detailed analysis of the accretion flow. Here we report the presence of X-ray emission lines arising from highly ionized iron, oxygen and neon with a cumulative significance in excess of five standard deviations, together with blueshifted (about 0.2 times light velocity) absorption lines of similar significance, in the high-resolution X-ray spectra of the ultraluminous X-ray sources NGC 1313 X-1 and NGC 5408 X-1. The blueshifted absorption lines must occur in a fast-outflowing gas, whereas the emission lines originate in slow-moving gas around the source. We conclude that the compact object in each source is surrounded by powerful winds with an outflow velocity of about 0.2 times that of light, as predicted by models of accreting supermassive black holes and hyper-accreting stellar-mass black holes. PMID:27120159

  16. Plant Genetic Archaeology: Whole-Genome Sequencing Reveals the Pedigree of a Classical Trisomic Line

    PubMed Central

    Salomé, Patrice A.; Weigel, Detlef

    2014-01-01

    The circadian oscillator is astonishingly robust to changes in the environment but also to genomic changes that alter the copy number of its components through genome duplication, gene duplication, and homeologous gene loss. While studying the potential effect of aneuploidy on the Arabidopsis thaliana circadian clock, we discovered that a line thought to be trisomic for chromosome 3 also bears the gi-1 mutation, resulting in a short period and late flowering. With the help of whole-genome sequencing, we uncovered the unexpected complexity of this trisomic stock’s history, as its genome shows evidence of past outcrossing with another A. thaliana accession. Our study indicates that although historical aneuploidy lines exist and are available, it might be safer to generate new individuals and confirm their genomes and karyotypes by sequencing. PMID:25524155

  17. Integrated analysis of breast cancer cell lines reveals unique signaling pathways

    SciTech Connect

    Heiser, Laura M.; Wang, Nicholas J.; Talcott, Carolyn L.; Laderoute, Keith R.; Knapp, Merrill; Guan, Yinghui; Hu, Zhi; Ziyad, Safiyyah; Weber, Barbara L.; Laquerre, Sylvie; Jackson, Jeffrey R.; Wooster, Richard F.; Kuo, Wen-Lin; Gray, Joe W.; Spellman, Paul T.

    2009-03-31

    Cancer is a heterogeneous disease resulting from the accumulation of genetic defects that negatively impact control of cell division, motility, adhesion and apoptosis. Deregulation in signaling along the EGFR-MAPK pathway is common in breast cancer, though the manner in which deregulation occurs varies between both individuals and cancer subtypes. We were interested in identifying subnetworks within the EGFR-MAPK pathway that are similarly deregulated across subsets of breast cancers. To that end, we mapped genomic, transcriptional and proteomic profiles for 30 breast cancer cell lines onto a curated Pathway Logic symbolic systems model of EGFR-MEK signaling. This model was comprised of 539 molecular states and 396 rules governing signaling between active states. We analyzed these models and identified several subtype specific subnetworks, including one that suggested PAK1 is particularly important in regulating the MAPK cascade when it is over-expressed. We hypothesized that PAK1 overexpressing cell lines would have increased sensitivity to MEK inhibitors. We tested this experimentally by measuring quantitative responses of 20 breast cancer cell lines to three MEK inhibitors. We found that PAK1 over-expressing luminal breast cancer cell lines are significantly more sensitive to MEK inhibition as compared to those that express PAK1 at low levels. This indicates that PAK1 over-expression may be a useful clinical marker to identify patient populations that may be sensitive to MEK inhibitors. All together, our results support the utility of symbolic system biology models for identification of therapeutic approaches that will be effective against breast cancer subsets.

  18. RNA-seq reveals determinants for irinotecan sensitivity/resistance in colorectal cancer cell lines

    PubMed Central

    Li, Xin-Xiang; Zheng, Hong-Tu; Peng, Jun-Jie; Huang, Li-Yong; Shi, De-Bing; Liang, Lei; Cai, San-Jun

    2014-01-01

    Irinotecan is a topoisomerase I inhibitor approved worldwide as a first- and second-line chemotherapy for advanced or recurrent colorectal cancer (CRC). Although irinotecan showed significant survival advantage for patients, a relatively low response rate and severe adverse effects demonstrated the urgent need for biomarkers searching to select the suitable patients who can benefit from irinotecan-based therapy and avoid the adverse effects. In present work, the irinotecan response (IC50 doses) of 20 CRC cell lines were correlated with the basal expression profiles investigated by RNA-seq to figure out genes responsible for irinotecan sensitivity/resistance. Genes negatively or positively correlated to irinotecan sensitivity were given after biocomputation, and 7 (CDC20, CTNNAL1, FZD7, CITED2, ABR, ARHGEF7, and RNMT) of them were validated in two CRC cell lines by quantitative real-time PCR, several of these 7 genes has been proposed to promote cancer cells proliferation and hence may confer CRC cells resistance to irinotecan. Our work might provide potential biomarkers and therapeutic targets for irinotecan sensitivity in CRC cells. PMID:24966994

  19. Representational difference analysis of a committed myeloid progenitor cell line reveals evidence for bilineage potential.

    PubMed

    Lawson, N D; Berliner, N

    1998-08-18

    In this study we have sought to characterize a committed myeloid progenitor cell line in an attempt to isolate general factors that may promote differentiation. We used cDNA representational difference analysis (RDA), which allows analysis of differential gene expression, to compare EML and EPRO cells. We have isolated nine differentially expressed cDNA fragments as confirmed by slot blot, Northern, and PCR analysis. Three of nine sequences appear to be novel whereas the identity of the remaining fragments suggested that the EPRO cell line is multipotent. Among the isolated sequences were eosinophilic, monocytic, and neutrophilic specific genes. Therefore, we tested the ability of EPRO cells to differentiate along multiple myeloid lineages and found that EPRO cells exhibited morphologic maturation into either monocyte/macrophages or neutrophils, but not eosinophils. Furthermore, when EPRO cells were exposed to ATRA, neutrophil specific genes were induced, whereas monocytic markers were induced by phorbol ester treatment. This study highlights the use of cDNA RDA in conjunction with the EML/EPRO cell line to isolate markers associated with macrophage and neutrophil differentiation and establishes the usefulness of this system in the search for factors involved in myeloid commitment. PMID:9707612

  20. Information Gathering Revealed within the Social Network of Line-Managers.

    ERIC Educational Resources Information Center

    Mackenzie, Maureen L.

    2003-01-01

    Results of this study revealed that relationship, more than knowledge, may be the reason a manager is sought as an information source within a business environment. Social network mapping was used to capture a more intimate view of the information relationships within a business environment. Content analysis was used to analyze the data and to…

  1. Revealing the Nature of Extreme Coronal-line Emitter SDSS J095209.56+214313.3

    NASA Astrophysics Data System (ADS)

    Palaversa, Lovro; Gezari, Suvi; Sesar, Branimir; Stuart, J. Scott; Wozniak, Przemyslaw; Holl, Berry; Ivezić, Željko

    2016-03-01

    Extreme coronal-line emitter (ECLE) SDSS J095209.56+214313.3, known by its strong, fading, high-ionization lines, has been a long-standing candidate for a tidal disruption event however, a supernova (SN) origin has not yet been ruled out. Here we add several new pieces of information to the puzzle of the nature of the transient that powered its variable coronal lines: (1) an optical light curve from the Lincoln Near Earth Asteroid Research (LINEAR) survey that serendipitously catches the optical flare, and (2) late-time observations of the host galaxy with the Swift Ultraviolet and Optical Telescope (UVOT) and X-ray telescope (XRT) and the ground-based Mercator telescope. The well-sampled, ˜10 yr long, unfiltered LINEAR light curve constrains the onset of the flare to a precision of ±5 days and enables us to place a lower limit on the peak optical magnitude. Difference imaging allows us to estimate the location of the flare in proximity of the host galaxy core. Comparison of the GALEX data (early 2006) with the recently acquired Swift UVOT (2015 June) and Mercator observations (2015 April) demonstrates a decrease in the UV flux over a ˜10 yr period, confirming that the flare was UV-bright. The long-lived UV-bright emission, detected 1.8 rest-frame years after the start of the flare, strongly disfavors an SN origin. These new data allow us to conclude that the flare was indeed powered by the tidal disruption of a star by a supermassive black hole and that tidal disruption events are in fact capable of powering the enigmatic class of ECLEs.

  2. Germ-line deletion in DICER1 revealed by a novel MLPA assay using synthetic oligonucleotides.

    PubMed

    Sabbaghian, Nelly; Srivastava, Archana; Hamel, Nancy; Plourde, François; Gajtko-Metera, Malgorzata; Niedziela, Marek; Foulkes, William D

    2014-04-01

    DICER1 is an endoribonuclease responsible for the production of mature microRNAs which are small, single-stranded RNA molecules that regulate gene expression post-transcriptionally by binding to mRNA and repressing the expression of target genes. Germ-line mutations in DICER1 are responsible for a rare cancer syndrome, including tumors that can co-occur with multinodular goiter (MNG). Using Sanger sequencing, we screened all DICER1 exons and intron boundaries in 20 suspected mutation carriers: nine with ovarian sex cord-stromal tumors (including Sertoli-Leydig cell tumors (SLCTs)), five with pleuropulmonary blastoma, one with cystic nephroma, one with nasal chondromesenchymal hamartoma and four with more than one manifestation suggestive of a germ-line DICER1 mutation. All were negative for any apparently deleterious variants. We developed a Multiplex Ligation-based Probe Amplification assay for DICER1 to screen for large deletions or duplications. Synthetic oligonucleotides were designed to cover all exons in three probe-mixes. In a child with a SLCT and MNG, and in her mother and brother (both diagnosed with MNG), we identified a heterozygous germ-line deletion of approximately 3 kilobases that eliminates exon 21 of DICER1 and two-thirds of intron 21, accompanied by an insertion of a G nucleotide at the 3' end of the deletion (c.3270-6_4051-1280delinsG). This allele is expressed in the patient's cDNA, creating an out-of-frame deletion predicted to result in a truncated protein (r.3270_4050del; p.Tyr1091Ser*28). Our novel finding of a disease-causing large deletion in DICER1 emphasizes the need to include assays that can detect rearrangements, duplications and deletions in any DICER1 screening protocol. PMID:24065110

  3. Germ-line deletion in DICER1 revealed by a novel MLPA assay using synthetic oligonucleotides

    PubMed Central

    Sabbaghian, Nelly; Srivastava, Archana; Hamel, Nancy; Plourde, François; Gajtko-Metera, Malgorzata; Niedziela, Marek; Foulkes, William D

    2014-01-01

    DICER1 is an endoribonuclease responsible for the production of mature microRNAs which are small, single-stranded RNA molecules that regulate gene expression post-transcriptionally by binding to mRNA and repressing the expression of target genes. Germ-line mutations in DICER1 are responsible for a rare cancer syndrome, including tumors that can co-occur with multinodular goiter (MNG). Using Sanger sequencing, we screened all DICER1 exons and intron boundaries in 20 suspected mutation carriers: nine with ovarian sex cord-stromal tumors (including Sertoli–Leydig cell tumors (SLCTs)), five with pleuropulmonary blastoma, one with cystic nephroma, one with nasal chondromesenchymal hamartoma and four with more than one manifestation suggestive of a germ-line DICER1 mutation. All were negative for any apparently deleterious variants. We developed a Multiplex Ligation-based Probe Amplification assay for DICER1 to screen for large deletions or duplications. Synthetic oligonucleotides were designed to cover all exons in three probe-mixes. In a child with a SLCT and MNG, and in her mother and brother (both diagnosed with MNG), we identified a heterozygous germ-line deletion of approximately 3 kilobases that eliminates exon 21 of DICER1 and two-thirds of intron 21, accompanied by an insertion of a G nucleotide at the 3′ end of the deletion (c.3270-6_4051-1280delinsG). This allele is expressed in the patient's cDNA, creating an out-of-frame deletion predicted to result in a truncated protein (r.3270_4050del; p.Tyr1091Ser*28). Our novel finding of a disease-causing large deletion in DICER1 emphasizes the need to include assays that can detect rearrangements, duplications and deletions in any DICER1 screening protocol. PMID:24065110

  4. Actin marker lines in grapevine reveal a gatekeeper function of guard cells.

    PubMed

    Guan, Xin; Buchholz, Günther; Nick, Peter

    2014-08-15

    Resistance to abiotic and biotic stress is a central topic for sustainable agriculture, especially in grapevine, one of the field crops with the highest economic output per acreage. As early cellular factors for plant defense, actin microfilaments (AF) are of high relevance. We therefore generated a transgenic actin marker line for grapevine by expressing a fusion protein between green fluorescent protein and the second actin-binding domain of Arabidopsis (Arabidopsis thaliana) fimbrin, AtFIM1. Based on this first cytoskeletal-marker line in grapevine, the response of AFs to phytopathogenic microorganisms could be followed in vivo. Upon inoculation with fluorescently labeled strains of phytopathogenic bacteria, actin responses were confined to the guard cells. In contrast, upon contact with zoospores of Plasmopara viticola, not only the guard cells, but also epidermal pavement cells, where no zoospores had attached responded with the formation of a perinuclear actin basket. Our data support the hypothesis that guard cells act as pacemakers of defense, dominating the responses of the remaining epidermal cells. PMID:24973589

  5. Exome sequencing reveals recurrent germ line variants in patients with familial Waldenström macroglobulinemia.

    PubMed

    Roccaro, Aldo M; Sacco, Antonio; Shi, Jiantao; Chiarini, Marco; Perilla-Glen, Adriana; Manier, Salomon; Glavey, Siobhan; Aljawai, Yosra; Mishima, Yuji; Kawano, Yawara; Moschetta, Michele; Correll, Mick; Improgo, Ma Reina; Brown, Jennifer R; Imberti, Luisa; Rossi, Giuseppe; Castillo, Jorge J; Treon, Steven P; Freedman, Matthew L; Van Allen, Eliezer M; Hide, Winston; Hiller, Elaine; Rainville, Irene; Ghobrial, Irene M

    2016-05-26

    Familial aggregation of Waldenström macroglobulinemia (WM) cases, and the clustering of B-cell lymphoproliferative disorders among first-degree relatives of WM patients, has been reported. Nevertheless, the possible contribution of inherited susceptibility to familial WM remains unrevealed. We performed whole exome sequencing on germ line DNA obtained from 4 family members in which coinheritance for WM was documented in 3 of them, and screened additional independent 246 cases by using gene-specific mutation sequencing. Among the shared germ line variants, LAPTM5(c403t) and HCLS1(g496a) were the most recurrent, being present in 3/3 affected members of the index family, detected in 8% of the unrelated familial cases, and present in 0.5% of the nonfamilial cases and in <0.05 of a control population. LAPTM5 and HCLS1 appeared as relevant WM candidate genes that characterized familial WM individuals and were also functionally relevant to the tumor clone. These findings highlight potentially novel contributors for the genetic predisposition to familial WM and indicate that LAPTM5(c403t) and HCLS1(g496a) may represent predisposition alleles in patients with familial WM. PMID:26903547

  6. Secretome analysis of multiple pancreatic cancer cell lines reveals perturbations of key functional networks.

    PubMed

    Schiarea, Silvia; Solinas, Graziella; Allavena, Paola; Scigliuolo, Graziana Maria; Bagnati, Renzo; Fanelli, Roberto; Chiabrando, Chiara

    2010-09-01

    The cancer secretome is a rich repository in which to mine useful information for both cancer biology and clinical oncology. To help understand the mechanisms underlying the progression of pancreatic cancer, we characterized the secretomes of four human pancreatic ductal adenocarcinoma (PDAC) cell lines versus a normal counterpart. To this end, we used a proteomic workflow based on high-confidence protein identification by mass spectrometry, semiquantitation by a label-free approach, and network enrichment analysis by a system biology tool. Functional networks significantly enriched with PDAC-dysregulated proteins included not only expected alterations within key mechanisms known to be relevant for tumor progression (e.g., cell-cell/cell-matrix adhesion, extracellular matrix remodeling, and cytoskeleton rearrangement), but also other extensive, coordinated perturbations never observed in pancreatic cancer. In particular, we highlighted perturbations possibly favoring tumor progression through immune escape (i.e., inhibition of the complement system, deficiency of selected proteasome components within the antigen-presentation machinery, and inhibition of T cell cytoxicity), and a defective protein folding machinery. Among the proteins found concordantly oversecreted in all of our PDAC cell lines, many are reportedly overexpressed in pancreatic cancer (e.g., CD9 and Vimentin), while others (PLOD3, SH3L3, PCBP1, and SFRS1) represent novel PDAC-secreted proteins that may be worth investigating. PMID:20687567

  7. Standardized orthotopic xenografts in zebrafish reveal glioma cell-line-specific characteristics and tumor cell heterogeneity

    PubMed Central

    Welker, Alessandra M.; Jaros, Brian D.; Puduvalli, Vinay K.; Imitola, Jaime; Kaur, Balveen; Beattie, Christine E.

    2016-01-01

    ABSTRACT Glioblastoma (GBM) is a deadly brain cancer, for which few effective drug treatments are available. Several studies have used zebrafish models to study GBM, but a standardized approach to modeling GBM in zebrafish was lacking to date, preventing comparison of data across studies. Here, we describe a new, standardized orthotopic xenotransplant model of GBM in zebrafish. Dose-response survival assays were used to define the optimal number of cells for tumor formation. Techniques to measure tumor burden and cell spread within the brain over real time were optimized using mouse neural stem cells as control transplants. Applying this standardized approach, we transplanted two patient-derived GBM cell lines, serum-grown adherent cells and neurospheres, into the midbrain region of embryonic zebrafish and analyzed transplanted larvae over time. Progressive brain tumor growth and premature larval death were observed using both cell lines; however, fewer transplanted neurosphere cells were needed for tumor growth and lethality. Tumors were heterogeneous, containing both cells expressing stem cell markers and cells expressing markers of differentiation. A small proportion of transplanted neurosphere cells expressed glial fibrillary acidic protein (GFAP) or vimentin, markers of more differentiated cells, but this number increased significantly during tumor growth, indicating that these cells undergo differentiation in vivo. By contrast, most serum-grown adherent cells expressed GFAP and vimentin at the earliest times examined post-transplant. Both cell types produced brain tumors that contained Sox2+ cells, indicative of tumor stem cells. Transplanted larvae were treated with currently used GBM therapeutics, temozolomide or bortezomib, and this resulted in a reduction in tumor volume in vivo and an increase in survival. The standardized model reported here facilitates robust and reproducible analysis of glioblastoma tumor cells in real time and provides a platform for

  8. Comparative study of human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) as a treatment for retinal dystrophies

    PubMed Central

    Riera, Marina; Fontrodona, Laura; Albert, Silvia; Ramirez, Diana Mora; Seriola, Anna; Salas, Anna; Muñoz, Yolanda; Ramos, David; Villegas-Perez, Maria Paz; Zapata, Miguel Angel; Raya, Angel; Ruberte, Jesus; Veiga, Anna; Garcia-Arumi, Jose

    2016-01-01

    Retinal dystrophies (RD) are major causes of familial blindness and are characterized by progressive dysfunction of photoreceptor and/or retinal pigment epithelium (RPE) cells. In this study, we aimed to evaluate and compare the therapeutic effects of two pluripotent stem cell (PSC)-based therapies. We differentiated RPE from human embryonic stem cells (hESCs) or human-induced pluripotent stem cells (hiPSCs) and transplanted them into the subretinal space of the Royal College of Surgeons (RCS) rat. Once differentiated, cells from either source of PSC resembled mature RPE in their morphology and gene expression profile. Following transplantation, both hESC- and hiPSC-derived cells maintained the expression of specific RPE markers, lost their proliferative capacity, established tight junctions, and were able to perform phagocytosis of photoreceptor outer segments. Remarkably, grafted areas showed increased numbers of photoreceptor nuclei and outer segment disk membranes. Regardless of the cell source, human transplants protected retina from cell apoptosis, glial stress and accumulation of autofluorescence, and responded better to light stimuli. Altogether, our results show that hESC- and hiPSC-derived cells survived, migrated, integrated, and functioned as RPE in the RCS rat retina, providing preclinical evidence that either PSC source could be of potential benefit for treating RD. PMID:27006969

  9. Comparative study of human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) as a treatment for retinal dystrophies.

    PubMed

    Riera, Marina; Fontrodona, Laura; Albert, Silvia; Ramirez, Diana Mora; Seriola, Anna; Salas, Anna; Muñoz, Yolanda; Ramos, David; Villegas-Perez, Maria Paz; Zapata, Miguel Angel; Raya, Angel; Ruberte, Jesus; Veiga, Anna; Garcia-Arumi, Jose

    2016-01-01

    Retinal dystrophies (RD) are major causes of familial blindness and are characterized by progressive dysfunction of photoreceptor and/or retinal pigment epithelium (RPE) cells. In this study, we aimed to evaluate and compare the therapeutic effects of two pluripotent stem cell (PSC)-based therapies. We differentiated RPE from human embryonic stem cells (hESCs) or human-induced pluripotent stem cells (hiPSCs) and transplanted them into the subretinal space of the Royal College of Surgeons (RCS) rat. Once differentiated, cells from either source of PSC resembled mature RPE in their morphology and gene expression profile. Following transplantation, both hESC- and hiPSC-derived cells maintained the expression of specific RPE markers, lost their proliferative capacity, established tight junctions, and were able to perform phagocytosis of photoreceptor outer segments. Remarkably, grafted areas showed increased numbers of photoreceptor nuclei and outer segment disk membranes. Regardless of the cell source, human transplants protected retina from cell apoptosis, glial stress and accumulation of autofluorescence, and responded better to light stimuli. Altogether, our results show that hESC- and hiPSC-derived cells survived, migrated, integrated, and functioned as RPE in the RCS rat retina, providing preclinical evidence that either PSC source could be of potential benefit for treating RD. PMID:27006969

  10. Induction of primitive streak and mesendoderm formation in monolayer hESC culture by activation of TGF-β signaling pathway by Activin B.

    PubMed

    Mahmood, Amer; Aldahmash, Abdullah

    2015-11-01

    Human embryonic stem cells (hESCs) have the ability to differentiate into all human cells, however controlling the differentiation has always been a challenge. In the present study we have investigated the direct differentiation of hESCs on MEFs by using TGF-β signaling pathway activators Activin A and Activin B. Activation of the TGF-β pathway with Activin B in low serum highly induced primitive streak and mesendoderm formation after 24 h, which included up-regulation of SOX 17 and BRACHYURY protein and gene expression. Continuous stimulation with Activin B in 2% serum further induced mesendoderm formation by increased gene expression of Brachyury, SOX17, MEOX and FOX at the same time we found down-regulation of neuroectodermal marker genes. Further, by stimulating the mesodermal cells by BMP-2 we succeeded to induce mesenchymal like cells with high expression of mesenchymal markers including; MEOX, FOX, RUNX2, COL1 and OSTEOPONTIN. In conclusion we have directed the differentiation of hESCs as monolayer to primitive streak like cells with Activin B and further into pure mesoderm and mesenchymal like cells by BMP-2. PMID:26586995

  11. A Panel of Embryonic Stem Cell Lines Reveals the Variety and Dynamic of Pluripotent States in Rabbits.

    PubMed

    Osteil, Pierre; Moulin, Anaïs; Santamaria, Claire; Joly, Thierry; Jouneau, Luc; Aubry, Maxime; Tapponnier, Yann; Archilla, Catherine; Schmaltz-Panneau, Barbara; Lecardonnel, Jérôme; Barasc, Harmonie; Mouney-Bonnet, Nathalie; Genthon, Clémence; Roulet, Alain; Donnadieu, Cécile; Acloque, Hervé; Gocza, Elen; Duranthon, Véronique; Afanassieff, Marielle; Savatier, Pierre

    2016-09-13

    Conventional rabbit embryonic stem cell (ESC) lines are derived from the inner cell mass (ICM) of pre-implantation embryos using methods and culture conditions that are established for primate ESCs. In this study, we explored the capacity of the rabbit ICM to give rise to ESC lines using conditions similar to those utilized to generate naive ESCs in mice. On single-cell dissociation and culture in fibroblast growth factor 2 (FGF2)-free, serum-supplemented medium, rabbit ICMs gave rise to ESC lines lacking the DNA-damage checkpoint in the G1 phase like mouse ESCs, and with a pluripotency gene expression profile closer to the rabbit ICM/epiblast profiles. These cell lines can be converted to FGF2-dependent ESCs after culture in conventional conditions. They can also colonize the rabbit pre-implantation embryo. These results indicate that rabbit epiblast cells can be coaxed toward different types of pluripotent stem cells and reveal the dynamics of pluripotent states in rabbit ESCs. PMID:27594588

  12. Mapping the LINE1 ORF1 protein interactome reveals associated inhibitors of human retrotransposition

    PubMed Central

    Goodier, John L.; Cheung, Ling E.; Kazazian, Haig H.

    2013-01-01

    LINE1s occupy 17% of the human genome and are its only active autonomous mobile DNA. L1s are also responsible for genomic insertion of processed pseudogenes and >1 million non-autonomous retrotransposons (Alus and SVAs). These elements have significant effects on gene organization and expression. Despite the importance of retrotransposons for genome evolution, much about their biology remains unknown, including cellular factors involved in the complex processes of retrotransposition and forming and transporting L1 ribonucleoprotein particles. By co-immunoprecipitation of tagged L1 constructs and mass spectrometry, we identified proteins associated with the L1 ORF1 protein and its ribonucleoprotein. These include RNA transport proteins, gene expression regulators, post-translational modifiers, helicases and splicing factors. Many cellular proteins co-localize with L1 ORF1 protein in cytoplasmic granules. We also assayed the effects of these proteins on cell culture retrotransposition and found strong inhibiting proteins, including some that control HIV and other retroviruses. These data suggest candidate cofactors that interact with the L1 to modulate its activity and increase our understanding of the means by which the cell coexists with these genomic ‘parasites’. PMID:23749060

  13. Chromatin states reveal functional associations for globally defined transcription start sites in four human cell lines

    PubMed Central

    2014-01-01

    Background Deciphering the most common modes by which chromatin regulates transcription, and how this is related to cellular status and processes is an important task for improving our understanding of human cellular biology. The FANTOM5 and ENCODE projects represent two independent large scale efforts to map regulatory and transcriptional features to the human genome. Here we investigate chromatin features around a comprehensive set of transcription start sites in four cell lines by integrating data from these two projects. Results Transcription start sites can be distinguished by chromatin states defined by specific combinations of both chromatin mark enrichment and the profile shapes of these chromatin marks. The observed patterns can be associated with cellular functions and processes, and they also show association with expression level, location relative to nearby genes, and CpG content. In particular we find a substantial number of repressed inter- and intra-genic transcription start sites enriched for active chromatin marks and Pol II, and these sites are strongly associated with immediate-early response processes and cell signaling. Associations between start sites with similar chromatin patterns are validated by significant correlations in their global expression profiles. Conclusions The results confirm the link between chromatin state and cellular function for expressed transcripts, and also indicate that active chromatin states at repressed transcripts may poise transcripts for rapid activation during immune response. PMID:24669905

  14. Proteomic comparison reveals the contribution of chloroplast to salt tolerance of a wheat introgression line.

    PubMed

    Xu, Wenjing; Lv, Hongjun; Zhao, Mingming; Li, Yongchao; Qi, Yueying; Peng, Zhenying; Xia, Guangmin; Wang, Mengcheng

    2016-01-01

    We previously bred a salt tolerant wheat cv. SR3 with bread wheat cv. JN177 as the parent via asymmetric somatic hybridization, and found that the tolerance is partially attributed to the superior photosynthesis capacity. Here, we compared the proteomes of two cultivars to unravel the basis of superior photosynthesis capacity. In the maps of two dimensional difference gel electrophoresis (2D-DIGE), there were 26 differentially expressed proteins (DEPs), including 18 cultivar-based and 8 stress-responsive ones. 21 of 26 DEPs were identified and classified into four categories, including photosynthesis, photosynthesis system stability, linolenic acid metabolism, and protein synthesis in chloroplast. The chloroplast localization of some DEPs confirmed that the identified DEPs function in the chloroplast. The overexpression of a DEP enhanced salt tolerance in Arabidopsis thaliana. In line with these data, it is concluded that the contribution of chloroplast to high salinity tolerance of wheat cv. SR3 appears to include higher photosynthesis efficiency by promoting system protection and ROS clearance, stronger production of phytohormone JA by enhancing metabolism activity, and modulating the in chloroplast synthesis of proteins. PMID:27562633

  15. Genetic Variability and Selection Criteria in Rice Mutant Lines as Revealed by Quantitative Traits

    PubMed Central

    Oladosu, Yusuff; Rafii, M. Y.; Abdullah, Norhani; Abdul Malek, Mohammad; Rahim, H. A.; Hussin, Ghazali; Abdul Latif, Mohammad; Kareem, Isiaka

    2014-01-01

    Genetic based knowledge of different vegetative and yield traits play a major role in varietal improvement of rice. Genetic variation gives room for recombinants which are essential for the development of a new variety in any crop. Based on this background, this work was carried out to evaluate genetic diversity of derived mutant lines and establish relationships between their yield and yield components using multivariate analysis. To achieve this objective, two field trials were carried out on 45 mutant rice genotypes to evaluate their growth and yield traits. Data were taken on vegetative traits and yield and its components, while genotypic and phenotypic coefficients, variance components, expected genetic advance, and heritability were calculated. All the genotypes showed variations for vegetative traits and yield and its components. Also, there was positive relationship between the quantitative traits and the final yield with the exception of number of tillers. Finally, the evaluated genotypes were grouped into five major clusters based on the assessed traits with the aid of UPGMA dendrogram. So hybridization of group I with group V or group VI could be used to attain higher heterosis or vigour among the genotypes. Also, this evaluation could be useful in developing reliable selection indices for important agronomic traits in rice. PMID:25431777

  16. Proteomic comparison reveals the contribution of chloroplast to salt tolerance of a wheat introgression line

    PubMed Central

    Xu, Wenjing; Lv, Hongjun; Zhao, Mingming; Li, Yongchao; Qi, Yueying; Peng, Zhenying; Xia, Guangmin; Wang, Mengcheng

    2016-01-01

    We previously bred a salt tolerant wheat cv. SR3 with bread wheat cv. JN177 as the parent via asymmetric somatic hybridization, and found that the tolerance is partially attributed to the superior photosynthesis capacity. Here, we compared the proteomes of two cultivars to unravel the basis of superior photosynthesis capacity. In the maps of two dimensional difference gel electrophoresis (2D-DIGE), there were 26 differentially expressed proteins (DEPs), including 18 cultivar-based and 8 stress-responsive ones. 21 of 26 DEPs were identified and classified into four categories, including photosynthesis, photosynthesis system stability, linolenic acid metabolism, and protein synthesis in chloroplast. The chloroplast localization of some DEPs confirmed that the identified DEPs function in the chloroplast. The overexpression of a DEP enhanced salt tolerance in Arabidopsis thaliana. In line with these data, it is concluded that the contribution of chloroplast to high salinity tolerance of wheat cv. SR3 appears to include higher photosynthesis efficiency by promoting system protection and ROS clearance, stronger production of phytohormone JA by enhancing metabolism activity, and modulating the in chloroplast synthesis of proteins. PMID:27562633

  17. A germ cell determinant reveals parallel pathways for germ line development in Caenorhabditis elegans.

    PubMed

    Mainpal, Rana; Nance, Jeremy; Yanowitz, Judith L

    2015-10-15

    Despite the central importance of germ cells for transmission of genetic material, our understanding of the molecular programs that control primordial germ cell (PGC) specification and differentiation are limited. Here, we present findings that X chromosome NonDisjunction factor-1 (XND-1), known for its role in regulating meiotic crossover formation, is an early determinant of germ cell fates in Caenorhabditis elegans. xnd-1 mutant embryos display a novel 'one PGC' phenotype as a result of G2 cell cycle arrest of the P4 blastomere. Larvae and adults display smaller germ lines and reduced brood size consistent with a role for XND-1 in germ cell proliferation. Maternal XND-1 proteins are found in the P4 lineage and are exclusively localized to the nucleus in PGCs, Z2 and Z3. Zygotic XND-1 turns on shortly thereafter, at the ∼300-cell stage, making XND-1 the earliest zygotically expressed gene in worm PGCs. Strikingly, a subset of xnd-1 mutants lack germ cells, a phenotype shared with nos-2, a member of the conserved Nanos family of germline determinants. We generated a nos-2 null allele and show that nos-2; xnd-1 double mutants display synthetic sterility. Further removal of nos-1 leads to almost complete sterility, with the vast majority of animals without germ cells. Sterility in xnd-1 mutants is correlated with an increase in transcriptional activation-associated histone modification and aberrant expression of somatic transgenes. Together, these data strongly suggest that xnd-1 defines a new branch for PGC development that functions redundantly with nos-2 and nos-1 to promote germline fates by maintaining transcriptional quiescence and regulating germ cell proliferation. PMID:26395476

  18. Gene and microRNA expression reveals sensitivity to paclitaxel in laryngeal cancer cell line

    PubMed Central

    Xu, Cheng-Zhi; Xie, Jin; Jin, Bin; Chen, Xin-Wei; Sun, Zhen-Feng; Wang, Bao-Xing; Dong, Pin

    2013-01-01

    Paclitaxel is a widely used chemotherapy drug for advanced laryngeal cancer patients. However, the fact that there are 20-40% of advanced laryngeal cancer patients do not response to paclitaxel makes it necessary to figure out potential biomarkers for paclitaxel sensitivity prediction. In this work, Hep2, a laryngeal cancer cell line, untreated or treated with lower dose of paclitaxel for 24 h, was applied to DNA microarray chips for gene and miR expression profile analysis. Expression of eight genes altered significantly following paclitaxel treatment, which was further validated by quantitative real-time PCR. Four up-regulated genes were ID2, BMP4, CCL4 and ACTG2, in which ID2 and BMP4 were implicated to be involved in several drugs sensitivity. While the down-regulated four genes, MAPK4, FASN, INSIG1 and SCD, were mainly linked to the endoplasmic reticulum and fatty acid biosynthesis, these two cell processes that are associated with drug sensitivity by increasing evidences. After paclitaxel treatment, expression of 49 miRs was significantly altered. Within these miRs, the most markedly expression-changed were miR-31-star, miR-1264, miR-3150b-5p and miR-210. While the miRs putatively modulated the mRNA expression of the most significantly expression-altered genes were miR-1264, miR-130a, miR-27b, miR-195, miR-1291, miR-214, miR-1277 and miR-1265, which were obtained by miR target prediction and miRNA target correlation. Collectively, our study might provide potential biomarkers for paclitaxel sensitivity prediction and drug resistance targets in laryngeal cancer patients. PMID:23826416

  19. Boolean ErbB network reconstructions and perturbation simulations reveal individual drug response in different breast cancer cell lines

    PubMed Central

    2014-01-01

    Background Despite promising progress in targeted breast cancer therapy, drug resistance remains challenging. The monoclonal antibody drugs trastuzumab and pertuzumab as well as the small molecule inhibitor erlotinib were designed to prevent ErbB-2 and ErbB-1 receptor induced deregulated protein signalling, contributing to tumour progression. The oncogenic potential of ErbB receptors unfolds in case of overexpression or mutations. Dimerisation with other receptors allows to bypass pathway blockades. Our intention is to reconstruct the ErbB network to reveal resistance mechanisms. We used longitudinal proteomic data of ErbB receptors and downstream targets in the ErbB-2 amplified breast cancer cell lines BT474, SKBR3 and HCC1954 treated with erlotinib, trastuzumab or pertuzumab, alone or combined, up to 60 minutes and 30 hours, respectively. In a Boolean modelling approach, signalling networks were reconstructed based on these data in a cell line and time course specific manner, including prior literature knowledge. Finally, we simulated network response to inhibitor combinations to detect signalling nodes reflecting growth inhibition. Results The networks pointed to cell line specific activation patterns of the MAPK and PI3K pathway. In BT474, the PI3K signal route was favoured, while in SKBR3, novel edges highlighted MAPK signalling. In HCC1954, the inferred edges stimulated both pathways. For example, we uncovered feedback loops amplifying PI3K signalling, in line with the known trastuzumab resistance of this cell line. In the perturbation simulations on the short-term networks, we analysed ERK1/2, AKT and p70S6K. The results indicated a pathway specific drug response, driven by the type of growth factor stimulus. HCC1954 revealed an edgetic type of PIK3CA-mutation, contributing to trastuzumab inefficacy. Drug impact on the AKT and ERK1/2 signalling axes is mirrored by effects on RB and RPS6, relating to phenotypic events like cell growth or proliferation

  20. Germ line knockout of IGFBP-3 reveals influences of the gene on mammary gland neoplasia.

    PubMed

    Blouin, Marie-José; Bazile, Miguel; Birman, Elena; Zakikhani, Mahvash; Florianova, Livia; Aleynikova, Olga; Powell, David R; Pollak, Michael

    2015-02-01

    Insulin-like growth factor binding protein-3 (IGFBP-3) is an important carrier protein for insulin-like growth factors (IGFs) in the circulation. IGFBP-3 antagonizes the growth-promoting and anti-apoptotic activities of IGFs in experimental systems, but in certain contexts can increase IGF bioactivity, probably by increasing its half-life. The goal of this study was to investigate the role of IGFBP-3 in breast carcinogenesis and breast cancer metastasis. In the first part of the study, we exposed IGFBP-3 knockout and wild-type female mice to dimethylbenz[a]anthracene (DMBA) and followed them for appearance of primary tumors for up to 13 months. In the second part, mice of each genotype received an IV injection of 4T1 mammary carcinoma cells and then lung nodules were counted. Our results show that IGFBP-3 knockout mice developed breast tumors significantly earlier than the wild-type (13.9 ± 1.1 versus 22.5 ± 3.3 weeks, respectively, P = 0.0144), suggesting tumor suppression activity of IGFBP-3. In tumors of IGFBP-3 knockout mice, levels of phospho-AKT(Ser473) were increased compared to wild-type mice. The lung metastasis assay showed significantly more and larger lung nodules in IGFBP-3 knockout mice than in wild-type mice. While we observed increased levels of IGFBP-5 protein in the IGFBP-3 knockout mice, our findings suggest that this was not sufficient to completely compensate for the absence of IGFBP-3. Even though knockout of IGFBP-3 is associated with only a subtle phenotype under control conditions, our results reveal that loss of this gene has measurable effects on breast carcinogenesis and breast cancer metastasis. PMID:25614235

  1. Lines

    ERIC Educational Resources Information Center

    Mires, Peter B.

    2006-01-01

    National Geography Standards for the middle school years generally stress the teaching of latitude and longitude. There are many creative ways to explain the great grid that encircles our planet, but the author has found that students in his college-level geography courses especially enjoy human-interest stories associated with lines of latitude…

  2. Morphological characterization of a newly established human osteosarcoma cell line, HS-Os-1, revealing its distinct osteoblastic nature.

    PubMed

    Sonobe, H; Mizobuchi, H; Manabe, Y; Furihata, M; Iwata, J; Hikita, T; Oka, T; Ohtsuki, Y; Goto, T

    1991-01-01

    A newly established human osteosarcoma cell line, HS-Os-1, from an osteoblastic tumor arising in the left humerus of an 11-year-old girl was morphologically characterized in vitro and in vivo. HS-Os-1 cells in a monolayer have been maintained for more than 2 years since the initial cultivation, and were round or polygonal in shape with marked pleomorphism. Their cytoplasm was strongly positive for specific markers of osteoblasts, such as alkaline phosphatase and osteocalcin. Tumors induced in nude mice by HS-Os-1 cell inoculation at passage 12 or 23 revealed typical histological features of osteoblastic osteosarcoma, similar to those observed in the original tumor, producing prominent osteoid matrix with calcification. Ultrastructurally, HS-Os-1 cells in vitro and tumor cells in vivo showed similar well-developed, markedly dilated rough endoplasmic reticulum, polysomes and microfilaments in their cytoplasm. Additionally, many collagen fibers associated with deposition of electron-dense material were detected in the stroma featuring osteoid matrix. Thus, the HS-Os-1 cell line was shown to exhibit its osteoblastic nature in vitro and in vivo, and therefore might become an extremely useful tool for various pathomorphological investigations on human osteosarcomas. PMID:1679269

  3. Replication Fork Polarity Gradients Revealed by Megabase-Sized U-Shaped Replication Timing Domains in Human Cell Lines

    PubMed Central

    Baker, Antoine; Audit, Benjamin; Chen, Chun-Long; Moindrot, Benoit; Leleu, Antoine; Guilbaud, Guillaume; Rappailles, Aurélien; Vaillant, Cédric; Goldar, Arach; Mongelard, Fabien; d'Aubenton-Carafa, Yves; Hyrien, Olivier; Thermes, Claude; Arneodo, Alain

    2012-01-01

    In higher eukaryotes, replication program specification in different cell types remains to be fully understood. We show for seven human cell lines that about half of the genome is divided in domains that display a characteristic U-shaped replication timing profile with early initiation zones at borders and late replication at centers. Significant overlap is observed between U-domains of different cell lines and also with germline replication domains exhibiting a N-shaped nucleotide compositional skew. From the demonstration that the average fork polarity is directly reflected by both the compositional skew and the derivative of the replication timing profile, we argue that the fact that this derivative displays a N-shape in U-domains sustains the existence of large-scale gradients of replication fork polarity in somatic and germline cells. Analysis of chromatin interaction (Hi-C) and chromatin marker data reveals that U-domains correspond to high-order chromatin structural units. We discuss possible models for replication origin activation within U/N-domains. The compartmentalization of the genome into replication U/N-domains provides new insights on the organization of the replication program in the human genome. PMID:22496629

  4. The Swift-BAT monitoring reveals a long term decay of the cyclotron line energy in Vela X-1

    NASA Astrophysics Data System (ADS)

    La Parola, V.; Cusumano, G.; Segreto, A.; D'Aì, A.

    2016-08-01

    We study the behaviour of the cyclotron resonant scattering feature (CRSF) of the high mass X-ray binary Vela X-1 using the long-term hard X-ray monitoring performed by the Burst Alert Telescope (BAT) on board Swift. High statistics, intensity selected spectra were built along 11 years of BAT survey. While the fundamental line is not revealed, the second harmonic of the CRSF can be clearly detected in all the spectra, at an energy varying between ˜53 keV and ˜58 keV, directly correlated with the luminosity. We have further investigated the evolution of the CRSF in time, by studying the intensity selected spectra built along four 33-month time intervals along the survey. For the first time we find in this source a secular variation in the CRSF energy: independent of the source luminosity, the CRSF second harmonic energy decreases by ˜0.36 keV/year between the first and the third time interval, corresponding to an apparent decay of the magnetic field of ˜3 × 1010 G/year. The intensity-cyclotron energy pattern is consistent between the third and the last time intervals. A possible interpretation for this decay could be the settling of an accreted mound that produces either a distortion of the poloidal magnetic field on the polar cap or a geometrical displacement of the line forming region. This hypothesis seems supported by the correspondance between the rate of the line shift per unit accreted mass and the mass accreted on the polar cap per unit area in Vela X-1 and Her X-1, respectively.

  5. Biophysical characterization of MDR breast cancer cell lines reveals the cytoplasm is critical in determining drug sensitivity.

    PubMed

    Coley, Helen M; Labeed, Fatima H; Thomas, Hilary; Hughes, Michael P

    2007-04-01

    Dielectrophoresis (DEP) was used to examine a panel of MCF-7 cell lines comprising parental MCF-7 cells and MDR derivatives: MCF-7TaxR (paclitaxel-resistant, P-glycoprotein (P-gp) positive), MCF-7DoxR (doxorubicin-resistant MRP2 positive) plus MCF-7MDR1 (MDR1 transfected, P-gp positive). MCF-7DoxR and MCF-7MDR1 were broadly cross-resistant to natural product anticancer agents, whereas MCF-7TaxR cells were not, contrary to P-gp expression. Whilst DEP revealed modest membrane changes in MDR sub-lines, we saw significant changes in their cytoplasmic conductivity: MCF-7TaxRlines can be delineated by assessment of cytoplasmic biophysical properties using DEP. PMID:17270349

  6. Development, expansion, and in vivo monitoring of human NK cells from human embryonic stem cells (hESCs) and and induced pluripotent stem cells (iPSCs).

    PubMed

    Bock, Allison M; Knorr, David; Kaufman, Dan S

    2013-01-01

    We present a method for deriving natural killer (NK) cells from undifferentiated hESCs and iPSCs using a feeder-free approach. This method gives rise to high levels of NK cells after 4 weeks culture and can undergo further 2-log expansion with artificial antigen presenting cells. hESC- and iPSC-derived NK cells developed in this system have a mature phenotype and function. The production of large numbers of genetically modifiable NK cells is applicable for both basic mechanistic as well as anti-tumor studies. Expression of firefly luciferase in hESC-derived NK cells allows a non-invasive approach to follow NK cell engraftment, distribution, and function. We also describe a dual-imaging scheme that allows separate monitoring of two different cell populations to more distinctly characterize their interactions in vivo. This method of derivation, expansion, and dual in vivo imaging provides a reliable approach for producing NK cells and their evaluation which is necessary to improve current NK cell adoptive therapies. PMID:23644738

  7. On-line monitoring of Soxhlet extraction by chromatography and mass spectrometry to reveal temporal extract profiles.

    PubMed

    Chen, Ssu-Ying; Urban, Pawel L

    2015-06-30

    Soxhlet extraction is a popular sample preparation technique used in chemical analysis. It enables liberation of molecules embedded in complex matrices (for example, plant tissues, foodstuffs). In most protocols, samples are analyzed after the extraction process is complete. However, in order to optimize extraction conditions and enable comparisons between different types of extraction, it would be desirable to monitor it in real time. The main development of this work is the design and construction of the interface between Soxhlet extractor and GC-MS as well as ESI-MS system. The temporal extract profiles, obtained in the course of real-time GC-MS monitoring, have been fitted with mathematical functions to analyze extraction kinetics of different analytes. For example, the mass transfer coefficients of pinene, limonene and terpinene in lemon sample, estimated using the first-order kinetic model, are 0.540h(-1), 0.507h(-1) and 0.722h(-1), respectively. On the other hand, the Peleg model provides the following extraction rates of pinene, limonene and terpinene: 0.370nMh(-1), 0.216nMh(-1) and 0.596nMh(-1), respectively. The results suggest that both first-order kinetic and Peleg equations can be used to describe the progress of Soxhlet extraction. On-line monitoring of Soxhlet extraction reveals extractability of various analytes present in natural samples (plant tissue), and can potentially facilitate optimization of the extraction process. PMID:26041522

  8. Molecular response to the pathogen Phytophthora sojae among ten soybean near isogenic lines revealed by comparative transcriptomics

    PubMed Central

    2014-01-01

    Background Phytophthora root and stem rot (PRR) of soybean, caused by Phytophthora sojae, is controlled by Rps genes. However, little is known regarding the Rps-induced molecular responses to P. sojae and how they actually overlap. We thus sequenced, analyzed, and compared the transcriptomes of 10 near isogenic lines (NILs), each with a unique Rps gene/allele, and the susceptible parent Williams, pre- and post-inoculation with the pathogen. Results A total of 4,330 differentially expressed genes (DEGs) were identified in Williams versus 2,014 to 5,499 DEGs in individual NILs upon inoculation with the pathogen. Comparisons of the DEGs between the NILs and Williams identified incompatible interaction genes (IIGs) and compatible interaction genes (CIGs). Hierarchical cluster and heatmap analyses consistently grouped the NILs into three clusters: Cluster I (Rps1-a), Cluster II (Rps1-b, 1-c and 1-k) and Cluster III (Rps3-a, 3-b, 3-c, 4, 5, and 6), suggesting an overlap in Rps-induced defense signaling among certain NILs. Gene ontology (GO) analysis revealed associations between members of the WRKY family and incompatible reactions and between a number of phytohormone signaling pathways and incompatible/compatible interactions. These associations appear to be distinguished according to the NIL clusters. Conclusions This study characterized genes and multiple branches of putative regulatory networks associated with resistance to P. sojae in ten soybean NILs, and depicted functional “fingerprints” of individual Rps-mediated resistance responses through comparative transcriptomic analysis. Of particular interest are dramatic variations of detected DEGs, putatively involved in ethylene (ET)-, jasmonic acid (JA)-, (reactive oxygen species) ROS-, and (MAP-kinase) MAPK- signaling, among these soybean NILs, implicating their important roles of these signaling in differentiating molecular defense responses. We hypothesize that different timing and robustness in defense

  9. Genetic profiling reveals an alarming rate of cross-contamination among human cell lines used in China.

    PubMed

    Ye, Fang; Chen, Chuguang; Qin, Jian; Liu, Jie; Zheng, Congyi

    2015-10-01

    Cell lines are widely used as in vitro model systems in biologic and medical research. However, much of the research has been invalidated by the unwitting use of false cell lines. A significant proportion of the research involving human cell lines was initiated in China. Paradoxically, the cell lines used in China have never been authenticated. Here, we present a comprehensive survey of cross-contamination in 380 samples from 113 independent sources in China using short tandem repeat profiling methods. High levels of cross-contamination were uncovered (95 of 380, 25%). Notable false cell lines (e.g., KB and WISH) are still actively used under their false identity and tissue attributions. Most strikingly, 85.51% of lines established in China were misidentified (59 of 69) and accounted for over half of the misidentifications (59 of 95, 62.11%). Further, 93.22% of the contaminants in cell lines established in laboratories of China were HeLa cells or a possible hybrid of HeLa with an unknown cell line. Results from these misidentified lines have been published in thousands of potentially erroneous articles and may have distorted the findings visible to the scientific community. False lines have been used in drug screening, potentially leading to unusable or even harmful therapeutic strategies. We also noted the causes of contamination and provided suggestions for remediation. PMID:26116706

  10. Fresh embryo donation for human embryonic stem cell (hESC) research: the experiences and values of IVF couples asked to be embryo donors

    PubMed Central

    Haimes, E.; Taylor, K.

    2009-01-01

    BACKGROUND This article reports on an investigation of the views of IVF couples asked to donate fresh embryos for research and contributes to the debates on: the acceptability of human embryonic stem cell (hESC) research, the moral status of the human embryo and embryo donation for research. METHODS A hypothesis-generating design was followed. All IVF couples in one UK clinic who were asked to donate embryos in 1 year were contacted 6 weeks after their pregnancy result. Forty four in-depth interviews were conducted. RESULTS Interviewees were preoccupied with IVF treatment and the request to donate was a secondary consideration. They used a complex and dynamic system of embryo classification. Initially, all embryos were important but then their focus shifted to those that had most potential to produce a baby. At that point, ‘other’ embryos were less important though they later realise that they did not know what happened to them. Guessing that these embryos went to research, interviewees preferred not to contemplate what that might entail. The embryos that caused interviewees most concern were good quality embryos that might have produced a baby but went to research instead. ‘The’ embryo, the morally laden, but abstract, entity, did not play a central role in their decision-making. CONCLUSIONS This study, despite missing those who refuse to donate embryos, suggests that debates on embryo donation for hESC research should include the views of embryo donors and should consider the social, as well as the moral, status of the human embryo. PMID:19502616

  11. Quantitative proteomic analysis of wheat grain proteins reveals differential effects of silencing of omega-5 gliadin genes in transgenic lines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Novel wheat lines with altered flour compositions can be used to decipher the roles of specific gluten proteins in flour quality. Grain proteins from transgenic wheat lines in which genes encoding the omega-5 gliadins were silenced by RNA interference (RNAi) were analyzed in detail by quantitative 2...

  12. The different origins of high- and low-ionization broad emission lines revealed by gravitational microlensing in the Einstein cross

    NASA Astrophysics Data System (ADS)

    Braibant, L.; Hutsemékers, D.; Sluse, D.; Anguita, T.

    2016-07-01

    We investigate the kinematics and ionization structure of the broad emission line region of the gravitationally lensed quasar QSO2237+0305 (the Einstein cross) using differential microlensing in the high- and low-ionization broad emission lines. We combine visible and near-infrared spectra of the four images of the lensed quasar and detect a large-amplitude microlensing effect distorting the high-ionization CIV and low-ionization Hα line profiles in image A. While microlensing only magnifies the red wing of the Balmer line, it symmetrically magnifies the wings of the CIV emission line. Given that the same microlensing pattern magnifies both the high- and low-ionization broad emission line regions, these dissimilar distortions of the line profiles suggest that the high- and low-ionization regions are governed by different kinematics. Since this quasar is likely viewed at intermediate inclination, we argue that the differential magnification of the blue and red wings of Hα favors a flattened, virialized, low-ionization region whereas the symmetric microlensing effect measured in CIV can be reproduced by an emission line formed in a polar wind, without the need of fine-tuned caustic configurations. Based on observations made with the ESO-VLT, Paranal, Chile; Proposals 076.B-0197 and 076.B-0607 (PI: Courbin).

  13. Registration of Human Embryonic Stem Cell Lines: Korea, 2010

    PubMed Central

    Lee, Ji-Yoon; Lee, Dae-Yeon; Choi, Young-Sil; Lee, Kyoung-Jae; Kim, Yong-Ou

    2011-01-01

    In an effort to increase the credibility of human embryonic stem cell (hESC) lines established in Korea, obligatory registration was introduced by the Bioethics and Safety Act 2008, effective as of January 1, 2010. The DNA fingerprint, chromosome stability, expression of pluripotency markers, and contamination of mycoplasma of the submitted lines were analyzed by Korea Centers for Disease Control and Prevention (KCDC). The characterization data and ethical aspects, such as informed consent for donation of surplus embryos, were reviewed by a 10-member advisory review committee for stem cell registry. A total of 55 domestic hESC lines were submitted for registration in 2010; among them 51 were registered. Among these submitted lines, 26 were additionally characterized by KCDC, while 25 lines previously characterized by the Ministry of Education, Science and Technology were not additionally analyzed by KCDC. Registration completed an oversight system for embryo research by registering the products of licensed embryo research projects, making embryo research more transparent in Korea. Information about hESC lines is available at the website of the Korea Stem Cell Registry (kscr.nih.go.kr). PMID:24159464

  14. N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression.

    PubMed

    Holst, Stephanie; Deuss, Anna J M; van Pelt, Gabi W; van Vliet, Sandra J; Garcia-Vallejo, Juan J; Koeleman, Carolien A M; Deelder, André M; Mesker, Wilma E; Tollenaar, Rob A; Rombouts, Yoann; Wuhrer, Manfred

    2016-01-01

    Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. However, differences between CRC cell lines with regard to their glycosylation have hitherto been largely neglected. Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation

  15. N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression*

    PubMed Central

    Holst, Stephanie; Deuss, Anna J. M.; van Pelt, Gabi W.; van Vliet, Sandra J.; Garcia-Vallejo, Juan J.; Koeleman, Carolien A. M.; Deelder, André M.; Mesker, Wilma E.; Tollenaar, Rob A.; Rombouts, Yoann; Wuhrer, Manfred

    2016-01-01

    Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. However, differences between CRC cell lines with regard to their glycosylation have hitherto been largely neglected. Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation

  16. Supernovae. ⁴⁴Ti gamma-ray emission lines from SN1987A reveal an asymmetric explosion.

    PubMed

    Boggs, S E; Harrison, F A; Miyasaka, H; Grefenstette, B W; Zoglauer, A; Fryer, C L; Reynolds, S P; Alexander, D M; An, H; Barret, D; Christensen, F E; Craig, W W; Forster, K; Giommi, P; Hailey, C J; Hornstrup, A; Kitaguchi, T; Koglin, J E; Madsen, K K; Mao, P H; Mori, K; Perri, M; Pivovaroff, M J; Puccetti, S; Rana, V; Stern, D; Westergaard, N J; Zhang, W W

    2015-05-01

    In core-collapse supernovae, titanium-44 ((44)Ti) is produced in the innermost ejecta, in the layer of material directly on top of the newly formed compact object. As such, it provides a direct probe of the supernova engine. Observations of supernova 1987A (SN1987A) have resolved the 67.87- and 78.32-kilo-electron volt emission lines from decay of (44)Ti produced in the supernova explosion. These lines are narrow and redshifted with a Doppler velocity of ~700 kilometers per second, direct evidence of large-scale asymmetry in the explosion. PMID:25954004

  17. Derivation of HVR1, HVR2 and HVR3 human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis (PGD) for monogenic disorder.

    PubMed

    Hmadcha, Abdelkrim; Aguilera, Yolanda; Lozano-Arana, Maria Dolores; Mellado, Nuria; Sánchez, Javier; Moya, Cristina; Sánchez-Palazón, Luis; Palacios, Jose; Antiñolo, Guillermo; Soria, Bernat

    2016-05-01

    From 106 human blastocyts donate for research after in vitro fertilization (IVF) and preimplantation genetic diagnosis (PGD) for monogenetic disorder, 3 human embryonic stem cells (hESCs) HVR1, HVR2 and HVR3 were successfully derived. HVR1 was assumed to be genetically normal, HVR2 carrying Becker muscular dystrophy and HVR3 Hemophilia B. Despite the translocation t(9;15)(q34.3;q14) detected in HVR2, all the 3 cell lines were characterised in vitro and in vivo as normal hESCs lines and were registered in the Spanish Stem Cell Bank. PMID:27346196

  18. MiRNA-Target Interaction Reveals Cell-Specific Post-Transcriptional Regulation in Mammalian Cell Lines

    PubMed Central

    Kulkarni, Varun; Naqvi, Afsar Raza; Uttamani, Juhi Raju; Nares, Salvador

    2016-01-01

    MicroRNAs are 18–22 nucleotides long, non-coding RNAs that bind transcripts with complementary sequences leading to either mRNA degradation or translational suppression. However, the inherent differences in preferred mode of miRNA regulation among cells of different origin have not been examined. In our previous transcriptome profiling studies, we observed that post-transcriptional regulation can differ substantially depending on the cell in context. Here we examined mechanistic differences in the regulation of a let-7a targeted (wild type) or resistant (mutant) engineered renilla transcript across various mammalian cell lines of diverse origin. Dual luciferase assays show that compared to mutant (mut), the reporter gene containing wild type (wt) let-7a binding sites was efficiently suppressed upon transfection in various cell lines. Importantly, the strength of miRNA regulation varied across the cell lines. Total RNA analysis demonstrates that wt renilla mRNA was expressed to similar or higher levels compared to mut suggesting that translation repression is a predominant mode of miRNA regulation. Nonetheless, transcript degradation was observed in some cell lines. Ago-2 immunoprecipitation show that miRNA repressed renilla mRNA are associated with functional mi-RISC (miRNA-RNA induced silencing complex). Given the immense potential of miRNA as a therapeutic option, these findings highlight the necessity to thoroughly examine the mode of mRNA regulation in order to achieve the beneficial effects in targeting cells. PMID:26761000

  19. Sex determination by SRY PCR and sequencing of Tasmanian devil facial tumour cell lines reveals non-allograft transmission.

    PubMed

    Cui, Xianlan; Wang, Yunfeng; Hua, Bobby; Miller, Webb; Zhao, Yan; Cui, Hongyu; Kong, Xiangang

    2016-05-20

    Devil facial tumour disease (DFTD) is an infectious tumour disease and was hypothesised to be transmitted by allograft during biting based on two cytogenetic findings of DFTD tumours in 2006. It was then believed that DFTD tumours were originally from a female devil. In this study the devil sex-determining region Y (SRY) gene was PCR amplified and sequenced, and six pairs of devil SRY PCR primers were used for detection of devil SRY gene fragments in purified DFTD tumour cell lines. Using three pairs of devil SRY PCR primers, devil SRY gene sequence was detected by PCR and sequencing in genomic DNA of DFTD tumour cell lines from six male devils, but not from six female devils. Four out of six DFTD tumour cell lines from male devils contained nucleotides 288-482 of the devil SRY gene, and another two DFTD tumour cell lines contained nucleotides 381-577 and 493-708 of the gene, respectively. These results indicate that the different portions of the SRY gene in the DFTD tumours of the male devils were originally from the male hosts, rejecting the currently believed DFTD allograft transmission theory. The reasons why DFTD transmission was incorrectly defined as allograft are discussed. PMID:27084454

  20. Single-cell lineage tracking analysis reveals that an established cell line comprises putative cancer stem cells and their heterogeneous progeny.

    PubMed

    Sato, Sachiko; Rancourt, Ann; Sato, Yukiko; Satoh, Masahiko S

    2016-01-01

    Mammalian cell culture has been used in many biological studies on the assumption that a cell line comprises putatively homogeneous clonal cells, thereby sharing similar phenotypic features. This fundamental assumption has not yet been fully tested; therefore, we developed a method for the chronological analysis of individual HeLa cells. The analysis was performed by live cell imaging, tracking of every single cell recorded on imaging videos, and determining the fates of individual cells. We found that cell fate varied significantly, indicating that, in contrast to the assumption, the HeLa cell line is composed of highly heterogeneous cells. Furthermore, our results reveal that only a limited number of cells are immortal and renew themselves, giving rise to the remaining cells. These cells have reduced reproductive ability, creating a functionally heterogeneous cell population. Hence, the HeLa cell line is maintained by the limited number of immortal cells, which could be putative cancer stem cells. PMID:27003384

  1. Revealing the second harmonic generation in a femtosecond laser-driven cluster-based plasma by analyzing shapes of Ar XVII spectral lines.

    PubMed

    Oks, Eugene; Dalimier, Elisabeth; Faenov, Anatoly; Pikuz, Tatiana; Fukuda, Yuji; Andreev, Alexander; Koga, James; Sakaki, Hironao; Kotaki, Hideyuki; Pirozhkov, Alexander; Hayashi, Yukio; Skobelev, Igor; Pikuz, Sergei; Kawachi, Tetsuya; Kando, Masaki; Kondo, Kiminori; Zhidkov, Alexei; Kodama, Ryosuke

    2015-12-14

    We present experiments dealing with a femtosecond laser-driven cluster-based plasma, where by analyzing the nonlinear phenomenon of satellites of spectral lines of Ar XVII, we revealed the nonlinear phenomenon of the generation of the second harmonic of the laser frequency. For performing this analysis we developed new results in the theory of satellites of spectral lines. From such lineshape analysis we found, in particular, that the efficiency of converting the short (40 fs) intense (3x10¹⁸ W/cm²) incident laser light into the second harmonic was 2%. This result is in the excellent agreement with the 2-Dimensional Particle-In-Cell (2D PIC) simulation that we also performed. There is also an order of magnitude agreement between the thresholds for the SHG found from the line shape analysis and from the 2D PIC simulations. PMID:26698990

  2. Single-cell lineage tracking analysis reveals that an established cell line comprises putative cancer stem cells and their heterogeneous progeny

    PubMed Central

    Sato, Sachiko; Rancourt, Ann; Sato, Yukiko; Satoh, Masahiko S.

    2016-01-01

    Mammalian cell culture has been used in many biological studies on the assumption that a cell line comprises putatively homogeneous clonal cells, thereby sharing similar phenotypic features. This fundamental assumption has not yet been fully tested; therefore, we developed a method for the chronological analysis of individual HeLa cells. The analysis was performed by live cell imaging, tracking of every single cell recorded on imaging videos, and determining the fates of individual cells. We found that cell fate varied significantly, indicating that, in contrast to the assumption, the HeLa cell line is composed of highly heterogeneous cells. Furthermore, our results reveal that only a limited number of cells are immortal and renew themselves, giving rise to the remaining cells. These cells have reduced reproductive ability, creating a functionally heterogeneous cell population. Hence, the HeLa cell line is maintained by the limited number of immortal cells, which could be putative cancer stem cells. PMID:27003384

  3. Reprogramming somatic cells into iPS cells activates LINE-1 retroelement mobility

    PubMed Central

    Wissing, Silke; Muñoz-Lopez, Martin; Macia, Angela; Yang, Zhiyuan; Montano, Mauricio; Collins, William; Garcia-Perez, Jose Luis; Moran, John V.; Greene, Warner C.

    2012-01-01

    Long interspersed element-1 (LINE-1 or L1) retrotransposons account for nearly 17% of human genomic DNA and represent a major evolutionary force that has reshaped the structure and function of the human genome. However, questions remain concerning both the frequency and the developmental timing of L1 retrotransposition in vivo and whether the mobility of these retroelements commonly results in insertional and post-insertional mechanisms of genomic injury. Cells exhibiting high rates of L1 retrotransposition might be especially at risk for such injury. We assessed L1 mRNA expression and L1 retrotransposition in two biologically relevant cell types, human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs), as well as in control parental human dermal fibroblasts (HDFs). Full-length L1 mRNA and the L1 open reading frame 1-encoded protein (ORF1p) were readily detected in hESCs and iPSCs, but not in HDFs. Sequencing analysis proved the expression of human-specific L1 element mRNAs in iPSCs. Bisulfite sequencing revealed that the increased L1 expression observed in iPSCs correlates with an overall decrease in CpG methylation in the L1 promoter region. Finally, retrotransposition of an engineered human L1 element was ∼10-fold more efficient in iPSCs than in parental HDFs. These findings indicate that somatic cell reprogramming is associated with marked increases in L1 expression and perhaps increases in endogenous L1 retrotransposition, which could potentially impact the genomic integrity of the resultant iPSCs. PMID:21989055

  4. Small RNA Profiles of the Rice PTGMS Line Wuxiang S Reveal miRNAs Involved in Fertility Transition

    PubMed Central

    Zhang, Hongyuan; Hu, Jihong; Qian, Qian; Chen, Hao; Jin, Jing; Ding, Yi

    2016-01-01

    MicroRNAs (miRNAs) play key roles in the regulation of plant growth and developmental processes. In this study, RNA-seq was used to examine the expression profiles of miRNAs in a novel, photo-thermosensitive genic male sterile (PTGMS) rice line, Wuxiang S (WXS), during fertility transition. A total of 497 known miRNAs and 273 novel miRNAs were identified. In a differential expression analysis, 26 miRNAs exhibited significant differential expression between WXS (Sterile, S) and WXS (Fertile, F). Some of these miRNAs were validated by quantitative real-time PCR. Among these miRNAs, 11 showed decreased expression levels, and 15 showed increased expression levels in WXS (S) compared to WXS (F). Some of these miRNAs, such as osa-miR156a-j, osa-miR164d, and osa-miR528, were shown to be negatively correlated with their targets. These targets have previously been reported to be related to pollen development and male sterility, suggesting that these miRNAs may be involved in the regulation of pollen development in the rice PTGMS line WXS. Furthermore, miRNA-mediated editing events were also observed. In this study, a possible model for the control of signaling pathways during the process of fertility transition in the rice PTGMS line WXS by miRNAs was developed. These findings contribute to our understanding of the roles of miRNAs during anther development in PTGMS lines in rice. PMID:27148335

  5. Small RNA Profiles of the Rice PTGMS Line Wuxiang S Reveal miRNAs Involved in Fertility Transition.

    PubMed

    Zhang, Hongyuan; Hu, Jihong; Qian, Qian; Chen, Hao; Jin, Jing; Ding, Yi

    2016-01-01

    MicroRNAs (miRNAs) play key roles in the regulation of plant growth and developmental processes. In this study, RNA-seq was used to examine the expression profiles of miRNAs in a novel, photo-thermosensitive genic male sterile (PTGMS) rice line, Wuxiang S (WXS), during fertility transition. A total of 497 known miRNAs and 273 novel miRNAs were identified. In a differential expression analysis, 26 miRNAs exhibited significant differential expression between WXS (Sterile, S) and WXS (Fertile, F). Some of these miRNAs were validated by quantitative real-time PCR. Among these miRNAs, 11 showed decreased expression levels, and 15 showed increased expression levels in WXS (S) compared to WXS (F). Some of these miRNAs, such as osa-miR156a-j, osa-miR164d, and osa-miR528, were shown to be negatively correlated with their targets. These targets have previously been reported to be related to pollen development and male sterility, suggesting that these miRNAs may be involved in the regulation of pollen development in the rice PTGMS line WXS. Furthermore, miRNA-mediated editing events were also observed. In this study, a possible model for the control of signaling pathways during the process of fertility transition in the rice PTGMS line WXS by miRNAs was developed. These findings contribute to our understanding of the roles of miRNAs during anther development in PTGMS lines in rice. PMID:27148335

  6. Comprehensive High-Throughput RNA Sequencing Analysis Reveals Contamination of Multiple Nasopharyngeal Carcinoma Cell Lines with HeLa Cell Genomes

    PubMed Central

    Strong, Michael J.; Baddoo, Melody; Nanbo, Asuka; Xu, Miao; Puetter, Adriane

    2014-01-01

    ABSTRACT In an attempt to explore infectious agents associated with nasopharyngeal carcinomas (NPCs), we employed our high-throughput RNA sequencing (RNA-seq) analysis pipeline, RNA CoMPASS, to investigate the presence of ectopic organisms within a number of NPC cell lines commonly used by NPC and Epstein-Barr virus (EBV) researchers. Sequencing data sets from both CNE1 and HONE1 were found to contain reads for human papillomavirus 18 (HPV-18). Subsequent real-time reverse transcription-PCR (RT-PCR) analysis on a panel of NPC cell lines identified HPV-18 in CNE1 and HONE1 as well as three additional NPC cell lines (CNE2, AdAH, and NPC-KT). Further analysis of the chromosomal integration arrangement of HPV-18 in NPCs revealed patterns identical to those observed in HeLa cells. Clustering based on human single nucleotide variation (SNV) analysis of two separate HeLa cell lines and several NPC cell lines demonstrated two distinct clusters with CNE1, as well as HONE1 clustering with the two HeLa cell lines. In addition, duplex-PCR-based genotyping showed that CNE1, CNE2, and HONE1 do not have a HeLa cell-specific L1 retrotransposon insertion, suggesting that these three HPV-18+ NPC lines are likely products of a somatic hybridization with HeLa cells, which is also consistent with our RNA-seq-based gene level SNV analysis. Taking all of these findings together, we conclude that a widespread HeLa contamination may exist in many NPC cell lines, and authentication of these cell lines is recommended. Finally, we provide a proof of concept for the utility of an RNA-seq-based approach for cell authentication. IMPORTANCE Nasopharyngeal carcinoma (NPC) cell lines are important model systems for analyzing the complex life cycle and pathogenesis of Epstein-Barr virus (EBV). Using an RNA-seq-based approach, we found HeLa cell contamination in several NPC cell lines that are commonly used in the EBV and related fields. Our data support the notion that contamination resulted from

  7. A Deep X-Ray View of the Bare AGN Ark 120. I. Revealing the Soft X-Ray Line Emission

    NASA Astrophysics Data System (ADS)

    Reeves, J. N.; Porquet, D.; Braito, V.; Nardini, E.; Lobban, A.; Turner, T. J.

    2016-09-01

    The Seyfert 1 galaxy Ark 120 is a prototype example of the so-called class of bare nucleus active galactic nuclei (AGNs), whereby there is no known evidence for the presence of ionized gas along the direct line of sight. Here deep (>400 ks exposure), high-resolution X-ray spectroscopy of Ark 120 is presented from XMM-Newton observations that were carried out in 2014 March, together with simultaneous Chandra/High Energy Transmission Grating exposures. The high-resolution spectra confirmed the lack of intrinsic absorbing gas associated with Ark 120, with the only X-ray absorption present originating from the interstellar medium (ISM) of our own Galaxy, with a possible slight enhancement of the oxygen abundance required with respect to the expected ISM values in the solar neighborhood. However, the presence of several soft X-ray emission lines are revealed for the first time in the XMM-Newton RGS spectrum, associated with the AGN and arising from the He- and H-like ions of N, O, Ne, and Mg. The He-like line profiles of N, O, and Ne appear velocity broadened, with typical FWHMs of ∼5000 km s‑1, whereas the H-like profiles are unresolved. From the clean measurement of the He-like triplets, we deduce that the broad lines arise from a gas of density n e ∼ 1011 cm‑3, while the photoionization calculations infer that the emitting gas covers at least 10% of 4π steradian. Thus the broad soft X-ray profiles appear coincident with an X-ray component of the optical–UV broad-line region on sub-parsec scales, whereas the narrow profiles originate on larger parsec scales, perhaps coincident with the AGN narrow-line region. The observations show that Ark 120 is not intrinsically bare and substantial X-ray-emitting gas exists out of our direct line of sight toward this AGN.

  8. Assessment of small RNA sorting into different extracellular fractions revealed by high-throughput sequencing of breast cell lines

    PubMed Central

    Tosar, Juan Pablo; Gámbaro, Fabiana; Sanguinetti, Julia; Bonilla, Braulio; Witwer, Kenneth W.; Cayota, Alfonso

    2015-01-01

    Intercellular communication can be mediated by extracellular small regulatory RNAs (sRNAs). Circulating sRNAs are being intensively studied for their promising use as minimally invasive disease biomarkers. To date, most attention is centered on exosomes and microRNAs as the vectors and the secreted species, respectively. However, this field would benefit from an increased understanding of the plethora of sRNAs secreted by different cell types in different extracellular fractions. It is still not clear if specific sRNAs are selected for secretion, or if sRNA secretion is mostly passive. We sequenced the intracellular sRNA content (19–60 nt) of breast epithelial cell lines (MCF-7 and MCF-10A) and compared it with extracellular fractions enriched in microvesicles, exosomes and ribonucleoprotein complexes. Our results are consistent with a non-selective secretion model for most microRNAs, although a few showed secretion patterns consistent with preferential secretion. On the contrary, 5′ tRNA halves and 5′ RNA Y4-derived fragments of 31–33 were greatly and significantly enriched in the extracellular space (even in non-mammary cell lines), where tRNA halves were detected as part of ∼45 kDa ribonucleoprotein complexes. Overall, we show that different sRNA families have characteristic secretion patterns and open the question of the role of these sRNAs in the extracellular space. PMID:25940616

  9. First-generation HapMap in Cajanus spp. reveals untapped variations in parental lines of mapping populations.

    PubMed

    Kumar, Vinay; Khan, Aamir W; Saxena, Rachit K; Garg, Vanika; Varshney, Rajeev K

    2016-08-01

    Whole genome re-sequencing (WGRS) was conducted on a panel of 20 Cajanus spp. accessions (crossing parentals of recombinant inbred lines, introgression lines, multiparent advanced generation intercross and nested association mapping population) comprising of two wild species and 18 cultivated species accessions. A total of 791.77 million paired-end reads were generated with an effective mapping depth of ~12X per accession. Analysis of WGRS data provided 5 465 676 genome-wide variations including 4 686 422 SNPs and 779 254 InDels across the accessions. Large structural variations in the form of copy number variations (2598) and presence and absence variations (970) were also identified. Additionally, 2 630 904 accession-specific variations comprising of 2 278 571 SNPs (86.6%), 166 243 deletions (6.3%) and 186 090 insertions (7.1%) were also reported. Identified polymorphic sites in this study provide the first-generation HapMap in Cajanus spp. which will be useful in mapping the genomic regions responsible for important traits. PMID:26821983

  10. Revealing the broad iron Kα line in Cygnus X-1 through simultaneous XMM-Newton, RXTE, and INTEGRAL observations

    NASA Astrophysics Data System (ADS)

    Duro, Refiz; Dauser, Thomas; Grinberg, Victoria; Miškovičová, Ivica; Rodriguez, Jérôme; Tomsick, John; Hanke, Manfred; Pottschmidt, Katja; Nowak, Michael A.; Kreykenbohm, Sonja; Cadolle Bel, Marion; Bodaghee, Arash; Lohfink, Anne; Reynolds, Christopher S.; Kendziorra, Eckhard; Kirsch, Marcus G. F.; Staubert, Rüdiger; Wilms, Jörn

    2016-05-01

    We report on the analysis of the broad Fe Kα line feature of Cyg X-1 in the spectra of four simultaneous hard intermediate state observations made with the X-ray Multiple Mirror mission (XMM-Newton), the Rossi X-ray Timing Explorer (RXTE), and the International Gamma-Ray Astrophysics Laboratory (INTEGRAL). The high quality of the XMM-Newton data taken in the Modified Timing Mode of the EPIC-pn camera provides a great opportunity to investigate the broadened Fe Kα reflection line at 6.4 keV with a very high signal to noise ratio. The 4-500 keV energy range is used to constrain the underlying continuum and the reflection at higher energies. We first investigate the data by applying a phenomenological model that consists of the sum of an exponentially cutoff power law and relativistically smeared reflection. Additionally, we apply a more physical approach and model the irradiation of the accretion disk directly from the lamp post geometry. All four observations show consistent values for the black hole parameters with a spin of a ~ 0.9, in agreement with recent measurements from reflection and disk continuum fitting. The inclination is found to be i ~ 30°, consistent with the orbital inclination and different from inclination measurements made during the soft state, which show a higher inclination. We speculate that the difference between the inclination measurements is due to changes in the inner region of the accretion disk.

  11. Revealing the properties of the weak-lined TTauri binary HDE 245059 with Chandra HETGS and Keck

    NASA Astrophysics Data System (ADS)

    Baldovin Saavedra, C.

    2009-09-01

    We present the Chandra (HETGS) and Keck observations of the young weak-lined T Tauri star HDE 245059 located in the Lambda-Ori star forming region. Our observations in both wavelength regimes have shown that the star is in fact a binary separated by 0.87 arcsec. In the X-rays the plasma properties of both binary components are similar; they show a wide range of plasma temperatures from about 6 to 40 MK dominated by plasma between 8 to 15 MK. The hydrogen column density is low (NH ~ 8x10^19 cm-2), probably due to the clearing of the region by a supernova explosion 1 Myr ago. The coronal abundances show an inverse FIP effect, which is also observed in other young active stars. We have obtained line fluxes from the He-like triplets finding no evidence of high density plasma, which is consistent with the absence of accretion in the WTT binary. Furthermore, using the near-infrared photometry and evolutionary models we have also obtained the first estimates for the masses, effective temperatures, and radii for the binary components. According to our study HDE 245059 is 2-3 Myr old, accounting on our estimates the binary has experienced the supernova explosion leaving unchanged its coronal properties.

  12. The large scale structure of the Universe revealed with high redshift emission-line galaxies: implications for future surveys

    NASA Astrophysics Data System (ADS)

    Antonino Orsi, Alvaro

    2015-08-01

    Nebular emission in galaxies trace their star-formation activity within the last 10 Myr or so. Hence, these objects are typically found in the outskirts of massive clusters, where otherwise environmental effects can effectively stop the star formation process. In this talk I discuss the nature of emission-line galaxies (ELGs) and its implications for their clustering properties. To account for the relevant physical ingredients that produce nebular emission, I combine semi-analytical models of galaxy formation with a radiative transfer code of Ly-alpha photons, and the photoionzation and shock code MAPPINGS-III. As a result, the clustering strength of ELGs is found to correlate weakly with the line luminosities. Also, their 2-d clustering displays a weak finger-of-god effect, and the clustering in linear scales is affected by assembly bias. I review the impact of the nature of this galaxy population for future spectroscopic large surveys targeting ELGs to extract cosmological results. In particular, I present forecasts for the ELG population in J-PAS, an 8000 deg^2 survey with 54 narrow-band filters covering the optical range, expected to start in 2016.

  13. A genome-wide association analysis of temozolomide response using lymphoblastoid cell lines reveals a clinically relevant association with MGMT

    PubMed Central

    Brown, Chad C.; Havener, Tammy M.; Medina, Marisa Wong; Auman, J. Todd; Mangravite, Lara M.; Krauss, Ronald M.; McLeod, Howard L.; Motsinger-Reif, Alison A.

    2013-01-01

    Recently, lymphoblastoid cell lines (LCLs) have emerged as an innovative model system for mapping gene variants that predict dose response to chemotherapy drugs. In the current study, this strategy was expanded to the in vitro genome-wide association approach, using 516 LCLs derived from a Caucasian cohort to assess cytotoxic response to temozolomide. Genome-wide association analysis using approximately 2.1 million quality controlled single-nucleotide polymorphisms (SNPs) identified a statistically significant association (p < 10−8) with SNPs in the O6-methylguanine–DNA methyltransferase (MGMT) gene. We also demonstrate that the primary SNP in this region is significantly associated with differential gene expression of MGMT (p< 10−26) in LCLs, and differential methylation in glioblastoma samples from The Cancer Genome Atlas. The previously documented clinical and functional relationships between MGMT and temozolomide response highlight the potential of well-powered GWAS of the LCL model system to identify meaningful genetic associations. PMID:23047291

  14. Nustar Reveals an Intrinsically X-ray Weak Broad Absorption Line Quasar in the Ultraluminous Infrared Galaxy Markarian 231

    NASA Technical Reports Server (NTRS)

    Teng, Stacy H.; Brandt. W. N.; Harrison, F. A.; Luo, B.; Alexander, D. M.; Bauer, F. E.; Boggs, S. E.; Christensen, F. E.; Comastri, A.; Craig, W. W.; Fabian, A. C.; Farrah, D.; Fiore, F.; Gandhi, P.; Grefenstette, B. W.; Hailey, C. J.; Hickox, R. C.; Madsen, K. K.; Ptak, A. F.; Rigby, Jane Rebecca; Risaliti, G.; Saz, C.; Stern, D.; Veilleux, S.; Walton, D. J.; Wik, D. R.; Zhang, W. W.

    2014-01-01

    We present high-energy (3-30 keV) NuSTAR observations of the nearest quasar, the ultraluminous infrared galaxy (ULIRG) Markarian 231 (Mrk 231), supplemented with new and simultaneous low-energy (0.5-8 keV) data from Chandra. The source was detected, though at much fainter levels than previously reported, likely due to contamination in the large apertures of previous non-focusing hard X-ray telescopes. The full band (0.5-30 keV) X-ray spectrum suggests the active galactic nucleus (AGN) in Mrk 231 is absorbed by a patchy and Compton-thin N(sub H) approx. 1.2(sup +0.3) sub-0.3) x 10(exp 23) / sq cm) column. The intrinsic X-ray luminosity L(sub 0.5-30 Kev) approx. 1.0 x 10(exp 43) erg /s) is extremely weak relative to the bolometric luminosity where the 2-10 keV to bolometric luminosity ratio is approx. 0.03% compared to the typical values of 2-15%. Additionally, Mrk 231 has a low X-ray-to-optical power law slope alpha(sub 0X) approx. -1.7. It is a local example of a low-ionization broad absorption line (LoBAL) quasar that is intrinsically X-ray weak. The weak ionizing continuum may explain the lack of mid-infrared [O IV], [Ne V], and [Ne VI] fine-structure emission lines which are present in sources with otherwise similar AGN properties. We argue that the intrinsic X-ray weakness may be a result of the super-Eddington accretion occurring in the nucleus of this ULIRG, and may also be naturally related to the powerful wind event seen in Mrk 231, a merger remnant escaping from its dusty cocoon.

  15. NuSTAR reveals the Comptonizing corona of the broad-line radio galaxy 3C 382

    SciTech Connect

    Ballantyne, D. R.; Bollenbacher, J. M.; Brenneman, L. W.; Madsen, K. K.; Baloković, M.; Harrison, F. A.; Walton, D. J.; Boggs, S. E.; Christensen, F. E.; Craig, W. W.; Gandhi, P.; Hailey, C. J.; Lohfink, A. M.; Marinucci, A.; Markwardt, C. B.; Zhang, W. W.; Stern, D.

    2014-10-10

    Broad-line radio galaxies (BLRGs) are active galactic nuclei that produce powerful, large-scale radio jets, but appear as Seyfert 1 galaxies in their optical spectra. In the X-ray band, BLRGs also appear like Seyfert galaxies, but with flatter spectra and weaker reflection features. One explanation for these properties is that the X-ray continuum is diluted by emission from the jet. Here, we present two NuSTAR observations of the BLRG 3C 382 that show clear evidence that the continuum of this source is dominated by thermal Comptonization, as in Seyfert 1 galaxies. The two observations were separated by over a year and found 3C 382 in different states separated by a factor of 1.7 in flux. The lower flux spectrum has a photon-index of Γ=1.68{sub −0.02}{sup +0.03}, while the photon-index of the higher flux spectrum is Γ=1.78{sub −0.03}{sup +0.02}. Thermal and anisotropic Comptonization models provide an excellent fit to both spectra and show that the coronal plasma cooled from kT{sub e} = 330 ± 30 keV in the low flux data to 231{sub −88}{sup +50} keV in the high flux observation. This cooling behavior is typical of Comptonizing corona in Seyfert galaxies and is distinct from the variations observed in jet-dominated sources. In the high flux observation, simultaneous Swift data are leveraged to obtain a broadband spectral energy distribution and indicates that the corona intercepts ∼10% of the optical and ultraviolet emitting accretion disk. 3C 382 exhibits very weak reflection features, with no detectable relativistic Fe Kα line, that may be best explained by an outflowing corona combined with an ionized inner accretion disk.

  16. Close women, distant men: line bisection reveals sex-dimorphic patterns of visuomotor performance in near and far space.

    PubMed

    Stancey, Helen; Turner, Mark

    2010-05-01

    The mid-points of a series of lines which were positioned both within hand-reach (near space) and beyond hand-reach (far space) were estimated by 24 women and 24 men. When using a laser pointer to perform estimations, women were more accurate in the near condition than the far, whereas men were more accurate in the far condition than the near. When using a stick pointer for the far condition, women were more accurate than when using the laser, whereas men were more accurate using the laser pointer than the stick for the far condition. There was no difference between near and far accuracy scores for either sex using the stick. These results suggest that use of a tool which provides proprioceptive feedback causes the brain to remap far-space stimuli as if situated in near space. Possible origins and neural bases for these differences are considered. Finally, the study found evidence for pseudoneglect, but no evidence for pseudoneglect shift. PMID:19646327

  17. Protein Profiles Reveal Diverse Responsive Signaling Pathways in Kernels of Two Maize Inbred Lines with Contrasting Drought Sensitivity

    PubMed Central

    Yang, Liming; Jiang, Tingbo; Fountain, Jake C.; Scully, Brian T.; Lee, Robert D.; Kemerait, Robert C.; Chen, Sixue; Guo, Baozhu

    2014-01-01

    Drought stress is a major factor that contributes to disease susceptibility and yield loss in agricultural crops. To identify drought responsive proteins and explore metabolic pathways involved in maize tolerance to drought stress, two maize lines (B73 and Lo964) with contrasting drought sensitivity were examined. The treatments of drought and well water were applied at 14 days after pollination (DAP), and protein profiles were investigated in developing kernels (35 DAP) using iTRAQ (isobaric tags for relative and absolute quantitation). Proteomic analysis showed that 70 and 36 proteins were significantly altered in their expression under drought treatments in B73 and Lo964, respectively. The numbers and levels of differentially expressed proteins were generally higher in the sensitive genotype, B73, implying an increased sensitivity to drought given the function of the observed differentially expressed proteins, such as redox homeostasis, cell rescue/defense, hormone regulation and protein biosynthesis and degradation. Lo964 possessed a more stable status with fewer differentially expressed proteins. However, B73 seems to rapidly initiate signaling pathways in response to drought through adjusting diverse defense pathways. These changes in protein expression allow for the production of a drought stress-responsive network in maize kernels. PMID:25334062

  18. Protein profiles reveal diverse responsive signaling pathways in kernels of two maize inbred lines with contrasting drought sensitivity.

    PubMed

    Yang, Liming; Jiang, Tingbo; Fountain, Jake C; Scully, Brian T; Lee, Robert D; Kemerait, Robert C; Chen, Sixue; Guo, Baozhu

    2014-01-01

    Drought stress is a major factor that contributes to disease susceptibility and yield loss in agricultural crops. To identify drought responsive proteins and explore metabolic pathways involved in maize tolerance to drought stress, two maize lines (B73 and Lo964) with contrasting drought sensitivity were examined. The treatments of drought and well water were applied at 14 days after pollination (DAP), and protein profiles were investigated in developing kernels (35 DAP) using iTRAQ (isobaric tags for relative and absolute quantitation). Proteomic analysis showed that 70 and 36 proteins were significantly altered in their expression under drought treatments in B73 and Lo964, respectively. The numbers and levels of differentially expressed proteins were generally higher in the sensitive genotype, B73, implying an increased sensitivity to drought given the function of the observed differentially expressed proteins, such as redox homeostasis, cell rescue/defense, hormone regulation and protein biosynthesis and degradation. Lo964 possessed a more stable status with fewer differentially expressed proteins. However, B73 seems to rapidly initiate signaling pathways in response to drought through adjusting diverse defense pathways. These changes in protein expression allow for the production of a drought stress-responsive network in maize kernels. PMID:25334062

  19. DNA microarray reveals different pathways responding to paclitaxel and docetaxel in non-small cell lung cancer cell line

    PubMed Central

    Che, Chun-Li; Zhang, Yi-Mei; Zhang, Hai-Hong; Sang, Yu-Lan; Lu, Ben; Dong, Fu-Shi; Zhang, Li-Juan; Lv, Fu-Zhen

    2013-01-01

    The wide use of paclitaxel and docetaxel in NSCLC clinical treatment makes it necessary to find biomarkers for identifying patients who can benefit from paclitaxel or docetaxel. In present study, NCI-H460, a NSCLC cell line with different sensitivity to paclitaxel and docetaxel, was applied to DNA microarray expression profiling analysis at different time points of lower dose treatment with paclitaxel or docetaxel. And the complex signaling pathways regulating the drug response were identified, and several novel sensitivity-realted markers were biocomputated.The dynamic changes of responding genes showed that paclitaxel effect is acute but that of docetaxel is durable at least for 48 hours in NCI-H460 cells. Functional annotation of the genes with altered expression showed that genes/pathways responding to these two drugs were dramatically different. Gene expression changes induced by paclitaxel treatment were mainly enriched in actin cytoskeleton (ACTC1, MYL2 and MYH2), tyrosine-protein kinases (ERRB4, KIT and TIE1) and focal adhesion pathway (MYL2, IGF1 and FLT1), while the expression alterations responding to docetaxel were highly co-related to cell surface receptor linked signal transduction (SHH, DRD5 and ADM2), cytokine-cytokine receptor interaction (IL1A and IL6) and cell cycleregulation (CCNB1, CCNE2 and PCNA). Moreover, we also confirmed some different expression patterns with real time PCR. Our study will provide the potential biomarkers for paclitaxel and docetaxel-selection therapy in clinical application. PMID:23923072

  20. On-line stable isotope gas exchange reveals an inducible but leaky carbon concentrating mechanism in Nannochloropsis salina.

    PubMed

    Hanson, David T; Collins, Aaron M; Jones, Howland D T; Roesgen, John; Lopez-Nieves, Samuel; Timlin, Jerilyn A

    2014-09-01

    Carbon concentrating mechanisms (CCMs) are common among microalgae, but their regulation and even existence in some of the most promising biofuel production strains is poorly understood. This is partly because screening for new strains does not commonly include assessment of CCM function or regulation despite its fundamental role in primary carbon metabolism. In addition, the inducible nature of many microalgal CCMs means that environmental conditions should be considered when assessing CCM function and its potential impact on biofuels. In this study, we address the effect of environmental conditions by combining novel, high frequency, on-line (13)CO2 gas exchange screen with microscope-based lipid characterization to assess CCM function in Nannochloropsis salina and its interaction with lipid production. Regulation of CCM function was explored by changing the concentration of CO2 provided to continuous cultures in airlift bioreactors where cell density was kept constant across conditions by controlling the rate of media supply. Our isotopic gas exchange results were consistent with N. salina having an inducible "pump-leak" style CCM similar to that of Nannochloropsis gaditana. Though cells grew faster at high CO2 and had higher rates of net CO2 uptake, we did not observe significant differences in lipid content between conditions. Since the rate of CO2 supply was much higher for the high CO2 conditions, we calculated that growing cells bubbled with low CO2 is about 40 % more efficient for carbon capture than bubbling with high CO2. We attribute this higher efficiency to the activity of a CCM under low CO2 conditions. PMID:24844569

  1. Genomewide Variation in an Introgression Line of Rice-Zizania Revealed by Whole-Genome re-Sequencing

    PubMed Central

    Bai, Yan; Zhang, Yun-Hong; Liu, Ying; Wu, Ying; Lin, Xiu-Yun; Wen, Jia-Wei; Xu, Chun-Ming; Li, Lin-Feng; Liu, Bao

    2013-01-01

    Background Hybridization between genetically diverged organisms is known as an important avenue that drives plant genome evolution. The possible outcomes of hybridization would be the occurrences of genetic instabilities in the resultant hybrids. It remained under-investigated however whether pollination by alien pollens of a closely related but sexually "incompatible" species could evoke genomic changes and to what extent it may result in phenotypic novelties in the derived progenies. Methodology/Principal Findings In this study, we have re-sequenced the genomes of Oryza sativa ssp. japonica cv. Matsumae and one of its derived introgressant RZ35 that was obtained from an introgressive hybridization between Matsumae and Zizanialatifolia Griseb. in general, 131 millions 90 base pair (bp) paired-end reads were generated which covered 13.2 and 21.9 folds of the Matsumae and RZ35 genomes, respectively. Relative to Matsumae, a total of 41,724 homozygous single nucleotide polymorphisms (SNPs) and 17,839 homozygous insertions/deletions (indels) were identified in RZ35, of which 3,797 SNPs were nonsynonymous mutations. Furthermore, rampant mobilization of transposable elements (TEs) was found in the RZ35 genome. The results of pathogen inoculation revealed that RZ35 exhibited enhanced resistance to blast relative to Matsumae. Notably, one nonsynonymous mutation was found in the known blast resistance gene Pid3/Pi25 and real-time quantitative (q) RT-PCR analysis revealed constitutive up-regulation of its expression, suggesting both altered function and expression of Pid3/Pi25 may be responsible for the enhanced resistance to rice blast by RZ35. Conclusions/Significance Our results demonstrate that introgressive hybridization by Zizania has provoked genomewide, extensive genomic changes in the rice genome, and some of which have resulted in important phenotypic novelties. These findings suggest that introgressive hybridization by alien pollens of even a sexually incompatible

  2. Analysis of photosynthetic picoeukaryote community structure along an extended Ellett Line transect in the northern North Atlantic reveals a dominance of novel prymnesiophyte and prasinophyte phylotypes

    NASA Astrophysics Data System (ADS)

    Kirkham, Amy R.; Jardillier, Ludwig E.; Holland, Ross; Zubkov, Mikhail V.; Scanlan, Dave J.

    2011-07-01

    Photosynthetic picoeukaryotes (PPEs) of a size <3 μm can contribute significantly to primary production. Here, PPE community structure was analysed along an extended Ellett Line transect, an area in the North Atlantic well studied by physical oceanographers but largely neglected in the field of microalgal ecology. Distribution patterns of specific PPE classes were determined using dot-blot hybridization analysis, while the taxonomic composition of specific PPE classes was revealed by phylogenetic analysis of plastid 16S rRNA gene sequences. In addition, we performed fluorescent in situ hybridization (FISH) analysis of seawater samples collected along the transect to provide a PCR-independent survey of class level PPE distribution patterns. We found the PPE community was dominated by members of the Prymnesiophyceae, Prasinophyceae and Mamiellophyceae. Interestingly, phylogenetic analysis revealed several novel Prymnesiophyceae and Prasinophyceae phylotypes (with only 85-96% identity to neighbouring sequences) within lineages for which cultured counterparts are unknown.

  3. Water-rock interactions in the Median Tectonic Line fault zone, SW Japan revealed by core sample analysis

    NASA Astrophysics Data System (ADS)

    Fujimoto, K.; Tanaka, N.; Shigematsu, N.

    2012-12-01

    Alteration minerals along the fault zone of the Median Tectonic Line (MTL), the Japan's largest onshore fault, are characterized to understand the crustal faulting at various depth conditions by describing a borehole penetrating the MTL. The borehole was drilled by the Geological Survey of Japan, AIST (GSJ, AIST) to predict the forthcoming Nankai-Tonankai Earthquake at Matsusaka-Iitaka (ITA), SW Japan. The drilling length is 600m. It penetrates the MTL at the depth of 473.9m. The fault plane has an almost E-W strike and dips at 56° to the north. Hangingwall of the MTL consists of Ryoke-derived tonalitic mylonite and footwall of the MTL consists of fractured rocks derived from Sanbagawa metamorphic rocks. Ryoke-derived tonalitic mylonite in the hangingwall suffered more or less later cataclastic deformations. X-ray diffraction patterns of powdered samples derived from ITA borehole were obtained to identify alteration mineral assemblages and estimate alteration conditions. Chlorite, laumontite, white-mica, prehnite and calcite are main alteration minerals in the Ryoke belt above 400m depth, while analcine and stilbite occur as zeolite minerals instead of laumontite between 400m and 473.9m depth. Laumontite is preferentially occurred in the cataclastically deformed zones and calcite often fills fractures and cavities. The fault plane and fractures in the fault zone provide fluid conduit. The presence of laumontite and prehnite indicates that the alteration temperature was more than 200°C, while analcime and stilbite formed at lower temperature than laumontite. These suggest that the zone of hydrothermal alteration less than 200°C was restricted below 400m depth in the ITA borehole. We also applied chlorite geothermometry (Inoue et al., 2009), which yeileds temperatures of about 300°C for mylonitic rocks and those of about 200°C for cataclastic rocks. Immediately beneath the MTL, approximately 40m thick fractured rocks form the major strand of the MTL fault zone

  4. Genome Wide Expression Profiling of Cancer Cell Lines Cultured in Microgravity Reveals Significant Dysregulation of Cell Cycle and MicroRNA Gene Networks

    PubMed Central

    Vidyasekar, Prasanna; Shyamsunder, Pavithra; Arun, Rajpranap; Santhakumar, Rajalakshmi; Kapadia, Nand Kishore; Kumar, Ravi; Verma, Rama Shanker

    2015-01-01

    Zero gravity causes several changes in metabolic and functional aspects of the human body and experiments in space flight have demonstrated alterations in cancer growth and progression. This study reports the genome wide expression profiling of a colorectal cancer cell line-DLD-1, and a lymphoblast leukemic cell line-MOLT-4, under simulated microgravity in an effort to understand central processes and cellular functions that are dysregulated among both cell lines. Altered cell morphology, reduced cell viability and an aberrant cell cycle profile in comparison to their static controls were observed in both cell lines under microgravity. The process of cell cycle in DLD-1 cells was markedly affected with reduced viability, reduced colony forming ability, an apoptotic population and dysregulation of cell cycle genes, oncogenes, and cancer progression and prognostic markers. DNA microarray analysis revealed 1801 (upregulated) and 2542 (downregulated) genes (>2 fold) in DLD-1 cultures under microgravity while MOLT-4 cultures differentially expressed 349 (upregulated) and 444 (downregulated) genes (>2 fold) under microgravity. The loss in cell proliferative capacity was corroborated with the downregulation of the cell cycle process as demonstrated by functional clustering of DNA microarray data using gene ontology terms. The genome wide expression profile also showed significant dysregulation of post transcriptional gene silencing machinery and multiple microRNA host genes that are potential tumor suppressors and proto-oncogenes including MIR22HG, MIR17HG and MIR21HG. The MIR22HG, a tumor-suppressor gene was one of the highest upregulated genes in the microarray data showing a 4.4 log fold upregulation under microgravity. Real time PCR validated the dysregulation in the host gene by demonstrating a 4.18 log fold upregulation of the miR-22 microRNA. Microarray data also showed dysregulation of direct targets of miR-22, SP1, CDK6 and CCNA2. PMID:26295583

  5. Genome Wide Expression Profiling of Cancer Cell Lines Cultured in Microgravity Reveals Significant Dysregulation of Cell Cycle and MicroRNA Gene Networks.

    PubMed

    Vidyasekar, Prasanna; Shyamsunder, Pavithra; Arun, Rajpranap; Santhakumar, Rajalakshmi; Kapadia, Nand Kishore; Kumar, Ravi; Verma, Rama Shanker

    2015-01-01

    Zero gravity causes several changes in metabolic and functional aspects of the human body and experiments in space flight have demonstrated alterations in cancer growth and progression. This study reports the genome wide expression profiling of a colorectal cancer cell line-DLD-1, and a lymphoblast leukemic cell line-MOLT-4, under simulated microgravity in an effort to understand central processes and cellular functions that are dysregulated among both cell lines. Altered cell morphology, reduced cell viability and an aberrant cell cycle profile in comparison to their static controls were observed in both cell lines under microgravity. The process of cell cycle in DLD-1 cells was markedly affected with reduced viability, reduced colony forming ability, an apoptotic population and dysregulation of cell cycle genes, oncogenes, and cancer progression and prognostic markers. DNA microarray analysis revealed 1801 (upregulated) and 2542 (downregulated) genes (>2 fold) in DLD-1 cultures under microgravity while MOLT-4 cultures differentially expressed 349 (upregulated) and 444 (downregulated) genes (>2 fold) under microgravity. The loss in cell proliferative capacity was corroborated with the downregulation of the cell cycle process as demonstrated by functional clustering of DNA microarray data using gene ontology terms. The genome wide expression profile also showed significant dysregulation of post transcriptional gene silencing machinery and multiple microRNA host genes that are potential tumor suppressors and proto-oncogenes including MIR22HG, MIR17HG and MIR21HG. The MIR22HG, a tumor-suppressor gene was one of the highest upregulated genes in the microarray data showing a 4.4 log fold upregulation under microgravity. Real time PCR validated the dysregulation in the host gene by demonstrating a 4.18 log fold upregulation of the miR-22 microRNA. Microarray data also showed dysregulation of direct targets of miR-22, SP1, CDK6 and CCNA2. PMID:26295583

  6. An Induced Pluripotent Stem Cell Model of Hypoplastic Left Heart Syndrome (HLHS) Reveals Multiple Expression and Functional Differences in HLHS-Derived Cardiac Myocytes

    PubMed Central

    Jiang, Yan; Habibollah, Saba; Tilgner, Katarzyna; Collin, Joseph; Barta, Tomas; Al-Aama, Jumana Yousuf; Tesarov, Lenka; Hussain, Rafiqul; Trafford, Andrew W.; Kirkwood, Graham; Sernagor, Evelyne; Eleftheriou, Cyril G.; Przyborski, Stefan; Stojković, Miodrag; Lako, Majlinda; Keavney, Bernard

    2014-01-01

    Hypoplastic left heart syndrome (HLHS) is a serious congenital cardiovascular malformation resulting in hypoplasia or atresia of the left ventricle, ascending aorta, and aortic and mitral valves. Diminished flow through the left side of the heart is clearly a key contributor to the condition, but any myocardial susceptibility component is as yet undefined. Using recent advances in the field of induced pluripotent stem cells (iPSCs), we have been able to generate an iPSC model of HLHS malformation and characterize the properties of cardiac myocytes (CMs) differentiated from these and control-iPSC lines. Differentiation of HLHS-iPSCs to cardiac lineages revealed changes in the expression of key cardiac markers and a lower ability to give rise to beating clusters when compared with control-iPSCs and human embryonic stem cells (hESCs). HLHS-iPSC-derived CMs show a lower level of myofibrillar organization, persistence of a fetal gene expression pattern, and changes in commitment to ventricular versus atrial lineages, and they display different calcium transient patterns and electrophysiological responses to caffeine and β-adrenergic antagonists when compared with hESC- and control-iPSC-derived CMs, suggesting that alternative mechanisms to release calcium from intracellular stores such as the inositol trisphosphate receptor may exist in HLHS in addition to the ryanodine receptor thought to function in control-iPSC-derived CMs. Together our findings demonstrate that CMs derived from an HLHS patient demonstrate a number of marker expression and functional differences to hESC/control iPSC-derived CMs, thus providing some evidence that cardiomyocyte-specific factors may influence the risk of HLHS. PMID:24591732

  7. An induced pluripotent stem cell model of hypoplastic left heart syndrome (HLHS) reveals multiple expression and functional differences in HLHS-derived cardiac myocytes.

    PubMed

    Jiang, Yan; Habibollah, Saba; Tilgner, Katarzyna; Collin, Joseph; Barta, Tomas; Al-Aama, Jumana Yousuf; Tesarov, Lenka; Hussain, Rafiqul; Trafford, Andrew W; Kirkwood, Graham; Sernagor, Evelyne; Eleftheriou, Cyril G; Przyborski, Stefan; Stojković, Miodrag; Lako, Majlinda; Keavney, Bernard; Armstrong, Lyle

    2014-04-01

    Hypoplastic left heart syndrome (HLHS) is a serious congenital cardiovascular malformation resulting in hypoplasia or atresia of the left ventricle, ascending aorta, and aortic and mitral valves. Diminished flow through the left side of the heart is clearly a key contributor to the condition, but any myocardial susceptibility component is as yet undefined. Using recent advances in the field of induced pluripotent stem cells (iPSCs), we have been able to generate an iPSC model of HLHS malformation and characterize the properties of cardiac myocytes (CMs) differentiated from these and control-iPSC lines. Differentiation of HLHS-iPSCs to cardiac lineages revealed changes in the expression of key cardiac markers and a lower ability to give rise to beating clusters when compared with control-iPSCs and human embryonic stem cells (hESCs). HLHS-iPSC-derived CMs show a lower level of myofibrillar organization, persistence of a fetal gene expression pattern, and changes in commitment to ventricular versus atrial lineages, and they display different calcium transient patterns and electrophysiological responses to caffeine and β-adrenergic antagonists when compared with hESC- and control-iPSC-derived CMs, suggesting that alternative mechanisms to release calcium from intracellular stores such as the inositol trisphosphate receptor may exist in HLHS in addition to the ryanodine receptor thought to function in control-iPSC-derived CMs. Together our findings demonstrate that CMs derived from an HLHS patient demonstrate a number of marker expression and functional differences to hESC/control iPSC-derived CMs, thus providing some evidence that cardiomyocyte-specific factors may influence the risk of HLHS. PMID:24591732

  8. A new sensitive indicator cell line reveals cross-transactivation of the viral LTR by gorilla and chimpanzee simian foamy viruses.

    PubMed

    Lambert, Caroline; Rua, Réjane; Gessain, Antoine; Buseyne, Florence

    2016-09-01

    The majority of currently identified simian foamy virus (SFV)-infected Cameroonian and Gabonese individuals harbor SFV from the gorilla lineage. We constructed an indicator cell line for the quantification of gorilla SFVs, in which the U3 sequence of a gorilla SFV directs the expression of the β-galactosidase protein. The gorilla foamy virus activated β-galactosidase (GFAB) cells efficiently quantified two zoonotic primary gorilla isolates and SFVs from three chimpanzee subspecies. Primary gorilla SFVs replicated more slowly and at lower levels than primary chimpanzee SFVs. Analysis of previously described motifs of Tas proteins and U3 LTRs involved in viral gene synthesis revealed conservation of such motifs in Tas proteins from gorilla and chimpanzee SFVs, but little sequence homology in the LTR regions previously shown to interact with viral and cellular factors. PMID:27348053

  9. Large scale integration of drug-target information reveals poly-pharmacological drug action mechanisms in tumor cell line growth inhibition assays

    PubMed Central

    Knight, Richard A.; Gostev, Mikhail; Ilisavskii, Sergei; Willis, Anne E.; Melino, Gerry; Antonov, Alexey V.

    2014-01-01

    Understanding therapeutic mechanisms of drug anticancer cytotoxicity represents a key challenge in preclinical testing. Here we have performed a meta-analysis of publicly available tumor cell line growth inhibition assays (~ 70 assays from 6 independent experimental groups covering ~ 500 000 molecules) with the primary goal of understanding molecular therapeutic mechanisms of cancer cytotoxicity. To implement this we have collected currently available information on protein targets for molecules that were tested in the assays. We used a statistical methodology to identify protein targets overrepresented among molecules exhibiting cancer cytotoxicity with the particular focus of identifying overrepresented patterns consisting of several proteins (i.e. proteins “A” and “B” and “C”). Our analysis demonstrates that targeting individual proteins can result in a significant increase (up to 50-fold) of the observed odds for a molecule to be an efficient inhibitor of tumour cell line growth. However, further insight into potential molecular mechanisms reveals a multi-target mode of action: targeting a pattern of several proteins drastically increases the observed odds (up to 500-fold) for a molecule to be tumour cytotoxic. In contrast, molecules targeting only one protein but not targeting an additional set of proteins tend to be nontoxic. Our findings support a poly-pharmacology drug discovery paradigm, demonstrating that anticancer cytotoxicity is a product, in most cases, of multi-target mode of drug action PMID:24553133

  10. Widely Extended [O III] 88μm Line Emission around the 30 Doradus Region Revealed with AKARI FIS-FTS

    NASA Astrophysics Data System (ADS)

    Kawada, Mitsunobu; Takahashi, Ai; Yasuda, Akiko; Kiriyama, Yuichi; Mori, Tatsuya; Mouri, Akio; Kaneda, Hidehiro; Okada, Yoko; Takahashi, Hidenori; Murakami, Noriko

    2011-08-01

    We present a distribution map of the far-infrared [O II] 88 μm line emission around the 30 Doradus (30 Dor) region in the Large Magellanic Cloud obtained with the Fourier Transform Spectrometer of the Far-Infrared Surveyor on board AKARI. The map reveals that the [O III] emission is widely distributed by more than 10' around the super star cluster R 136, implying that the 30 Dor region is affluent with interstellar radiation field that is hard enough to ionize O2+. The observed [O III] line intensities are as high as (1-2) × 10-6 W m-2 sr-1 on the peripheral regions 4'-5' away from the center of 30 Dor, which requires gas densities of 60-100 cm-3. However, the observed size of the distribution of the [O III] emission is too large to be explained by massive stars in the 30 Dor region enshrouded by clouds with a constant gas density of 102 cm-3. Therefore, the surrounding structure is likely to be highly clumpy. We also find a global correlation between the [O III] and the far-infrared continuum emission, suggesting that the gas and dust are well mixed in the highly ionized region where the dust survives in clumpy dense clouds shielded from energetic photons.

  11. Genetic Basis of Heterosis for Growth-Related Traits in Arabidopsis Investigated by Testcross Progenies of Near-Isogenic Lines Reveals a Significant Role of Epistasis

    PubMed Central

    Melchinger, Albrecht E.; Piepho, Hans-Peter; Utz, H. Friedrich; Muminović, Jasmina; Wegenast, Thilo; Törjék, Otto; Altmann, Thomas; Kusterer, Barbara

    2007-01-01

    Epistasis seems to play a significant role in the manifestation of heterosis. However, the power of detecting epistatic interactions among quantitative trait loci (QTL) in segregating populations is low. We studied heterosis in Arabidopsis thaliana hybrid C24 × Col-0 by testing near-isogenic lines (NILs) and their triple testcross (TTC) progenies. Our objectives were to (i) provide the theoretical basis for estimating different types of genetic effects with this experimental design, (ii) determine the extent of heterosis for seven growth-related traits, (iii) map the underlying QTL, and (iv) determine their gene action. Two substitution libraries, each consisting of 28 NILs and covering ∼61 and 39% of the Arabidopsis genome, were assayed by 110 single-nucleotide polymorphism (SNP) markers. With our novel generation means approach 38 QTL were detected, many of which confirmed heterotic QTL detected previously in the same cross with TTC progenies of recombinant inbred lines. Furthermore, many of the QTL were common for different traits and in common with the 58 QTL detected by a method that compares triplets consisting of a NIL, its recurrent parent, and their F1 cross. While the latter approach revealed mostly (75%) overdominant QTL, the former approach allowed separation of dominance and epistasis by analyzing all materials simultaneously and yielded substantial positive additive × additive effects besides directional dominance. Positive epistatic effects reduced heterosis for growth-related traits in our materials. PMID:18039884

  12. The 5-HT{sub 2A} serotoninergic receptor is expressed in the MCF-7 human breast cancer cell line and reveals a mitogenic effect of serotonin

    SciTech Connect

    Sonier, Brigitte; Arseneault, Madeleine; Lavigne, Carole; Ouellette, Rodney J.; Vaillancourt, Cathy . E-mail: cathy.vaillancourt@iaf.inrs.ca

    2006-05-19

    Serotonin (5-hydroxytryptamine, 5-HT) has been described as a mitogen in a variety of cell types and carcinomas. It exerts its mitogenic effect by interacting with a wide range of 5-HT receptor types. Certain studies suggest that some selective serotonin re-uptake inhibitors promote breast cancer in animals and humans. This study attempts to clarify the role of serotonin in promoting the growth of neoplastic mammary cells. Expression of the 5-HT{sub 2A} serotoninergic receptor subtype in MCF-7 cells was determined by RT-PCR, Western blotting, and immunofluorescence analysis. The mitogenic effect of 5-HT on MCF-7 cells was determined by means of the MTT proliferation assay. We have demonstrated that the 5-HT{sub 2A} receptor subtype is fully expressed in the MCF-7 human breast cancer cell line, in terms of encoding mRNA and receptor protein. Automated sequencing has confirmed that the 5-HT{sub 2A} receptor present in this cell line is identical to the 5-HT{sub 2A} receptor found in human platelets and in human cerebral cortex. Furthermore, this receptor was found by immunofluorescence to be on the plasma membrane. MTT proliferation assays revealed that 5-HT and DOI, a selective 5-HT{sub 2A} receptor subtype agonist, stimulated MCF-7 cell. These results indicate that 5-HT plays a mitogenic role in neoplastic mammary cells. Our data also indicate that 5-HT exerts this positive growth effect on MCF-7 cells through, in part, the 5-HT{sub 2A} receptor subtype, which is fully expressed in this cell line.

  13. Isogenic human pluripotent stem cell pairs reveal the role of a KCNH2 mutation in long-QT syndrome

    PubMed Central

    Bellin, Milena; Casini, Simona; Davis, Richard P; D'Aniello, Cristina; Haas, Jessica; Ward-van Oostwaard, Dorien; Tertoolen, Leon G J; Jung, Christian B; Elliott, David A; Welling, Andrea; Laugwitz, Karl-Ludwig; Moretti, Alessandra; Mummery, Christine L

    2013-01-01

    Patient-specific induced pluripotent stem cells (iPSCs) will assist research on genetic cardiac maladies if the disease phenotype is recapitulated in vitro. However, genetic background variations may confound disease traits, especially for disorders with incomplete penetrance, such as long-QT syndromes (LQTS). To study the LQT2-associated c.A2987T (N996I) KCNH2 mutation under genetically defined conditions, we derived iPSCs from a patient carrying this mutation and corrected it. Furthermore, we introduced the same point mutation in human embryonic stem cells (hESCs), generating two genetically distinct isogenic pairs of LQTS and control lines. Correction of the mutation normalized the current (IKr) conducted by the HERG channel and the action potential (AP) duration in iPSC-derived cardiomyocytes (CMs). Introduction of the same mutation reduced IKr and prolonged the AP duration in hESC-derived CMs. Further characterization of N996I-HERG pathogenesis revealed a trafficking defect. Our results demonstrated that the c.A2987T KCNH2 mutation is the primary cause of the LQTS phenotype. Precise genetic modification of pluripotent stem cells provided a physiologically and functionally relevant human cellular context to reveal the pathogenic mechanism underlying this specific disease phenotype. PMID:24213244

  14. Lines of Evidence–Incremental Markings in Molar Enamel of Soay Sheep as Revealed by a Fluorochrome Labeling and Backscattered Electron Imaging Study

    PubMed Central

    Kierdorf, Horst; Kierdorf, Uwe; Frölich, Kai; Witzel, Carsten

    2013-01-01

    We studied the structural characteristics and periodicities of regular incremental markings in sheep enamel using fluorochrome injections for vital labeling of forming enamel and backscattered electron imaging in the scanning electron microscope. Microscopic analysis of mandibular first molars revealed the presence of incremental markings with a daily periodicity (laminations) that indicated successive positions of the forming front of interprismatic enamel. In addition to the laminations, incremental markings with a sub-daily periodicity were discernible both in interprismatic enamel and in enamel prisms. Five sub-daily increments were present between two consecutive laminations. Backscattered electron imaging revealed that each sub-daily growth increment consisted of a broader and more highly mineralized band and a narrower and less mineralized band (line). The sub-daily markings in the prisms of sheep enamel morphologically resembled the (daily) prisms cross striations seen in primate enamel. Incremental markings with a supra-daily periodicity were not observed in sheep enamel. Based on the periodicity of the incremental markings, maximum mean daily apposition rates of 17.0 µm in buccal enamel and of 13.4 µm in lingual enamel were recorded. Enamel extension rates were also high, with maximum means of 180 µm/day and 217 µm/day in upper crown areas of buccal and lingual enamel, respectively. Values in more cervical crown portions were markedly lower. Our results are in accordance with previous findings in other ungulate species. Using the incremental markings present in primate enamel as a reference could result in a misinterpretation of the incremental markings in ungulate enamel. Thus, the sub-daily growth increments in the prisms of ungulate enamel might be mistaken as prism cross striations with a daily periodicity, and the laminations misidentified as striae of Retzius with a supra-daily periodicity. This would lead to a considerable overestimation of

  15. Comparison of Cardiomyogenic Potential among Human ESC and iPSC Lines

    PubMed Central

    Sepac, Ana; Si-Tayeb, Karim; Sedlic, Filip; Barrett, Sara; Canfield, Scott; Duncan, Stephen A.; Bosnjak, Zeljko J.; Lough, John

    2012-01-01

    We recently reported that following induction of clumps of pluripotent H1 human embryonic stem cells (hESCs) with activin-A and Bmp4 in defined medium for five days, widespread differentiation of rhythmically contracting cardiomyocytes occurs within 3–4 weeks. In this study the same approach was used to assess whether human induced pluripotent cells (hiPSCs), which may theoretically provide an unlimited source of patient-matched cells for transplantation therapy, can similarly undergo cardiomyocyte differentiation. Differentiation of four pluripotent cell-lines – H1 and H9 hESCs, and C2a and C6a hiPSCs – was compared in parallel by monitoring rhythmic contraction, morphologic differentiation, and expression of cardiomyogenic genes. Based on expression of the cardiomyogenic lineage markers MESP1, ISL1 and NKX2-5, all four cell-lines were induced into the cardiomyogenic lineage. However, in contrast to the widespread appearance of striations and rhythmic contractility seen in H9 and especially in H1 hESCs, both hiPSC lines exhibited poor terminal differentiation. These findings suggest that refined modes of generating hiPSCs, as well as of inducing cardiomyogenesis in them, may be required to fulfill their potential as agents of cardiac regeneration. PMID:22863088

  16. On-line species-unspecific isotope dilution analysis in the picomolar range reveals the time- and species-depending mercury uptake in human astrocytes.

    PubMed

    Wehe, Christoph A; Pieper, Imke; Holtkamp, Michael; Thyssen, Georgina M; Sperling, Michael; Schwerdtle, Tanja; Karst, Uwe

    2014-03-01

    In order to reveal the time-depending mercury species uptake by human astrocytes, a novel approach for total mercury analysis is presented, which uses an accelerated sample introduction system combined on-line with an inductively coupled plasma mass spectrometer equipped with a collision/reaction cell. Human astrocyte samples were incubated with inorganic mercury (HgCl2), methylmercury chloride (MeHgCl), and thimerosal. After 1-h incubation with Hg(2+), cellular concentrations of 3 μM were obtained, whereas for organic species, concentrations of 14-18 μM could be found. After 24 h, a cellular accumulation factor of 0.3 was observed for the cells incubated with Hg(2+), whereas the organic species both showed values of about 5. Due to the obtained steady-state signals, reliable results with relative standard deviations of well below 5 % and limits of detection in the concentration range of 1 ng L(-1) were obtained using external calibration and species-unspecific isotope dilution analysis approaches. The results were further validated using atomic fluorescence spectrometry. PMID:24442014

  17. Nature of the boundary between open and closed magnetic field line regions at the Sun revealed by composition data and numerical models

    NASA Astrophysics Data System (ADS)

    Posner, Arik; Zurbuchen, Thomas H.; Schwadron, Nathan A.; Fisk, Lennard A.; Gloeckler, George; Linker, Jon A.; Mikić, Zoran; Riley, Pete

    2001-08-01

    Recently, Fisk et al. [1999] have presented a theory that describes a number of features of the large-scale coronal and heliospheric magnetic field. This theory predicts large-scale transport of magnetic flux across the boundaries of the polar coronal holes, which leads to reconnection processes of open field lines with preliminary closed magnetic structures. Reconnection processes reveal themselves in solar wind composition data: Plasma released out of previously closed magnetic field structures exhibits hotter charge state distributions and has a tendency to be enriched by elements with low first ionization potentials. The idea of reconnection at the boundaries of coronal holes is not new. For example, Wang and Sheeley [1993] and Luhmann et al. [1999] found evidence for that mechanism by comparison of observations of the rotation and evolution of coronal holes with potential field models of the solar corona. We use Ulysses Solar Wind Ion Composition Spectrometer composition measurements and sophisticated numerical models [Linker et al., 1999; Riley et al., 1999] to accurately map these observations back to the solar surface. We then constrain the thickness of the stream interface at the Sun and compare the location of the source region with SOHO observations of the low corona. The results are discussed in the context of the global structure of the heliospheric magnetic field.

  18. Analysis of Glu-1 deletion lines reveals the importance of high molecular weight glutenin subunits 7+9 Glu-B1 in wheat flour tortilla making

    Technology Transfer Automated Retrieval System (TEKTRAN)

    High molecular weight glutenin subunits (HMW-GS) play a significant role in the functional properties of wheat flour. Wheat lines in which one or more of the HMW-GS alleles were absent from Glu-A1, Glu-B1 or Glu-D1 loci (deletion lines) were compared with non-deletion lines for dough and tortilla ma...

  19. Genome-wide SNP scan of pooled DNA reveals nonsense mutation in FGF20 in the scaleless line of featherless chickens

    PubMed Central

    2012-01-01

    Background Scaleless (sc/sc) chickens carry a single recessive mutation that causes a lack of almost all body feathers, as well as foot scales and spurs, due to a failure of skin patterning during embryogenesis. This spontaneous mutant line, first described in the 1950s, has been used extensively to explore the tissue interactions involved in ectodermal appendage formation in embryonic skin. Moreover, the trait is potentially useful in tropical agriculture due to the ability of featherless chickens to tolerate heat, which is at present a major constraint to efficient poultry meat production in hot climates. In the interests of enhancing our understanding of feather placode development, and to provide the poultry industry with a strategy to breed heat-tolerant meat-type chickens (broilers), we mapped and identified the sc mutation. Results Through a cost-effective and labour-efficient SNP array mapping approach using DNA from sc/sc and sc/+ blood sample pools, we map the sc trait to chromosome 4 and show that a nonsense mutation in FGF20 is completely associated with the sc/sc phenotype. This mutation, common to all sc/sc individuals and absent from wild type, is predicted to lead to loss of a highly conserved region of the FGF20 protein important for FGF signalling. In situ hybridisation and quantitative RT-PCR studies reveal that FGF20 is epidermally expressed during the early stages of feather placode patterning. In addition, we describe a dCAPS genotyping assay based on the mutation, developed to facilitate discrimination between wild type and sc alleles. Conclusions This work represents the first loss of function genetic evidence supporting a role for FGF ligand signalling in feather development, and suggests FGF20 as a novel central player in the development of vertebrate skin appendages, including hair follicles and exocrine glands. In addition, this is to our knowledge the first report describing the use of the chicken SNP array to map genes based on

  20. Characterization of pathogenic and resistant genome repertoire reveals two clonal lines in Salmonella enterica subsp. enterica serovar Paratyphi B (+)-tartrate positive.

    PubMed

    Huehn, Stephan; Helmuth, Reiner; Bunge, Cornelia; Guerra, Beatriz; Junker, Ernst; Davies, Rob H; Wattiau, Pierre; van Pelt, Wilfrid; Malorny, Burkhard

    2009-05-01

    A total of 36 contemporary human, animal, and environmental (+)-tartrate-fermenting (dT+) Salmonella enterica serovar Paratyphi B isolates, formerly called Salmonella serovar Java, and five related monophasic S. enterica serovar 4,5,12:b:- isolates from Belgium, Germany, the Netherlands, and the United Kingdom were investigated for clonality and antimicrobial resistance profiles, as well as their virulence and resistance gene repertoire. Two major clonal lines, which could be phenotypically differentiated by the expression of the O:5 antigen, were identified. All O:5 antigen negative strains were multidrug resistant and originated (with two exceptions) from Belgian, Dutch, or German poultry. Strains exhibiting the O:5 antigen encoded by the oafA gene revealed a more heterogeneous group including multidrug-resistant and susceptible strains. Compared to O:5 antigen negative isolates, Salmonella Paratyphi B dT+ O:5 positive strains possessed additional virulence determinants. The Salmonella genomic island 1 was only found in O:5 positive strains. Five monophasic Salmonella 4,5,12:b:- lacking the phase-2 flagellar antigen were assigned to Salmonella Paratyphi B dT+ isolates of the O:5 positive group. The conclusion of the analysis is that Salmonella Paratyphi B dT+ O:5 negative and O:5 positive isolates evolved from a different lineage. Salmonella Paratyphi B dT+ O:5 positive strains possess additional fimbrial and virulence genes that probably enable this clone to interact with a broader range of hosts and the environment. Salmonella Paratyphi B dT+ O:5 negative continuously persists in poultry across Western Europe, especially Belgium, the Netherlands, and Germany. PMID:19292689

  1. An Engineered Cardiac Reporter Cell Line Identifies Human Embryonic Stem Cell-Derived Myocardial Precursors

    PubMed Central

    Mihardja, Shirley S.; Liszewski, Walter; Erle, David J.; Lee, Randall J.; Bernstein, Harold S.

    2011-01-01

    Unlike some organs, the heart is unable to repair itself after injury. Human embryonic stem cells (hESCs) grow and divide indefinitely while maintaining the potential to develop into many tissues of the body. As such, they provide an unprecedented opportunity to treat human diseases characterized by tissue loss. We have identified early myocardial precursors derived from hESCs (hMPs) using an α-myosin heavy chain (αMHC)-GFP reporter line. We have demonstrated by immunocytochemistry and quantitative real-time PCR (qPCR) that reporter activation is restricted to hESC-derived cardiomyocytes (CMs) differentiated in vitro, and that hMPs give rise exclusively to muscle in an in vivo teratoma formation assay. We also demonstrate that the reporter does not interfere with hESC genomic stability. Importantly, we show that hMPs give rise to atrial, ventricular and specialized conduction CM subtypes by qPCR and microelectrode array analysis. Expression profiling of hMPs over the course of differentiation implicate Wnt and transforming growth factor-β signaling pathways in CM development. The identification of hMPs using this αMHC-GFP reporter line will provide important insight into the pathways regulating human myocardial development, and may provide a novel therapeutic reagent for the treatment of cardiac disease. PMID:21245908

  2. A Human Pluripotent Stem Cell Surface N-Glycoproteome Resource Reveals Markers, Extracellular Epitopes, and Drug Targets

    PubMed Central

    Boheler, Kenneth R.; Bhattacharya, Subarna; Kropp, Erin M.; Chuppa, Sandra; Riordon, Daniel R.; Bausch-Fluck, Damaris; Burridge, Paul W.; Wu, Joseph C.; Wersto, Robert P.; Chan, Godfrey Chi Fung; Rao, Sridhar; Wollscheid, Bernd; Gundry, Rebekah L.

    2014-01-01

    Summary Detailed knowledge of cell-surface proteins for isolating well-defined populations of human pluripotent stem cells (hPSCs) would significantly enhance their characterization and translational potential. Through a chemoproteomic approach, we developed a cell-surface proteome inventory containing 496 N-linked glycoproteins on human embryonic (hESCs) and induced PSCs (hiPSCs). Against a backdrop of human fibroblasts and 50 other cell types, >100 surface proteins of interest for hPSCs were revealed. The >30 positive and negative markers verified here by orthogonal approaches provide experimental justification for the rational selection of pluripotency and lineage markers, epitopes for cell isolation, and reagents for the characterization of putative hiPSC lines. Comparative differences between the chemoproteomic-defined surfaceome and the transcriptome-predicted surfaceome directly led to the discovery that STF-31, a reported GLUT-1 inhibitor, is toxic to hPSCs and efficient for selective elimination of hPSCs from mixed cultures. PMID:25068131

  3. Derivation of Huntington Disease affected Genea091 human embryonic stem cell line.

    PubMed

    Dumevska, Biljana; Schaft, Julia; McKernan, Robert; Hu, Jesselyn; Schmidt, Uli

    2016-03-01

    The Genea091 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying Htt gene CAG expansion of 40 repeats, indicative of Huntington Disease. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female Allele pattern. The hESC line had pluripotent cell morphology, 92% of cells expressed Nanog, 97% Oct4, 79% Tra1-60 and 98% SSEA4 and gave a Pluritest pluripotency score of 38.36, Novelty of 1.35. The cell line was negative for Mycoplasma and visible contamination. PMID:27346013

  4. Derivation of DM1 affected human embryonic stem cell line Genea067.

    PubMed

    Dumevska, Biljana; Main, Heather; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea067 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying expansion of CTG repeats in the DMPK gene, indicative of Myotonic Dystrophy Type 1 (DM1). Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XY and STR analysis demonstrated a male Allele pattern. The hESC line had pluripotent cell morphology, 85% of cells expressed Nanog, 97% Oct4, 73% Tra1-60 and 98% SSEA4 and gave a Pluritest Pluripotency score of 25.75, Novelty of 1.46. The cell line was negative for Mycoplasma and visible contamination. PMID:27346009

  5. Derivation of DM2 affected human embryonic stem cell line Genea066.

    PubMed

    Dumevska, Biljana; Schaft, Julia; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea066 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying expansion of CCTG repeats in exon 1 of the ZNF9 gene, indicative of Myotonic Dystrophy Type 2 (DM2). Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XY and STR analysis demonstrated a male Allele pattern. The hESC line had pluripotent cell morphology, 88% of cells expressed Nanog, 97% Oct4, 80% Tra1-60 and 99% SSEA4 and gave a Pluritest Pluripotency score of 31.3, Novelty of 1.22. The cell line was negative for Mycoplasma and visible contamination. PMID:27346023

  6. Derivation of Huntington Disease affected Genea046 human embryonic stem cell line.

    PubMed

    Dumevska, Biljana; Chami, Omar; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea046 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying HTT gene CAG expansion of 45 repeats, indicative of Huntington Disease. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female Allele pattern. The hESC line had pluripotent cell morphology, 85% of cells expressed Nanog, 92% Oct4, 75% Tra1-60 and 99% SSEA4 and demonstrated Alkaline Phosphatase activity. The cell line was negative for Mycoplasma and visible contamination. PMID:27346012

  7. Derivation of Huntington Disease affected Genea089 human embryonic stem cell line.

    PubMed

    Dumevska, Biljana; McKernan, Robert; Hu, Jesselyn; Schmidt, Uli

    2016-03-01

    The Genea089 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying Htt gene CAG expansion of 41 repeats, indicative of Huntington Disease. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female Allele pattern. The hESC line had pluripotent cell morphology, 91% of cells expressed Nanog, 95% Oct4, 90% Tra1-60 and 100% SSEA4 and gave a PluriTest Pluripotency score of 39.28, Novelty of 1.2. The cell line was negative for Mycoplasma and visible contamination. PMID:27346008

  8. Derivation of Huntington Disease affected Genea090 human embryonic stem cell line.

    PubMed

    Dumevska, Biljana; Schaft, Julia; McKernan, Robert; Hu, Jesselyn; Schmidt, Uli

    2016-03-01

    The Genea090 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying Htt gene CAG expansion of 45 repeats, indicative of Huntington Disease. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female allele pattern. The hESC line had pluripotent cell morphology, 91% of cells expressed Nanog, 95% Oct4, 90% Tra1-60 and 100% SSEA4 and gave a pluritest pluripotency score of 30.91, novelty of 1.23. The cell line was negative for Mycoplasma and visible contamination. PMID:27346026

  9. Derivation of Huntington disease affected Genea020 human embryonic stem cell line.

    PubMed

    Dumevska, Biljana; Peura, Teija; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea020 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying Htt gene CAG expansion of 48 repeats, indicative of Huntington disease. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female allele pattern. The hESC line had pluripotent cell morphology, 89% of cells expressed Nanog, 95% Oct4, 29% Tra1-60 and 99% SSEA4, gave a Pluritest pluripotency score of 27.51, novelty of 1.43 and demonstrated alkaline phosphatase activity. The cell line was negative for Mycoplasma and visible contamination. PMID:27346007

  10. Derivation of Trisomy 21 affected human embryonic stem cell line Genea053.

    PubMed

    Dumevska, Biljana; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea053 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying Trisomy 21, indicative of Down Syndrome. Following ICM outgrowth on inactivated human feeders, CGH and STR analysis demonstrated a 47, XY, +21 karyotype and male allele pattern. The hESC line had pluripotent cell morphology and expressed pluripotent cell markers including 83% Nanog positive, 87% Oct4, 88% Tra1-60 and 98% SSEA4. The cell line was negative for Mycoplasma and visible contamination. PMID:27346024

  11. Derivation of Trisomy 21 affected human embryonic stem cell line Genea021.

    PubMed

    Dumevska, Biljana; Bosman, Alexis; McKernan, Robert; Main, Heather; Schmidt, Uli; Peura, Teija

    2016-03-01

    The Genea021 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying Trisomy 21, indicative of Down Syndrome. Following ICM outgrowth on inactivated human feeders, CGH and STR analyses demonstrated a 47, XY, +21 karyotype and male allele pattern. The hESC line had pluripotent cell morphology, 71% of cells expressed Nanog, 84% Oct4, 23% Tra1-60 and 95% SSEA4, gave a Pluritest Pluripotency score of 21.85, Novelty of 1.42, demonstrated Alkaline Phosphatase activity and tri-lineage teratoma formation. The cell line was negative for Mycoplasma and visible contamination. PMID:27346003

  12. Reversal of infall in SgrB2(M) revealed by Herschel/HIFI observations of HCN lines at THz frequencies

    NASA Astrophysics Data System (ADS)

    Rolffs, R.; Schilke, P.; Comito, C.; Bergin, E. A.; van der Tak, F. F. S.; Lis, D. C.; Qin, S.-L.; Menten, K. M.; Güsten, R.; Bell, T. A.; Blake, G. A.; Caux, E.; Ceccarelli, C.; Cernicharo, J.; Crockett, N. R.; Daniel, F.; Dubernet, M.-L.; Emprechtinger, M.; Encrenaz, P.; Gerin, M.; Giesen, T. F.; Goicoechea, J. R.; Goldsmith, P. F.; Gupta, H.; Herbst, E.; Joblin, C.; Johnstone, D.; Langer, W. D.; Latter, W. D.; Lord, S. D.; Maret, S.; Martin, P. G.; Melnick, G. J.; Morris, P.; Müller, H. S. P.; Murphy, J. A.; Ossenkopf, V.; Pearson, J. C.; Pérault, M.; Phillips, T. G.; Plume, R.; Schlemmer, S.; Stutzki, J.; Trappe, N.; Vastel, C.; Wang, S.; Yorke, H. W.; Yu, S.; Zmuidzinas, J.; Diez-Gonzalez, M. C.; Bachiller, R.; Martin-Pintado, J.; Baechtold, W.; Olberg, M.; Nordh, L. H.; Gill, J. J.; Chattopadhyay, G.

    2010-10-01

    Aims: To investigate the accretion and feedback processes in massive star formation, we analyze the shapes of emission lines from hot molecular cores, whose asymmetries trace infall and expansion motions. Methods: The high-mass star forming region SgrB2(M) was observed with Herschel/HIFI (HEXOS key project) in various lines of HCN and its isotopologues, complemented by APEX data. The observations are compared to spherically symmetric, centrally heated models with density power-law gradient and different velocity fields (infall or infall+expansion), using the radiative transfer code RATRAN. Results: The HCN line profiles are asymmetric, with the emission peak shifting from blue to red with increasing J and decreasing line opacity (HCN to H13CN). This is most evident in the HCN 12-11 line at 1062 GHz. These line shapes are reproduced by a model whose velocity field changes from infall in the outer part to expansion in the inner part. Conclusions: The qualitative reproduction of the HCN lines suggests that infall dominates in the colder, outer regions, but expansion dominates in the warmer, inner regions. We are thus witnessing the onset of feedback in massive star formation, starting to reverse the infall and finally disrupting the whole molecular cloud. To obtain our result, the THz lines uniquely covered by HIFI were critically important. Herschel is an ESA space observatory with science instruments provided by European-led Principal Investigator consortia and with important participation from NASA.

  13. Human induced pluripotent stem cell lines show stress defense mechanisms and mitochondrial regulation similar to those of human embryonic stem cells.

    PubMed

    Armstrong, Lyle; Tilgner, Katarzyna; Saretzki, Gabriele; Atkinson, Stuart P; Stojkovic, Miodrag; Moreno, Ruben; Przyborski, Stefan; Lako, Majlinda

    2010-04-01

    The generation of induced pluripotent stem cells (iPSC) has enormous potential for the development of patient-specific regenerative medicine. Human embryonic stem cells (hESC) are able to defend their genomic integrity by maintaining low levels of reactive oxygen species (ROS) through a combination of enhanced removal capacity and limited production of these molecules. Such limited ROS production stems partly from the small number of mitochondria present in hESC; thus, it was important to determine that human iPSC (hiPSC) generation is able to eliminate the extra mitochondria present in the parental fibroblasts (reminiscent of "bottleneck" situation after fertilization) and to show that hiPSC have antioxidant defenses similar to hESC. We were able to generate seven hiPSC lines from adult human dermal fibroblasts and have fully characterized two of those clones. Both hiPSC clones express pluripotency markers and are able to differentiate in vitro into cells belonging to all three germ layers. One of these clones is able to produce fully differentiated teratoma, whereas the other hiPSC clone is unable to silence the viral expression of OCT4 and c-MYC, produce fully differentiated teratoma, and unable to downregulate the expression of some of the pluripotency genes during the differentiation process. In spite of these differences, both clones show ROS stress defense mechanisms and mitochondrial biogenesis similar to hESC. Together our data suggest that, during the reprogramming process, certain cellular mechanisms are in place to ensure that hiPSC are provided with the same defense mechanisms against accumulation of ROS as the hESC. PMID:20073085

  14. Mutational analysis of genes coding for cell surface proteins in colorectal cancer cell lines reveal novel altered pathways, druggable mutations and mutated epitopes for targeted therapy

    PubMed Central

    Correa, Bruna R.; Bettoni, Fabiana; Koyama, Fernanda C.; Navarro, Fabio C.P.; Perez, Rodrigo O.; Mariadason, John; Sieber, Oliver M.; Strausberg, Robert L.; Simpson, Andrew J.G.; Jardim, Denis L.F.; Reis, Luiz Fernando L.; Parmigiani, Raphael B.; Galante, Pedro A.F.; Camargo, Anamaria A.

    2014-01-01

    We carried out a mutational analysis of 3,594 genes coding for cell surface proteins (Surfaceome) in 23 colorectal cancer cell lines, searching for new altered pathways, druggable mutations and mutated epitopes for targeted therapy in colorectal cancer. A total of 3,944 somatic non-synonymous substitutions and 595 InDels, occurring in 2,061 (57%) Surfaceome genes were catalogued. We identified 48 genes not previously described as mutated in colorectal tumors in the TCGA database, including genes that are mutated and expressed in >10% of the cell lines (SEMA4C, FGFRL1, PKD1, FAM38A, WDR81, TMEM136, SLC36A1, SLC26A6, IGFLR1). Analysis of these genes uncovered important roles for FGF and SEMA4 signaling in colorectal cancer with possible therapeutic implications. We also found that cell lines express on average 11 druggable mutations, including frequent mutations (>20%) in the receptor tyrosine kinases AXL and EPHA2, which have not been previously considered as potential targets for colorectal cancer. Finally, we identified 82 cell surface mutated epitopes, however expression of only 30% of these epitopes was detected in our cell lines. Notwithstanding, 92% of these epitopes were expressed in cell lines with the mutator phenotype, opening new venues for the use of “general” immune checkpoint drugs in this subset of patients. PMID:25193853

  15. A Euploid Line of Human Embryonic Stem Cells Derived from a 43,XX,dup(9q),+12,-14,-15,-18,-21 Embryo

    PubMed Central

    Fonseca, Simone Aparecida Siqueira; Costas, Roberta Montero; Morato-Marques, Mariana; Costa, Silvia; Alegretti, Jose Roberto; Rosenberg, Carla; da Motta, Eduardo Leme Alves; Serafini, Paulo C.; Pereira, Lygia V.

    2015-01-01

    Aneuploid embryos diagnosed by FISH-based preimplantation genetic screening (PGS) have been shown to yield euploid lines of human embryonic stem cells (hESCs) with a relatively high frequency. Given that the diagnostic procedure is usually based on the analysis of 1–2 blastomeres of 5 to 10-cell cleavage-stage embryos, mosaicism has been a likely explanation for the phenomena. However, FISH-based PGS can have a significant rate of misdiagnosis, and therefore some of those lines may have been derived from euploid embryos misdiagnosed as aneuploid. More recently, coupling of trophectoderm (TE) biopsy at the blastocyst stage and array-CGH lead to a more informative form of PGS. Here we describe the establishment of a new line of hESCs from an embryo with a 43,XX,dup(9q),+12,-14,-15,-18,-21 chromosomal content based on array-CGH of TE biopsy. We show that, despite the complex chromosomal abnormality, the corresponding hESC line BR-6 is euploid (46,XX). Single nucleotide polymorphism analysis showed that the embryo´s missing chromosomes were not duplicated in BR-6, suggesting the existence of extensive mosaicism in the TE lineage. PMID:26540511

  16. Spatio-Temporal Differences in Dystrophin Dynamics at mRNA and Protein Levels Revealed by a Novel FlipTrap Line

    PubMed Central

    Fraser, Scott E.; Trinh, Le A.

    2015-01-01

    Dystrophin (Dmd) is a structural protein that links the extracellular matrix to actin filaments in muscle fibers and is required for the maintenance of muscles integrity. Mutations in Dmd lead to muscular dystrophies in humans and other vertebrates. Here, we report the characterization of a zebrafish gene trap line that fluorescently labels the endogenous Dmd protein (Dmd-citrine, Gt(dmd-citrine) ct90a). We show that the Dmd-citrine line recapitulates endogenous dmd transcript expression and Dmd protein localization. Using this Dmd-citrine line, we follow Dmd localization to the myosepta in real-time using time-lapse microscopy, and find that the accumulation of Dmd protein at the transverse myosepta coincides with the onset of myotome formation, a critical stage in muscle maturation. We observed that Dmd protein localizes specifically to the myosepta prior to dmd mRNA localization. Additionally, we demonstrate that the Dmd-citrine line can be used to assess muscular dystrophy following both genetic and physical disruptions of the muscle. PMID:26083378

  17. A large number of tetraploid Arabidopsis thaliana lines, generated by a rapid strategy, reveal high stability of neo-tetraploids during consecutive generations.

    PubMed

    Yu, Zheng; Haage, Kristina; Streit, Verena E; Gierl, Alfons; Ruiz, Ramón A Torres

    2009-04-01

    Arabidopsis thaliana has, in conjunction with A. arenosa, developed into a system for the molecular analysis of alloplolyploidy. However, there are very few Arabidopsis lines available to study autopolyploidy. In order to investigate polyploidy on a reliable basis, we have optimised conventional methodologies and developed a novel strategy for the rapid generation and identification of polyploids based on trichome branching patterns. The analysis of more than two dozen independently induced Arabidopsis lines has led to interesting observations concerning the relationship between cell size and ploidy levels and on the relative stability of tetraploidy in Arabidopsis over at least three consecutive generations. The most important finding of this work is that neo-tetraploid lines exhibit considerable stability through all the generations tested. The systematic generation of tetraploid collections through this strategy as well as the lines generated in this work will help to unravel the consequences of polyploidy, particularly tetraploidy, on the genome, on gene expression and on natural diversity in Arabidopsis. PMID:19205656

  18. Expression Patterns of Three UGT Genes in Different Chemotype Safflower Lines and under MeJA Stimulus Revealed Their Potential Role in Flavonoid Biosynthesis.

    PubMed

    Guo, Dan-Dan; Liu, Fei; Tu, Yan-Hua; He, Bei-Xuan; Gao, Yue; Guo, Mei-Li

    2016-01-01

    Safflower (Carthamus tinctorius L.) has received a significant amount of attention as a medicinal plant in China. Flavonoids are the dominant active medical compounds. UDP-glycosyltransferase plays an essential role in the biosynthesis and storage of flavonoids in safflower. In this study, 45 UGT unigenes were screened from our transcriptomic database of safflower. Among them, 27 UGT unigenes were predicted to own a complete open reading frame with various pI and Mw. The phylogenetic tree showed that CtUGT3 and CtUGT16 were classified under the UGT71 subfamily involved in metabolite process, whereas CtUGT25 has high identities with PoUGT both catalyzing the glycosylation of flavonoids and belonging to the UGT90 subfamily. cDNA microarray exhibited that the three UGT genes displayed temporal difference in two chemotype safflower lines. To functionally characterize UGT in safflower, CtUGT3, CtUGT16 and CtUGT25 were cloned and analyzed. Subcellular localization suggested that the three UGTs might be located in the cell cytoplasm and chloroplast. The expression pattern showed that the three UGTs were all suppressed in two lines responsive to methyl jasmonate induction. The co-expression relation of expression pattern and metabolite accumulation demonstrated that CtUGT3 and CtUGT25 were positively related to kaempferol-3-O-β-D-glucoside and CtUGT16 was positively related to quercetin-3-O-β-D-glucoside in yellow line, whereas CtUGT3 and CtUGT25 were positively related to quercetin-3-O-β-D-glucoside in white line. This study indicates that the three CtUGTs play a significant and multiple role in flavonoids biosynthesis with presenting different functional characterization in two safflower lines. PMID:27391785

  19. Expression Patterns of Three UGT Genes in Different Chemotype Safflower Lines and under MeJA Stimulus Revealed Their Potential Role in Flavonoid Biosynthesis

    PubMed Central

    Guo, Dan-Dan; Liu, Fei; Tu, Yan-Hua; He, Bei-Xuan; Gao, Yue; Guo, Mei-Li

    2016-01-01

    Safflower (Carthamus tinctorius L.) has received a significant amount of attention as a medicinal plant in China. Flavonoids are the dominant active medical compounds. UDP-glycosyltransferase plays an essential role in the biosynthesis and storage of flavonoids in safflower. In this study, 45 UGT unigenes were screened from our transcriptomic database of safflower. Among them, 27 UGT unigenes were predicted to own a complete open reading frame with various pI and Mw. The phylogenetic tree showed that CtUGT3 and CtUGT16 were classified under the UGT71 subfamily involved in metabolite process, whereas CtUGT25 has high identities with PoUGT both catalyzing the glycosylation of flavonoids and belonging to the UGT90 subfamily. cDNA microarray exhibited that the three UGT genes displayed temporal difference in two chemotype safflower lines. To functionally characterize UGT in safflower, CtUGT3, CtUGT16 and CtUGT25 were cloned and analyzed. Subcellular localization suggested that the three UGTs might be located in the cell cytoplasm and chloroplast. The expression pattern showed that the three UGTs were all suppressed in two lines responsive to methyl jasmonate induction. The co-expression relation of expression pattern and metabolite accumulation demonstrated that CtUGT3 and CtUGT25 were positively related to kaempferol-3-O-β-D-glucoside and CtUGT16 was positively related to quercetin-3-O-β-D-glucoside in yellow line, whereas CtUGT3 and CtUGT25 were positively related to quercetin-3-O-β-D-glucoside in white line. This study indicates that the three CtUGTs play a significant and multiple role in flavonoids biosynthesis with presenting different functional characterization in two safflower lines. PMID:27391785

  20. Immunophenotyping of Waldenströms Macroglobulinemia Cell Lines Reveals Distinct Patterns of Surface Antigen Expression: Potential Biological and Therapeutic Implications

    PubMed Central

    Paulus, Aneel; Chitta, Kasyapa S.; Wallace, Paul K.; Advani, Pooja P.; Akhtar, Sharoon; Kuranz-Blake, Maja; Ailawadhi, Sikander; Chanan-Khan, Asher A.

    2015-01-01

    Waldenströms macroglobulinemia (WM) is a subtype of Non-Hodgkin’s lymphoma in which the tumor cell population is markedly heterogeneous, consisting of immunoglobulin-M secreting B-lymphocytes, plasmacytoid lymphocytes and plasma cells. Due to rarity of disease and scarcity of reliable preclinical models, many facets of WM molecular and phenotypic architecture remain incompletely understood. Currently, there are 3 human WM cell lines that are routinely used in experimental studies, namely, BCWM.1, MWCL-1 and RPCI-WM1. During establishment of RPCI-WM1, we observed loss of the CD19 and CD20 antigens, which are typically present on WM cells. Intrigued by this observation and in an effort to better define the immunophenotypic makeup of this cell line, we conducted a more comprehensive analysis for the presence or absence of other cell surface antigens that are present on the RPCI-WM1 model, as well as those on the two other WM cell lines, BCWM.1 and MWCL-1. We examined expression of 65 extracellular and 4 intracellular antigens, comprising B-cell, plasma cell, T-cell, NK-cell, myeloid and hematopoietic stem cell surface markers by flow cytometry analysis. RPCI-WM1 cells demonstrated decreased expression of CD19, CD20, and CD23 with enhanced expression of CD28, CD38 and CD184, antigens that were differentially expressed on BCWM.1 and MWCL-1 cells. Due to increased expression of CD184/CXCR4 and CD38, RPCI-WM1 represents a valuable model in which to study the effects anti-CXCR4 or anti-CD38 targeted therapies that are actively being developed for treatment of hematologic cancers. Overall, differences in surface antigen expression across the 3 cell lines may reflect the tumor clone population predominant in the index patients, from whom the cell lines were developed. Our analysis defines the utility of the most commonly employed WM cell lines as based on their immunophenotype profiles, highlighting unique differences that can be further studied for therapeutic exploit

  1. Revealing discriminating power of the elements in edible sea salts: Line-intensity correlation analysis from laser-induced plasma emission spectra

    NASA Astrophysics Data System (ADS)

    Lee, Yonghoon; Ham, Kyung-Sik; Han, Song-Hee; Yoo, Jonghyun; Jeong, Sungho

    2014-11-01

    We have investigated the discriminating power of the elements in edible sea salts using Laser-Induced Breakdown Spectroscopy (LIBS). For the ten different sea salts from South Korea, China, Japan, France, Mexico and New Zealand, LIBS spectra were recorded in the spectral range between 190 and 1040 nm, identifying the presence of Na, Cl, K, Ca, Mg, Li, Sr, Al, Si, Ti, Fe, C, O, N, and H. Intensity correlation analysis of the observed emission lines provided a valuable insight into the discriminating power of the different elements in the sea salts. The correlation analysis suggests that the elements with independent discrimination power can be categorized into three groups; those that represent dissolved ions in seawater (K, Li, and Mg), those that are associated with calcified particles (Ca and Sr), and those that are present in soils contained in the sea salts (Al, Si, Ti, and Fe). Classification models using a few emission lines selected based on the results from intensity correlation analysis and full broadband LIBS spectra were developed based on Principal Component Analysis (PCA) and Partial Least Squares-Discriminant Analysis (PLS-DA) and their performances were compared. Our results indicate that effective combination of a few emission lines can provide a dependable model for discriminating the edible sea salts and the performance is not much degraded from that based on the full broadband spectra. This can be rationalized by the intensity correlation results.

  2. Heterogeneity of functional responses in differentiated myeloid cell lines reveals EPRO cells as a valid model of murine neutrophil functional activation.

    PubMed

    Gaines, Peter; Chi, Jeffrey; Berliner, Nancy

    2005-05-01

    Mature neutrophils display multiple functional responses upon activation that include chemotaxis, adhesion to and transmigration across endothelial cells, phagocytosis, and pathogen destruction via potent microbicidal enzymes and reactive oxygen species. We are using myeloid cell line models to investigate the signaling pathways that govern neutrophil functional activation. To facilitate these studies, we have performed a direct comparison of functional responses of human and murine myeloid cell line models upon neutrophil differentiation. Our results show that EPRO cells, promyelocytes that undergo complete neutrophil maturation, demonstrate a full spectrum of functional responses, including respiratory burst, chemotaxis toward two murine chemokines, and phagocytosis. We also extend previous studies of granulocyte-colony stimulating factor-induced 32Dcl3 cells, showing they demonstrate chemotaxis and phogocytosis but completely lack a respiratory burst as a result of the absent expression of a critical oxidase subunit, gp91(phox). Induced human leukemic NB4 and HL-60 cells display a respiratory burst and phagocytosis but have defective chemotaxis to multiple chemoattractants. We also tested each cell line for the ability to up-regulate cell-surface membrane-activated complex-1 (Mac-1) expression upon activation, a response mediating neutrophil adhesion and a surrogate marker for degranulation. We show that EPRO cells, but not 32Dcl3 or NB4, significantly increase Mac-1 surface expression upon functional activation. Together, these data show that EPRO and MPRO cells demonstrate complete, functional activation upon neutrophil differentiation, suggesting these promyelocytic models accurately reflect the functional capacity of mature murine neutrophils. PMID:15673544

  3. MicroRNA profiling of novel African American and Caucasian Prostate Cancer cell lines reveals a reciprocal regulatory relationship of miR-152 and DNA methyltranferase 1

    PubMed Central

    Theodore, Shaniece C.; Davis, Melissa; Zhao, Fu; Wang, Honghe; Chen, Dongquan; Rhim, Johng; Dean-Colomb, Windy; Turner, Timothy; Ji, Weidong; Zeng, Guohua; Grizzle, William; Yates, Clayton

    2014-01-01

    miRNA expression in African American compared to Caucasian PCa patients has not been widely explored. Herein, we probed the miRNA expression profile of novel AA and CA derived prostate cancer cell lines. We found a unique miRNA signature associated with AA cell lines, independent of tumor status. Evaluation of the most differentially expressed miRNAs showed that miR-132, miR-367b, miR-410, and miR-152 were decreased in more aggressive cells, and this was reversed after treatment of the cells with 5-aza-2′-deoxycytidine. Sequencing of the miR-152 promoter confirmed that it was highly methylated. Ectopic expression of miR-152 resulted in decreased growth, migration, and invasion. Informatics analysis of a large patient cohort showed that decreased miR-152 expression correlated with increased metastasis and a decrease in biochemical recurrence free survival. Analysis of 39 prostate cancer tissues with matched controls (20 AA and 19 CA), showed that 50% of AA patients had statistically significant lower miR-152 expression compared to only 35% of CA patients. Ectopic expression of miR-152 in LNCaP, PC-3, and MDA-PCa-2b cells down-regulated DNA (cytosine-5)-methyltransferase 1 (DNMT1) through direct binding in the DNMT1 3'UTR. There appeared to be a reciprocal regulatory relationship of miR-152/DNMT1 expression, as cells treated with siRNA DNMT1 caused miR-152 to be re-expressed in all cell lines. In summary, these results demonstrate that epigenetic regulation of miR-152/DNMT1 may play an important role in multiple events that contribute to the aggressiveness of PCa tumors, with an emphasis on AA PCa patients. PMID:25004396

  4. The mid-infrared emission of narrow-line active galactic nuclei: Star formation, nuclear activity, and two populations revealed by WISE

    SciTech Connect

    Rosario, David J.; Burtscher, Leonard; Davies, Richard; Genzel, Reinhard; Lutz, Dieter; Tacconi, Linda J.

    2013-12-01

    We explore the nature of the long-wavelength mid-infrared (MIR) emission of a sample of 13,000 local Type II (narrow-line) active galactic nuclei (AGNs) from the Sloan Digital Sky Survey (SDSS) using 12 μm and 22 μm photometry from the WISE all-sky survey. In combination with FIRST 1.4 GHz photometry, we show that AGNs divide into two relatively distinct populations or 'branches' in the plane of MIR and radio luminosity. Seyfert galaxies lie almost exclusively on an MIR-bright branch (Branch A), while low-ionization nuclear emission line galaxies (LINERs) are split evenly into Branch A and the MIR-faint Branch B. We devise various tests to constrain the processes that define the branches, including a comparison to the properties of pure star-forming inactive galaxies on the MIR-radio plane. We demonstrate that the total MIR emission of objects on Branch A, including most Seyfert galaxies, is governed primarily by host star formation, with ≈15% of the 22 μm luminosity coming from AGN-heated dust. This implies that ongoing dusty star formation is a general property of Seyfert host galaxies. We show that the 12 μm broadband luminosity of AGNs on Branch A is suppressed with respect to star-forming galaxies, possibly due to the destruction of PAHs or deeper 10 μm Si absorption in AGNs. We uncover a correlation between the MIR luminosity and [O III] λ5007 luminosity in AGNs. This suggests a relationship between the star formation rate and nuclear luminosity in the AGN population, but we caution on the importance of selection effects inherent to such AGN-dominated emission-line galaxies in driving such a correlation. We highlight the MIR-radio plane as a useful tool in comparative studies of star formation and nuclear activity in AGNs.

  5. Whole genome and normalized mRNA sequencing reveal genetic status of TK6, WTK1, and NH32 human B-lymphoblastoid cell lines.

    PubMed

    Revollo, Javier; Petibone, Dayton M; McKinzie, Page; Knox, Bridgett; Morris, Suzanne M; Ning, Baitang; Dobrovolsky, Vasily N

    2016-01-01

    Closely related TK6, WTK1, and NH32 human B-lymphoblastoid cell lines differ in their p53 functional status. These lines are used frequently in genotoxicity studies and in studies aimed at understanding the role of p53 in DNA repair. Despite their routine use, little is known about the genetic status of these cells. To provide insight into their genetic composition, we sequenced and analyzed the entire genome of TK6 cells, as well as the normalized transcriptomes of TK6, WTK1, and NH32 cells. Whole genome sequencing (WGS) identified 21,561 genes and 5.17×10(6) small variants. Within the small variants, 50.54% were naturally occurring single nucleotide polymorphisms (SNPs) and 49.46% were mutations. The mutations were comprised of 92.97% single base-pair substitutions and 7.03% insertions or deletions (indels). The number of predicted genes, SNPs, and small mutations are similar to frequencies observed in the human population in general. Normalized mRNA-seq analysis identified the expression of transcripts bearing SNPs or mutations for TK6, WTK1, and NH32 as 2.88%, 2.04%, and 1.71%, respectively, and several of the variant transcripts identified appear to have important implications in genetic toxicology. These include a single base deletion mutation in the ferritin heavy chain gene (FTH1) resulting in a frame shift and protein truncation in TK6 that impairs iron metabolism. SNPs in the thiopurine S-methyltransferase (TPMT) gene (TPMT*3A SNP), and in the xenobiotic metabolizing enzyme, NADPH quinine oxidoreductase 1 (NQO1) gene (NQO1*2 SNP), are both associated with decreased enzyme activity. The clinically relevant TPMT*3A and NQO1*2 SNPs can make these cell lines useful in pharmacogenetic studies aimed at improving or tailoring drug treatment regimens that minimize toxicity and enhance efficacy. PMID:26774668

  6. Proteomic Analysis Reveals the Molecular Underpinnings of Mandibular Gland Development and Lipid Metabolism in Two Lines of Honeybees (Apis mellifera ligustica).

    PubMed

    Huo, Xinmei; Wu, Bin; Feng, Mao; Han, Bin; Fang, Yu; Hao, Yue; Meng, Lifeng; Wubie, Abebe Jenberie; Fan, Pei; Hu, Han; Qi, Yuping; Li, Jianke

    2016-09-01

    The mandibular glands (MGs) of honeybee workers are vital for the secretion of lipids, for both larval nutrition and pheromones. However, knowledge of how the proteome controls MG development and functionality at the different physiological stages of worker bees is still lacking. We characterized and compared the proteome across different ages of MGs in Italian bees (ITBs) and Royal Jelly (RJ) bees (RJBs), the latter being a line bred for increasing RJ yield, originating from the ITB. All 2000 proteins that were shared by differently aged MGs in both bee lines (>4000 proteins identified in all) were strongly enriched in metabolizing protein, nucleic acid, small molecule, and lipid functional groups. The fact that these shared proteins are enriched in similar groups in both lines suggests that they are essential for basic cellular maintenance and MG functions. However, great differences were found when comparing the proteome across different MG phases in each line. In newly emerged bees (NEBs), the unique and highly abundant proteins were enriched in protein synthesis, cytoskeleton, and development related functional groups, suggesting their importance to initialize young MG development. In nurse bees (NBs), specific and highly abundant proteins were mainly enriched in substance transport and lipid synthesis, indicating their priority may be in priming high secretory activity in lipid synthesis as larval nutrition. The unique and highly abundant proteins in forager bees (FBs) were enriched in lipid metabolism, small molecule, and carbohydrate metabolism. This indicates their emphasis on 2-heptanone synthesis as an alarm pheromone to enhance colony defense or scent marker for foraging efficiency. Furthermore, a wide range of different biological processes was observed between ITBs and RJBs at different MG ages. Both bee stocks may adapt different proteome programs to drive gland development and functionality. The RJB nurse bee has reshaped its proteome by enhancing

  7. Phenotypic, genomic and functional characterization reveals no differences between CD138++ and CD138low subpopulations in multiple myeloma cell lines.

    PubMed

    Paíno, Teresa; Sarasquete, María E; Paiva, Bruno; Krzeminski, Patryk; San-Segundo, Laura; Corchete, Luis A; Redondo, Alba; Garayoa, Mercedes; García-Sanz, Ramón; Gutiérrez, Norma C; Ocio, Enrique M; San-Miguel, Jesús F

    2014-01-01

    Despite recent advances in the treatment of multiple myeloma (MM), it remains an incurable disease potentially due to the presence of resistant myeloma cancer stem cells (MM-CSC). Although the presence of clonogenic cells in MM was described three decades ago, the phenotype of MM-CSC is still controversial, especially with respect to the expression of syndecan-1 (CD138). Here, we demonstrate the presence of two subpopulations--CD138++ (95-99%) and CD138low (1-5%)--in eight MM cell lines. To find out possible stem-cell-like features, we have phenotypically, genomic and functionally characterized the two subpopulations. Our results show that the minor CD138low subpopulation is morphologically identical to the CD138++ fraction and does not represent a more immature B-cell compartment (with lack of CD19, CD20 and CD27 expression). Moreover, both subpopulations have similar gene expression and genomic profiles. Importantly, both CD138++ and CD138low subpopulations have similar sensitivity to bortezomib, melphalan and doxorubicin. Finally, serial engraftment in CB17-SCID mice shows that CD138++ as well as CD138low cells have self-renewal potential and they are phenotypically interconvertible. Overall, our results differ from previously published data in MM cell lines which attribute a B-cell phenotype to MM-CSC. Future characterization of clonal plasma cell subpopulations in MM patients' samples will guarantee the discovery of more reliable markers able to discriminate true clonogenic myeloma cells. PMID:24658332

  8. Likelihood and Bayesian analyses reveal major genes affecting body composition, carcass, meat quality and the number of false teats in a Chinese European pig line

    PubMed Central

    Marie-Pierre, Sanchez; Bidanel, Jean-Pierre; Zhang, Siqing; Naveau, Jean; Burlot, Thierry; Le Roy, Pascale

    2003-01-01

    Segregation analyses were performed using both maximum likelihood – via a Quasi Newton algorithm – (ML-QN) and Bayesian – via Gibbs sampling – (Bayesian-GS) approaches in the Chinese European Tiameslan pig line. Major genes were searched for average ultrasonic backfat thickness (ABT), carcass fat (X2 and X4) and lean (X5) depths, days from 20 to 100 kg (D20100), Napole technological yield (NTY), number of false (FTN) and good (GTN) teats, as well as total teat number (TTN). The discrete nature of FTN was additionally considered using a threshold model under ML methodology. The results obtained with both methods consistently suggested the presence of major genes affecting ABT, X2, NTY, GTN and FTN. Major genes were also suggested for X4 and X5 using ML-QN, but not the Bayesian-GS, approach. The major gene affecting FTN was confirmed using the threshold model. Genetic correlations as well as gene effect and genotype frequency estimates suggested the presence of four different major genes. The first gene would affect fatness traits (ABT, X2 and X4), the second one a leanness trait (X5), the third one NTY and the last one GTN and FTN. Genotype frequencies of breeding animals and their evolution over time were consistent with the selection performed in the Tiameslan line. PMID:12927073

  9. Generation of Pet1210-Cre Transgenic Mouse Line Reveals Non-Serotonergic Expression Domains of Pet1 Both in CNS and Periphery

    PubMed Central

    Pelosi, Barbara; Migliarini, Sara; Pacini, Giulia; Pratelli, Marta; Pasqualetti, Massimo

    2014-01-01

    Neurons producing serotonin (5-hydroxytryptamine, 5-HT) constitute one of the most widely distributed neuronal networks in the mammalian central nervous system (CNS) and exhibit a profuse innervation throughout the CNS already at early stages of development. Serotonergic neuron specification is controlled by a combination of secreted molecules and transcription factors such as Shh, Fgf4/8, Nkx2.2, Lmx1b and Pet1. In the mouse, Pet1 mRNA expression appears between 10 and 11 days post coitum (dpc) in serotonergic post-mitotic precursors and persists in serotonergic neurons up to adulthood, where it promotes the expression of genes defining the mature serotonergic phenotype such as tryptophan hydroxylase 2 (Tph2) and serotonin transporter (SERT). Hence, the generation of genetic tools based on Pet1 specific expression represents a valuable approach to study the development and function of the serotonergic system. Here, we report the generation of a Pet1210-Cre transgenic mouse line in which the Cre recombinase is expressed under the control of a 210 kb fragment from the Pet1 genetic locus to ensure a reliable and faithful control of somatic recombination in Pet1 cell lineage. Besides Cre-mediated recombination accurately occurred in the serotonergic system as expected and according to previous studies, Pet1210-Cre transgenic mouse line allowed us to identify novel, so far uncharacterized, Pet1 expression domains. Indeed, we showed that in the raphe Pet1 is expressed also in a non-serotonergic neuronal population intermingled with Tph2-expressing cells and mostly localized in the B8 and B9 nuclei. Moreover, we detected Cre-mediated recombination also in the developing pancreas and in the ureteric bud derivatives of the kidney, where it reflected a specific Pet1 expression. Thus, Pet1210-Cre transgenic mouse line faithfully drives Cre-mediated recombination in all Pet1 expression domains representing a valuable tool to genetically manipulate serotonergic and non

  10. NuSTAR reveals an intrinsically X-ray weak broad absorption line quasar in the ultraluminous infrared galaxy Markarian 231

    SciTech Connect

    Teng, Stacy H.; Rigby, J. R.; Brandt, W. N.; Luo, B.; Harrison, F. A.; Grefenstette, B. W.; Madsen, K. K.; Alexander, D. M.; Gandhi, P.; Bauer, F. E.; Boggs, S. E.; Craig, W. W.; Christensen, F. E.; Comastri, A.; Fabian, A. C.; Farrah, D.; Fiore, F.; Hailey, C. J.; Hickox, R. C.; Ptak, A. F.; and others

    2014-04-10

    We present high-energy (3-30 keV) NuSTAR observations of the nearest quasar, the ultraluminous infrared galaxy (ULIRG) Markarian 231 (Mrk 231), supplemented with new and simultaneous low-energy (0.5-8 keV) data from Chandra. The source was detected, though at much fainter levels than previously reported, likely due to contamination in the large apertures of previous non-focusing hard X-ray telescopes. The full band (0.5-30 keV) X-ray spectrum suggests the active galactic nucleus (AGN) in Mrk 231 is absorbed by a patchy and Compton-thin (N{sub H}∼1.2{sub −0.3}{sup +0.3}×10{sup 23} cm{sup –2}) column. The intrinsic X-ray luminosity (L {sub 0.5–30} {sub keV} ∼ 1.0 × 10{sup 43} erg s{sup –1}) is extremely weak relative to the bolometric luminosity where the 2-10 keV to bolometric luminosity ratio is ∼0.03% compared to the typical values of 2%-15%. Additionally, Mrk 231 has a low X-ray-to-optical power law slope (α{sub OX} ∼ –1.7). It is a local example of a low-ionization broad absorption line quasar that is intrinsically X-ray weak. The weak ionizing continuum may explain the lack of mid-infrared [O IV], [Ne V], and [Ne VI] fine-structure emission lines which are present in sources with otherwise similar AGN properties. We argue that the intrinsic X-ray weakness may be a result of the super-Eddington accretion occurring in the nucleus of this ULIRG, and may also be naturally related to the powerful wind event seen in Mrk 231, a merger remnant escaping from its dusty cocoon.

  11. A coronagraphic absorbing cloud reveals the narrow-line region and extended Lyman α emission of QSO J0823+0529

    NASA Astrophysics Data System (ADS)

    Fathivavsari, H.; Petitjean, P.; Noterdaeme, P.; Pâris, I.; Finley, H.; López, S.; Srianand, R.; Sánchez, P.

    2015-11-01

    We report long-slit spectroscopic observations of the quasar SDSS J082303.22+052907.6 (z_{C IV}} ˜ 3.1875), whose broad-line region (BLR) is partly eclipsed by a strong damped Lyman α (DLA; logN(H I) = 21.7) cloud. This allows us to study the narrow-line region (NLR) of the quasar and the Lyman α emission from the host galaxy. Using CLOUDY models that explain the presence of strong N V and P V absorption together with the detection of Si II* and O I** absorption in the DLA, we show that the density and the distance of the cloud to the quasar are in the ranges 180 < nH < 710 cm-3 and 580 > r0 > 230 pc, respectively. Sizes of the neutral (˜2-9pc) and highly ionized phases (˜3-80pc) are consistent with the partial coverage of the C IV BLR by the C IV absorption from the DLA (covering factor of ˜0.85). We show that the residuals are consistent with emission from the NLR with C IV/Lyman α ratios varying from 0 to 0.29 through the profile. Remarkably, we detect extended Lyman α emission up to 25 kpc to the north and west directions and 15 kpc to the south and east. We interpret the emission as the superposition of strong emission in the plane of the galaxy up to 10 kpc with emission in a wind of projected velocity ˜500 km s-1 which is seen up to 25 kpc. The low metallicity of the DLA (0.27 solar) argues for at least part of this gas being infalling towards the active galactic nucleus and possibly being located where accretion from cold streams ends up.

  12. A new method for detection of tumor driver-dependent changes of protein sialylation in a colon cancer cell line reveals nectin-3 as TGFBR2 target.

    PubMed

    Lee, Jennifer; Warnken, Uwe; Schnölzer, Martina; Gebert, Johannes; Kopitz, Jürgen

    2015-10-01

    Protein-linked glycans play key roles in cell differentiation, cell-cell interactions, cell growth, adhesion and immune response. Aberrant glycosylation is a characteristic feature of tumor cells and is involved in tumor growth, escape from apoptosis, metastasis formation, and resistance to therapy. It can serve as cancer biomarker and treatment target. To enable comprehensive screening for the impact of tumor driving mutations in colorectal cancer cells we present a method for specific analysis of tumor driver-induced glycome changes. The strategy is based on a combination of three technologies, that is recombinase-mediated cassette exchange (RMCE), Click-It chemistry and mass spectrometry. The new method is exemplified by the analysis of the impact of inactivating mutations of the TGF-ß-receptor type II (TGFBR2) on sialic acid incorporation into protein-linked glycans of the colon cancer cell line HCT116. Overall, 70 proteins were found to show de novo sialic acid incorporation exclusively upon TGFBR2 expression whereas 7 proteins lost sialylation upon TGFBR2 reconstitution. Validation of detected candidate glycoproteins is demonstrated with the cell surface glycoprotein nectin-3 known to be involved in metastasis, invasion and prognosis of various cancers. Altogether, our new approach can help to systematically puzzle out the influence of tumor-specific mutations in a major signaling pathway, as exemplified by the TGFBR2 tumor suppressor, on the tumor glycome. It facilitates the identification of glycan-based tumor markers that could be used for diagnostic and therapeutic applications. In principle the outlined strategy can be adapted to any cancer cell line, tumor driver mutation and several glycan-building blocks. PMID:26177744

  13. In Situ Observations of Line-of-Sight Extinction Reveals the Depth of the Planetary Boundary Layer within Gale Crater, Mars

    NASA Astrophysics Data System (ADS)

    Moore, Casey A.; Moores, John E.; Lemmon, Mark T.; Van Beek, Jason

    2015-11-01

    Imagers onboard the Mars Science Laboratory have been used to derive line-of-sight extinction coefficients within Gale Crater, Mars. The images in use have similar pointings and look at roughly the same section of the north crater rim some 30 km away. The observations are taken, on average, once every seven sols and have spanned more than an entire martian year. Another longstanding and more frequent observation is one in which the total column optical depth above Gale Crater can be derived. When comparing the two datasets, one can conclude that the air within the northern part of the crater supports less dust per volume than the bulk atmosphere above the crater. As such, little to no mixing has been observed between the two air masses. More recently, line-of-sight extinction variations with height have allowed the depth of the planetary boundary layer (PBL) to be examined by comparing morning and noon-time imaging. The top of the PBL can be seen as a change in the slope of the extinction. Below this change of slope, we observe that the PBL is well mixed with near constant extinction. The reduced extinction within the PBL may indicate that, in contrast to other Mars landing sites where mixing is implicated in dust lifting, that the relatively mild dynamics of mixing at Gale contribute to clearing of the PBL by enhancing the delivery of dust to the surface. Above the PBL, extinction increases rapidly. The PBL is typically seen to be shallower than the height of the crater rim, some 2 km above the crater floor. Not only was a suppressed PBL within Gale Crater predicted before landing, the fact that extinction is seen to increase above the PBL provides more evidence that the air mass above the crater initially supports more dust on average than the air lower down in the crater.

  14. Metabolic Profiling Reveals Sphingosine-1-Phosphate Kinase 2 and Lyase as Key Targets of (Phyto-) Estrogen Action in the Breast Cancer Cell Line MCF-7 and Not in MCF-12A

    PubMed Central

    Engel, Nadja; Lisec, Jan; Piechulla, Birgit; Nebe, Barbara

    2012-01-01

    To search for new targets of anticancer therapies using phytoestrogens we performed comparative metabolic profiling of the breast cancer cell line MCF-7 and the non-tumorigenic breast cell line MCF-12A. Application of gas chromatography-mass spectrometry (GC-MS) revealed significant differences in the metabolic levels after exposure with 17ß-estradiol, genistein or a composition of phytoestrogens within a native root flax extract. We observed the metabolites 3-(4-hydroxyphenyl)-lactic acid, cis-aconitic acid, 11-beta-hydroxy-progesterone, chenodeoxycholic acid and triacontanoic acid with elevated levels due to estrogen action. Particularly highlighted were metabolites of the sphingolipid metabolism. Sphingosine and its dihydro derivate as well as ethanolaminephosphate were significantly altered after exposure with 1 nM 17ß-estradiol in the cell line MCF-7, while MCF-12A was not affected. Treatment with genistein and the flax extract normalized the sphingosine concentrations to the basic levels found in MCF-12A cells. We could further demonstrate that the expression levels of the sphingosine metabolizing enzymes: sphingosine-1-phosphate kinase (Sphk) and lyase (S1P lyase) were significantly influenced by estrogens as well as phytoestrogens. The isoform Sphk2 was overexpressed in the tumorigenic cell line MCF-7, while S1P lyase was predominantly expressed in the non-tumorigenic cell line MCF-12A. Importantly, in MCF-7 the weak S1P lyase expression could be significantly increased after exposure with 10 µM genistein and 1 µg/ml root flax extract. Here, we present, for the first time, an analysis of metabolic response of phytoestrogens to breast cancer cell lines. The contrasting regulation of sphingolipid enzymes in MCF-7 and MCF-12A render them as preferred targets for future anticancer strategies. PMID:23112854

  15. Emergent Self-Organized Criticality in Gene Expression Dynamics: Temporal Development of Global Phase Transition Revealed in a Cancer Cell Line

    PubMed Central

    Tsuchiya, Masa; Giuliani, Alessandro; Hashimoto, Midori; Erenpreisa, Jekaterina; Yoshikawa, Kenichi

    2015-01-01

    Background The underlying mechanism of dynamic control of the genome-wide expression is a fundamental issue in bioscience. We addressed it in terms of phase transition by a systemic approach based on both density analysis and characteristics of temporal fluctuation for the time-course mRNA expression in differentiating MCF-7 breast cancer cells. Methodology In a recent work, we suggested criticality as an essential aspect of dynamic control of genome-wide gene expression. Criticality was evident by a unimodal-bimodal transition through flattened unimodal expression profile. The flatness on the transition suggests the existence of a critical transition at which up- and down-regulated expression is balanced. Mean field (averaging) behavior of mRNAs based on the temporal expression changes reveals a sandpile type of transition in the flattened profile. Furthermore, around the transition, a self-similar unimodal-bimodal transition of the whole expression occurs in the density profile of an ensemble of mRNA expression. These singular and scaling behaviors identify the transition as the expression phase transition driven by self-organized criticality (SOC). Principal Findings Emergent properties of SOC through a mean field approach are revealed: i) SOC, as a form of genomic phase transition, consolidates distinct critical states of expression, ii) Coupling of coherent stochastic oscillations between critical states on different time-scales gives rise to SOC, and iii) Specific gene clusters (barcode genes) ranging in size from kbp to Mbp reveal similar SOC to genome-wide mRNA expression and ON-OFF synchronization to critical states. This suggests that the cooperative gene regulation of topological genome sub-units is mediated by the coherent phase transitions of megadomain-scaled conformations between compact and swollen chromatin states. Conclusion and Significance In summary, our study provides not only a systemic method to demonstrate SOC in whole-genome expression

  16. Examining Two Sets of Introgression Lines in Rice (Oryza sativa L.) Reveals Favorable Alleles that Improve Grain Zn and Fe Concentrations

    PubMed Central

    Hu, Xia; Cheng, Li-Rui; Xu, Jian-Long; Shi, Yu-Min; Li, Zhi-Kang

    2015-01-01

    In the modern world, the grain mineral concentration (GMC) in rice (Oryza sativa L.) not only includes important micronutrient elements such as iron (Fe) and zinc (Zn), but it also includes toxic heavy metal elements, especially cadmium (Cd) and lead (Pb). To date, the genetic mechanisms underlying the regulation of GMC, especially the genetic background and G × E effects of GMC, remain largely unknown. In this study, we adopted two sets of backcross introgression lines (BILs) derived from IR75862 (a Zn-dense rice variety) as the donor parent and two elite indica varieties, Ce258 and Zhongguangxiang1, as recurrent parents to detect QTL affecting GMC traits including Fe, Zn, Cd and Pb concentrations in two environments. We detected a total of 22 loci responsible for GMC traits, which are distributed on all 12 rice chromosomes except 5, 9 and 10. Six genetic overlap (GO) regions affecting multiple elements were found, in which most donor alleles had synergistic effects on GMC. Some toxic heavy metal-independent loci (such as qFe1, qFe2 and qZn12) and some regions that have opposite genetic effects on micronutrient (Fe and Zn) and heavy metal element (Pb) concentrations (such as GO-IV) may be useful for marker-assisted biofortification breeding in rice. We discuss three important points affecting biofortification breeding efforts in rice, including correlations between different GMC traits, the genetic background effect and the G × E effect. PMID:26161553

  17. Molecular analysis of genetic mutations among cross-resistant second-line injectable drugs reveals a new resistant mutation in Mycobacterium tuberculosis.

    PubMed

    Malinga, Lesibana; Brand, Jeannette; Olorunju, Steve; Stoltz, Anton; van der Walt, Martie

    2016-08-01

    Mutations causing mono and cross-resistance among amikacin, kanamycin and capreomycin of second-line injectable drugs (SLIDs) namely are not well understood. We investigated 124 isolates of Mycobacterium tuberculosis for mutations within rrs, eis, tlyA and efflux pump (Rv1258c and Rv0194) genes involved in resistance towards SLIDs. The distribution of mutations across these genes were significantly different in strains with mono-resistance or cross-resistance. A new mutation G878A was found in rrs gene, among strains with capreomycin mono-resistant, or in strains with cross-resistance of capreomycin, kanamycin and amikacin. This mutation was associated with the Euro-American X3 lineage (P < 0.0001). Mutations in the two efflux genes Rv1258c and Rv0194 were confined to strains with only capreomycin/amikacin/kanamycin cross-resistance. We further investigated the minimum inhibitory concentration of capreomycin on isolates with new G878A mutation ranging from 8 μg/mL to 64 μg/mL. Inclusion of G878A on new molecular assays could increase the sensitivity of capreomycin resistance detection. PMID:27298046

  18. Deep geometry and evolution of the northern part of Itoigwa-Shizuoka Tectonic Line active fault system, Central Japan, revealed by Seismic profiling

    NASA Astrophysics Data System (ADS)

    Sato, H.; Ikeda, Y.; Iwasaki, T.; Matsuta, N.; Takeda, T.; Kawasaki, S.; Kozawa, T.; Elouai, D.; Hirata, N.; Kawanaka, T.

    2003-12-01

    The northern Fossa Magna (NFM) is a Miocene rift system produced in the final stages of the opening of the Sea of Japan. It divides the major structure of Japan into SW and NE portions. The Itoigawa-Shizuoka Tectonic Line (ISTL) bounds the western part of the northern Fossa Magna and forms an active fault system showing the one of the largest slip rates in the Japanese islands. Based on the paleo-seismological data, the ISTL active fault system was evaluated to have the highest seismic risk among active faults within inland Japan. A quantitative understanding of active tectonic processes, including crustal deformation and related destructive earthquakes, is important in reducing seismic hazards through precise estimation of strong ground motions. The structure of the crust, especially the deep geometry of active fault systems, is the most important piece information required to construct such a dynamic model. In this context, the seismic reflection profiling was performed across the northern part of the ISTL active fault system by three seismic lines. Obtained seismic sections are interpreted based on the pattern of reflectors, surface geology and velocity model by refraction analysis, using the balanced cross section technique. The 68-km-long Itoshizu 2002 seismic section across the northern middle part of the ISTL active fault system suggest that the Miocene NFM basin was formed by an east dipping normal fault with shallow flat (6 km), deeper ramp (6 15 km) and deeper flat at 15 km in depth. This unique geometry is interpreted that this low-angle normal fault was produced by Miocene high thermal regime, estimated from the thick volcanic rocks at the base of the basin fill. Namely, the normal fault reflects the brittle-ductile boundary in Miocene. Consequently, since the Pliocene, the basin fill was strongly folded by the reverse faulting along the pre-existing normal faults in the Pre-Neogene rocks. The reverse faults in the basin fill produced fault

  19. Differential increase in taurine levels by low-dose ethanol in the dorsal and ventral striatum revealed by microdialysis with on-line capillary electrophoresis.

    PubMed

    Smith, A; Watson, C J; Frantz, K J; Eppler, B; Kennedy, R T; Peris, J

    2004-07-01

    Ethanol increases taurine efflux in the nucleus accumbens or ventral striatum (VS), a dopaminergic terminal region involved in positive reinforcement. However, this has been found only at ethanol doses above 1 g/kg intraperitoneally, which is higher than what most rats will self-administer. We used a sensitive on-line assay of microdialysate content to test whether lower doses of ethanol selectively increase taurine efflux in VS as opposed to other dopaminergic regions not involved in reinforcement (e.g., dorsal striatum; DS). Adult male rats with microdialysis probes in VS or DS were injected with ethanol (0, 0.5, 1, and 2 g/kg intraperitoneally), and the amino acid content of the dialysate was measured every 11 sec using capillary electrophoresis and laser-induced fluorescence detection. In VS, 0.5 g/kg ethanol significantly increased taurine levels by 20% for 10 min. A similar increase was seen after 1 g/kg ethanol, which lasted for about 20 min after injection. A two-phased taurine efflux was observed with the 2.0 g/kg dose, where taurine was increased by 2-fold after 5 min but it remained elevated by 30% for at least 60 min. In contrast, DS exhibited much smaller dose-related increases in taurine. Glycine, glutamate, serine, and gamma-aminobutyric acid were not systematically affected by lower doses of ethanol; however, 2 g/kg slowly decreased these amino acids in both brain regions during the hour after injection. These data implicate a possible role of taurine in the mechanism of action of ethanol in the VS. The high sensitivity and time resolution afforded by capillary electrophoresis and laser-induced fluorescence detection will be useful for detecting subtle changes of neuronally active amino acids levels due to low doses of ethanol. PMID:15252289

  20. Seed colour loci, homoeology and linkage groups of the C genome chromosomes revealed in Brassica rapa–B. oleracea monosomic alien addition lines

    PubMed Central

    Heneen, Waheeb K.; Geleta, Mulatu; Brismar, Kerstin; Xiong, Zhiyong; Pires, J. Chris; Hasterok, Robert; Stoute, Andrew I.; Scott, Roderick J.; King, Graham J.; Kurup, Smita

    2012-01-01

    Background and Aims Brassica rapa and B. oleracea are the progenitors of oilseed rape B. napus. The addition of each chromosome of B. oleracea to the chromosome complement of B. rapa results in a series of monosomic alien addition lines (MAALs). Analysis of MAALs determines which B. oleracea chromosomes carry genes controlling specific phenotypic traits, such as seed colour. Yellow-seeded oilseed rape is a desirable breeding goal both for food and livestock feed end-uses that relate to oil, protein and fibre contents. The aims of this study included developing a missing MAAL to complement an available series, for studies on seed colour control, chromosome homoeology and assignment of linkage groups to B. oleracea chromosomes. Methods A new batch of B. rapa–B. oleracea aneuploids was produced to generate the missing MAAL. Seed colour and other plant morphological features relevant to differentiation of MAALs were recorded. For chromosome characterization, Snow's carmine, fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) were used. Key Results The final MAAL was developed. Morphological traits that differentiated the MAALs comprised cotyledon number, leaf morphology, flower colour and seed colour. Seed colour was controlled by major genes on two B. oleracea chromosomes and minor genes on five other chromosomes of this species. Homoeologous pairing was largely between chromosomes with similar centromeric positions. FISH, GISH and a parallel microsatellite marker analysis defined the chromosomes in terms of their linkage groups. Conclusions A complete set of MAALs is now available for genetic, genomic, evolutionary and breeding perspectives. Defining chromosomes that carry specific genes, physical localization of DNA markers and access to established genetic linkage maps contribute to the integration of these approaches, manifested in the confirmed correspondence of linkage groups with specific chromosomes. Applications include marker

  1. Generation of iPSC line iPSC-FH2.1 in hypoxic conditions from human foreskin fibroblasts.

    PubMed

    Questa, María; Romorini, Leonardo; Blüguermann, Carolina; Solari, Claudia María; Neiman, Gabriel; Luzzani, Carlos; Scassa, María Élida; Sevlever, Gustavo Emilio; Guberman, Alejandra Sonia; Miriuka, Santiago Gabriel

    2016-03-01

    Human foreskin fibroblasts were used to generate the iPSC line iPSC-FH2.1 using the EF1a-hSTEMCCA-loxP vector expressing OCT4, SOX2, c-MYC and KLF4, in 5% O2 culture conditions. Stemness was confirmed, as was pluripotency both in vivo and in vitro, in normoxia and hypoxia. Human Embryonic Stem Cell (hESC) line WA-09 and reprogrammed fibroblast primary culture HFF-FM were used as controls. PMID:27345989

  2. Suberoylanilide Hydroxamic Acid Treatment Reveals Crosstalks among Proteome, Ubiquitylome and Acetylome in Non-Small Cell Lung Cancer A549 Cell Line

    PubMed Central

    Wu, Quan; Cheng, Zhongyi; Zhu, Jun; Xu, Weiqing; Peng, Xiaojun; Chen, Chuangbin; Li, Wenting; Wang, Fengsong; Cao, Lejie; Yi, Xingling; Wu, Zhiwei; Li, Jing; Fan, Pingsheng

    2015-01-01

    Suberoylanilide hydroxamic acid (SAHA) is a well-known histone deacetylase (HDAC) inhibitor and has been used as practical therapy for breast cancer and non-small cell lung cancer (NSCLC). It is previously demonstrated that SAHA treatment could extensively change the profile of acetylome and proteome in cancer cells. However, little is known about the impact of SAHA on other protein modifications and the crosstalks among different modifications and proteome, hindering the deep understanding of SAHA-mediated cancer therapy. In this work, by using SILAC technique, antibody-based affinity enrichment and high-resolution LC-MS/MS analysis, we investigated quantitative proteome, acetylome and ubiquitylome as well as crosstalks among the three datasets in A549 cells toward SAHA treatment. In total, 2968 proteins, 1099 acetylation sites and 1012 ubiquitination sites were quantified in response to SAHA treatment, respectively. With the aid of intensive bioinformatics, we revealed that the proteome and ubiquitylome were negatively related upon SAHA treatment. Moreover, the impact of SAHA on acetylome resulted in 258 up-regulated and 99 down-regulated acetylation sites at the threshold of 1.5 folds. Finally, we identified 55 common sites with both acetylation and ubiquitination, among which ubiquitination level in 43 sites (78.2%) was positive related to acetylation level. PMID:25825284

  3. Analysis of Magnaporthe oryzae Genome Reveals a Fungal Effector, Which Is Able to Induce Resistance Response in Transgenic Rice Line Containing Resistance Gene, Pi54

    PubMed Central

    Ray, Soham; Singh, Pankaj K.; Gupta, Deepak K.; Mahato, Ajay K.; Sarkar, Chiranjib; Rathour, Rajeev; Singh, Nagendra K.; Sharma, Tilak R.

    2016-01-01

    Rice blast caused by Magnaporthe oryzae is one of the most important diseases of rice. Pi54, a rice gene that imparts resistance to M. oryzae isolates prevalent in India, was already cloned but its avirulent counterpart in the pathogen was not known. After decoding the whole genome of an avirulent isolate of M. oryzae, we predicted 11440 protein coding genes and then identified four candidate effector proteins which are exclusively expressed in the infectious structure, appresoria. In silico protein modeling followed by interaction analysis between Pi54 protein model and selected four candidate effector proteins models revealed that Mo-01947_9 protein model encoded by a gene located at chromosome 4 of M. oryzae, interacted best at the Leucine Rich Repeat domain of Pi54 protein model. Yeast-two-hybrid analysis showed that Mo-01947_9 protein physically interacts with Pi54 protein. Nicotiana benthamiana leaf infiltration assay confirmed induction of hypersensitive response in the presence of Pi54 gene in a heterologous system. Genetic complementation test also proved that Mo-01947_9 protein induces avirulence response in the pathogen in presence of Pi54 gene. Here, we report identification and cloning of a new fungal effector gene which interacts with blast resistance gene Pi54 in rice. PMID:27551285

  4. Suberoylanilide hydroxamic acid treatment reveals crosstalks among proteome, ubiquitylome and acetylome in non-small cell lung cancer A549 cell line.

    PubMed

    Wu, Quan; Cheng, Zhongyi; Zhu, Jun; Xu, Weiqing; Peng, Xiaojun; Chen, Chuangbin; Li, Wenting; Wang, Fengsong; Cao, Lejie; Yi, Xingling; Wu, Zhiwei; Li, Jing; Fan, Pingsheng

    2015-01-01

    Suberoylanilide hydroxamic acid (SAHA) is a well-known histone deacetylase (HDAC) inhibitor and has been used as practical therapy for breast cancer and non-small cell lung cancer (NSCLC). It is previously demonstrated that SAHA treatment could extensively change the profile of acetylome and proteome in cancer cells. However, little is known about the impact of SAHA on other protein modifications and the crosstalks among different modifications and proteome, hindering the deep understanding of SAHA-mediated cancer therapy. In this work, by using SILAC technique, antibody-based affinity enrichment and high-resolution LC-MS/MS analysis, we investigated quantitative proteome, acetylome and ubiquitylome as well as crosstalks among the three datasets in A549 cells toward SAHA treatment. In total, 2968 proteins, 1099 acetylation sites and 1012 ubiquitination sites were quantified in response to SAHA treatment, respectively. With the aid of intensive bioinformatics, we revealed that the proteome and ubiquitylome were negatively related upon SAHA treatment. Moreover, the impact of SAHA on acetylome resulted in 258 up-regulated and 99 down-regulated acetylation sites at the threshold of 1.5 folds. Finally, we identified 55 common sites with both acetylation and ubiquitination, among which ubiquitination level in 43 sites (78.2%) was positive related to acetylation level. PMID:25825284

  5. Analysis of Magnaporthe oryzae Genome Reveals a Fungal Effector, Which Is Able to Induce Resistance Response in Transgenic Rice Line Containing Resistance Gene, Pi54.

    PubMed

    Ray, Soham; Singh, Pankaj K; Gupta, Deepak K; Mahato, Ajay K; Sarkar, Chiranjib; Rathour, Rajeev; Singh, Nagendra K; Sharma, Tilak R

    2016-01-01

    Rice blast caused by Magnaporthe oryzae is one of the most important diseases of rice. Pi54, a rice gene that imparts resistance to M. oryzae isolates prevalent in India, was already cloned but its avirulent counterpart in the pathogen was not known. After decoding the whole genome of an avirulent isolate of M. oryzae, we predicted 11440 protein coding genes and then identified four candidate effector proteins which are exclusively expressed in the infectious structure, appresoria. In silico protein modeling followed by interaction analysis between Pi54 protein model and selected four candidate effector proteins models revealed that Mo-01947_9 protein model encoded by a gene located at chromosome 4 of M. oryzae, interacted best at the Leucine Rich Repeat domain of Pi54 protein model. Yeast-two-hybrid analysis showed that Mo-01947_9 protein physically interacts with Pi54 protein. Nicotiana benthamiana leaf infiltration assay confirmed induction of hypersensitive response in the presence of Pi54 gene in a heterologous system. Genetic complementation test also proved that Mo-01947_9 protein induces avirulence response in the pathogen in presence of Pi54 gene. Here, we report identification and cloning of a new fungal effector gene which interacts with blast resistance gene Pi54 in rice. PMID:27551285

  6. Rapid micropatterning of cell lines and human pluripotent stem cells on elastomeric membranes.

    PubMed

    Paik, Isha; Scurr, David J; Morris, Bryan; Hall, Graham; Denning, Chris; Alexander, Morgan R; Shakesheff, Kevin M; Dixon, James E

    2012-10-01

    Tissue function during development and in regenerative medicine completely relies on correct cell organization and patterning at micro and macro scales. We describe a rapid method for patterning mammalian cells including human embryonic stem cells (HESCs) and induced pluripotent stem cells (iPSCs) on elastomeric membranes such that micron-scale control of cell position can be achieved over centimeter-length scales. Our method employs surface engineering of hydrophobic polydimethylsiloxane (PDMS) membranes by plasma polymerization of allylamine. Deposition of plasma polymerized allylamine (ppAAm) using our methods may be spatially restricted using a micro-stencil leaving faithful hydrophilic ppAAm patterns. We employed airbrushing to create aerosols which deposit extracellular matrix (ECM) proteins (such as fibronectin and Matrigel™) onto the same patterned ppAAm rich regions. Cell patterns were created with a variety of well characterized cell lines (e.g., NIH-3T3, C2C12, HL1, BJ6, HESC line HUES7, and HiPSC line IPS2). Individual and multiple cell line patterning were also achieved. Patterning remains faithful for several days and cells are viable and proliferate. To demonstrate the utility of our technique we have patterned cells in a variety of configurations. The ability to rapidly pattern cells at high resolution over macro scales should aid future tissue engineering efforts for regenerative medicine applications and in creating in vitro stem cell niches. PMID:22511037

  7. Conservative site-specific and single-copy transgenesis in human LINE-1 elements

    PubMed Central

    Vijaya Chandra, Shree Harsha; Makhija, Harshyaa; Peter, Sabrina; Myint Wai, Cho Mar; Li, Jinming; Zhu, Jindong; Ren, Zhonglu; D'Alcontres, Martina Stagno; Siau, Jia Wei; Chee, Sharon; Ghadessy, Farid John; Dröge, Peter

    2016-01-01

    Genome engineering of human cells plays an important role in biotechnology and molecular medicine. In particular, insertions of functional multi-transgene cassettes into suitable endogenous sequences will lead to novel applications. Although several tools have been exploited in this context, safety issues such as cytotoxicity, insertional mutagenesis and off-target cleavage together with limitations in cargo size/expression often compromise utility. Phage λ integrase (Int) is a transgenesis tool that mediates conservative site-specific integration of 48 kb DNA into a safe harbor site of the bacterial genome. Here, we show that an Int variant precisely recombines large episomes into a sequence, termed attH4X, found in 1000 human Long INterspersed Elements-1 (LINE-1). We demonstrate single-copy transgenesis through attH4X-targeting in various cell lines including hESCs, with the flexibility of selecting clones according to transgene performance and downstream applications. This is exemplified with pluripotency reporter cassettes and constitutively expressed payloads that remain functional in LINE1-targeted hESCs and differentiated progenies. Furthermore, LINE-1 targeting does not induce DNA damage-response or chromosomal aberrations, and neither global nor localized endogenous gene expression is substantially affected. Hence, this simple transgene addition tool should become particularly useful for applications that require engineering of the human genome with multi-transgenes. PMID:26673710

  8. Conservative site-specific and single-copy transgenesis in human LINE-1 elements.

    PubMed

    Vijaya Chandra, Shree Harsha; Makhija, Harshyaa; Peter, Sabrina; Myint Wai, Cho Mar; Li, Jinming; Zhu, Jindong; Ren, Zhonglu; D'Alcontres, Martina Stagno; Siau, Jia Wei; Chee, Sharon; Ghadessy, Farid John; Dröge, Peter

    2016-04-01

    Genome engineering of human cells plays an important role in biotechnology and molecular medicine. In particular, insertions of functional multi-transgene cassettes into suitable endogenous sequences will lead to novel applications. Although several tools have been exploited in this context, safety issues such as cytotoxicity, insertional mutagenesis and off-target cleavage together with limitations in cargo size/expression often compromise utility. Phage λ integrase (Int) is a transgenesis tool that mediates conservative site-specific integration of 48 kb DNA into a safe harbor site of the bacterial genome. Here, we show that an Int variant precisely recombines large episomes into a sequence, termedattH4X, found in 1000 human Long INterspersed Elements-1 (LINE-1). We demonstrate single-copy transgenesis throughattH4X-targeting in various cell lines including hESCs, with the flexibility of selecting clones according to transgene performance and downstream applications. This is exemplified with pluripotency reporter cassettes and constitutively expressed payloads that remain functional inLINE1-targeted hESCs and differentiated progenies. Furthermore,LINE-1targeting does not induce DNA damage-response or chromosomal aberrations, and neither global nor localized endogenous gene expression is substantially affected. Hence, this simple transgene addition tool should become particularly useful for applications that require engineering of the human genome with multi-transgenes. PMID:26673710

  9. A common promoter hypomethylation signature in invasive breast, liver and prostate cancer cell lines reveals novel targets involved in cancer invasiveness

    PubMed Central

    Yi, Cao; Li, Chen Chen; Yu, Patricia; Arakelian, Ani; Tanvir, Imrana; Khan, Haseeb Ahmed; Rabbani, Shafaat

    2015-01-01

    Cancer invasion and metastasis is the most morbid aspect of cancer and is governed by different cellular mechanisms than those driving the deregulated growth of tumors. We addressed here the question of whether a common DNA methylation signature of invasion exists in cancer cells from different origins that differentiates invasive from non-invasive cells. We identified a common DNA methylation signature consisting of hyper- and hypomethylation and determined the overlap of differences in DNA methylation with differences in mRNA expression using expression array analyses. A pathway analysis reveals that the hypomethylation signature includes some of the major pathways that were previously implicated in cancer migration and invasion such as TGF beta and ERBB2 triggered pathways. The relevance of these hypomethylation events in human tumors was validated by identification of the signature in several publicly available databases of human tumor transcriptomes. We shortlisted novel invasion promoting candidates and tested the role of four genes in cellular invasiveness from the list C11orf68, G0S2, SHISA2 and TMEM156 in invasiveness using siRNA depletion. Importantly these genes are upregulated in human cancer specimens as determined by immunostaining of human normal and cancer breast, liver and prostate tissue arrays. Since these genes are activated in cancer they constitute a group of targets for specific pharmacological inhibitors of cancer invasiveness. SUMMARY Our study provides evidence that common DNA hypomethylation signature exists between cancer cells derived from different tissues, pointing to a common mechanism of cancer invasiveness in cancer cells from different origins that could serve as drug targets. PMID:26427334

  10. Brief report: human pluripotent stem cell models of fanconi anemia deficiency reveal an important role for fanconi anemia proteins in cellular reprogramming and survival of hematopoietic progenitors.

    PubMed

    Yung, Sun K; Tilgner, Katarzyna; Ledran, Maria H; Habibollah, Saba; Neganova, Irina; Singhapol, Chatchawan; Saretzki, Gabriele; Stojkovic, Miodrag; Armstrong, Lyle; Przyborski, Stefan; Lako, Majlinda

    2013-05-01

    Fanconi anemia (FA) is a genomic instability disorder caused by mutations in genes involved in replication-dependant-repair and removal of DNA cross-links. Mouse models with targeted deletions of FA genes have been developed; however, none of these exhibit the human bone marrow aplasia. Human embryonic stem cell (hESC) differentiation recapitulates many steps of embryonic hematopoietic development and is a useful model system to investigate the early events of hematopoietic progenitor specification. It is now possible to derive patient-specific human-induced pluripotent stem cells (hiPSC); however, this approach has been rather difficult to achieve in FA cells due to a requirement for activation of FA pathway during reprogramming process which can be bypassed either by genetic complementation or reprogramming under hypoxic conditions. In this study, we report that FA-C patient-specific hiPSC lines can be derived under normoxic conditions, albeit at much reduced efficiency. These disease-specific hiPSC lines and hESC with stable knockdown of FANCC display all the in vitro hallmarks of pluripotency. Nevertheless, the disease-specific hiPSCs show a much higher frequency of chromosomal abnormalities compared to parent fibroblasts and are unable to generate teratoma composed of all three germ layers in vivo, likely due to increased genomic instability. Both FANCC-deficient hESC and hiPSC lines are capable of undergoing hematopoietic differentiation, but the hematopoietic progenitors display an increased apoptosis in culture and reduced clonogenic potential. Together these data highlight the critical requirement for FA proteins in survival of hematopoietic progenitors, cellular reprogramming, and maintenance of genomic stability. PMID:23280624

  11. Gamma ray line astronomy

    NASA Technical Reports Server (NTRS)

    Ramaty, R.

    1984-01-01

    The interpretations and implications of the astrophysical observations of gamma-ray lines are reviewed. At the Galactic Center e(+)-e(-) pairs from a compact object produce an annihilation line that shows no redshift, indicating an annihilation site far removed from this object. In the jets of SS433, gamma-ray lines are produced by inelastic excitations, probably in dust grains, although line emission from fusion reactions has also been considered. Observations of diffuse galactic line emission reveal recently synthesized radioactive aluminum in the interstellar medium. In gamma-ray bursts, redshifted pair annihilation lines are consistent with a neutron star origin for the bursts. In solar flares, gamma-ray line emission reveals the prompt acceleration of protons and nuclei, in close association with the flare energy release mechanism.

  12. Direct comparison between genomic constitution and flavonoid contents in Allium multiple alien addition lines reveals chromosomal locations of genes related to biosynthesis from dihydrokaempferol to quercetin glucosides in scaly leaf of shallot (Allium cepa L.).

    PubMed

    Masuzaki, S; Shigyo, M; Yamauchi, N

    2006-02-01

    The extrachromosome 5A of shallot (Allium cepa L., genomes AA) has an important role in flavonoid biosynthesis in the scaly leaf of Allium fistulosum-shallot monosomic addition lines (FF+nA). This study deals with the production and biochemical characterisation of A. fistulosum-shallot multiple alien addition lines carrying at least 5A to determine the chromosomal locations of genes for quercetin formation. The multiple alien additions were selected from the crossing between allotriploid FFA (female symbol) and A. fistulosum (male symbol). The 113 plants obtained from this cross were analysed by a chromosome 5A-specific PGI isozyme marker of shallot. Thirty plants were preliminarily selected for an alien addition carrying 5A. The chromosome numbers of the 30 plants varied from 18 to 23. The other extrachromosomes in 19 plants were completely identified by using seven other chromosome markers of shallot. High-performance liquid chromatography analyses of the 19 multiple additions were conducted to identify the flavonoid compounds produced in the scaly leaves. Direct comparisons between the chromosomal constitution and the flavonoid contents of the multiple alien additions revealed that a flavonoid 3'-hydroxylase (F3'H) gene for the synthesis of quercetin from kaempferol was located on 7A and that an anonymous gene involved in the glucosidation of quercetin was on 3A or 4A. As a result of supplemental SCAR analyses by using genomic DNAs from two complete sets of A. fistulosum-shallot monosomic additions, we have assigned F3'H to 7A and flavonol synthase to 4A. PMID:16411131

  13. Derivation of NEM2 affected human embryonic stem cell line Genea080.

    PubMed

    Dumevska, Biljana; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea080 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying compound heterozygous mutations in the NEB gene, exon 55 deletion & c.15110dupA, indicative of Nemaline Myopathy Type 2 (NEM2). Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XY and STR analysis demonstrated a male allele pattern. The hESC line had pluripotent cell morphology, 90% of cells expressed Nanog, 95% Oct4, 54% Tra1-60 and 99% SSEA4 and gave a PluriTest Pluripotency score of 32.08, Novelty of 1.3. The cell line was negative for Mycoplasma and visible contamination. PMID:27346011

  14. Derivation of FSHD1 affected human embryonic stem cell line Genea049.

    PubMed

    Dumevska, Biljana; Chami, Omar; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea049 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying a deletion in 4q35 with only 5 D4Z4 repeats by PGD linkage analysis, indicative of FSHD1. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female Allele pattern. The hESC line had pluripotent cell morphology, 90% of cells expressed Nanog, 96% Oct4, 80% Tra1-60 and 99% SSEA4, gave a Pluritest Pluripotency score of 23.16, Novelty of 1.43 and demonstrated Alkaline Phosphatase activity. The cell line was negative for Mycoplasma and visible contamination. PMID:27346016

  15. Derivation of NEM2 affected human embryonic stem cell line Genea078.

    PubMed

    Dumevska, Biljana; Main, Heather; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea078 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying compound heterozygous mutations in the NEB gene, exon 55 deletion & c.15110dupA, indicative of Nemaline Myopathy Type 2 (NEM2). Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX and STR analysis demonstrated a female Allele pattern. The hESC line had pluripotent cell morphology, 76% of cells expressed Nanog, 93% Oct4, 67% Tra1-60 and 97% SSEA4 and gave a Pluritest Pluripotency score of 42.18, Novelty of 1.37. The cell line was negative for Mycoplasma and visible contamination. PMID:27346006

  16. Derivation of Huntington Disease affected Genea017 human embryonic stem cell line.

    PubMed

    Dumevska, Biljana; McKernan, Robert; Goel, Divya; Schmidt, Uli; Peura, Teija

    2016-03-01

    The Genea017 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying Htt gene CAG expansion of 40 repeats, indicative of Huntington Disease. Following ICM outgrowth on inactivated human feeders, genetic analysis confirmed a 46, XY karyotype and male allele pattern through CGH and STR analysis. The hESC line had pluripotent cell morphology, 87% of cells expressed Nanog, 95% Oct4, 88% Tra1-60 and 99% SSEA4, gave a PluriTest pluripotency score of 34.74, novelty of 1.27, demonstrated alkaline phosphatase activity and tri-lineage teratoma formation. The cell line was negative for Mycoplasma and visible contamination. PMID:27346022

  17. Derivation of FSHD1 affected human embryonic stem cell line Genea050.

    PubMed

    Dumevska, Biljana; Chami, Omar; Main, Heather; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea050 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying a deletion in 4q35 with only 5 D4Z4 repeats by PGD linkage analysis, indicative of FSHD1. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XY and STR analysis demonstrated a male Allele pattern. The hESC line had pluripotent cell morphology, 92% of cells expressed Nanog, 97% Oct4, 79% Tra1-60 and 99% SSEA4, gave a Pluritest Pluripotency score of 25.45, Novelty of 1.45 demonstrated Alkaline Phosphatase activity and tri-lineage teratoma formation. The cell line was negative for Mycoplasma and visible contamination. PMID:27346025

  18. Derivation of FSHD1 affected human embryonic stem cell line Genea096.

    PubMed

    Dumevska, Biljana; Schaft, Julia; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea096 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying a deletion in 4q35 with only 6 D4Z4 repeats by PGD linkage analysis, indicative of FSHD1. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female Allele pattern. The hESC line had pluripotent cell morphology, 64% of cells expressed Nanog, 93% Oct4, 58% Tra1-60 and 93% SSEA4 and a Pluritest Pluripotency score of 39.41, Novelty of 1.25. The cell line was negative for Mycoplasma and visible contamination. PMID:27346027

  19. Derivation of Huntington Disease affected Genea018 human embryonic stem cell line.

    PubMed

    Dumevska, Biljana; Main, Heather; McKernan, Robert; Goel, Divya; Schmidt, Uli; Peura, Teija

    2016-03-01

    The Genea018 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying Htt gene CAG expansion of 46 repeats, indicative of Huntington Disease. Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XX by CGH and STR analysis demonstrated a female Allele pattern. The hESC line had pluripotent cell morphology, 75% of cells expressed Nanog, 91% Oct4, 73% Tra1-60 and 96% SSEA4, gave a Pluritest pluripotency score of 31.12, Novelty of 1.45, demonstrated Alkaline Phosphatase activity and tri-lineage teratoma formation. The cell line was negative for Mycoplasma and visible contamination. PMID:27346005

  20. Derivation of NEM2 affected human embryonic stem cell line Genea079.

    PubMed

    Dumevska, Biljana; McKernan, Robert; Goel, Divya; Schmidt, Uli

    2016-03-01

    The Genea079 human embryonic stem cell line was derived from a donated, fully commercially consented ART blastocyst, carrying compound heterozygous mutations in the NEB gene, exon 55 deletion & c.15110dupA, indicative of Nemaline Myopathy Type 2 (NEM2). Following ICM outgrowth on inactivated human feeders, karyotype was confirmed as 46, XY and STR analysis demonstrated a male Allele pattern. The hESC line had pluripotent cell morphology, 86% of cells expressed Nanog, 95% Oct4, 54% Tra1-60 and 98% SSEA4 and gave a PluriTest Pluripotency score of 30.25, Novelty of 1.21. The cell line was negative for Mycoplasma and visible contamination. PMID:27346010

  1. CYR61 is a novel gene associated with temperature-dependent changes in fish metabolism as revealed by cDNA microarray analysis on a medaka Oryzias latipes cell line.

    PubMed

    Hirayama, Makoto; Ahsan, Md Nazmul; Mitani, Hiroshi; Watabe, Shugo

    2008-07-01

    A microarray comprising 3,514 cDNAs was constructed from a medaka EST library to elucidate the transcriptional responses associated with temperature shift from 25 to 15 degrees C in a medaka cell line. Microarray analysis revealed that the mRNA levels of 313 clones were significantly different in at least one combination of different incubation periods up to 7 days at a given incubation temperature or between 25 and 15 degrees C at a given incubation period (P < 0.05). These genes are known to be associated with various biological processes including morphogenesis, cell proliferation and response to stress. A number of genes encoding proteins which localize in extracellular areas were apparently up-regulated at 15 degrees C, whereas those localizing in intracellular areas were down-regulated at this temperature. In addition, while a number of genes represented long-term expression changes, only a few responded to short-term inductions. A typical example was CYR61, a multifunctional matricellular signaling modulator, the mRNA levels of which increased after temperature shift from 25 to 15 degrees C in 3 h, and then decreased rapidly to near the original level within 12 h. Another series of analyses by quantitative reverse transcription-PCR revealed that the mRNA levels of CYR61 at 5 degrees C were significantly higher even at 24 h after temperature shift compared to those of the cells successively maintained at 25 degrees C. These analyses suggest that remodeling and reorganizing of extracellular structure of cells are important to offset the low temperature effect and CYR61 is considered to be a novel gene associated with temperature response in poikilotherms. PMID:18286541

  2. Divergence and long-distance overseas dispersals of island populations of the Ryukyu five-lined skink, Plestiodon marginatus (Scincidae: Squamata), in the Ryukyu Archipelago, Japan, as revealed by mitochondrial DNA phylogeography.

    PubMed

    Kurita, Kazuki; Hikida, Tsutomu

    2014-04-01

    We assessed the historical biogeography of the Ryukyu five-lined skink, Plestiodon marginatus, and related species (P. stimpsonii and P. elegans). Our specific aims were to reveal the origin, tim- ing, and route of the colonization to three volcanic islands in the northern Tokara Group of the northern Ryukyus: Kuchinoshima, Nakanoshima, and Suwanosejima. We conducted phylogenetic analyses and divergence time estimation using a partial sequence of the mitochondrial cytochrome b gene for P. marginatus collected from across its whole range (the northern and central Ryukyus), and for P. stimpsonii (from the Yaeyama Group of the southern Ryukyus) and P. elegans (from Taiwan). Our results suggest three major clades (A, B, and C). Clades A and B consist of P. marginatus, excluding the Kuchinoshima population, and Clade C consisted of the Kuchinoshima population, P. stimpsonii, and P. elegans. These clades are estimated to have diverged during the Late Miocene to the Late Pliocene. Among the three examined northern Tokara populations, the Kuchinoshima population was shown to be a sister group of P. stimpsonii. The two other populations from Nakanoshima and Suwanosejima Islands were closely related to P. marginatus from the northern part of the Okinawa Group and that from Kodakarajima Island in the southern Tokara Group, respectively. These populations are estimated to have diverged from their respective related spe cies in various ages of the Early to Late Pleistocene, suggesting that they colonized the islands by independent overseas dispersals of approximately 50-850 km via the Kuroshio Current. Taxonomic implications for P. marginatus are also discussed. PMID:24694220

  3. Cell Lines

    PubMed Central

    Cherbas, Lucy; Gong, Lei

    2014-01-01

    We review the properties and uses of cell lines in Drosophila research, emphasizing the variety of lines, the large body of genomic and transcriptional data available for many of the lines, and the variety of ways the lines have been used to provide tools for and insights into the developmental, molecular, and cell biology of Drosophila and mammals. PMID:24434506

  4. Coseismic fault zone deformation caused by the 2014 Mw=6.2 Nagano-ken-hokubu, Japan, earthquake on the Itoigawa-Shizuoka Tectonic Line revealed with differential LiDAR

    NASA Astrophysics Data System (ADS)

    Toda, S.; Ishimura, D.; Homma, S.; Mukoyama, S.; Niwa, Y.

    2015-12-01

    The Mw = 6.2 Nagano-ken-hokubu earthquake struck northern Nagano, central Japan, on November 22, 2014, and accompanied a 9-km-long surface rupture mostly along the previously mapped N-NW trending Kamishiro fault, one of the segments of the 150-km-long Itoigawa-Shizuoka Tectonic Line active fault system. While we mapped the rupture and measured vertical displacement of up to 80 cm at the field, interferometric synthetic aperture radar (InSAR) shows densely spaced fringes on the hanging wall side, suggesting westward or uplift movement associated with thrust faulting. The mainshock focal mechanism and aftershock hypocenters indicate the source fault dips to the east but the InSAR images cannot exactly differentiate between horizontal and vertical movements and also lose coherence within and near the fault zone itself. To reveal near-field deformation and shallow fault slip, here we demonstrate a differential LiDAR analysis using a pair of 1 m-resolution pre-event and post-event bare Earth digital terrain models (DTMs) obtained from commercial LiDAR provider. We applied particle image velocity (PIV) method incorporating elevation change to obtain 3-D vectors of coseismic displacements (Mukoyama, 2011, J. Mt. Sci). Despite sporadic noises mostly due to local landslides, we detected up to 1.5 m net movement at the tip of the hanging wall, more than the field measurement of 80 cm. Our result implies that a 9-km-long rupture zone is not a single continuous fault but composed of two bow-shaped fault strands, suggesting a combination of shallow fault dip and modest amount (< 1.5 m) of slip. Eastward movement without notable subsidence on the footwall also supports the low angle fault dip near the surface, and significant fault normal contraction, observed as buckled cultural features across the fault zone. Secondary features, such as subsidiary back-thrust faults confirmed at the field, are also visible as a significant contrast of vector directions and slip amounts.

  5. Clinically failed eggs as a source of normal human embryo stem cells.

    PubMed

    De Sousa, Paul A; Gardner, John; Sneddon, Sharon; Pells, Steve; Tye, Britt Jorgensen; Dand, Pawlina; Collins, Daniel M; Stewart, Karen; Shaw, Lisa; Przyborski, Stefan; Cooke, Michael; McLaughlin, K John; Kimber, Susan J; Lieberman, Brian A; Wilmut, Ian; Brison, Daniel R

    2009-05-01

    The promise of human embryo stem cells (hESCs) for regenerative medicine is offset by the ethical and practical challenges involved in sourcing eggs and embryos for this objective. In this study we sought to isolate an hESC line from clinically failed eggs, the usage of which would not conflict with donor interests to conceive. A total of 8 blastocysts were allocated for hESC derivation from a pool of 579 eggs whose fertilization had been clinically assessed to have occurred abnormally (i.e., three pronuclei) or failed (i.e., no pronuclei) following in vitro insemination or intracytoplasmic sperm injection (ICSI). The latter were subjected to a recovery intervention consisting of either reinsemination by ICSI or parthenogenetic stimulation. One hESC line (RCM1) was obtained from a failed-to-fertilize inseminated egg recovered by parthenogenetic activation. Standard in vitro and in vivo characterization revealed this line to possess all of the properties attributed to a normal euploid hESC line. Whole-genome single-nucleotide polymorphism analysis further revealed that the line was biparental, indicating that sperm penetration had occurred, although parthenogenetic stimulation was required for activation. Our results demonstrate the viability of an alternative strategy to generate normal hESC lines from clinically failed eggs, thereby further minimizing the potential to conflict with donor reproductive interest to conceive. PMID:19393594

  6. A clumpy stellar wind and luminosity-dependent cyclotron line revealed by the first Suzaku observation of the high-mass X-ray binary 4U 1538–522

    SciTech Connect

    Hemphill, Paul B.; Rothschild, Richard E.; Markowitz, Alex; Fürst, Felix; Pottschmidt, Katja; Wilms, Jörn

    2014-09-01

    We present results from the first Suzaku observation of the high-mass X-ray binary 4U 1538–522. The broadband spectral coverage of Suzaku allows for a detailed spectral analysis, characterizing the cyclotron resonance scattering feature at 23.0 ± 0.4 keV and the iron Kα line at 6.426 ± 0.008 keV, as well as placing limits on the strengths of the iron Kβ line and the iron K edge. We track the evolution of the spectral parameters both in time and in luminosity, notably finding a significant positive correlation between cyclotron line energy and luminosity. A dip and spike in the light curve is shown to be associated with an order-of-magnitude increase in column density along the line of sight, as well as significant variation in the underlying continuum, implying the accretion of a overdense region of a clumpy stellar wind. We also present a phase-resolved analysis, with most spectral parameters of interest showing significant variation with phase. Notably, both the cyclotron line energy and the iron Kα line intensity vary significantly with phase, with the iron line intensity significantly out of phase with the pulse profile. We discuss the implications of these findings in the context of recent work in the areas of accretion column physics and cyclotron resonance scattering feature formation.

  7. Measure Lines

    ERIC Educational Resources Information Center

    Crissman, Sally

    2011-01-01

    One tool for enhancing students' work with data in the science classroom is the measure line. As a coteacher and curriculum developer for The Inquiry Project, the author has seen how measure lines--a number line in which the numbers refer to units of measure--help students not only represent data but also analyze it in ways that generate…

  8. Revealing Roosevelt

    NASA Technical Reports Server (NTRS)

    2006-01-01

    This image mosaic from the microscopic imager aboard NASA's Mars Exploration Rover Opportunity shows detailed structure of a small fin-like structure dubbed 'Roosevelt,' which sticks out from the outcrop pavement at the edge of 'Erebus Crater.'

    Roosevelt lines a fracture in the local pavement and scientists hypothesize that it is a fracture fill, formed by water that percolated through the fracture. This would mean the feature is younger than surrounding rocks and, therefore, might provide evidence of water that was present some time after the formation of Meridiani Planum sedimentary rocks.

    The image shows fine laminations (layers about 1 millimeter or .04 inch thick) that run parallel to the axis of the fin. Some of the textures visible in the image likely indicate that minerals precipitated from the outcrop rocks, but sediment grains are also apparent.

    The three frames combined into this mosaic were taken during Opportunity's 727th Martian day, or sol (Feb. 8, 2006). In subsequent days, the rover completed textural and chemical inspection of Roosevelt to help the science team understand this structure's significance for Martian history.

  9. Revealing Rembrandt

    PubMed Central

    Parker, Andrew J.

    2014-01-01

    The power and significance of artwork in shaping human cognition is self-evident. The starting point for our empirical investigations is the view that the task of neuroscience is to integrate itself with other forms of knowledge, rather than to seek to supplant them. In our recent work, we examined a particular aspect of the appreciation of artwork using present-day functional magnetic resonance imaging (fMRI). Our results emphasized the continuity between viewing artwork and other human cognitive activities. We also showed that appreciation of a particular aspect of artwork, namely authenticity, depends upon the co-ordinated activity between the brain regions involved in multiple decision making and those responsible for processing visual information. The findings about brain function probably have no specific consequences for understanding how people respond to the art of Rembrandt in comparison with their response to other artworks. However, the use of images of Rembrandt's portraits, his most intimate and personal works, clearly had a significant impact upon our viewers, even though they have been spatially confined to the interior of an MRI scanner at the time of viewing. Neuroscientific studies of humans viewing artwork have the capacity to reveal the diversity of human cognitive responses that may be induced by external advice or context as people view artwork in a variety of frameworks and settings. PMID:24795552

  10. Multivariate Calibration Approach for Quantitative Determination of Cell-Line Cross Contamination by Intact Cell Mass Spectrometry and Artificial Neural Networks

    PubMed Central

    Prokeš, Lubomír; Amato, Filippo; Pivetta, Tiziana; Hampl, Aleš; Havel, Josef; Vaňhara, Petr

    2016-01-01

    Cross-contamination of eukaryotic cell lines used in biomedical research represents a highly relevant problem. Analysis of repetitive DNA sequences, such as Short Tandem Repeats (STR), or Simple Sequence Repeats (SSR), is a widely accepted, simple, and commercially available technique to authenticate cell lines. However, it provides only qualitative information that depends on the extent of reference databases for interpretation. In this work, we developed and validated a rapid and routinely applicable method for evaluation of cell culture cross-contamination levels based on mass spectrometric fingerprints of intact mammalian cells coupled with artificial neural networks (ANNs). We used human embryonic stem cells (hESCs) contaminated by either mouse embryonic stem cells (mESCs) or mouse embryonic fibroblasts (MEFs) as a model. We determined the contamination level using a mass spectra database of known calibration mixtures that served as training input for an ANN. The ANN was then capable of correct quantification of the level of contamination of hESCs by mESCs or MEFs. We demonstrate that MS analysis, when linked to proper mathematical instruments, is a tangible tool for unraveling and quantifying heterogeneity in cell cultures. The analysis is applicable in routine scenarios for cell authentication and/or cell phenotyping in general. PMID:26821236

  11. Multivariate Calibration Approach for Quantitative Determination of Cell-Line Cross Contamination by Intact Cell Mass Spectrometry and Artificial Neural Networks.

    PubMed

    Valletta, Elisa; Kučera, Lukáš; Prokeš, Lubomír; Amato, Filippo; Pivetta, Tiziana; Hampl, Aleš; Havel, Josef; Vaňhara, Petr

    2016-01-01

    Cross-contamination of eukaryotic cell lines used in biomedical research represents a highly relevant problem. Analysis of repetitive DNA sequences, such as Short Tandem Repeats (STR), or Simple Sequence Repeats (SSR), is a widely accepted, simple, and commercially available technique to authenticate cell lines. However, it provides only qualitative information that depends on the extent of reference databases for interpretation. In this work, we developed and validated a rapid and routinely applicable method for evaluation of cell culture cross-contamination levels based on mass spectrometric fingerprints of intact mammalian cells coupled with artificial neural networks (ANNs). We used human embryonic stem cells (hESCs) contaminated by either mouse embryonic stem cells (mESCs) or mouse embryonic fibroblasts (MEFs) as a model. We determined the contamination level using a mass spectra database of known calibration mixtures that served as training input for an ANN. The ANN was then capable of correct quantification of the level of contamination of hESCs by mESCs or MEFs. We demonstrate that MS analysis, when linked to proper mathematical instruments, is a tangible tool for unraveling and quantifying heterogeneity in cell cultures. The analysis is applicable in routine scenarios for cell authentication and/or cell phenotyping in general. PMID:26821236

  12. Revealing Mercury

    NASA Astrophysics Data System (ADS)

    Prockter, L. M.; Solomon, S. C.; Head, J. W.; Watters, T. R.; Murchie, S. L.; Robinson, M. S.; Chapman, C. R.; McNutt, R. L.

    2009-04-01

    The MErcury Surface, Space ENvironment, GEochemistry, and Ranging (MESSENGER) spacecraft, developed under NASA's Discovery Program, launched in August 2004. En route to insertion into orbit about Mercury in 2011, MESSENGER flies by Mercury three times. The first and second of these encounters were accomplished in January and October of 2008. These flybys viewed portions of Mercury's surface that were not observed by Mariner 10 during its reconnaissance of somewhat less than half of the planet in 1974-1975. All MESSENGER instruments operated during each flyby and returned a wealth of new data. Many of the new observations were focused on the planet's geology, including monochrome imaging at resolutions as high as 100 m/pixel, multispectral imaging in 11 filters at resolutions as high as 500 m/pixel, laser altimetry tracks extending over several thousands of kilometers, and high-resolution spectral measurements of several types of terrain. Here we present an overview of the first inferences on the global geology of Mercury from the MESSENGER observations. Whereas evidence for volcanism was equivocal from Mariner 10 data, the new MESSENGER images and altimetry provide compelling evidence that volcanism was widespread and protracted on Mercury. Color imaging reveals three common spectral units on the surface: a higher-reflectance, relatively red material occurring as a distinct class of smooth plains, typically with distinct embayment relationships interpreted to indicate volcanic emplacement; a lower-reflectance, relatively blue material typically excavated by impact craters and therefore inferred to be more common at depth; and a spectrally intermediate terrain that constitutes much of the uppermost crust. Three more minor spectral units are also seen: fresh crater ejecta, reddish material associated with rimless depressions interpreted to be volcanic centers, and high-reflectance deposits seen in some crater floors. Preliminary measurements of crater size

  13. Familial Dysautonomia (FD) Human Embryonic Stem Cell Derived PNS Neurons Reveal that Synaptic Vesicular and Neuronal Transport Genes Are Directly or Indirectly Affected by IKBKAP Downregulation

    PubMed Central

    Kantor, Gal; Cheishvili, David; Even, Aviel; Birger, Anastasya; Turetsky, Tikva; Gil, Yaniv; Even-Ram, Sharona; Aizenman, Einat; Bashir, Nibal; Maayan, Channa; Razin, Aharon; Reubinoff, Benjamim E.; Weil, Miguel

    2015-01-01

    A splicing mutation in the IKBKAP gene causes Familial Dysautonomia (FD), affecting the IKAP protein expression levels and proper development and function of the peripheral nervous system (PNS). Here we found new molecular insights for the IKAP role and the impact of the FD mutation in the human PNS lineage by using a novel and unique human embryonic stem cell (hESC) line homozygous to the FD mutation originated by pre implantation genetic diagnosis (PGD) analysis. We found that IKBKAP downregulation during PNS differentiation affects normal migration in FD-hESC derived neural crest cells (NCC) while at later stages the PNS neurons show reduced intracellular colocalization between vesicular proteins and IKAP. Comparative wide transcriptome analysis of FD and WT hESC-derived neurons together with the analysis of human brains from FD and WT 12 weeks old embryos and experimental validation of the results confirmed that synaptic vesicular and neuronal transport genes are directly or indirectly affected by IKBKAP downregulation in FD neurons. Moreover we show that kinetin (a drug that corrects IKBKAP alternative splicing) promotes the recovery of IKAP expression and these IKAP functional associated genes identified in the study. Altogether, these results support the view that IKAP might be a vesicular like protein that might be involved in neuronal transport in hESC derived PNS neurons. This function seems to be mostly affected in FD-hESC derived PNS neurons probably reflecting some PNS neuronal dysfunction observed in FD. PMID:26437462

  14. Efficient Recombinase-Mediated Cassette Exchange in hPSCs to Study the Hepatocyte Lineage Reveals AAVS1 Locus-Mediated Transgene Inhibition.

    PubMed

    Ordovás, Laura; Boon, Ruben; Pistoni, Mariaelena; Chen, Yemiao; Wolfs, Esther; Guo, Wenting; Sambathkumar, Rangarajan; Bobis-Wozowicz, Sylwia; Helsen, Nicky; Vanhove, Jolien; Berckmans, Pieter; Cai, Qing; Vanuytsel, Kim; Eggermont, Kristel; Vanslembrouck, Veerle; Schmidt, Béla Z; Raitano, Susanna; Van Den Bosch, Ludo; Nahmias, Yaakov; Cathomen, Toni; Struys, Tom; Verfaillie, Catherine M

    2015-11-10

    Tools for rapid and efficient transgenesis in "safe harbor" loci in an isogenic context remain important to exploit the possibilities of human pluripotent stem cells (hPSCs). We created hPSC master cell lines suitable for FLPe recombinase-mediated cassette exchange (RMCE) in the AAVS1 locus that allow generation of transgenic lines within 15 days with 100% efficiency and without random integrations. Using RMCE, we successfully incorporated several transgenes useful for lineage identification, cell toxicity studies, and gene overexpression to study the hepatocyte lineage. However, we observed unexpected and variable transgene expression inhibition in vitro, due to DNA methylation and other unknown mechanisms, both in undifferentiated hESC and differentiating hepatocytes. Therefore, the AAVS1 locus cannot be considered a universally safe harbor locus for reliable transgene expression in vitro, and using it for transgenesis in hPSC will require careful assessment of the function of individual transgenes. PMID:26455413

  15. A field-grown transgenic tomato line expressing higher levels of polyamines reveals legume cover crop mulch-specific perturbations in fruit phenotype at the levels of metabolite profiles, gene expression, and agronomic characteristics

    PubMed Central

    Neelam, Anil; Cassol, Tatiana; Mehta, Roshni A.; Abdul-Baki, Aref A.; Sobolev, Anatoli P.; Goyal, Ravinder K.; Abbott, Judith; Segre, Anna L.; Handa, Avtar K.; Mattoo, Autar K.

    2008-01-01

    Genetic modification of crop plants to introduce desirable traits such as nutritional enhancement, disease and pest resistance, and enhanced crop productivity is increasingly seen as a promising technology for sustainable agriculture and boosting food production in the world. Independently, cultural practices that utilize alternative agriculture strategies including organic cultivation subscribe to sustainable agriculture by limiting chemical usage and reduced tillage. How the two together affect fruit metabolism or plant growth in the field or whether they are compatible has not yet been tested. Fruit-specific yeast S-adenosylmethionine decarboxylase (ySAMdc) line 579HO, and a control line 556AZ were grown in leguminous hairy vetch (Vicia villosa Roth) (HV) mulch and conventional black polyethylene (BP) mulch, and their fruit analysed. Significant genotype×mulch-dependent interactions on fruit phenotype were exemplified by differential profiles of 20 fruit metabolites such as amino acids, sugars, and organic acids. Expression patterns of the ySAMdc transgene, and tomato SAMdc, E8, PEPC, and ICDHc genes were compared between the two lines as a function of growth on either BP or HV mulch. HV mulch significantly stimulated the accumulation of asparagine, glutamate, glutamine, choline, and citrate concomitant with a decrease in glucose in the 556AZ fruits during ripening as compared to BP. It enables a metabolic system in tomato somewhat akin to the one in higher polyamine-accumulating transgenic fruit that have higher phytonutrient content. Finally, synergism was found between HV mulch and transgenic tomato in up-regulating N:C indicator genes PEPC and ICDHc in the fruit. PMID:18469323

  16. A field-grown transgenic tomato line expressing higher levels of polyamines reveals legume cover crop mulch-specific perturbations in fruit phenotype at the levels of metabolite profiles, gene expression, and agronomic characteristics.

    PubMed

    Neelam, Anil; Cassol, Tatiana; Mehta, Roshni A; Abdul-Baki, Aref A; Sobolev, Anatoli P; Goyal, Ravinder K; Abbott, Judith; Segre, Anna L; Handa, Avtar K; Mattoo, Autar K

    2008-01-01

    Genetic modification of crop plants to introduce desirable traits such as nutritional enhancement, disease and pest resistance, and enhanced crop productivity is increasingly seen as a promising technology for sustainable agriculture and boosting food production in the world. Independently, cultural practices that utilize alternative agriculture strategies including organic cultivation subscribe to sustainable agriculture by limiting chemical usage and reduced tillage. How the two together affect fruit metabolism or plant growth in the field or whether they are compatible has not yet been tested. Fruit-specific yeast S-adenosylmethionine decarboxylase (ySAMdc) line 579HO, and a control line 556AZ were grown in leguminous hairy vetch (Vicia villosa Roth) (HV) mulch and conventional black polyethylene (BP) mulch, and their fruit analysed. Significant genotypexmulch-dependent interactions on fruit phenotype were exemplified by differential profiles of 20 fruit metabolites such as amino acids, sugars, and organic acids. Expression patterns of the ySAMdc transgene, and tomato SAMdc, E8, PEPC, and ICDHc genes were compared between the two lines as a function of growth on either BP or HV mulch. HV mulch significantly stimulated the accumulation of asparagine, glutamate, glutamine, choline, and citrate concomitant with a decrease in glucose in the 556AZ fruits during ripening as compared to BP. It enables a metabolic system in tomato somewhat akin to the one in higher polyamine-accumulating transgenic fruit that have higher phytonutrient content. Finally, synergism was found between HV mulch and transgenic tomato in up-regulating N:C indicator genes PEPC and ICDHc in the fruit. PMID:18469323

  17. The L1TD1 Protein Interactome Reveals the Importance of Post-transcriptional Regulation in Human Pluripotency

    PubMed Central

    Emani, Maheswara Reddy; Närvä, Elisa; Stubb, Aki; Chakroborty, Deepankar; Viitala, Miro; Rokka, Anne; Rahkonen, Nelly; Moulder, Robert; Denessiouk, Konstantin; Trokovic, Ras; Lund, Riikka; Elo, Laura L.; Lahesmaa, Riitta

    2015-01-01

    Summary The RNA-binding protein L1TD1 is one of the most specific and abundant proteins in pluripotent stem cells and is essential for the maintenance of pluripotency in human cells. Here, we identify the protein interaction network of L1TD1 in human embryonic stem cells (hESCs) and provide insights into the interactome network constructed in human pluripotent cells. Our data reveal that L1TD1 has an important role in RNA splicing, translation, protein traffic, and degradation. L1TD1 interacts with multiple stem-cell-specific proteins, many of which are still uncharacterized in the context of development. Further, we show that L1TD1 is a part of the pluripotency interactome network of OCT4, SOX2, and NANOG, bridging nuclear and cytoplasmic regulation and highlighting the importance of RNA biology in pluripotency. PMID:25702638

  18. Analysis of Genes Induced by Sendai Virus Infection of Mutant Cell Lines Reveals Essential Roles of Interferon Regulatory Factor 3, NF-κB, and Interferon but Not Toll-Like Receptor 3†

    PubMed Central

    Elco, Christopher P.; Guenther, Jeanna M.; Williams, Bryan R. G.; Sen, Ganes C.

    2005-01-01

    Sendai virus (SeV) infection causes the transcriptional induction of many cellular genes that are also induced by interferon (IFN) or double-stranded RNA (dsRNA). We took advantage of various mutant cell lines to investigate the putative roles of the components of the IFN and dsRNA signaling pathways in the induction of those genes by SeV. Profiling the patterns of gene expression in SeV-infected cells demonstrated that Toll-like receptor 3, although essential for gene induction by dsRNA, was dispensable for gene induction by SeV. In contrast, Jak1, which mediates IFN signaling, was required for the induction of a small subset of genes by SeV. NF-κB and interferon regulatory factor 3 (IRF-3), the two major transcription factors activated by virus infection, were essential for the induction of two sets of genes by SeV. As expected, some of the IRF-3-dependent genes, such as ISG56, were more strongly induced by SeV in IRF-3-overexpressing cells. Surprisingly, in those cells, a number of NF-κB-dependent genes, such as the A20 gene, were induced poorly. Using a series of cell lines expressing increasing levels of IRF-3, we demonstrated that the degree of induction of A20 mRNA, upon SeV infection, was inversely proportional to the cellular level of IRF-3, whereas that of ISG56 mRNA was directly proportional. Thus, IRF-3 can suppress the expression of NF-κB-dependent genes in SeV-infected cells. PMID:15767394

  19. Comparison of biomaterials and extracellular matrices as a culture platform for multiple, independently derived human embryonic stem cell lines.

    PubMed

    Hakala, Heidi; Rajala, Kristiina; Ojala, Marisa; Panula, Sarita; Areva, Sami; Kellomäki, Minna; Suuronen, Riitta; Skottman, Heli

    2009-07-01

    Long-term in vitro culture of undifferentiated human embryonic stem cells (hESCs) traditionally requires a fibroblast feeder cell layer. Using feeder cells in hESC cultures is highly laborious and limits large-scale hESC production for potential application in regenerative medicine. Replacing feeder cells with defined human extracellular matrix (ECM) components or synthetic biomaterials would be ideal for large-scale production of clinical-grade hESCs. We tested and compared different feeder cell-free hESC culture methods based on different human ECM proteins, human and animal sera matrices, and a Matrigel matrix. Also selected biomaterials were tested for feeder cell-free propagation of undifferentiated hESCs. The matrices were tested together with conventional and modified hESC culture media, human foreskin fibroblast-conditioned culture medium, chemically defined medium, TeSR1, and modified TeSR1 media. The results showed the undefined, xenogeneic Matrigel to be a superior matrix for hESC culture compared with the purified human ECM proteins, serum matrices, and the biomaterials tested. A long-term, feeder cell-free culture system was successful on Matrigel in combination with mTeSR1 culture medium, but a xeno-free, fully defined, and reproducible feeder cell-free hESC culture method still remains to be developed. PMID:19132919

  20. Three Years After Transplants in Human Mandibles, Histological and In-Line Holotomography Revealed That Stem Cells Regenerated a Compact Rather Than a Spongy Bone: Biological and Clinical Implications

    PubMed Central

    Giuliani, Alessandra; Manescu, Adrian; Langer, Max; Rustichelli, Franco; Desiderio, Vincenzo; Paino, Francesca; De Rosa, Alfredo; Laino, Luigi; d'Aquino, Riccardo; Tirino, Virginia

    2013-01-01

    Mesenchymal stem cells deriving from dental pulp differentiate into osteoblasts capable of producing bone. In previous studies, we extensively demonstrated that, when seeded on collagen I scaffolds, these cells can be conveniently used for the repair of human mandible defects. Here, we assess the stability and quality of the regenerated bone and vessel network 3 years after the grafting intervention, with conventional procedures and in-line holotomography, an advanced phase-imaging method using synchrotron radiation that offers improved sensitivity toward low-absorbing structures. We found that the regenerated tissue from the graft sites was composed of a fully compact bone with a higher matrix density than control human alveolar spongy bone from the same patient. Thus, the regenerated bone, being entirely compact, is completely different from normal alveolar bone. Although the bone regenerated at the graft sites is not of the proper type found in the mandible, it does seem to have a positive clinical impact. In fact, it creates steadier mandibles, may well increase implant stability, and, additionally, may improve resistance to mechanical, physical, chemical, and pharmacological agents. PMID:23502599

  1. The Differential Gene Expression Pattern of Mycobacterium tuberculosis in Response to Capreomycin and PA-824 versus First-Line TB Drugs Reveals Stress- and PE/PPE-Related Drug Targets

    PubMed Central

    Fu, Li M.; Tai, Shu C.

    2009-01-01

    Tuberculosis is a leading infectious disease causing millions of deaths each year. How to eradicate mycobacterial persistence has become a central research focus for developing next-generation TB drugs. Yet, the knowledge in this area is fundamentally limited and only a few drugs, notably capreomycin and PA-824, have been shown to be active against non-replicating persistent TB bacilli. In this study, we performed a new bioinformatics analysis on microarray-based gene expression data obtained from the public domain to explore genes that were differentially induced by drugs between the group of capreomycin and PA-824 and the group of mainly the first-line TB drugs. Our study has identified 42 genes specifically induced by capreomycin and PA-824. Many of these genes are related to stress responses. In terms of the distribution of identified genes in a specific category relative to the whole genome, only the categories of PE/PPE and conserved hypotheticals have statistical significance. Six among the 42 genes identified in this study are on the list of the top 100 persistence targets selected by the TB Structural Genomics Consortium. Further biological elucidation of their roles in mycobacterial persistence is warranted. PMID:20016672

  2. REVEALING THE PHYSICAL PROPERTIES OF MOLECULAR GAS IN ORION WITH A LARGE-SCALE SURVEY IN J = 2-1 LINES OF {sup 12}CO, {sup 13}CO, AND C{sup 18}O

    SciTech Connect

    Nishimura, Atsushi; Tokuda, Kazuki; Kimura, Kimihiro; Muraoka, Kazuyuki; Maezawa, Hiroyuki; Ogawa, Hideo; Onishi, Toshikazu; Dobashi, Kazuhito; Shimoikura, Tomomi; Mizuno, Akira; Fukui, Yasuo

    2015-01-01

    We present fully sampled ∼3' resolution images of {sup 12}CO(J = 2-1), {sup 13}CO(J = 2-1), and C{sup 18}O(J = 2-1) emission taken with the newly developed 1.85 m millimeter-submillimeter telescope over the entire area of the Orion A and B giant molecular clouds. The data were compared with J = 1-0 of the {sup 12}CO, {sup 13}CO, and C{sup 18}O data taken with the Nagoya 4 m telescope and the NANTEN telescope at the same angular resolution to derive the spatial distributions of the physical properties of the molecular gas. We explore the large velocity gradient formalism to determine the gas density and temperature using line combinations of {sup 12}CO(J = 2-1), {sup 13}CO(J = 2-1), and {sup 13}CO(J = 1-0) assuming a uniform velocity gradient and abundance ratio of CO. The derived gas density is in the range of 500 to 5000 cm{sup –3}, and the derived gas temperature is mostly in the range of 20 to 50 K along the cloud ridge with a temperature gradient depending on the distance from the star forming region. We found that the high-temperature region at the cloud edge faces the H II region, indicating that the molecular gas is interacting with the stellar wind and radiation from the massive stars. In addition, we compared the derived gas properties with the young stellar objects distribution obtained with the Spitzer telescope to investigate the relationship between the gas properties and the star formation activity therein. We found that the gas density and star formation efficiency are positively well correlated, indicating that stars form effectively in the dense gas region.

  3. A novel lineage restricted, pericyte-like cell line isolated from human embryonic stem cells.

    PubMed

    Greenwood-Goodwin, Midori; Yang, Jiwei; Hassanipour, Mohammad; Larocca, David

    2016-01-01

    Pericytes (PCs) are endothelium-associated cells that play an important role in normal vascular function and maintenance. We developed a method comparable to GMP quality protocols for deriving self-renewing perivascular progenitors from the human embryonic stem cell (hESC), line ESI-017. We identified a highly scalable, perivascular progenitor cell line that we termed PC-A, which expressed surface markers common to mesenchymal stromal cells. PC-A cells were not osteogenic or adipogenic under standard differentiation conditions and showed minimal angiogenic support function in vitro. PC-A cells were capable of further differentiation to perivascular progenitors with limited differentiation capacity, having osteogenic potential (PC-O) or angiogenic support function (PC-M), while lacking adipogenic potential. Importantly, PC-M cells expressed surface markers associated with pericytes. Moreover, PC-M cells had pericyte-like functionality being capable of co-localizing with human umbilical vein endothelial cells (HUVECs) and enhancing tube stability up to 6 days in vitro. We have thus identified a self-renewing perivascular progenitor cell line that lacks osteogenic, adipogenic and angiogenic potential but is capable of differentiation toward progenitor cell lines with either osteogenic potential or pericyte-like angiogenic function. The hESC-derived perivascular progenitors described here have potential applications in vascular research, drug development and cell therapy. PMID:27109637

  4. A novel lineage restricted, pericyte-like cell line isolated from human embryonic stem cells

    PubMed Central

    Greenwood-Goodwin, Midori; Yang, Jiwei; Hassanipour, Mohammad; Larocca, David

    2016-01-01

    Pericytes (PCs) are endothelium-associated cells that play an important role in normal vascular function and maintenance. We developed a method comparable to GMP quality protocols for deriving self-renewing perivascular progenitors from the human embryonic stem cell (hESC), line ESI-017. We identified a highly scalable, perivascular progenitor cell line that we termed PC-A, which expressed surface markers common to mesenchymal stromal cells. PC-A cells were not osteogenic or adipogenic under standard differentiation conditions and showed minimal angiogenic support function in vitro. PC-A cells were capable of further differentiation to perivascular progenitors with limited differentiation capacity, having osteogenic potential (PC-O) or angiogenic support function (PC-M), while lacking adipogenic potential. Importantly, PC-M cells expressed surface markers associated with pericytes. Moreover, PC-M cells had pericyte-like functionality being capable of co-localizing with human umbilical vein endothelial cells (HUVECs) and enhancing tube stability up to 6 days in vitro. We have thus identified a self-renewing perivascular progenitor cell line that lacks osteogenic, adipogenic and angiogenic potential but is capable of differentiation toward progenitor cell lines with either osteogenic potential or pericyte-like angiogenic function. The hESC-derived perivascular progenitors described here have potential applications in vascular research, drug development and cell therapy. PMID:27109637

  5. Sourcing human embryos for embryonic stem cell lines: problems & perspectives.

    PubMed

    Mehta, Rajvi H

    2014-11-01

    The ability to successfully derive human embryonic stem cells (hESC) lines from human embryos following in vitro fertilization (IVF) opened up a plethora of potential applications of this technique. These cell lines could have been successfully used to increase our understanding of human developmental biology, transplantation medicine and the emerging science of regenerative medicine. The main source for human embryos has been 'discarded' or 'spare' fresh or frozen human embryos following IVF. It is a common practice to stimulate the ovaries of women undergoing any of the assisted reproductive technologies (ART) and retrieve multiple oocytes which subsequently lead to multiple embryos. Of these, only two or maximum of three embryos are transferred while the rest are cryopreserved as per the decision of the couple. in case a couple does not desire to 'cryopreserve' their embryos then all the embryos remaining following embryo transfer can be considered 'spare' or if a couple is no longer in need of the 'cryopreserved' embryos then these also can be considered as 'spare'. But, the question raised by the ethicists is, "what about 'slightly' over-stimulating a woman to get a few extra eggs and embryos? The decision becomes more difficult when it comes to 'discarded' embryos. As of today, the quality of the embryos is primarily assessed based on morphology and the rate of development mainly judged by single point assessment. Despite many criteria described in the literature, the quality assessment is purely subjective. The question that arises is on the decision of 'discarding' embryos. What would be the criteria for discarding embryos and the potential 'use' of ESC derived from the 'abnormal appearing' embryos? This paper discusses some of the newer methods to procure embryos for the derivation of embryonic stem cell lines which will respect the ethical concerns but still provide the source material. PMID:25673530

  6. Sourcing human embryos for embryonic stem cell lines: Problems & perspectives

    PubMed Central

    Mehta, Rajvi H.

    2014-01-01

    The ability to successfully derive human embryonic stem cells (hESC) lines from human embryos following in vitro fertilization (IVF) opened up a plethora of potential applications of this technique. These cell lines could have been successfully used to increase our understanding of human developmental biology, transplantation medicine and the emerging science of regenerative medicine. The main source for human embryos has been ‘discarded’ or ‘spare’ fresh or frozen human embryos following IVF. It is a common practice to stimulate the ovaries of women undergoing any of the assisted reproductive technologies (ART) and retrieve multiple oocytes which subsequently lead to multiple embryos. Of these, only two or maximum of three embryos are transferred while the rest are cryopreserved as per the decision of the couple. In case a couple does not desire to ‘cryopreserve’ their embryos then all the embryos remaining following embryo transfer can be considered ‘spare’ or if a couple is no longer in need of the ‘cryopreserved’ embryos then these also can be considered as ‘spare’. But, the question raised by the ethicists is, “what about ‘slightly’ over-stimulating a woman to get a few extra eggs and embryos? The decision becomes more difficult when it comes to ‘discarded’ embryos. As of today, the quality of the embryos is primarily assessed based on morphology and the rate of development mainly judged by single point assessment. Despite many criteria described in the literature, the quality assessment is purely subjective. The question that arises is on the decision of ‘discarding’ embryos. What would be the criteria for discarding embryos and the potential ‘use’ of ESC derived from the ‘abnormal appearing’ embryos? This paper discusses some of the newer methods to procure embryos for the derivation of embryonic stem cell lines which will respect the ethical concerns but still provide the source material. PMID:25673530

  7. Prevention of lysosomal storage diseases and derivation of mutant stem cell lines by preimplantation genetic diagnosis.

    PubMed

    Altarescu, Gheona; Beeri, Rachel; Eiges, Rachel; Epsztejn-Litman, Silvina; Eldar-Geva, Talia; Elstein, Deborah; Zimran, Ari; Margalioth, Ehud J; Levy-Lahad, Ephrat; Renbaum, Paul

    2012-01-01

    Preimplantation genetic diagnosis (PGD) allows birth of unaffected children for couples at risk for a genetic disorder. We present the strategy and outcome of PGD for four lysosomal storage disorders (LSD): Tay-Sachs disease (TSD), Gaucher disease (GD), Fabry disease (FD), and Hunter syndrome (HS), and subsequent development of stem cell lines. For each disease, we developed a family-specific fluorescent multiplex single-cell PCR protocol that included the familial mutation and informative markers surrounding the mutation. Embryo biopsy and PGD analysis were performed on either oocytes (polar bodies one and two) or on single blastomeres from a six-cell embryo. We treated twenty families carrying mutations in these lysosomal storage disorders, including 3 couples requiring simultaneous analysis for two disorders (TSD/GD, TSD/balanced Robertsonian translocation 45XYder(21;14), and HS/oculocutaneus albinism). These analyses led to an overall pregnancy rate/embryo transfer of 38% and the birth of 20 unaffected children from 17 families. We have found that PGD for lysosomal disorders is a safe and effective method to prevent birth of affected children. In addition, by using mutant embryos for the derivation of stem cell lines, we have successfully established GD and HS hESC lines for use as valuable models in LSD research. PMID:23320174

  8. Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and Quantitative Comparison of the Membrane Proteomes of Self-renewing and Differentiating Human Embryonic Stem Cells*S⃞

    PubMed Central

    Prokhorova, Tatyana A.; Rigbolt, Kristoffer T. G.; Johansen, Pia T.; Henningsen, Jeanette; Kratchmarova, Irina; Kassem, Moustapha; Blagoev, Blagoy

    2009-01-01

    Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful quantitative proteomics platform for comprehensive characterization of complex biological systems. However, the potential of SILAC-based approaches has not been fully utilized in human embryonic stem cell (hESC) research mainly because of the complex nature of hESC culture conditions. Here we describe complete SILAC labeling of hESCs with fully preserved pluripotency, self-renewal capabilities, and overall proteome status that was quantitatively analyzed to a depth of 1556 proteins and 527 phosphorylation events. SILAC-labeled hESCs appear to be perfectly suitable for functional studies, and we exploited a SILAC-based proteomics strategy for discovery of hESC-specific surface markers. We determined and quantitatively compared the membrane proteomes of the self-renewing versus differentiating cells of two distinct human embryonic stem cell lines. Of the 811 identified membrane proteins, six displayed significantly higher expression levels in the undifferentiated state compared with differentiating cells. This group includes the established marker CD133/Prominin-1 as well as novel candidates for hESC surface markers: Glypican-4, Neuroligin-4, ErbB2, receptor-type tyrosine-protein phosphatase ζ (PTPRZ), and Glycoprotein M6B. Our study also revealed 17 potential markers of hESC differentiation as their corresponding protein expression levels displayed a dramatic increase in differentiated embryonic stem cell populations. PMID:19151416

  9. Switching of the core structures of glycosphingolipids from globo- and lacto- to ganglio-series upon human embryonic stem cell differentiation

    PubMed Central

    Liang, Yuh-Jin; Kuo, Huan-Hsien; Chen, Yen-Ying; Yang, Bei-Chia; Cheng, Yuan-Yuan; Yu, Alice L.; Khoo, Kay-Hooi; Yu, John

    2010-01-01

    A systematic survey of expression profiles of glycosphingolipids (GSLs) in two hESC lines and their differentiated embryoid body (EB) outgrowth with three germ layers was carried out using immunofluorescence, flow cytometry, and MALDI-MS and MS/MS analyses. In addition to the well-known hESC-specific markers stage-specific embryonic antigen 3 (SSEA-3) and SSEA-4, we identified several globosides and lacto-series GSLs, previously unrevealed in hESCs, including Gb4Cer, Lc4Cer, fucosyl Lc4Cer, Globo H, and disialyl Gb5Cer. During hESC differentiation into EBs, MS analysis revealed a clear-cut switch in the core structures of GSLs from globo- and lacto- to ganglio-series, which was not as evident by immunostaining with antibodies against SSEA-3 and SSEA-4, owing to their cross-reactivities with various glycosphingolipids. Such a switch was attributable to altered expression of key glycosyltransferases (GTs) in the biosynthetic pathways by the up-regulation of ganglio-series–related GTs with simultaneous down-regulation of globo- and lacto-series–related GTs. Thus, these results provide insights into the unique stage-specific transition and mechanism for alterations of GSL core structures during hESC differentiation. In addition, unique glycan structures uncovered by MS analyses may serve as surface markers for further delineation of hESCs and help identify of their functional roles not only in hESCs but also in cancers. PMID:21149694

  10. Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling

    PubMed Central

    Wang, Linlin; Schulz, Thomas C.; Sherrer, Eric S.; Dauphin, Derek S.; Shin, Soojung; Nelson, Angelique M.; Ware, Carol B.; Zhan, Mei; Song, Chao-Zhong; Chen, Xiaoji; Brimble, Sandii N.; McLean, Amanda; Galeano, Maria J.; Uhl, Elizabeth W.; D'Amour, Kevin A.; Chesnut, Jonathan D.; Rao, Mahendra S.

    2007-01-01

    Despite progress in developing defined conditions for human embryonic stem cell (hESC) cultures, little is known about the cell-surface receptors that are activated under conditions supportive of hESC self-renewal. A simultaneous interrogation of 42 receptor tyrosine kinases (RTKs) in hESCs following stimulation with mouse embryonic fibroblast (MEF) conditioned medium (CM) revealed rapid and prominent tyrosine phosphorylation of insulin receptor (IR) and insulin-like growth factor-1 receptor (IGF1R); less prominent tyrosine phosphorylation of epidermal growth factor receptor (EGFR) family members, including ERBB2 and ERBB3; and trace phosphorylation of fibroblast growth factor receptors. Intense IGF1R and IR phosphorylation occurred in the absence of MEF conditioning (NCM) and was attributable to high concentrations of insulin in the proprietary KnockOut Serum Replacer (KSR). Inhibition of IGF1R using a blocking antibody or lentivirus-delivered shRNA reduced hESC self-renewal and promoted differentiation, while disruption of ERBB2 signaling with the selective inhibitor AG825 severely inhibited hESC proliferation and promoted apoptosis. A simple defined medium containing an IGF1 analog, heregulin-1β (a ligand for ERBB2/ERBB3), fibroblast growth factor-2 (FGF2), and activin A supported long-term growth of multiple hESC lines. These studies identify previously unappreciated RTKs that support hESC proliferation and self-renewal, and provide a rationally designed medium for the growth and maintenance of pluripotent hESCs. PMID:17761519

  11. Central line infections - hospitals

    MedlinePlus

    ... infection; CVC - infection; Central venous device - infection; Infection control - central line infection; Nosocomial infection - central line infection; Hospital acquired infection - central line infection; Patient safety - central ...

  12. The ribosomal subunit assembly line

    PubMed Central

    Dlakić, Mensur

    2005-01-01

    Recent proteomic studies in Saccharomyces cerevisiae have identified nearly 200 proteins, other than the structural ribosomal proteins, that participate in the assembly of ribosomal subunits and their transport from the nucleus. In a separate line of research, proteomic studies of mature plant ribosomes have revealed considerable variability in the protein composition of individual ribosomes. PMID:16207363

  13. Robust Expansion of Human Pluripotent Stem Cells: Integration of Bioprocess Design With Transcriptomic and Metabolomic Characterization

    PubMed Central

    Silva, Marta M.; Rodrigues, Ana F.; Correia, Cláudia; Sousa, Marcos F.Q.; Brito, Catarina; Coroadinha, Ana S.

    2015-01-01

    Human embryonic stem cells (hESCs) have an enormous potential as a source for cell replacement therapies, tissue engineering, and in vitro toxicology applications. The lack of standardized and robust bioprocesses for hESC expansion has hindered the application of hESCs and their derivatives in clinical settings. We developed a robust and well-characterized bioprocess for hESC expansion under fully defined conditions and explored the potential of transcriptomic and metabolomic tools for a more comprehensive assessment of culture system impact on cell proliferation, metabolism, and phenotype. Two different hESC lines (feeder-dependent and feeder-free lines) were efficiently expanded on xeno-free microcarriers in stirred culture systems. Both hESC lines maintained the expression of stemness markers such as Oct-4, Nanog, SSEA-4, and TRA1-60 and the ability to spontaneously differentiate into the three germ layers. Whole-genome transcriptome profiling revealed a phenotypic convergence between both hESC lines along the expansion process in stirred-tank bioreactor cultures, providing strong evidence of the robustness of the cultivation process to homogenize cellular phenotype. Under low-oxygen tension, results showed metabolic rearrangement with upregulation of the glycolytic machinery favoring an anaerobic glycolysis Warburg-effect-like phenotype, with no evidence of hypoxic stress response, in contrast to two-dimensional culture. Overall, we report a standardized expansion bioprocess that can guarantee maximal product quality. Furthermore, the “omics” tools used provided relevant findings on the physiological and metabolic changes during hESC expansion in environmentally controlled stirred-tank bioreactors, which can contribute to improved scale-up production systems. Significance The clinical application of human pluripotent stem cells (hPSCs) has been hindered by the lack of robust protocols able to sustain production of high cell numbers, as required for

  14. Potential flow inside an evaporating cylindrical line.

    PubMed

    Petsi, A J; Burganos, V N

    2005-10-01

    An analytical solution to the problem of potential flow inside an evaporating line is obtained. The line is shaped as a half-cylinder lying on a substrate, and evaporates with either pinned or depinned contact lines. The solution is provided through the technique of separation of variables in the velocity potential and stream function formulations. Based on the flow field calculations, it is estimated that the coffee-stain phenomenon should be expected even for uniform evaporation flux throughout the cylindrical surface, provided that the contact lines remain anchored. A simple expression for the velocity potential is also suggested, which reproduces the local velocity vector with excellent accuracy. The vertically averaged velocity is calculated also for other contact line values, revealing for any value an outward liquid flow for pinned lines as opposed to inward flow for depinned lines. PMID:16383581

  15. Electric Field Lines

    NASA Astrophysics Data System (ADS)

    Arribas, E.; Gallardo, C.; Molina, M.; Sanjosé, V.

    We present the computer program called LINES which is able to calculate and visualize the electric field lines due to seven different discrete configurations of electric point charges. Also we show two examples of the graphic screens generated by LINES.

  16. Effects of Pulsed Electromagnetic Field on Differentiation of HUES-17 Human Embryonic Stem Cell Line

    PubMed Central

    Wu, Yi-Lin; Ma, Shi-Rong; Peng, Tao; Teng, Zeng-Hui; Liang, Xiang-Yan; Guo, Guo-Zhen; Zhang, Hai-Feng; Li, Kang-Chu

    2014-01-01

    Electromagnetic fields are considered to potentially affect embryonic development, but the mechanism is still unknown. In this study, human embryonic stem cell (hESC) line HUES-17 was applied to explore the mechanism of exposure on embryonic development to pulsed electromagnetic field (PEMF) for 400 pulses at different electric field intensities and the differentiation of HUES-17 cells was observed after PEMF exposure. The expression of alkaline phosphatase (AP), stage-specific embryonic antigen-3 (SSEA-3), SSEA-4 and the mRNA level and protein level of Oct4, Sox2 and Nanog in HUES-17 cells remained unchanged after PEMF exposure at the electric field intensities of 50, 100, 200 or 400 kV/m. Four hundred pulses PEMF exposure at the electric field intensities of 50, 100, 200 or 400 kV/m did not affect the differentiation of HUES-17 cells. The reason why electromagnetic fields affect embryonic development may be due to other mechanisms rather than affecting the differentiation of embryonic stem cells. PMID:25196518

  17. Effects of pulsed electromagnetic field on differentiation of HUES-17 human embryonic stem cell line.

    PubMed

    Wu, Yi-Lin; Ma, Shi-Rong; Peng, Tao; Teng, Zeng-Hui; Liang, Xiang-Yan; Guo, Guo-Zhen; Zhang, Hai-Feng; Li, Kang-Chu

    2014-01-01

    Electromagnetic fields are considered to potentially affect embryonic development, but the mechanism is still unknown. In this study, human embryonic stem cell (hESC) line HUES-17 was applied to explore the mechanism of exposure on embryonic development to pulsed electromagnetic field (PEMF) for 400 pulses at different electric field intensities and the differentiation of HUES-17 cells was observed after PEMF exposure. The expression of alkaline phosphatase (AP), stage-specific embryonic antigen-3 (SSEA-3), SSEA-4 and the mRNA level and protein level of Oct4, Sox2 and Nanog in HUES-17 cells remained unchanged after PEMF exposure at the electric field intensities of 50, 100, 200 or 400 kV/m. Four hundred pulses PEMF exposure at the electric field intensities of 50, 100, 200 or 400 kV/m did not affect the differentiation of HUES-17 cells. The reason why electromagnetic fields affect embryonic development may be due to other mechanisms rather than affecting the differentiation of embryonic stem cells. PMID:25196518

  18. How Are Preferences Revealed?

    PubMed Central

    Beshears, John; Choi, James J.; Laibson, David; Madrian, Brigitte C.

    2009-01-01

    Revealed preferences are tastes that rationalize an economic agent’s observed actions. Normative preferences represent the agent’s actual interests. It sometimes makes sense to assume that revealed preferences are identical to normative preferences. But there are many cases where this assumption is violated. We identify five factors that increase the likelihood of a disparity between revealed preferences and normative preferences: passive choice, complexity, limited personal experience, third-party marketing, and intertemporal choice. We then discuss six approaches that jointly contribute to the identification of normative preferences: structural estimation, active decisions, asymptotic choice, aggregated revealed preferences, reported preferences, and informed preferences. Each of these approaches uses consumer behavior to infer some property of normative preferences without equating revealed and normative preferences. We illustrate these issues with evidence from savings and investment outcomes. PMID:24761048

  19. Dynamic line rating in the operating environment

    SciTech Connect

    Foss, S.D. ); Maraio, R.A. )

    1990-04-01

    Much attention has been placed on the benefits derived by the dynamic rating of overhead conductors based on typical or single station weather data. Reported power line capacity show increases from 20 to 70% on the average over existing static rating practices. Because of the necessity of multistation monitoring and the implication with respect to critical span, Niagara Mohawk performed an extensive one year multistation power line study. The study revealed that the dynamic line rating was 20% lower than the average dynamic rating for the power line.

  20. Global transcriptome analysis of Human Bone Marrow Stromal Cells (BMSCs) reveals proliferative, mobile, and Interactive cells that produce abundant extracellular matrix proteins, some of which may affect BMSC Potency

    PubMed Central

    Ren, Jiaqiang; Jin, Ping; Sabatino, Marianna; Balakumaran, Arun; Feng, Ji; Kuznetsov, Sergei A.; Klein, Harvey G.; Robey, Pamela G.; Stroncek, David F.

    2012-01-01

    Background Bone marrow stromal cells (BMSCs) are being used for immune modulatory, anti-inflammatory and tissue engineering applications, but the properties responsible for these effects are not completely understood. Human BMSCs were characterized to identify factors that might be responsible for their clinical effects and biomarkers for assessing their quality. Methods Early passage BMSCs prepared from marrow aspirates of 4 healthy subjects were compared to 3 human embryonic stem cell (hESC) samples, CD34+ cells from 3 healthy subjects and 3 fibroblast cell lines. The cells were analyzed with oligonucleotide expression microarrays with more than 35,000 probes. Results BMSC gene expression signatures of BMSCs differed from those of hematopoietic stem cells (HSCs), hESCs and fibroblasts. Genes up-regulated in BMSCs were involved with cell movement, cell-to-cell signaling and interaction and proliferation. The up-regulated genes most likely belonged to pathways for integrin signaling, integrin linked kinase (ILK) signaling, NFR2-mediated oxidative stress response, regulation of actin-based motility by Rho, actin cytoskeletal signaling, caveolar-mediated endocytosis, clathrin-mediated endocytosis and Wnt/β catenin signaling. Among the most highly up-regulated genes were structural extracellular (ECM) proteins:α5 and β 5 integrin chains, fibronectin, collagen type IIIα1 and Vα1; and functional EMC proteins: connective tissue growth factor (CTGF), transforming growth factor beta induced protein (TGFBI) and ADAM12. Conclusions Global analysis of human BMSCs suggests that they are mobile, metabolically active, proliferative and interactive cells that make use of integrins and integrin signaling. They produce abundant ECM proteins that may contribute to their clinical immune modulatory and anti-inflammatory effects. PMID:21250865

  1. Abstract Line Designs

    ERIC Educational Resources Information Center

    Nevinskas, Nancy

    2011-01-01

    In this article, the author describes a unit on the exploration of line. The unit was composed of two individual line lessons. In the first lesson, students were introduced to line as an element of design. They were asked to describe different types of lines, and look for them in art reproductions. The second lesson in the unit involved painting…

  2. The Language of Lines.

    ERIC Educational Resources Information Center

    Breig-Allen, Cheryl; Hill, Janet; Geismar-Ryan, Lori; Cadwell, Louise Boyd

    1998-01-01

    Describes a project about lines in the environment used with 2- and 3-year olds and based on the Reggio Emilia approach. Activities included making tracks with riding toys, drawing lines on papers, seeing cloud lines, and making lines with yarn and Cuisenaire rods. Shows how young children's observations and ongoing discoveries can uncover their…

  3. Continuous Hypoxic Culturing of Human Embryonic Stem Cells Enhances SSEA-3 and MYC Levels

    PubMed Central

    Laiho, Asta; Rahkonen, Nelly; Emani, Maheswara Reddy; Viitala, Miro; Laurila, Kirsti; Sahla, Roosa; Lund, Riikka; Lähdesmäki, Harri; Jaakkola, Panu; Lahesmaa, Riitta

    2013-01-01

    Low oxygen tension (hypoxia) contributes critically to pluripotency of human embryonic stem cells (hESCs) by preventing spontaneous differentiation and supporting self-renewal. However, it is not well understood how hESCs respond to reduced oxygen availability and what are the molecular mechanisms maintaining pluripotency in these conditions. In this study we characterized the transcriptional and molecular responses of three hESC lines (H9, HS401 and HS360) on short (2 hours), intermediate (24 hours) and prolonged (7 days) exposure to low oxygen conditions (4% O2). In response to prolonged hypoxia the expression of pluripotency surface marker SSEA-3 was increased. Furthermore, the genome wide gene-expression analysis revealed that a substantial proportion (12%) of all hypoxia-regulated genes in hESCs, were directly linked to the mechanisms controlling pluripotency or differentiation. Moreover, transcription of MYC oncogene was induced in response to continuous hypoxia. At the protein level MYC was stabilized through phosphorylation already in response to a short hypoxic exposure. Total MYC protein levels remained elevated throughout all the time points studied. Further, MYC protein expression in hypoxia was affected by silencing HIF2α, but not HIF1α. Since MYC has a crucial role in regulating pluripotency we propose that induction of sustained MYC expression in hypoxia contributes to activation of transcriptional programs critical for hESC self-renewal and maintenance of enhanced pluripotent state. PMID:24236059

  4. Efficient Recombinase-Mediated Cassette Exchange in hPSCs to Study the Hepatocyte Lineage Reveals AAVS1 Locus-Mediated Transgene Inhibition

    PubMed Central

    Ordovás, Laura; Boon, Ruben; Pistoni, Mariaelena; Chen, Yemiao; Wolfs, Esther; Guo, Wenting; Sambathkumar, Rangarajan; Bobis-Wozowicz, Sylwia; Helsen, Nicky; Vanhove, Jolien; Berckmans, Pieter; Cai, Qing; Vanuytsel, Kim; Eggermont, Kristel; Vanslembrouck, Veerle; Schmidt, Béla Z.; Raitano, Susanna; Van Den Bosch, Ludo; Nahmias, Yaakov; Cathomen, Toni; Struys, Tom; Verfaillie, Catherine M.

    2015-01-01

    Summary Tools for rapid and efficient transgenesis in “safe harbor” loci in an isogenic context remain important to exploit the possibilities of human pluripotent stem cells (hPSCs). We created hPSC master cell lines suitable for FLPe recombinase-mediated cassette exchange (RMCE) in the AAVS1 locus that allow generation of transgenic lines within 15 days with 100% efficiency and without random integrations. Using RMCE, we successfully incorporated several transgenes useful for lineage identification, cell toxicity studies, and gene overexpression to study the hepatocyte lineage. However, we observed unexpected and variable transgene expression inhibition in vitro, due to DNA methylation and other unknown mechanisms, both in undifferentiated hESC and differentiating hepatocytes. Therefore, the AAVS1 locus cannot be considered a universally safe harbor locus for reliable transgene expression in vitro, and using it for transgenesis in hPSC will require careful assessment of the function of individual transgenes. PMID:26455413

  5. Reconnection of vorticity lines and magnetic lines

    NASA Technical Reports Server (NTRS)

    Greene, John M.

    1993-01-01

    Magnetic field and fluid vorticity share many features. First, as divergence-free vector fields they are conveniently visualized in terms of their field lines, curves that are everywhere tangent to the field. The lines indicate direction and their density indicates field strength. The question arises of the extent to which the evolution of the fields can be treated in terms of the evolution of their field lines. Newcomb (1958) derived the general conditions on the evolution of vector fields that permit the identification of field lines from one instant to the next. The equations of evolution of the vorticity field and the magnetic field fall within Newcomb's analysis. The dynamics of the flows differ between these two systems, so that geometrically similar phenomena happen in different ways in the two systems. In this paper the geometrical similarities are emphasized. Reconnection will be defined here as evolution in which it is not possible to preserve the global identification of some field lines. There is a close relation between reconnection and the topology of the vector field lines. Nontrivial topology occurs where the field has null points or there are field lines that are closed loops.

  6. Nuclear reactor overflow line

    DOEpatents

    Severson, Wayne J.

    1976-01-01

    The overflow line for the reactor vessel of a liquid-metal-cooled nuclear reactor includes means for establishing and maintaining a continuous bleed flow of coolant amounting to 5 to 10% of the total coolant flow through the overflow line to prevent thermal shock to the overflow line when the reactor is restarted following a trip. Preferably a tube is disposed concentrically just inside the overflow line extending from a point just inside the reactor vessel to an overflow tank and a suction line is provided opening into the body of liquid metal in the reactor vessel and into the annulus between the overflow line and the inner tube.

  7. US weapons secrets revealed

    SciTech Connect

    Norris, R.S.; Arkin, W.M.

    1993-03-01

    Extraordinary details have only recently been revealed about the struggle over the control of early U.S. nuclear weapons and their initial deployments abroad. The information comes from a newly declassified top secret report, part of a larger study, The History of the Strategic Arms Competition, 1945-1972, commissioned by Defense Secretary James R. Schlisinger in summer 1974.

  8. Peripheral arterial line - infants

    MedlinePlus

    PAL - infants; Art line - infants ... an "art line." This article addresses PALs in babies. Why is a PAL used? Doctors and nurses use a PAL to watch your baby's blood pressure. A PAL can also be use ...

  9. Reveal for Salmonella test system.

    PubMed

    Bird, C B; Miller, R L; Miller, B M

    1999-01-01

    The Reveal for Salmonella (RSS) test system is a presumptive qualitative test that detects the presence of Salmonella organisms in foods within 21 h total testing time, allowing the user to release negative products 24 h earlier than when using other rapid test kits. Foods are enriched with a proprietary resuscitation medium called Revive and then selectively enriched with either Selenite Cystine or Rappaport-Vassiliadis selective media. The enriched culture is used to inoculate the RSS detection device, which initiates a lateral flow through a reagent zone containing anti-Salmonella antibodies conjugated to colloidal gold particles that capture antigens present in the culture. The antigen-antibody complex migrates farther and is captured by an additional anti-Salmonella antibody, causing the colloidal gold to precipitate and form a visual line, indicating a positive result. A procedural control line also will form regardless of the presence of Salmonella organisms to indicate the test is working properly. Existing AOAC Official Methods for Salmonella organisms require a 48 h enrichment before testing. Hence, a food product has to be held before release, adding extra cost to the company and the consumer. The RSS test system was evaluated by quantitative spiking studies. Although AOAC encourages inclusion of naturally contaminated foods, almost all microbiological AOAC validation studies have been performed with artificially contaminated foods for absolute control over the study. The RSS test system is designed to test many food types for Salmonella organisms and has a limit of detection of 5-10 colony-forming units (cfu)/25 g with a false-negative rate of < 1% and a false-positive rate of < 5.0%. It showed an 81% overall agreement with the traditional procedure of the U.S. Department of Agriculture's Food Safety Inspection Service. PMID:10367381

  10. Heightened potency of human pluripotent stem cell lines created by transient BMP4 exposure

    PubMed Central

    Yang, Ying; Adachi, Katsuyuki; Sheridan, Megan A.; Alexenko, Andrei P.; Schust, Danny J.; Schulz, Laura C.; Ezashi, Toshihiko; Roberts, R. Michael

    2015-01-01

    Human pluripotent stem cells (PSCs) show epiblast-type pluripotency that is maintained with ACTIVIN/FGF2 signaling. Here, we report the acquisition of a unique stem cell phenotype by both human ES cells (hESCs) and induced pluripotent stem cells (iPSCs) in response to transient (24–36 h) exposure to bone morphogenetic protein 4 (BMP4) plus inhibitors of ACTIVIN signaling (A83-01) and FGF2 (PD173074), followed by trypsin dissociation and recovery of colonies capable of growing on a gelatin substratum in standard medium for human PSCs at low but not high FGF2 concentrations. The self-renewing cell lines stain weakly for CDX2 and strongly for NANOG, can be propagated clonally on either Matrigel or gelatin, and are morphologically distinct from human PSC progenitors on either substratum but still meet standard in vitro criteria for pluripotency. They form well-differentiated teratomas in immune-compromised mice that secrete human chorionic gonadotropin (hCG) into the host mouse and include small areas of trophoblast-like cells. The cells have a distinct transcriptome profile from the human PSCs from which they were derived (including higher expression of NANOG, LEFTY1, and LEFTY2). In nonconditioned medium lacking FGF2, the colonies spontaneously differentiated along multiple lineages, including trophoblast. They responded to PD173074 in the absence of both FGF2 and BMP4 by conversion to trophoblast, and especially syncytiotrophoblast, whereas an A83-01/PD173074 combination favored increased expression of HLA-G, a marker of extravillous trophoblast. Together, these data suggest that the cell lines exhibit totipotent potential and that BMP4 can prime human PSCs to a self-renewing alternative state permissive for trophoblast development. The results may have implications for regulation of lineage decisions in the early embryo. PMID:25870291

  11. Revealing power in truth

    PubMed Central

    Lee, Kelley

    2015-01-01

    Jeremy Shiffman’s editorial appropriately calls on making all forms of power more apparent and accountable, notably productive power derived from expertise and claims to moral authority. This commentary argues that relationships based on productive power can be especially difficult to reveal in global health policy because of embedded notions about the nature of power and politics. Yet, it is essential to recognize that global health is shot through with power relationships, that they can take many forms, and that their explicit acknowledgement should be part of, rather than factored out of, any reform of global health governance. PMID:25844390

  12. The Universe Revealed

    NASA Astrophysics Data System (ADS)

    Spence, Pam

    1998-10-01

    The Universe is a bewildering place to the uninitiated. The concepts and theories that govern space seem complex and often contradictory. The Universe Revealed provides the keys to unlocking the wonders of the cosmos. Elegantly written and lavishly illustrated, it begins with the Sun and stretches through our solar system into deepest space. Lucid prose, written by many of the people who have shaped our current thinking on space, and spectacular photographs make the physics of the Universe accessible and provide a solid background for understanding the most recent astronomical discoveries. Covering the most intriguing features of the cosmos, the topics discussed range from the Earth and global warming to cosmic collisions and the size of the Universe. Major sections examine the Solar System, stars, galaxies, cosmology, and the observational techniques used by astronomers, both amateur and professional. The Universe Revealed represents the collaboration of internationally renowned experts in astronomy and cosmology, with contributions from authors including David Malin, F. Duccio Macchetto, Iain Nicholson, Neil Bone, Ian Ridpath, Seth Shostak, Mike Lancaster, Steve Miller, Ken Croswell, Geoff McNamara, and Steven Young. This extraordinary blend of astronomy, astrophysics, and cosmology, will appeal to amateur and armchair astronomers alike.

  13. High energy beam lines

    NASA Astrophysics Data System (ADS)

    Marchetto, M.; Laxdal, R. E.

    2014-01-01

    The ISAC post accelerator comprises an RFQ, DTL and SC-linac. The high energy beam lines connect the linear accelerators as well as deliver the accelerated beams to two different experimental areas. The medium energy beam transport (MEBT) line connects the RFQ to the DTL. The high energy beam transport (HEBT) line connects the DTL to the ISAC-I experimental stations (DRAGON, TUDA-I, GPS). The DTL to superconducting beam (DSB) transport line connects the ISAC-I and ISAC-II linacs. The superconducting energy beam transport (SEBT) line connects the SC linac to the ISAC-II experimental station (TUDA-II, HERACLES, TIGRESS, EMMA and GPS). All these lines have the function of transporting and matching the beams to the downstream sections by manipulating the transverse and longitudinal phase space. They also contain diagnostic devices to measure the beam properties.

  14. Series Transmission Line Transformer

    DOEpatents

    Buckles, Robert A.; Booth, Rex; Yen, Boris T.

    2004-06-29

    A series transmission line transformer is set forth which includes two or more of impedance matched sets of at least two transmissions lines such as shielded cables, connected in parallel at one end ans series at the other in a cascading fashion. The cables are wound about a magnetic core. The series transmission line transformer (STLT) which can provide for higher impedance ratios and bandwidths, which is scalable, and which is of simpler design and construction.

  15. Gusev's Rim Revealed

    NASA Technical Reports Server (NTRS)

    2004-01-01

    NASA's Mars Exploration Rover Spirit took this panoramic camera image on sol 91 (April 5, 2004). Spirit is looking to the southeast, and through the martian haze has captured the rim of Gusev Crater approximately 80 kilometers (49.7 miles) away on the horizon.

    The right side of this image reveals the portion of the crater edge that descends into the mouth of Ma'adim Vallis, a channel that opens into Gusev Crater. Spirit is currently traveling toward the informally named 'Columbia Hills,' which lie to the left of the region pictured here.

    This image is similar to a panoramic camera image taken on sol 68, but Gusev's ridge is more visible here because the atmospheric dust caused by winter dust storms has settled. Scientists expect to get even clearer images than this one in upcoming sols.

    This image has been modified to make the crater rim more visible.

  16. Birds on power lines

    NASA Astrophysics Data System (ADS)

    Arnaldo Redinz, José

    2014-07-01

    Why can a bird safely rest on a high-voltage power line? We discuss three effects that can lead to the development of voltages and currents in the bird's body. To explain the absence of electric shocks, we give numerical estimates of these voltages and currents obtained from the standard solution for the voltage along a two-wire transmission line.

  17. Structuring a service line.

    PubMed

    Zablocki, E

    1997-05-01

    In 1993, Community Hospitals Indianapolis created 10 service lines--each unique to the needs of the patients it serves. Two service line leaders talk about the factors critical to successful restructuring, the working relationships established, and the importance of clear communication to ensure understanding and get buy-in to the process. PMID:10167712

  18. Tangent Lines without Calculus

    ERIC Educational Resources Information Center

    Rabin, Jeffrey M.

    2008-01-01

    This article presents a problem that can help high school students develop the concept of instantaneous velocity and connect it with the slope of a tangent line to the graph of position versus time. It also gives a method for determining the tangent line to the graph of a polynomial function at any point without using calculus. (Contains 1 figure.)

  19. Transmission Line Security Monitor

    SciTech Connect

    2011-01-01

    The Transmission Line Security Monitor is a multi-sensor monitor that mounts directly on high-voltage transmission lines to detect, characterize and communicate terrorist activity, human tampering and threatening conditions around support towers. For more information about INL's critical infrastructure protection research, visit http://www.facebook.com/idahonationallaboratory.

  20. Transmission Line Security Monitor

    ScienceCinema

    None

    2013-05-28

    The Transmission Line Security Monitor is a multi-sensor monitor that mounts directly on high-voltage transmission lines to detect, characterize and communicate terrorist activity, human tampering and threatening conditions around support towers. For more information about INL's critical infrastructure protection research, visit http://www.facebook.com/idahonationallaboratory.

  1. Airborne Fraunhofer Line Discriminator

    NASA Technical Reports Server (NTRS)

    Gabriel, F. C.; Markle, D. A.

    1969-01-01

    Airborne Fraunhofer Line Discriminator enables prospecting for fluorescent materials, hydrography with fluorescent dyes, and plant studies based on fluorescence of chlorophyll. Optical unit design is the coincidence of Fraunhofer lines in the solar spectrum occurring at the characteristic wavelengths of some fluorescent materials.

  2. Database of emission lines

    NASA Astrophysics Data System (ADS)

    Binette, L.; Ortiz, P.; Joguet, B.; Rola, C.

    1998-11-01

    A widely accessible data bank (available through Netscape) and consiting of all (or most) of the emission lines reported in the litterature is being built. It will comprise objects as diverse as HII regions, PN, AGN, HHO. One of its use will be to define/refine existing diagnostic emission line diagrams.

  3. Hubble Images Reveal Jupiter's Auroras

    NASA Technical Reports Server (NTRS)

    1996-01-01

    These images, taken by the Hubble Space Telescope, reveal changes in Jupiter's auroral emissions and how small auroral spots just outside the emission rings are linked to the planet's volcanic moon, Io. The images represent the most sensitive and sharply-detailed views ever taken of Jovian auroras.

    The top panel pinpoints the effects of emissions from Io, which is about the size of Earth's moon. The black-and-white image on the left, taken in visible light, shows how Io and Jupiter are linked by an invisible electrical current of charged particles called a 'flux tube.' The particles - ejected from Io (the bright spot on Jupiter's right) by volcanic eruptions - flow along Jupiter's magnetic field lines, which thread through Io, to the planet's north and south magnetic poles. This image also shows the belts of clouds surrounding Jupiter as well as the Great Red Spot.

    The black-and-white image on the right, taken in ultraviolet light about 15 minutes later, shows Jupiter's auroral emissions at the north and south poles. Just outside these emissions are the auroral spots. Called 'footprints,' the spots are created when the particles in Io's 'flux tube' reach Jupiter's upper atmosphere and interact with hydrogen gas, making it fluoresce. In this image, Io is not observable because it is faint in the ultraviolet.

    The two ultraviolet images at the bottom of the picture show how the auroral emissions change in brightness and structure as Jupiter rotates. These false-color images also reveal how the magnetic field is offset from Jupiter's spin axis by 10 to 15 degrees. In the right image, the north auroral emission is rising over the left limb; the south auroral oval is beginning to set. The image on the left, obtained on a different date, shows a full view of the north aurora, with a strong emission inside the main auroral oval.

    The images were taken by the telescope's Wide Field and Planetary Camera 2 between May 1994 and September 1995.

    This image and

  4. Canada Basin revealed

    USGS Publications Warehouse

    Mosher, David C.; Shimeld, John; Hutchinson, Deborah R.; Chian, D; Lebedeva-Ivanova, Nina; Jackson, Ruth

    2012-01-01

    More than 15,000 line-km of new regional seismic reflection and refraction data in the western Arctic Ocean provide insights into the tectonic and sedimentologic history of Canada Basin, permitting development of new geologic understanding in one of Earth's last frontiers. These new data support a rotational opening model for southern Canada Basin. There is a central basement ridge possibly representing an extinct spreading center with oceanic crustal velocities and blocky basement morphology characteristic of spreading centre crust surrounding this ridge. Basement elevation is lower in the south, mostly due to sediment loading subsidence. The sedimentary succession is thickest in the southern Beaufort Sea region, reaching more than 15 km, and generally thins to the north and west. In the north, grabens and half-grabens are indicative of extension. Alpha-Mendeleev Ridge is a large igneous province in northern Amerasia Basin, presumably emplaced synchronously with basin formation. It overprints most of northern Canada Basin structure. The seafloor and sedimentary succession of Canada Basin is remarkably flat-lying in its central region, with little bathymetric change over most of its extent. Reflections that correlate over 100s of kms comprise most of the succession and on-lap bathymetric and basement highs. They are interpreted as representing deposits from unconfined turbidity current flows. Sediment distribution patterns reflect changing source directions during the basin’s history. Initially, probably late Cretaceous to Paleocene synrift sediments sourced from the Alaska and Mackenzie-Beaufort margins. This unit shows a progressive series of onlap unconformities with a younging trend towards Alpha and Northwind ridges, likely a response to contemporaneous subsidence. Sediment source direction appeared to shift to the Canadian Arctic Archipelago margin for the Eocene and Oligocene, likely due to uplift of Arctic islands during the Eurekan Orogeny. The final

  5. Nebular UV Absorption Lines in Planetary Nebulae

    NASA Astrophysics Data System (ADS)

    Dinerstein, Harriet

    We propose to continue our Cycle 1 program of studying the Lyman and Werner bands of H_2, seen in absorption against the UV continua of planetary nebula central stars, which arise within neutral-molecular envelopes surrounding the ionized gas. These are the pump lines for a fluorescent cascade of near-infrared emission lines which are observed in many planetary nebulae. By observing the UV lines we can probe the chemical and thermal structure of the envelopes, as well as measure molecular column densities and clarify the excitation processes for the infrared lines. In Cycle 1 we were granted time for three targets, one of which was successfully observed shortly before submission of this proposal. Although the data were not yet available for examination, similar target observed by the project team revealed a rich set of H_2 circumstellar absorption features, demonstrating the feasibility of our program. FUSE spectra also include absorption features from atomic species such as O I and C II, which give rise to important far-infrared fine-structure cooling lines that likewise have been observed from planetary nebulae. In Cycle 2, we add as a secondary goal a search for nebular components of the O VI 032, 1038 AA absorption lines, which trace the presence of hot shocked gas, in nebulae with anomalously strong optical recombination lines of ions of oxygen and nitrogen. This will test a plausible hypothesis for the origin of this anomaly.

  6. Coherent soliton communication lines

    SciTech Connect

    Yushko, O. V. Redyuk, A. A.; Fedoruk, M. P.; Turitsyn, S. K.

    2014-11-15

    The data transmission in coherent fiber-optical communication lines using solitons with a variable phase is studied. It is shown that nonlinear coherent structures (solitons) can be applied for effective signal transmission over a long distance using amplitude and optical-phase keying of information. The optimum ratio of the pulse width to the bit slot at which the spectral efficiency (transmitted bits per second and hertz) is maximal is determined. It is shown that soliton fiber-optical communication lines can ensure data transmission at a higher spectral efficiency as compared to traditional communication lines and at a high signal-to-noise ratio.

  7. Flow line sampler

    DOEpatents

    Nicholls, Colin I.

    1992-07-14

    An on-line product sampling apparatus and method for measuring product samples from a product stream (12) in a flow line (14) having a sampling aperture (11), includes a sampling tube (18) for containing product samples removed from flow line (14). A piston (22) removes product samples from the product stream (12) through the sampling aperture (11) and returns samples to product stream (12). A sensor (20) communicates with sample tube (18), and senses physical properties of samples while the samples are within sample tube (18). In one embodiment, sensor (20) comprises a hydrogen transient nuclear magnetic resonance sensor for measuring physical properties of hydrogen molecules.

  8. Automatic 3D power line reconstruction of multi-angular imaging power line inspection system

    NASA Astrophysics Data System (ADS)

    Zhang, Wuming; Yan, Guangjian; Wang, Ning; Li, Qiaozhi; Zhao, Wei

    2007-06-01

    We develop a multi-angular imaging power line inspection system. Its main objective is to monitor the relative distance between high voltage power line and around objects, and alert if the warning threshold is exceeded. Our multi-angular imaging power line inspection system generates DSM of the power line passage, which comprises ground surface and ground objects, for example trees and houses, etc. For the purpose of revealing the dangerous regions, where ground objects are too close to the power line, 3D power line information should be extracted at the same time. In order to improve the automation level of extraction, reduce labour costs and human errors, an automatic 3D power line reconstruction method is proposed and implemented. It can be achieved by using epipolar constraint and prior knowledge of pole tower's height. After that, the proper 3D power line information can be obtained by space intersection using found homologous projections. The flight experiment result shows that the proposed method can successfully reconstruct 3D power line, and the measurement accuracy of the relative distance satisfies the user requirement of 0.5m.

  9. Expression of Ribosomal RNA and Protein Genes in Human Embryonic Stem Cells Is Associated With the Activating H3K4me3 Histone Mark.

    PubMed

    Zaidi, Sayyed K; Boyd, Joseph R; Grandy, Rodrigo A; Medina, Ricardo; Lian, Jane B; Stein, Gary S; Stein, Janet L

    2016-09-01

    Embryonic stem cells (ESCs) exhibit unrestricted and indefinite, but stringently controlled, proliferation, and can differentiate into any lineage in the body. In the current study, we test the hypothesis that expression of ribosomal RNA (rRNA) and ribosomal protein genes (RPGs) contribute to the ability of hESCs to proliferate indefinitely. Consistent with the accelerated growth rate of hESCs, we find that hESC lines H1 and H9 both exhibit significantly higher levels of rRNA when compared to a panel of normal and cancer human cell lines. Although many RPGs are expressed at levels that comparable to other human cell lines, a few RPGs also exhibit higher expression levels. In situ nuclear run-on assays reveal that both nucleoli in hESCs actively transcribe nascent rRNA. Employing genome-wide chromatin immunoprecipitation-deep sequencing and bioinformatics approaches, we discovered that, RPGs are dominantly marked by the activating H3K4me3 histone mark in the G1, M, and G2 phases of the cell cycle. Interestingly, the rDNA repeats are marked by the activating H3K4me3 only in the M phase, and repressive H3K27me3 histone mark in all three cell cycle phases. Bioinformatics analyses also reveal that Myc, a known regulator of cell growth and proliferation, occupies both the rRNA genes and RPGs. Functionally, down-regulation of Myc expression by siRNA results in a concomitant decrease in rRNA levels. Together, our results show that expression of rRNA, which is regulated by the Myc pluripotency transcription factor, and of RPGs in hESCs is associated with the activating H3K4me3 modification. J. Cell. Physiol. 231: 2007-2013, 2016. © 2016 Wiley Periodicals, Inc. PMID:26755341

  10. Power line detection system

    DOEpatents

    Latorre, Victor R.; Watwood, Donald B.

    1994-01-01

    A short-range, radio frequency (RF) transmitting-receiving system that provides both visual and audio warnings to the pilot of a helicopter or light aircraft of an up-coming power transmission line complex. Small, milliwatt-level narrowband transmitters, powered by the transmission line itself, are installed on top of selected transmission line support towers or within existing warning balls, and provide a continuous RF signal to approaching aircraft. The on-board receiver can be either a separate unit or a portion of the existing avionics, and can also share an existing antenna with another airborne system. Upon receipt of a warning signal, the receiver will trigger a visual and an audio alarm to alert the pilot to the potential power line hazard.

  11. Power line detection system

    DOEpatents

    Latorre, V.R.; Watwood, D.B.

    1994-09-27

    A short-range, radio frequency (RF) transmitting-receiving system that provides both visual and audio warnings to the pilot of a helicopter or light aircraft of an up-coming power transmission line complex. Small, milliwatt-level narrowband transmitters, powered by the transmission line itself, are installed on top of selected transmission line support towers or within existing warning balls, and provide a continuous RF signal to approaching aircraft. The on-board receiver can be either a separate unit or a portion of the existing avionics, and can also share an existing antenna with another airborne system. Upon receipt of a warning signal, the receiver will trigger a visual and an audio alarm to alert the pilot to the potential power line hazard. 4 figs.

  12. Line, Shape, Color.

    ERIC Educational Resources Information Center

    Greenman, Geri

    2002-01-01

    Describes an art project used with beginning high school art students that teaches them about continuous line drawing. Explains that the students create portraits of themselves, or another student, using glue, black construction paper, and chalk. (CMK)

  13. IRIS Spectrum Line Plot

    NASA Video Gallery

    This video shows a line plot of the spectrum. The spectra here are shown for various locations on the Sun. The changes in the movie are caused by differing physical conditions in the locations. Cre...

  14. Central line infections - hospitals

    MedlinePlus

    ... risk is higher if you: Are in the intensive care unit (ICU) Have a weakened immune system or serious ... unless you have washed your hands. Tell your nurse if your central line: Gets dirty Is coming ...

  15. Veterans Crisis Line

    MedlinePlus

    ... also access and download the Veterans Crisis Line Branding Guidelines for guidance on how to consistently apply ... Program ADMINISTRATION Veterans Health Administration Veterans Benefits Administration National Cemetery Administration U.S. Department of Veterans Affairs | 810 ...

  16. On-Line and Off-Line Assessment of Metacognition

    ERIC Educational Resources Information Center

    Saraç, Seda; Karakelle, Sema

    2012-01-01

    The study investigates the interrelationships between different on-line and off-line measures for assessing metacognition. The participants were 47 fifth grade elementary students. Metacognition was assessed through two off-line and two on-line measures. The off-line measures consisted of a teacher rating scale and a self-report questionnaire. The…

  17. PORTION OF A LINE (AT LEFT) AND B LINE (AT ...

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

    PORTION OF A LINE (AT LEFT) AND B LINE (AT RIGHT) ON GENTLE TERRACE SLOPE. A LINE POLE 75A (LEFT FOREGROUND) HAS ORIGINAL GLASS PIN-TYPE INSULATORS AND INTACT COMMUNICATION LINE CROSS ARM. VIEW TO WEST-SOUTHWEST. - Mystic Lake Hydroelectric Facility, Electric Transmission A Line, Along West Rosebud Creek, Fishtail, Stillwater County, MT

  18. Rapid bottom melting widespread near Antarctic ice sheet grounding lines

    NASA Technical Reports Server (NTRS)

    Rignot, E.; Jacobs, S.

    2002-01-01

    As continental ice from Antartica reaches the grounding line and begins to float, its underside melts into the ocean. Results obtained with satellite radar interferometry reveal that bottom melt rates experienced by large outlet glaciers near their grounding lines are far higher than generally assumed.

  19. Nasca Lines, Peru

    NASA Technical Reports Server (NTRS)

    2001-01-01

    The Nasca Lines are located in the Pampa region of Peru, the desolate plain of the Peruvian coast 400 km south of Lima. The Lines were first spotted when commercial airlines began flying across the Peruvian desert in the 1920's. Passengers reported seeing 'primitive landing strips' on the ground below. The Lines were made by removing the iron-oxide coated pebbles which cover the surface of the desert. When the gravel is removed, they contrast with the light color underneath. In this way the lines were drawn as furrows of a lighter color. On the pampa, south of the Nasca Lines, archaeologists have now uncovered the lost city of the line-builders, Cahuachi. It was built nearly two thousand years ago and was mysteriously abandoned 500 years later. This ASTER sub-image covers an area of 14 x 18 km, was acquired on December 22, 2000, and is located at 14.7 degrees south latitude and 75.1 degrees west longitude.

    The U.S. science team is located at NASA's Jet Propulsion Laboratory, Pasadena, Calif. The Terra mission is part of NASA's Science Mission Directorate.

  20. Wear-Lines and Split-Lines of Human Patellar Cartilage: Relation to Tensile Biomechanical Properties

    PubMed Central

    Bae, Won C.; Wong, Van W.; Hwang, Jennifer; Antonacci, Jennifer M.; Nugent-Derfus, Gayle E.; Blewis, Megan E.; Temple-Wong, Michele M.; Sah, Robert L.

    2008-01-01

    Background Articular cartilage undergoes age-associated degeneration, resulting in both structural and functional biomechanical changes. At early stages of degeneration, wear-lines develop in the general direction of joint movement. With aging, cartilage exhibits a decrease in tensile modulus. The tensile modulus of cartilage has also been related to the orientation of the collagen network, as revealed by split-lines. Objective To determine the relative contribution of wear-line and split-line orientation on the tensile biomechanical properties of human patellar cartilage from different depths. Methods In human patellar cartilage, wear- and split-lines are aligned parallel to each other at the proximal facet, and perpendicular to each other at the medial facet. Using superficial, middle, and deep cartilage sections from these two sites, tensile samples were prepared in two orthogonal orientations. Thus, for each depth, there were four groups of samples, with their long axes were aligned either parallel or perpendicular to wear-line direction and also aligned parallel or perpendicular to split-line direction. Uniaxial tensile tests were performed to assess equilibrium and ramp moduli. Results Tensile equilibrium modulus varied with wear-line orientation (p<0.05) and depth (p<0.001), in an interactive manner (p<0.05), and tended to vary with split-line orientation (p=0.16). In the superficial layer, equilibrium and ramp modulus was higher when the samples were loaded parallel to wear-lines (p<0.05). Conclusion These results indicate that mild wear (i.e., wear-line formation) at the articular surface has deleterious functional effects on articular cartilage and represents an early aging associated degenerative change. The identification and recognition of functional biomechanical consequences of wear-lines is useful for planning and interpreting tensile biomechanical tests in human articular cartilage. PMID:18248747

  1. Titan Casts Revealing Shadow

    NASA Astrophysics Data System (ADS)

    2004-05-01

    A rare celestial event was captured by NASA's Chandra X-ray Observatory as Titan -- Saturn's largest moon and the only moon in the Solar System with a thick atmosphere -- crossed in front of the X-ray bright Crab Nebula. The X-ray shadow cast by Titan allowed astronomers to make the first X-ray measurement of the extent of its atmosphere. On January 5, 2003, Titan transited the Crab Nebula, the remnant of a supernova explosion that was observed to occur in the year 1054. Although Saturn and Titan pass within a few degrees of the Crab Nebula every 30 years, they rarely pass directly in front of it. "This may have been the first transit of the Crab Nebula by Titan since the birth of the Crab Nebula," said Koji Mori of Pennsylvania State University in University Park, and lead author on an Astrophysical Journal paper describing these results. "The next similar conjunction will take place in the year 2267, so this was truly a once in a lifetime event." Animation of Titan's Shadow on Crab Nebula Animation of Titan's Shadow on Crab Nebula Chandra's observation revealed that the diameter of the X-ray shadow cast by Titan was larger than the diameter of its solid surface. The difference in diameters gives a measurement of about 550 miles (880 kilometers) for the height of the X-ray absorbing region of Titan's atmosphere. The extent of the upper atmosphere is consistent with, or slightly (10-15%) larger, than that implied by Voyager I observations made at radio, infrared, and ultraviolet wavelengths in 1980. "Saturn was about 5% closer to the Sun in 2003, so increased solar heating of Titan may account for some of this atmospheric expansion," said Hiroshi Tsunemi of Osaka University in Japan, one of the coauthors on the paper. The X-ray brightness and extent of the Crab Nebula made it possible to study the tiny X-ray shadow cast by Titan during its transit. By using Chandra to precisely track Titan's position, astronomers were able to measure a shadow one arcsecond in

  2. Line Lengths and Starch Scores.

    ERIC Educational Resources Information Center

    Moriarty, Sandra E.

    1986-01-01

    Investigates readability of different line lengths in advertising body copy, hypothesizing a normal curve with lower scores for shorter and longer lines, and scores above the mean for lines in the middle of the distribution. Finds support for lower scores for short lines and some evidence of two optimum line lengths rather than one. (SKC)

  3. Revealing the Beast Within

    NASA Astrophysics Data System (ADS)

    2003-07-01

    Deeply Embedded Massive Stellar Clusters Discovered in Milky Way Powerhouse Summary Peering into a giant molecular cloud in the Milky Way galaxy - known as W49 - astronomers from the European Southern Observatory (ESO) have discovered a whole new population of very massive newborn stars . This research is being presented today at the International Astronomical Union's 25th General Assembly held in Sydney, Australia, by ESO-scientist João Alves. With the help of infrared images obtained during a period of excellent observing conditions with the ESO 3.5-m New Technology Telescope (NTT) at the La Silla Observatory (Chile), the astronomers looked deep into this molecular cloud and discovered four massive stellar clusters, with hot and energetic stars as massive as 120 solar masses. The exceedingly strong radiation from the stars in the largest of these clusters is "powering" a 20 light-year diameter region of mostly ionized hydrogen gas (a "giant HII region"). W49 is one of the most energetic regions of star formation in the Milky Way. With the present discovery, the true sources of the enormous energy have now been revealed for the first time, finally bringing to an end some decades of astronomical speculations and hypotheses. PR Photo 21a/03 : Colour Composite of W49A (NTT+SOFI). PR Photo 21b/03 : Radio and Near-Infrared Composite of W49A Giant molecular clouds Stars form predominantly inside Giant Molecular Clouds which populate our Galaxy, the Milky Way. One of the most prominent of these is W49 , which has a mass of a million solar masses. It is located some 37,000 light-years away and is the most luminous star-forming region known in our home galaxy: its luminosity is several million times the luminosity of our Sun. A smaller region within this cloud is denoted W49A - this is one of the strongest radio-emitting areas known in the Galaxy . Massive stars are excessive in all ways. Compared to their smaller and ligther brethren, they form at an Olympic speed and

  4. EVOLUTION OF [O III] {lambda}5007 EMISSION-LINE PROFILES IN NARROW EMISSION-LINE GALAXIES

    SciTech Connect

    Wang, J.; Mao, Y. F.; Wei, J. Y.

    2011-11-01

    The active galactic nucleus (AGN)-host co-evolution issue is investigated here by focusing on the evolution of the [O III] {lambda}5007 emission-line profile. A large sample of narrow emission-line galaxies is selected from the Max-Planck Institute for Astrophysics/Johns Hopkins University Sloan Digital Sky Survey DR7 catalog to simultaneously measure both the [O III] line profile and circumnuclear stellar population in an individual spectrum. By requiring that (1) the [O III] line signal-to-noise ratio is larger than 30 and (2) the [O III] line width is larger than the instrumental resolution by a factor of two, our sample is narrowed down to 2333 Seyfert galaxies/LINERs (AGNs), 793 transition galaxies, and 190 star-forming galaxies. In addition to the commonly used profile parameters (i.e., line centroid, relative velocity shift, and velocity dispersion), two dimensionless shape parameters, skewness and kurtosis, are used to quantify the line shape deviation from a pure Gaussian function. We show that the transition galaxies are systematically associated with narrower line widths and weaker [O III] broad wings than the AGNs, which implies that the kinematics of emission-line gas are different in the two kinds of objects. By combining the measured host properties and line shape parameters, we find that the AGNs with stronger blue asymmetries tend to be associated with younger stellar populations. However, a similar trend is not identified in the transition galaxies. The failure likely results from a selection effect in which the transition galaxies are systematically associated with younger stellar populations than the AGNs. The evolutionary significance revealed here suggests that both narrow-line region kinematics and outflow feedback in AGNs co-evolve with their host galaxies.

  5. Evolution of [O III] λ5007 Emission-line Profiles in Narrow Emission-line Galaxies

    NASA Astrophysics Data System (ADS)

    Wang, J.; Mao, Y. F.; Wei, J. Y.

    2011-11-01

    The active galactic nucleus (AGN)-host co-evolution issue is investigated here by focusing on the evolution of the [O III] λ5007 emission-line profile. A large sample of narrow emission-line galaxies is selected from the Max-Planck Institute for Astrophysics/Johns Hopkins University Sloan Digital Sky Survey DR7 catalog to simultaneously measure both the [O III] line profile and circumnuclear stellar population in an individual spectrum. By requiring that (1) the [O III] line signal-to-noise ratio is larger than 30 and (2) the [O III] line width is larger than the instrumental resolution by a factor of two, our sample is narrowed down to 2333 Seyfert galaxies/LINERs (AGNs), 793 transition galaxies, and 190 star-forming galaxies. In addition to the commonly used profile parameters (i.e., line centroid, relative velocity shift, and velocity dispersion), two dimensionless shape parameters, skewness and kurtosis, are used to quantify the line shape deviation from a pure Gaussian function. We show that the transition galaxies are systematically associated with narrower line widths and weaker [O III] broad wings than the AGNs, which implies that the kinematics of emission-line gas are different in the two kinds of objects. By combining the measured host properties and line shape parameters, we find that the AGNs with stronger blue asymmetries tend to be associated with younger stellar populations. However, a similar trend is not identified in the transition galaxies. The failure likely results from a selection effect in which the transition galaxies are systematically associated with younger stellar populations than the AGNs. The evolutionary significance revealed here suggests that both narrow-line region kinematics and outflow feedback in AGNs co-evolve with their host galaxies.

  6. Blasted Cell Line Names

    PubMed Central

    Wang, Jing; Byers, Lauren A.; Yordy, John S.; Liu, Wenbin; Shen, Li; Baggerly, Keith A.; Giri, Uma; Myers, Jeffrey N.; Ang, K. Kian; Story, Michael D.; Girard, Luc; Minna, John D.; Heymach, John V.; Coombes, Kevin R.

    2010-01-01

    Background: While trying to integrate multiple data sets collected by different researchers, we noticed that the sample names were frequently entered inconsistently. Most of the variations appeared to involve punctuation, white space, or their absence, at the juncture between alphabetic and numeric portions of the cell line name. Results: Reasoning that the variant names could be described in terms of mutations or deletions of character strings, we implemented a simple version of the Needleman-Wunsch global sequence alignment algorithm and applied it to the cell line names. All correct matches were found by this procedure. Incorrect matches only occured when a cell line was present in one data set but not in the other. The raw match scores tended to be substantially worse for the incorrect matches. Conclusions: A simple application of the Needleman-Wunsch global sequence alignment algorithm provides a useful first pass at matching sample names from different data sets. PMID:21082038

  7. Circumstellar radio molecular lines

    NASA Technical Reports Server (NTRS)

    NGUYEN-QUANG-RIEU

    1987-01-01

    Radio molecular lines appear to be useful probes into the stellar environment. Silicon oxide masers provide information on the physical conditions in the immediate vicinity of the stellar photosphere. Valuable information on the physics operating in the envelope of IRC + 10216 was recently obtained by high sensitivity observations and detailed theoretical analyses. Infrared speckle interferometry in the molecular lines and in the continuum is helpful in the investigation of the inner region of the envelope. These techniques are discussed in terms of late-type star mass loss.

  8. Revealing Evidence of Spin in Galactic Black Holes

    NASA Astrophysics Data System (ADS)

    Miller, J. M.; Fabian, A. C.; Reynolds, C. S.; Nowak, M.; Wijnands, R.; Homan, J.; Belloni, T.; van der Klis, M.; Lewin, W. H. G.

    2002-12-01

    Our observations of Galactic black holes with Chandra and XMM-Newton have recently revealed broad, skewed Fe K-alpha emission lines similar to those seen in some AGN. If such lines are produced near the inner edge of the accretion disk, they can serve as an excellent spin diagnostic. Our analysis of the XMM-Newton spectrum of XTE J1650-500 reveals a strong, skewed Fe K-alpha emission line that requires near-maximal spin; with RXTE, we have discovered a 250 Hz QPO in this source which may also be produced near to the black hole. Although many Galactic black holes have been studied extensively, it is extremely rare that spectroscopic and timing studies both reveal features plausibly associated with the inner accretion disk. We will present the results of our XMM-Newton, Chandra, and RXTE observations of XTE J1650-500, as well as the results of our Chandra observations of Cygnus X-1 and preliminary results from our observations of GX 339-4 with XMM-Newton. We will discuss these results within the context of the relativistic Fe K lines reported by our group and others, and comment on the prospects for revealing evidence of spin in future observations. (JMM acknowledges support from the NSF through the Astronomy and Astrophysics Fellowship program.)

  9. Microwave spectral line listing

    NASA Technical Reports Server (NTRS)

    White, W. F., Jr.

    1975-01-01

    The frequency, intensity, and identification of 9615 spectral lines belonging to 75 molecules are tabulated in order of increasing frequency. Measurements for all 75 molecules were made in the frequency range from 26500 to 40000 MHz by a computer controlled spectrometer. Measurements were also made in the 18000 to 26500 MHz range for some of the molecules.

  10. Straight Line Graphs

    ERIC Educational Resources Information Center

    Krueger, Tom

    2010-01-01

    In this article, the author shares one effective lesson idea on straight line graphs that he applied in his lower ability Y9 class. The author wanted something interesting for his class to do, something that was fun and engaging with direct feedback, and something that worked because someone else had tried it before. In a word, the author admits…

  11. Cracks and Lines

    NASA Technical Reports Server (NTRS)

    2004-01-01

    6 June 2004 This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) picture shows an odd area of the south polar region that has sets of fine, nearly parallel lines running from the northeast (upper right) toward southwest (lower left) and a darker, wider set of cracks with a major trend running almost perpendicular to the finer lines. The appearance of these features is enhanced by seasonal frost. Dark areas have no frost, bright areas still have frozen carbon dioxide ice. In summer, the ice would be gone and the cracks and lines less obvious when viewed from orbit. Although unknown, wind might be responsible for forming the fine set of lines, and perhaps freeze-thaw cycles of ground ice or structural deformation would have contributed to formation of the wider cracks. The image is located near 85.0oS, 324.0oW, and covers an area about 1.5 km (nearly 1 mi) across. The scene is illuminated by sunlight from the upper left.

  12. Control line sealing connection

    SciTech Connect

    Tohill, H.O.

    1984-07-10

    A sealing connection for the passage of a fluid control line through adjoining members is claimed. The connection comprises a metallic tubular sealing element provided with tapered end portions, each end having internal frusto-conical surfaces and external frusto-conical surfaces in coaxial alignment with the bore through the element. The external frusto-conical surfaceas provide metal-to-metal sealing with aligned frusto-conical seating surfaces in opposed pockets formed in aligned portions of the control line at their respective openings at the adjoining surfaces of the adjoined member. The tubular sealing element is subjected to axial compression between the frusto-conical pocket seating surfaces when the adjoined members are bolted tightly together which results in deformation of its tapered end portions to provide metal-to-metal sealing with the adjoined members which is effective to seal against both internal and external fluid pressurization of the control line and without internally obstructing or restricting the control line. Secondary sealing is provided by elastomeric O-ring seals mounted in circumferential annular grooves formed in the exterior of the tubular sealing element.

  13. Building with a Line

    ERIC Educational Resources Information Center

    Hubbert, Beth

    2011-01-01

    Architecture is a versatile, multifaceted area to study in the artroom with multiple age levels. It can easily stimulate a study of basic line, shape, and various other art elements and principles. It can then be extended into a more extensive study of architectural elements, styles, specific architects, architecture of different cultures, and…

  14. The Front Line.

    ERIC Educational Resources Information Center

    Unks, Gerald

    1979-01-01

    The author draws an analogy between today's school system and an assembly line, deploring the notion that all children are taught the same thing at the same time, ending in humiliation, disgrace, and failure for some, and nonchallenging academic activities for others. (KC)

  15. Giant Broad Line Regions in Dwarf Seyferts

    NASA Astrophysics Data System (ADS)

    Devereux, Nicholas A.

    2016-01-01

    High angular resolution spectroscopy obtained with the Hubble Space Telescope has revealed a remarkable population of galaxies hosting dwarf Seyfert nuclei with an unusually large broad-line region (BLR). These objects are remarkable for two reasons. Firstly, the size of the BLR can, in some cases, rival those seen in the most luminous quasars. Secondly, the size of the BLR is not correlated with the central continuum luminosity, an observation that distinguishes them from their reverberating counterparts. Collectively, these early results suggest that non-reverberating dwarf Seyferts are a heterogeneous group and not simply scaled versions of each other. Careful inspection reveals broad H Balmer emission lines with single peaks, double peaks, and a combination of the two, suggesting that the broad emission lines are produced in kinematically distinct regions centered on the black hole (BH). Because the gravitational field strength is already known for these objects, by virtue of knowing their BH mass, the relationship between velocity and radius may be established, given a kinematic model for the BLR gas. In this way, one can determine the inner and outer radii of the BLRs by modeling the shape of their broad emission line profiles. In the present contribution, high quality spectra obtained with the Space Telescope Imaging Spectrograph are used to constrain the size of the BLR in the dwarf Seyfert nuclei of M81, NGC 3998, NGC 4203, NGC 3227, NGC 4051, and NGC 3516.

  16. Giant Broad Line Regions in Dwarf Seyferts

    NASA Astrophysics Data System (ADS)

    Devereux, Nick

    2015-12-01

    High angular resolution spectroscopy obtained with the Hubble Space Telescope (HST) has revealed a remarkable population of galaxies hosting dwarf Seyfert nuclei with an unusually large broad-line region (BLR). These objects are remarkable for two reasons. Firstly, the size of the BLR can, in some cases, rival those seen in the most luminous quasars. Secondly, the size of the BLR is not correlated with the central continuum luminosity, an observation that distinguishes them from their reverberating counterparts. Collectively, these early results suggest that non-reverberating dwarf Seyferts are a heterogeneous group, and not simply scaled versions of each other. Careful inspection reveals broad H Balmer emission lines with single peaks, double peaks, and a combination of the two, suggesting that the broad emission lines are produced in kinematically distinct regions centered on the black hole (BH). Because the gravitational field strength is already known for these objects, by virtue of knowing their BH mass, the relationship between velocity and radius may be established, given a kinematic model for the BLR gas. In this way, one can determine the inner and outer radii of the BLRs by modeling the shape of their broad emission line profiles. In the present contribution, high quality spectra obtained with the Space Telescope Imaging Spectrograph (STIS) are used to constrain the size of the BLR in the dwarf Seyfert nuclei of M81, NGC 3998, NGC 4203, NGC 3227, NGC 4051 and NGC 3516.

  17. A Quantitative Transmission Line Experiment

    ERIC Educational Resources Information Center

    Johnston, D. C.; Silbernagel, B. G.

    1969-01-01

    Describes modifications of a commercially available strip-type transmission line, which makes possible reproducible measurements of standing waves on the line. Experimental data yield values for the characteristic impedance, phase velocity and line wavelength of radiation in the transmission line, and the dielectric constant of material in the…

  18. The Bicycle Assembly Line Game

    ERIC Educational Resources Information Center

    Klotz, Dorothy

    2011-01-01

    "The Bicycle Assembly Line Game" is a team-based, in-class activity that helps students develop a basic understanding of continuously operating processes. Each team of 7-10 students selects one of seven prefigured bicycle assembly lines to operate. The lines are run in real-time, and the team that operates the line that yields the…

  19. Printed circuit dispersive transmission line

    DOEpatents

    Ikezi, H.; Lin-Liu, Y.R.; DeGrassie, J.S.

    1991-08-27

    A printed circuit dispersive transmission line structure is disclosed comprising an insulator, a ground plane formed on one surface of the insulator, a first transmission line formed on a second surface of the insulator, and a second transmission line also formed on the second surface of the insulator and of longer length than the first transmission line and periodically intersecting the first transmission line. In a preferred embodiment, the transmission line structure exhibits highly dispersive characteristics by designing the length of one of the transmission line between two adjacent periodic intersections to be longer than the other. 5 figures.

  20. Printed circuit dispersive transmission line

    DOEpatents

    Ikezi, Hiroyuki; Lin-Liu, Yuh-Ren; DeGrassie, John S.

    1991-01-01

    A printed circuit dispersive transmission line structure is disclosed comprising an insulator, a ground plane formed on one surface of the insulator, a first transmission line formed on a second surface of the insulator, and a second transmission line also formed on the second surface of the insulator and of longer length than the first transmission line and periodically intersecting the first transmission line. In a preferred embodiment, the transmission line structure exhibits highly dispersive characteristics by designing the length of one of the transmission line between two adjacent periodic intersections to be longer than the other.

  1. The human embryonic stem cell proteome revealed by multidimensional fractionation followed by tandem mass spectrometry

    PubMed Central

    Zhao, Peng; Schulz, Thomas C.; Sherrer, Eric S.; Weatherly, D. Brent; Robins, Allan J.; Wells, Lance

    2015-01-01

    Human embryonic stem cells (hESCs) have received considerable attention due to their therapeutic potential and usefulness in understanding early development and cell fate commitment. In order to appreciate the unique properties of these pluripotent, self-renewing cells, we have performed an in-depth multidimensional fractionation followed by LC-MS/MS analysis of the hESCs harvested from defined media to elucidate expressed, phosphorylated, O-linked β-N-acetylglucosamine (O-GlcNAc) modified, and secreted proteins. From the triplicate analysis, we were able to assign more than 3000 proteins with less than 1% false-discovery rate. This analysis also allowed us to identify nearly 500 phosphorylation sites and 68 sites of O-GlcNAc modification with the same high confidence. Investigation of the phosphorylation sites allowed us to deduce the set of kinases that are likely active in these cells. We also identified more than 100 secreted proteins of hESCs that likely play a role in extracellular matrix formation and remodeling, as well as autocrine signaling for self-renewal and maintenance of the undifferentiated state. Finally, by performing in-depth analysis in triplicate, spectral counts were obtained for these proteins and posttranslationally modified peptides, which will allow us to perform relative quantitative analysis between these cells and any derived cell type in the future. PMID:25367160

  2. The human embryonic stem cell proteome revealed by multidimensional fractionation followed by tandem mass spectrometry.

    PubMed

    Zhao, Peng; Schulz, Thomas C; Sherrer, Eric S; Weatherly, D Brent; Robins, Allan J; Wells, Lance

    2015-01-01

    Human embryonic stem cells (hESCs) have received considerable attention due to their therapeutic potential and usefulness in understanding early development and cell fate commitment. In order to appreciate the unique properties of these pluripotent, self-renewing cells, we have performed an in-depth multidimensional fractionation followed by LC-MS/MS analysis of the hESCs harvested from defined media to elucidate expressed, phosphorylated, O-linked β-N-acetylglucosamine (O-GlcNAc) modified, and secreted proteins. From the triplicate analysis, we were able to assign more than 3000 proteins with less than 1% false-discovery rate. This analysis also allowed us to identify nearly 500 phosphorylation sites and 68 sites of O-GlcNAc modification with the same high confidence. Investigation of the phosphorylation sites allowed us to deduce the set of kinases that are likely active in these cells. We also identified more than 100 secreted proteins of hESCs that likely play a role in extracellular matrix formation and remodeling, as well as autocrine signaling for self-renewal and maintenance of the undifferentiated state. Finally, by performing in-depth analysis in triplicate, spectral counts were obtained for these proteins and posttranslationally modified peptides, which will allow us to perform relative quantitative analysis between these cells and any derived cell type in the future. PMID:25367160

  3. Coolant line hydrometer

    SciTech Connect

    Barber, M.D.; Kipp, W.G.

    1987-03-17

    This patent describes a hydrometer unit for connection in an automobile coolant flow line comprising: a tubular fitting adapted to be connected to the coolant flow line; a coolant receiving chamber means connected to the tubular fitting for receiving coolant from the tubular fitting; and indicating float elements contained within the coolant receiving chamber means and adapted to rise therein individually as a function of the specific gravity of the coolant. The coolant receiving chamber means includes a closure cap which when connected to the tubular fitting forms a coolant receiving chamber, retaining means for retaining the indicating float elements within the coolant receiving chamber, a viewing window member of a substantially clear material through which the float elements can be visually observed within the coolant receiving chamber means, and air venturi means located within the coolant receiving chamber means for automatically removing air which may collect within the coolant chamber means.

  4. Central line complications

    PubMed Central

    Kornbau, Craig; Lee, Kathryn C; Hughes, Gwendolyn D; Firstenberg, Michael S

    2015-01-01

    Central venous access is a common procedure performed in many clinical settings for a variety of indications. Central lines are not without risk, and there are a multitude of complications that are associated with their placement. Complications can present in an immediate or delayed fashion and vary based on type of central venous access. Significant morbidity and mortality can result from complications related to central venous access. These complications can cause a significant healthcare burden in cost, hospital days, and patient quality of life. Advances in imaging, access technique, and medical devices have reduced and altered the types of complications encountered in clinical practice; but most complications still center around vascular injury, infection, and misplacement. Recognition and management of central line complications is important when caring for patients with vascular access, but prevention is the ultimate goal. This article discusses common and rare complications associated with central venous access, as well as techniques to recognize, manage, and prevent complications. PMID:26557487

  5. CRUCIBLE LINING METHOD

    DOEpatents

    Bone, W.H.; Schmidt, W.W.

    1958-11-01

    A method is presented for forming refractory liners in cylindrical reaction vessels used for the reductlon of uranium tetrafluoride to metallic uranium. A preliminary form, having positioning lugs attached thereto, is inserted into the reaction vessel and the refractory powder, usually CaO, is put in the annular space between the form and the inner wall of the reaction vessel. A jolting table is used to compact this charge of liner material ln place, and after thls has been done, the preliminary form is removed and the flnal form or plug is lnserted without disturbing the partially completed lining. The remainder of the lining charge is then introduced and compacted by jolting, after which the form is removed.

  6. Cryoresistive gas insulated line

    NASA Astrophysics Data System (ADS)

    Hidaka, K.; Matsumoto, S.; Kouno, T.

    An insulation strength of more than 16 kV mm -1 has been realized using nitrogen gas at 133 K and 0.4 MPa with the application of a.c. and impulse voltages. A cryoresistive gas insulated line (CRGIL) using nitrogen gas is proposed on the basis of the measured insulation strength. An efficient method for use of the cooling heat of liquefied natural gas is also proposed so that no refrigerator is required and the cost of cooling is reduced. The CRGIL has the advantages of having a large current carrying capacity, small power loss and small capacitance. Moreover, a given transmission capacity is achieved by a CRGIL whose rated voltage is about half that in a conventional gas insulated line.

  7. Drill string transmission line

    DOEpatents

    Hall, David R.; Hall, Jr., H. Tracy; Pixton, David S.; Bradford, Kline; Fox, Joe

    2006-03-28

    A transmission line assembly for transmitting information along a downhole tool comprising a pin end, a box end, and a central bore traveling between the pin end and the box end, is disclosed in one embodiment of the invention as including a protective conduit. A transmission line is routed through the protective conduit. The protective conduit is routed through the central bore and the ends of the protective conduit are routed through channels formed in the pin end and box end of the downhole tool. The protective conduit is elastically forced into a spiral or other non-linear path along the interior surface of the central bore by compressing the protective conduit to a length within the downhole tool shorter than the protective conduit.

  8. A comparison of fluoropolymer linings

    SciTech Connect

    Khaladkar, P.R. )

    1994-02-01

    Five established methods of applying fluoropolymer to metallic and fiber-reinforced plastic (FRP) substrates of barrier or lining applications include: (1) loose linings; (2) rotolining; (3) adhesively bonded linings; (4) spray and baked linings; and (5) dual laminate with FRP linings. With their low strength and high creep rates, fluoropolymers most often are used as barriers on substrates. The article offers some guidelines on the selection of suitable options for given applications.

  9. Linear Polarization Measurements of Chromospheric Emission Lines

    NASA Technical Reports Server (NTRS)

    Sheeley, N. R., Jr.; Keller, C. U.

    2003-01-01

    We have used the Zurich Imaging Stokes Polarimeter (ZIMPOL I) with the McMath-Pierce 1.5 m main telescope on Kitt Peak to obtain linear polarization measurements of the off-limb chromosphere with a sensitivity better than 1 x 10(exp -5). We found that the off-disk observations require a combination of good seeing (to show the emission lines) and a clean heliostat (to avoid contamination by scattered light from the Sun's disk). When these conditions were met, we obtained the following principal results: 1. Sometimes self-reversed emission lines of neutral and singly ionized metals showed linear polarization caused by the transverse Zeeman effect or by instrumental cross talk from the longitudinal Zeeman effect in chromospheric magnetic fields. Otherwise, these lines tended to depolarize the scattered continuum radiation by amounts that ranged up to 0.2%. 2. Lines previously known to show scattering polarization just inside the limb (such as the Na I lambda5889 D2 and the He I lambda5876 D3 lines) showed even more polarization above the Sun's limb, with values approaching 0.7%. 3. The O I triplet at lambda7772, lambda7774, and lambda7775 showed a range of polarizations. The lambda7775 line, whose maximum intrinsic polarizability, P(sub max), is less than 1%, revealed mainly Zeeman contributions from chromospheric magnetic fields. However, the more sensitive lambda7772 (P(sub max) = 19%) and lambda7774 (P(sub max) = 29%) lines had relatively strong scattering polarizations of approximately 0.3% in addition to their Zeeman polarizations. At times of good seeing, the polarization spectra resolve into fine structures that seem to be chromospheric spicules.

  10. Vertical bloch line memory

    NASA Technical Reports Server (NTRS)

    Katti, Romney R. (Inventor); Stadler, Henry L. (Inventor); Wu, Jiin-chuan (Inventor)

    1995-01-01

    A new read gate design for the vertical Bloch line (VBL) memory is disclosed which offers larger operating margin than the existing read gate designs. In the existing read gate designs, a current is applied to all the stripes. The stripes that contain a VBL pair are chopped, while the stripes that do not contain a VBL pair are not chopped. The information is then detected by inspecting the presence or absence of the bubble. The margin of the chopping current amplitude is very small, and sometimes non-existent. A new method of reading Vertical Bloch Line memory is also disclosed. Instead of using the wall chirality to separate the two binary states, the spatial deflection of the stripe head is used. Also disclosed herein is a compact memory which uses vertical Bloch line (VBL) memory technology for providing data storage. A three-dimensional arrangement in the form of stacks of VBL memory layers is used to achieve high volumetric storage density. High data transfer rate is achieved by operating all the layers in parallel. Using Hall effect sensing, and optical sensing via the Faraday effect to access the data from within the three-dimensional packages, an even higher data transfer rate can be achieved due to parallel operation within each layer.

  11. Line managers as marketers.

    PubMed

    Rynne, T J

    1986-09-01

    Many hospital administrators and boards of directors, having overcome their initial opposition to advertising and hired a marketing professional, are wondering why their marketing programs have not accomplished all that they had hoped. The answer lies in their perception of who is responsible for marketing. Marketing is not a specialty separate from the disciplines of management--it is a basic management skill. Its goal is to satisfy the customer by providing products and services that meet the customer's needs, wants, and preferences. Line managers must understand the hospital's customers and make the critical day-to-day differences in satisfying them. They are the hospital's front-line marketers, and the marketing department's role is to support them in this function. The marketing department should develop the line managers' marketing skills by furnishing them with the appropriate tools: formats, models, examples, and instructions. In addition, the marketing staff should provide such specialized services as marketing research, ad agency management, and development of communications. PMID:10277616

  12. 45. SELAH LINE, LOOKING NORTHEAST, SHOWING END OF LINE AT ...

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

    45. SELAH LINE, LOOKING NORTHEAST, SHOWING END OF LINE AT LARSON FRUIT COMPANY - Yakima Valley Transportation Company Interurban Railroad, Connecting towns of Yakima, Selah & Wiley City, Yakima, Yakima County, WA

  13. PEARS Emission Line Galaxies

    NASA Technical Reports Server (NTRS)

    Pirzkal, Nor; Rothberg, Barry; Ly, Chun; Rhoads, James E.; Malhotra, Sangeeta; Grogin, Norman A.; Dahlen, Tomas; Meurer, Gerhardt R.; Walsh, Jeremy; Hathi, Nimish P.; Cohen, Seth; Belini, Andrea; Holwerda, Benne W.; Straughn, Amber; Mechtley, Matthew

    2012-01-01

    We present a full analysis of the Probing Evolution And Reionization Spectroscopically (PEARS) slitless grism spectroscopic data obtained vl'ith the Advanced Camera for Surveys on HST. PEARS covers fields within both the Great Observatories Origins Deep Survey (GOODS) North and South fields, making it ideal as a random surveY of galaxies, as well as the availability of a wide variety of ancillary observations to support the spectroscopic results. Using the PEARS data we are able to identify star forming galaxies within the redshift volume 0 < z < 1.5. Star forming regions in the PEARS survey are pinpointed independently of the host galaxy. This method allOW8 us to detect the presence of multiple emission line regions (ELRs) within a single galaxy. 1162 [OII], [OIII] and/or H-alpha emission lines have been identified in the PEARS sample of approx 906 galaxies down to a limiting flux of approx 10 - 18 erg/s/sq cm . The ELRs have also been compared to the properties of the host galaxy, including morphology, luminosity, and mass. From this analysis we find three key results: 1) The computed line luminosities show evidence of a flattening in the luminosity function with increasing redshift; 2) The star forming systems show evidence of disturbed morphologies, with star formation occurring predominantly within one effective (half-light) radius. However, the morphologies show no correlation with host stellar mass; and 3) The number density of star forming galaxies with M(*) >= 10(exp 9) Solar M decreases by an order of magnitude at z<=0.5 relative to the number at 0.5 < z < 0.9 in support of the argument for galaxy downsizing.

  14. Vertical bloch line memory

    NASA Technical Reports Server (NTRS)

    Katti, R.; Wu, J.; Stadler, H.

    1990-01-01

    Vertical Bloch Line (VBL) memory is a recently conceived, integrated, solid-state, block-access, VLSI memory which offers the potential of 1Gbit/sq cm real storage density, gigabit per second data rates, and sub-millisecond average access times simultaneously at relatively low mass, volume, and power values when compared to alternative technologies. VBL's are micromagnetic structures within magnetic domain walls which can be manipulated using magnetic fields from integrated conductors. The presence or absence of VBL pairs are used to store binary information. At present, efforts are being directed at developing a single-chip memory using 25Mbit/sq cm technology in magnetic garnet material which integrates, at a single operating point, the writing, storage, reading, and amplification functions needed in a memory. This paper describes the current design architecture, functional elements, and supercomputer simulation results which are used to assist the design process. The current design architecture uses three metal layers, two ion implantation steps for modulating the thickness of the magnetic layer, one ion implantation step for assisting propagation in the major line track, one NiFe soft magnetic layer, one CoPt hard magnetic layer, and one reflective Cr layer for facilitating magneto-optic observation of magnetic structure. Data are stored in a series of elongated magnetic domains, called stripes, which serve as storage sites for arrays of VBL pairs. The ends of these stripes are placed near conductors which serve as VBL read/write gates. A major line track is present to provide a source and propagation path for magnetic bubbles. Writing and reading, respectively, are achieved by converting magnetic bubbles to VBL's and vice versa. The output function is effected by stretching a magnetic bubble and detecting it magnetoresistively. Experimental results from the past design cycle created four design goals for the current design cycle. First, the bias field ranges

  15. Read Between the Lines

    NASA Astrophysics Data System (ADS)

    Moore, Lynny

    2009-10-01

    When viewing it is best to have another person standing behind you reading the list of names of some of the women in the physical sciences that are placed ``between the lines''. It creates a profound contrast to the written words of the author of the letter, Robert A. Millikan. My mother-in-law, Dr. Isabelle Ganz, said she attended a physics conference during her freshman year of college at the University of Rochester and personally ``waited'' on Robert Oppenheimer. She noted that there were no women present. They were most likely in the lab.

  16. Diatomic predissociation line widths

    NASA Technical Reports Server (NTRS)

    Child, M. S.

    1973-01-01

    Predissociation by rotation and curve crossing in diatomic molecules is discussed. The pattern of predissociation line widths is seen as providing a highly sensitive yardstick for the determination of unknown potential curves. In addition, the computation of such a pattern for given potential curves is considered a matter of routine, unless the predissociation happens to occur from an adiabatic potential curve. Analytic formulas are used to provide physical insight into the details of the predissociation pattern, to the extent that a direct inversion procedure is developed for determination of the repulsive potential curves for Type 1 predissociations.

  17. Worldlines as Wilson Lines

    SciTech Connect

    Green, Daniel; /SLAC /Stanford U., Dept. Phys.

    2008-04-29

    Gravitational theories do not admit gauge invariant local operators. We study the limits under which there exists a quasi-local description for a class of non-local gravitational observables where a sum over worldlines plays the role of the Wilson line for gauge theory observables. We study non-local corrections to the local description and circumstances where these corrections become large. We find that these operators are quasi-local in at space and AdS, but fail to be quasi-local in de Sitter space.

  18. A short line segment squirms along a zigzag line.

    PubMed

    Ito, Hiroyuki; Yang, Xie

    2013-01-01

    When a short straight line segment moves across a zigzag line and is viewed in one's peripheral vision, it appears to exhibit nonrigid squirming motion (the squirm effect). This phenomenon demonstrates that the form, orientation, and motion direction of a short line are influenced by those of a longer one when they are viewed in one's peripheral vision. PMID:23755359

  19. Structure and form of grounding lines of modern ice sheets

    NASA Astrophysics Data System (ADS)

    Tinto, K. J.; Bell, R. E.; Cochran, J. R.; Boghosian, A.; Porter, D. F.

    2015-12-01

    The form of the bed at the grounding line of a glacier and the character of the underlying rock can be critical to the stability of the glacier. Aerogravity measurements offer a unique insight in to the character of the grounding line environment. By combining depth measurements from further onshore radar and geological information from magnetic surveys, gravity-based models can reveal both the depth and slope of the bed at the grounding line. Where bed elevation is known at the grounding line, gravity models can show the density structure of the underlying rock. Operation IceBridge has flown coincident radar, lidar, photography, gravity and magnetic airborne surveys along fjords and over ice shelves in both Greenland and Antarctica. Aerogravity measurements have been used extensively to model the bathymetry of the sea floor in front of the grounding line, and to identify the depth of the grounding line in areas where radar measurements have proven challenging. These models have also been used to reveal the range of conditions at present day grounding lines, as well as those experienced in the past and predicted for future grounding line positions. In some regions, we have identified low-density sediment accumulations, at both present day grounding lines and within fjords, that we interpret to be terminal moraines deposited by the glacier itself during hiatuses in retreat. In other regions, we find that the present day grounding line is stalled on a ridge of high-density rock. Ridges such as these remain in the same position through many cycles of advance and retreat of the glacier. Our synthesis of gravity data from a wide range of glacial environments can be used to identify likely drivers of change at the grounding line, whether this is the depth, the slope, or the geological character of the glacier bed.

  20. Bacterial populations on brewery filling hall surfaces as revealed by next-generation sequencing.

    PubMed

    Priha, Outi; Raulio, Mari; Maukonen, Johanna; Vehviläinen, Anna-Kaisa; Storgårds, Erna

    2016-01-01

    Due to the presence of moisture and nutrients, brewery filling line surfaces are susceptible to unwanted microbial attachment. Knowledge of the attaching microbes will aid in designing hygienic control of the process. In this study the bacterial diversity present on brewery filling line surfaces was revealed by next generation sequencing. The two filling lines studied maintained their characteristic bacterial community throughout three sampling times (13-163 days). On the glass bottle line, γ-proteobacteria dominated (35-82% of all OTUs), whereas on the canning line α-, β- and γ-proteobacteria and actinobacteria were most common. The most frequently detected genera were Acinetobacter, Propinobacterium and Pseudomonas. The halophilic genus Halomonas was commonly detected, which might be due to its tolerance to alkaline foam cleaners. This study has revealed a detailed overall picture of the bacterial groups present on filling line surfaces. Further effort should be given to determine the efficacy of washing procedures on different bacterial groups. PMID:27064426

  1. Iron Line Diagnostics of Narrow Emission Line Galaxies

    NASA Astrophysics Data System (ADS)

    Nousek, John A.

    1996-05-01

    This report describes the activities at Penn State University supported by NASA Grant NAG5-2528, 'Iron Line Diagnostics of Narrow Emission Line Galaxies'. The aim of this investigation was to accurately measure the iron (Fe K) line emission in two X-ray selected Seyfert 2 galaxies (NGC 2992 and MCG-5-23-16). The astrophysics being probed was to determine whether the Fe line was narrow, broad or both. The broad line component is very important as a probe of the nature of the innermost accretion onto the central engine in AGN's.

  2. A Fe K Line in GRB 970508

    NASA Astrophysics Data System (ADS)

    Protassov, R.; van Dyk, D.; Connors, A.; Kashyap, V.; Siemiginowska, A.

    2000-12-01

    We examine the x-ray spectrum of the afterglow of GRB 970508, analyzed for Fe line emission by Piro et al (1999, ApJL, 514, L73). This is a difficult and extremely important measurement: the detection of x-ray afterglows from γ -ray bursts is at best a tricky business, relying on near-real satellite time response to unpredictable events; and a great deal of luck in catching a burst bright enough for a useful spectral analysis. Detecting a clear atomic (or cyclotron) line in the generally smooth and featureless afterglow (or burst) emission not only gives one of the few very specific keys to the physics local to the emission region, but also provides clues or confirmation of its distance (via redshift). Unfortunately, neither the likelihood ratio test or the related F-statistic commonly used to detect spectral lines adhere to their nominal Chi square and F-distributions. Thus we begin by calibrating the F-statistic used in Piro et al (1999, ApJL, 514, L73) via a simulation study. The simulation study relies on a completely specified source model, i.e. we do Monte Carlo simulations with all model parameters fixed (so--called ``parametric bootstrapping''). Second, we employ the method of posterior predictive p-values to calibrate a LRT statistic while accounting for the uncertainty in the parameters of the source model. Our analysis reveals evidence for the Fe K line.

  3. Line tension of multicomponent bilayer membranes

    NASA Astrophysics Data System (ADS)

    Dehghan, Ashkan; Pastor, Kyle A.; Shi, An-Chang

    2015-02-01

    The line tension or edge energy of bilayer membranes self-assembled from binary amphiphilic molecules is studied using self-consistent-field theory (SCFT). Specifically, solutions of the SCFT equations corresponding to an infinite membrane with a circular pore, or an open membrane, are obtained for a coarse-grained model in which the amphiphilic species and hydrophilic solvents are represented by A B and E D diblock copolymers and C homopolymers, respectively. The edge energy of the membrane is extracted from the free energy of the open membranes. Results for membranes composed of mixtures of symmetric and cone- or inverse cone-shaped amphiphilic molecules with neutral and/or repulsive interactions are obtained and analyzed. It is observed that an increase in the concentration of the cone-shaped species leads to a decrease of the line tension. In contrast, adding inverse cone-shaped copolymers results in an increase of the line tension. Furthermore, the density profile of the copolymers reveals that the line tension is regulated by the distribution of the amphiphiles at the bilayer edge.

  4. TIGER/LINE FILES, 1998

    EPA Science Inventory

    TIGER, TIGER/Line, and Census TIGER are registered trademarks of the Bureau of the Census. The 1998 TIGER/Line files are an extract of selected geographic and cartographic information from the Census TIGER data base. The geographic coverage for a single TIGER/Line file is a cou...

  5. Atlas of Infrared Absorption Lines

    NASA Technical Reports Server (NTRS)

    Park, J. H.

    1977-01-01

    This atlas of infrared absorption line contains absorption line parameters (line strength vs. wavenumber) from 500 to 7000 cm(exp-1) for 15 gases: H2O, CO2, O3, N2O, CO, CH4, O2, SO2, NO, NO2, NH3, HCl, HF, HNO3 and CH3Cl.

  6. CORONAL EMISSION LINES AS THERMOMETERS

    SciTech Connect

    Judge, Philip G.

    2010-01-10

    Coronal emission-line intensities are commonly used to measure electron temperatures using emission measure and/or line ratio methods. In the presence of systematic errors in atomic excitation calculations and data noise, the information on underlying temperature distributions is fundamentally limited. Increasing the number of emission lines used does not necessarily improve the ability to discriminate between different kinds of temperature distributions.

  7. Bee Line BR-1 Racer

    NASA Technical Reports Server (NTRS)

    1926-01-01

    Bee Line BR-1 Racer: The Bee Line BR-1 was a racing aircraft used to compete in the 1922 Pulitzer Air Race. The aircraft and its sister ship, the Bee Line BR-2, came to Langley and the NACA in 1926. The BR-1 is shown in the NACA hangar at Langley Field in early 1926.

  8. Conservation with underground power lines

    SciTech Connect

    Graneau, P.

    1980-01-01

    The following aspects of underground power transmission lines are discussed: their contribution to area beautification; line losses and their causes; the energy conservation potential of large-conductor underground cables; reliability and outage advantages as compared with overhead lines; the history of underground systems; problems with polyethylene insulation; and the development and performance of sodium conductors for underground cables. (LCL)

  9. Bloch state tomography using Wilson lines

    NASA Astrophysics Data System (ADS)

    Li, Tracy; Duca, Lucia; Reitter, Martin; Grusdt, Fabian; Demler, Eugene; Endres, Manuel; Schleier-Smith, Monika; Bloch, Immanuel; Schneider, Ulrich

    2016-05-01

    Topology and geometry are essential to our understanding of modern physics, underlying many foundational concepts from high-energy theories, quantum information, and condensed-matter physics. In condensed-matter systems, a wide range of phenomena stem from the geometry of the band eigenstates, which is encoded in the matrix-valued Wilson line for general multiband systems. Using an ultracold gas of rubidium atoms loaded in a honeycomb optical lattice, we realize strong-force dynamics in Bloch bands that are described by Wilson lines and observe an evolution in the band populations that directly reveals the band geometry. Our technique enables a full determination of band eigenstates, Berry curvature, and topological invariants, including single- and multiband Chern and Z2 numbers.

  10. Bloch state tomography using Wilson lines.

    PubMed

    Li, Tracy; Duca, Lucia; Reitter, Martin; Grusdt, Fabian; Demler, Eugene; Endres, Manuel; Schleier-Smith, Monika; Bloch, Immanuel; Schneider, Ulrich

    2016-05-27

    Topology and geometry are essential to our understanding of modern physics, underlying many foundational concepts from high-energy theories, quantum information, and condensed-matter physics. In condensed-matter systems, a wide range of phenomena stem from the geometry of the band eigenstates, which is encoded in the matrix-valued Wilson line for general multiband systems. Using an ultracold gas of rubidium atoms loaded in a honeycomb optical lattice, we realize strong-force dynamics in Bloch bands that are described by Wilson lines and observe an evolution in the band populations that directly reveals the band geometry. Our technique enables a full determination of band eigenstates, Berry curvature, and topological invariants, including single- and multiband Chern and Z₂ numbers. PMID:27230376

  11. Open field lines instabilities

    SciTech Connect

    Pozzoli, R. |

    1995-09-01

    The results of some recent theoretical papers dealing with flute-like instabilities in the scrape-off layer of a tokamak with limiter configuration, where the magnetic field intersects conducting walls, are briefly recalled. Attention is then paid to the instability driven by the electron temperature gradient across the field in conjunction with the formation of the Debye sheath at the boundary, and to the effects due to the inclination of the end walls with respect to the magnetic field. When a divertor configuration is considered, important modifications are found owing to the strong deformations of the flux tubes passing near the {ital x}-point, which contrast the onset of flute-like perturbations, and to the stochasticity of field lines that can be excited by magnetic field perturbations. {copyright} {ital 1995 American Institute of Physics.}

  12. Continuous gutter lining

    SciTech Connect

    Butler, R.W.

    1988-05-03

    An apparatus for lining a gutter having a downspout and horizontal support members is described comprising: a strip of waterproof, normally flat, resilient material of substantially uniform cross section having a drain hole therein; the strip being deformed out of its flat shape to a U-shaped transverse cross-section and being located within the gutter beneath the horizontal support members, having a width dimension sufficient to traverse a cross-section of the gutter underneath the horizontal members having a first edge of the strip contacting a back wall of the gutter, a second edge of the strip contacting a front wall of the gutter and forming a U-shaped cross section; and the drain hole being located above the gutter downspout to facilitate draining.

  13. Understanding Slag Freeze Linings

    NASA Astrophysics Data System (ADS)

    Fallah-Mehrjardi, Ata; Hayes, Peter C.; Jak, Evgueni

    2014-09-01

    Slag freeze linings, the formation of protective deposit layers on the inner walls of furnaces and reactors, are increasingly used in industrial pyrometallurgical processes to ensure that furnace integrity is maintained in these aggressive, high-temperature environments. Most previous studies of freeze-linings have analyzed the formation of slag deposits based solely on heat transfer considerations. These thermal models have assumed that the interface between the stationary frozen layer and the agitated molten bath at steady-state deposit thickness consists of the primary phase, which stays in contact with the bulk liquid at the liquidus temperature. Recent experimental studies, however, have clearly demonstrated that the temperature of the deposit/liquid bath interface can be lower than the liquidus temperature of the bulk liquid. A conceptual framework has been proposed to explain the observations and the factors influencing the microstructure and the temperature of the interface at steady-state conditions. The observations are consistent with a dynamic steady state that is a balance between (I) the rate of nucleation and growth of solids on detached crystals in a subliquidus layer as this fluid material moves toward the stagnant deposit interface and (II) the dissolution of these detached crystals as they are transported away from the interface by turbulent eddies. It is argued that the assumption that the interface temperature is the liquidus of the bulk material represents only a limiting condition, and that the interface temperature can be between T liquidus and T solidus depending on the process conditions and bath chemistry. These findings have implications for the modeling approach and boundary conditions required to accurately describe these systems. They also indicate the opportunity to integrate considerations of heat and mass flows with the selection of melt chemistries in the design of future high temperature industrial reactors.

  14. Selection Components in Background Replacement Lines of Drosophila

    PubMed Central

    Clark, Andrew G.; Bundgaard, Jørgen

    1984-01-01

    Selection components analysis was performed in lines of Drosophila melanogaster at three times during substitution backcrossing. The initial two lines were inbred isofemale lines from natural populations in California, and one had the spread wing mutation eagle. The selection components analysis revealed aspects of the genetic structure of the determinants of fitness by demonstrating changes in the marginal fitnesses of the eagle locus. Differences among backgrounds essentially disappeared by the 20th generation of backcrossing, suggesting that the previously observed differences were attributable to linkage disequilibrium. The method of bootstrapping was used as a novel means of determining statistical confidence in selection components. PMID:17246226

  15. Global attentional neglect of segmented lines in Parkinson's disease.

    PubMed

    Falchook, Adam D; Salazar, Liliana; Neal, Dan; Kesayan, Tigran; Williamson, John B; Malaty, Irene A; McFarland, Nikolaus R; Okun, Michael S; Rodriguez, Ramon L; Wagle Shukla, Aparna; Heilman, Kenneth M

    2015-01-01

    Global attention requires disengagement from focal elements of stimuli. Since people with Parkinson's disease (PD) may reveal impaired disengagement, this study attempted to learn if people with PD may be impaired at allocating global attention. Healthy adults and people with PD attempted to bisect lines of uniform thickness and lines composed of two segments of unequal thickness and length. When the longer line segment was to the right of the shorter segment, the group with PD demonstrated an increased deviation toward the longer segment, supporting the postulate that people with PD have an impaired ability to disengage focal attention and engage global spatial attention. PMID:25073971

  16. Antiproliferative effect of isopentenylated coumarins on several cancer cell lines.

    PubMed

    Kawaii, S; Tomono, Y; Ogawa, K; Sugiura, M; Yano, M; Yoshizawa, Y; Ito, C; Furukawa, H

    2001-01-01

    33 coumarins, mainly the simple isopentenylated coumarins and derived pyrano- and furanocoumarins, were examined for their antiproliferative activity towards several cancer and normal human cell lines. The pyrano- and furanocoumarins showed strong activity against the cancer cell lines, whereas they had weak antiproliferative activity against the normal human cell lines. The decreasing rank order of potency was osthenone (10), clausarin (25), clausenidin (26), dentatin (24), nordentatin (23), imperatorin (29), seselin (27), xanthyletin (21), suberosin (17), phebalosin (8) and osthol (12). The structure-activity relationship established from the results revealed that the 1,1-dimethylallyl and isopentenyl groups have an important role for antiproliferative activity. PMID:11497276

  17. Rapid bottom melting widespread near Antarctic Ice Sheet grounding lines.

    PubMed

    Rignot, Eric; Jacobs, Stanley S

    2002-06-14

    As continental ice from Antarctica reaches the grounding line and begins to float, its underside melts into the ocean. Results obtained with satellite radar interferometry reveal that bottom melt rates experienced by large outlet glaciers near their grounding lines are far higher than generally assumed. The melting rate is positively correlated with thermal forcing, increasing by 1 meter per year for each 0.1 degrees C rise in ocean temperature. Where deep water has direct access to grounding lines, glaciers and ice shelves are vulnerable to ongoing increases in ocean temperature. PMID:12065835

  18. Electrocardiograph abnormalities revealed during laparoscopy

    PubMed Central

    Nijjer, Sukhjinder; Dubrey, Simon William

    2010-01-01

    This brief case presents a well patient in whom an electrocardiograph abnormality consistent with an accessory pathway was found during a routine procedure. We present the electrocardiographs, explain the underlying condition, and consider why the abnormality was revealed in this manner. PMID:22419949

  19. Ischemic Colitis Revealing Polyarteritis Nodosa

    PubMed Central

    Hamzaoui, Amira; Litaiem, Noureddine; Smiti Khanfir, M.; Ayadi, Sofiene; Nfoussi, Haifa; Houman, M. H.

    2013-01-01

    Ischemic colitis is one of the most common intestinal ischemic injuries. It results from impaired perfusion of blood to the bowel and is rarely caused by vasculitis. We report a case of ischemic colitis revealing polyarteritis nodosa (PAN) in a 55-year-old man. Histological examination of the resected colon led to the diagnosis of PAN. PMID:24382967

  20. 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

  1. ANALYSIS OF CORONAL GREEN LINE PROFILES: EVIDENCE OF EXCESS BLUESHIFTS

    SciTech Connect

    Raju, K. P.; Chandrasekhar, T.; Ashok, N. M. E-mail: chandra@prl.res.in

    2011-08-01

    Coronal green line (Fe XIV 5303 A) profiles were obtained from Fabry-Perot interferometric observations of the solar corona during the total solar eclipse of 2001 June 21 from Lusaka, Zambia. The instrumental width is about 0.2 A and the spectral resolution is about 26,000. About 300 line profiles were obtained within a radial range of 1.0-1.5 R{sub sun} and a position angle coverage of about 240{sup 0}. The line profiles were fitted with single Gaussians, and their intensities, Doppler velocities, and line widths were obtained. Also obtained were the centroids of the line profiles, which give a measure of line asymmetry. The histograms of Doppler velocity show excess blueshifts, while the centroids reveal a predominant blue wing in the line profiles. It was found that the centroids and the Doppler velocities are highly correlated. This points to the presence of multiple components in the line profiles, with an excess of blueshifted components. We then obtained the (Blue-Red) wing intensities, which clearly reveal the secondary components, the majority of which are blueshifted. This confirms that the coronal green line profiles often contain multicomponents with excess blueshifts, which also depend on the solar activity. The magnitude of the Doppler velocity of the secondary components is in the range of 20-40 km s{sup -1} and shows an increase toward the poles. Possible explanations of the multicomponents are the type II spicules that were recently found to be important to coronal heating or the nascent solar wind flow, but the cause of the blue asymmetry in the coronal lines above the limb remains unclear.

  2. Triploid human embryonic stem cells derived from tripronuclear zygotes displayed pluripotency and trophoblast differentiation ability similar to the diploid human embryonic stem cells.

    PubMed

    Rungsiwiwut, Ruttachuk; Numchaisrika, Pranee; Ahnonkitpanit, Vichuda; Virutamasen, Pramuan; Pruksananonda, Kamthorn

    2016-04-22

    Because the diploid human embryonic stem cells (hESCs) can be successfully derived from tripronuclear zygotes thus, they can serve as an alternative source of derivation of normal karyotype hESC lines. The aim of the present study was to compare the pluripotency and trophoblast differentiation ability of hESCs derived from tripronuclear zygotes and diploid hESCs. In the present study, a total of 20 tripronuclear zygotes were cultured; 8 zygotes developed to the blastocyst stage and 1 hESC line was generated. Unlike the previous studies, chromosomal correction of tripronuclear zygotes during derivation of hESCs did not occur. The established line carries 3 sets of chromosomes and showed a numerical aberration. Although the cell line displayed an abnormal chromosome number, it was found the cell line has been shown to be pluripotent with the ability to differentiate into 3 embryonic germ layers both in vitro and in vivo. The expression of X inactive specific transcript (XIST) in mid-passage (passage 42) of undifferentiated triploid hESCs was detected, indicating X chromosome inactivation of the cell line. Moreover, when this cell line was induced to differentiate toward the trophoblast lineage, morphological and functional trophoblast cells were observed, similar to the diploid hESC line. PMID:26821869

  3. Triploid human embryonic stem cells derived from tripronuclear zygotes displayed pluripotency and trophoblast differentiation ability similar to the diploid human embryonic stem cells

    PubMed Central

    RUNGSIWIWUT, Ruttachuk; NUMCHAISRIKA, Pranee; AHNONKITPANIT, Vichuda; VIRUTAMASEN, Pramuan; PRUKSANANONDA, Kamthorn

    2016-01-01

    Because the diploid human embryonic stem cells (hESCs) can be successfully derived from tripronuclear zygotes thus, they can serve as an alternative source of derivation of normal karyotype hESC lines. The aim of the present study was to compare the pluripotency and trophoblast differentiation ability of hESCs derived from tripronuclear zygotes and diploid hESCs. In the present study, a total of 20 tripronuclear zygotes were cultured; 8 zygotes developed to the blastocyst stage and 1 hESC line was generated. Unlike the previous studies, chromosomal correction of tripronuclear zygotes during derivation of hESCs did not occur. The established line carries 3 sets of chromosomes and showed a numerical aberration. Although the cell line displayed an abnormal chromosome number, it was found the cell line has been shown to be pluripotent with the ability to differentiate into 3 embryonic germ layers both in vitro and in vivo. The expression of X inactive specific transcript (XIST) in mid-passage (passage 42) of undifferentiated triploid hESCs was detected, indicating X chromosome inactivation of the cell line. Moreover, when this cell line was induced to differentiate toward the trophoblast lineage, morphological and functional trophoblast cells were observed, similar to the diploid hESC line. PMID:26821869

  4. Optimization of line configuration and balancing for flexible machining lines

    NASA Astrophysics Data System (ADS)

    Liu, Xuemei; Li, Aiping; Chen, Zurui

    2016-04-01

    Line configuration and balancing is to select the type of line and allot a given set of operations as well as machines to a sequence of workstations to realize high-efficiency production. Most of the current researches for machining line configuration and balancing problems are related to dedicated transfer lines with dedicated machine workstations. With growing trends towards great product variety and fluctuations in market demand, dedicated transfer lines are being replaced with flexible machining line composed of identical CNC machines. This paper deals with the line configuration and balancing problem for flexible machining lines. The objective is to assign operations to workstations and find the sequence of execution, specify the number of machines in each workstation while minimizing the line cycle time and total number of machines. This problem is subject to precedence, clustering, accessibility and capacity constraints among the features, operations, setups and workstations. The mathematical model and heuristic algorithm based on feature group strategy and polychromatic sets theory are presented to find an optimal solution. The feature group strategy and polychromatic sets theory are used to establish constraint model. A heuristic operations sequencing and assignment algorithm is given. An industrial case study is carried out, and multiple optimal solutions in different line configurations are obtained. The case studying results show that the solutions with shorter cycle time and higher line balancing rate demonstrate the feasibility and effectiveness of the proposed algorithm. This research proposes a heuristic line configuration and balancing algorithm based on feature group strategy and polychromatic sets theory which is able to provide better solutions while achieving an improvement in computing time.

  5. Optimization of line configuration and balancing for flexible machining lines

    NASA Astrophysics Data System (ADS)

    Liu, Xuemei; Li, Aiping; Chen, Zurui

    2016-05-01

    Line configuration and balancing is to select the type of line and allot a given set of operations as well as machines to a sequence of workstations to realize high-efficiency production. Most of the current researches for machining line configuration and balancing problems are related to dedicated transfer lines with dedicated machine workstations. With growing trends towards great product variety and fluctuations in market demand, dedicated transfer lines are being replaced with flexible machining line composed of identical CNC machines. This paper deals with the line configuration and balancing problem for flexible machining lines. The objective is to assign operations to workstations and find the sequence of execution, specify the number of machines in each workstation while minimizing the line cycle time and total number of machines. This problem is subject to precedence, clustering, accessibility and capacity constraints among the features, operations, setups and workstations. The mathematical model and heuristic algorithm based on feature group strategy and polychromatic sets theory are presented to find an optimal solution. The feature group strategy and polychromatic sets theory are used to establish constraint model. A heuristic operations sequencing and assignment algorithm is given. An industrial case study is carried out, and multiple optimal solutions in different line configurations are obtained. The case studying results show that the solutions with shorter cycle time and higher line balancing rate demonstrate the feasibility and effectiveness of the proposed algorithm. This research proposes a heuristic line configuration and balancing algorithm based on feature group strategy and polychromatic sets theory which is able to provide better solutions while achieving an improvement in computing time.

  6. EC Transmission Line Materials

    SciTech Connect

    Bigelow, Tim S

    2012-05-01

    The purpose of this document is to identify materials acceptable for use in the US ITER Project Office (USIPO)-supplied components for the ITER Electron cyclotron Heating and Current Drive (ECH&CD) transmission lines (TL), PBS-52. The source of material property information for design analysis shall be either the applicable structural code or the ITER Material Properties Handbook. In the case of conflict, the ITER Material Properties Handbook shall take precedence. Materials selection, and use, shall follow the guidelines established in the Materials Assessment Report (MAR). Materials exposed to vacuum shall conform to the ITER Vacuum Handbook. [Ref. 2] Commercial materials shall conform to the applicable standard (e.g., ASTM, JIS, DIN) for the definition of their grade, physical, chemical and electrical properties and related testing. All materials for which a suitable certification from the supplier is not available shall be tested to determine the relevant properties, as part of the procurement. A complete traceability of all the materials including welding materials shall be provided. Halogenated materials (example: insulating materials) shall be forbidden in areas served by the detritiation systems. Exceptions must be approved by the Tritium System and Safety Section Responsible Officers.

  7. Bipolar pulse forming line

    DOEpatents

    Rhodes, Mark A.

    2008-10-21

    A bipolar pulse forming transmission line module for linear induction accelerators having first, second, third, fourth, and fifth planar conductors which form an interleaved stack with dielectric layers between the conductors. Each conductor has a first end, and a second end adjacent an acceleration axis. The first and second planar conductors are connected to each other at the second ends, the fourth and fifth planar conductors are connected to each other at the second ends, and the first and fifth planar conductors are connected to each other at the first ends via a shorting plate adjacent the first ends. The third planar conductor is electrically connectable to a high voltage source, and an internal switch functions to short a high voltage from the first end of the third planar conductor to the first end of the fourth planar conductor to produce a bipolar pulse at the acceleration axis with a zero net time integral. Improved access to the switch is enabled by an aperture through the shorting plate and the proximity of the aperture to the switch.

  8. Urticarial vasculitis reveals unsuspected thyroiditis.

    PubMed

    Ferreira, Olga; Mota, Alberto; Baudrier, Teresa; Azevedo, Filomena

    2012-01-01

    A 38-year-old woman presented with erythematous, violaceous plaques with a serpiginous and unusual appearance located on the left shoulder, left thigh, and right buttock, evolving for 5 days, which eventually became generalized. A skin biopsy revealed leukocytoclastic vasculitis and a diagnosis of urticarial vasculitis was made. The complete blood count, biochemistry, complement levels, and other immunological test results were unremarkable. However, antithyroid antibody titers were increased. Despite having normal thyroid function tests and an absence of specific symptoms, the patient underwent a thyroid ultrasound, which revealed features of thyroiditis, and was subsequently referred to an endocrinologist. Several diseases can be associated with urticarial vasculitis, namely infections and autoimmune connective-tissue disorders such as systemic lupus erythematosus and Sjögren syndrome. Thyroiditis is an uncommon association. PMID:23000939

  9. Targeted and genome-scale methylomics reveals gene body signatures in human cell lines

    PubMed Central

    Ball, Madeleine Price; Li, Jin Billy; Gao, Yuan; Lee, Je-Hyuk; LeProust, Emily; Park, In-Hyun; Xie, Bin; Daley, George Q.; Church, George M.

    2012-01-01

    Cytosine methylation, an epigenetic modification of DNA, is a target of growing interest for developing high throughput profiling technologies. Here we introduce two new, complementary techniques for cytosine methylation profiling utilizing next generation sequencing technology: bisulfite padlock probes (BSPPs) and methyl sensitive cut counting (MSCC). In the first method, we designed a set of ~10,000 BSPPs distributed over the ENCODE pilot project regions to take advantage of existing expression and chromatin immunoprecipitation data. We observed a pattern of low promoter methylation coupled with high gene body methylation in highly expressed genes. Using the second method, MSCC, we gathered genome-scale data for 1.4 million HpaII sites and confirmed that gene body methylation in highly expressed genes is a consistent phenomenon over the entire genome. Our observations highlight the usefulness of techniques which are not inherently or intentionally biased in favor of only profiling particular subsets like CpG islands or promoter regions. PMID:19329998

  10. Molecular genetic variation in cultivated peanut cultivars and breeding lines revealed by highly informative SSR markers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Groundnut or peanut (Arachis hypogaea L.) is an economically important crop worldwide as a source of protein and cooking oil, particularly in developing countries. Because of its narrow genetic background and shortage of polymorphic genetic markers, molecular characterization of cultivated peanuts e...

  11. Motion along the mental number line reveals shared representations for numerosity and space

    PubMed Central

    Schwiedrzik, Caspar M; Bernstein, Benjamin; Melloni, Lucia

    2016-01-01

    Perception of number and space are tightly intertwined. It has been proposed that this is due to ‘cortical recycling’, where numerosity processing takes over circuits originally processing space. Do such ‘recycled’ circuits retain their original functionality? Here, we investigate interactions between numerosity and motion direction, two functions that both localize to parietal cortex. We describe a new phenomenon in which visual motion direction adapts nonsymbolic numerosity perception, giving rise to a repulsive aftereffect: motion to the left adapts small numbers, leading to overestimation of numerosity, while motion to the right adapts large numbers, resulting in underestimation. The reference frame of this effect is spatiotopic. Together with the tuning properties of the effect this suggests that motion direction-numerosity cross-adaptation may occur in a homolog of area LIP. ‘Cortical recycling’ thus expands but does not obliterate the functions originally performed by the recycled circuit, allowing for shared computations across domains. DOI: http://dx.doi.org/10.7554/eLife.10806.001 PMID:26771249

  12. Trans-saccadic repetition priming: ERPs reveal on-line integration of information across words.

    PubMed

    Grainger, Jonathan; Midgley, Katherine J; Holcomb, Phillip J

    2016-01-01

    We used a trans-saccadic priming paradigm combined with ERP recordings to track the time-course of integration of information across a prime word briefly presented at fixation and a subsequent target word presented 4 degrees to the right of fixation. Trans-saccadic repetition priming effects (Experiments 1 and 2) were compared with priming effects obtained with centrally located targets (Experiment 3). In Experiment 2, target stimuli were preceded by a 100ms forward mask at the target location, hence allowing an attention shift to the target location prior to target onset. Compared with centrally located targets, repetition priming effects were found to onset later in Experiment 2 and even later in Experiment 1, and the growth of priming effects was slower in both Experiments 1 and 2 compared with Experiment 3. The results demonstrate integration of information across spatially distinct primes and targets, with the time-course of trans-saccadic priming being determined by the speed with which attention can be allocated to peripheral targets plus the quality of information available in peripheral vision prior to fixation of target stimuli. PMID:26656872

  13. Nonintegral stoichiometry in CFTR gating revealed by a pore-lining mutation.

    PubMed

    Jih, Kang-Yang; Sohma, Yoshiro; Hwang, Tzyh-Chang

    2012-10-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is a unique member of the ATP-binding cassette (ABC) protein superfamily. Unlike most other ABC proteins that function as active transporters, CFTR is an ATP-gated chloride channel. The opening of CFTR's gate is associated with ATP-induced dimerization of its two nucleotide-binding domains (NBD1 and NBD2), whereas gate closure is facilitated by ATP hydrolysis-triggered partial separation of the NBDs. This generally held theme of CFTR gating-a strict coupling between the ATP hydrolysis cycle and the gating cycle-is put to the test by our recent finding of a short-lived, post-hydrolytic state that can bind ATP and reenter the ATP-induced original open state. We accidentally found a mutant CFTR channel that exhibits two distinct open conductance states, the smaller O1 state and the larger O2 state. In the presence of ATP, the transition between the two states follows a preferred O1→O2 order, a telltale sign of a violation of microscopic reversibility, hence demanding an external energy input likely from ATP hydrolysis, as such preferred gating transition was abolished in a hydrolysis-deficient mutant. Interestingly, we also observed a considerable amount of opening events that contain more than one O1→O2 transition, indicating that more than one ATP molecule may be hydrolyzed within an opening burst. We thus conclude a nonintegral stoichiometry between the gating cycle and ATP consumption. Our results lead to a six-state gating model conforming to the classical allosteric mechanism: both NBDs and transmembrane domains hold a certain degree of autonomy, whereas the conformational change in one domain will facilitate the conformational change in the other domain. PMID:22966014

  14. Nonintegral stoichiometry in CFTR gating revealed by a pore-lining mutation

    PubMed Central

    Jih, Kang-Yang; Sohma, Yoshiro

    2012-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is a unique member of the ATP-binding cassette (ABC) protein superfamily. Unlike most other ABC proteins that function as active transporters, CFTR is an ATP-gated chloride channel. The opening of CFTR’s gate is associated with ATP-induced dimerization of its two nucleotide-binding domains (NBD1 and NBD2), whereas gate closure is facilitated by ATP hydrolysis-triggered partial separation of the NBDs. This generally held theme of CFTR gating—a strict coupling between the ATP hydrolysis cycle and the gating cycle—is put to the test by our recent finding of a short-lived, post-hydrolytic state that can bind ATP and reenter the ATP-induced original open state. We accidentally found a mutant CFTR channel that exhibits two distinct open conductance states, the smaller O1 state and the larger O2 state. In the presence of ATP, the transition between the two states follows a preferred O1→O2 order, a telltale sign of a violation of microscopic reversibility, hence demanding an external energy input likely from ATP hydrolysis, as such preferred gating transition was abolished in a hydrolysis-deficient mutant. Interestingly, we also observed a considerable amount of opening events that contain more than one O1→O2 transition, indicating that more than one ATP molecule may be hydrolyzed within an opening burst. We thus conclude a nonintegral stoichiometry between the gating cycle and ATP consumption. Our results lead to a six-state gating model conforming to the classical allosteric mechanism: both NBDs and transmembrane domains hold a certain degree of autonomy, whereas the conformational change in one domain will facilitate the conformational change in the other domain. PMID:22966014

  15. Low Dose Vaccination Revealed An Interaction Between Vaccine and Chicken Line Affecting Protective Efficacy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Marek’s disease (MD), caused by a highly oncogenic and contagious cell-associated alpha-herpesvirus, also known as Marek’s disease virus (MDV), remains as a highly potential threat to poultry industry worldwide. This threat has been continuously sustained by persistent viral evolution with increasin...

  16. MicroRNA screen of human embryonic stem cell differentiation reveals miR-105 as an enhancer of megakaryopoiesis from adult CD34+ cells.

    PubMed

    Kamat, Viraj; Paluru, Prasuna; Myint, Melissa; French, Deborah L; Gadue, Paul; Diamond, Scott L

    2014-05-01

    MicroRNAs (miRNAs) can control stem cell differentiation by targeting mRNAs. Using 96-well plate electroporation, we screened 466 human miRNA mimics by four-color flow cytometry to explore differentiation of common myeloid progenitors (CMP) derived from human embryonic stem cells (hESCs). The transfected cells were then cultured in a cytokine cocktail that supported multiple hematopoietic lineages. At 4-5 days post-transfection, flow cytometry of erythroid (CD235(+)CD41(-)), megakaryocyte (CD41(+)CD42(+)), and myeloid (CD18(+)CD235(-)) lineages revealed miR-105 as a novel enhancer of megakaryocyte production during in vitro primitive hematopoiesis. In hESC-derived CMPs, miR-105 caused a sixfold enhancement in megakaryocyte production. miR-513a, miR-571, and miR-195 were found to be less potent megakaryocyte enhancers. We confirmed the relevance of miR-105 in adult megakaryopoiesis by demonstrating increased megakaryocyte yield and megakaryocyte colony forming potential in human adult CD34(+) cells derived from peripheral blood. In addition, adult CD34(+) cells express endogenous miR-105 during megakaryocyte differentiation. siRNA knockdown of the hematopoietic transcription factor c-Myb caused a similar enhancement of megakaryocyte production as miR-105. Finally, a luciferase/c-Myb-3'UTR construct and Western blot analysis demonstrated that the hematopoietic transcription factor c-Myb mRNA was a target of miR-105. We report a novel hESC-based miR screening platform and demonstrate that miR-105 is an enhancer of megakaryopoiesis in both primitive and definitive hematopoiesis. PMID:24446170

  17. Dissociation between line bisection and mental-number-line bisection in healthy adults.

    PubMed

    Rotondaro, Francesca; Merola, Sheila; Aiello, Marilena; Pinto, Mario; Doricchi, Fabrizio

    2015-08-01

    Healthy adults bisect visual horizontal lines slightly to the left of their true center. This bias has been termed "pseudoneglect" and is considered to reflect right hemisphere dominance in the orienting of spatial attention. A previous investigation reported a positive correlation between pseudoneglect and a corresponding negative bias towards numbers lower than the true midpoint, i.e. supposedly to the left of the midpoint, during the mental bisection of number intervals that were defined by two visual arabic digits presented one to the left and one to the right of a horizontal line (Longo and Lourenco, 2007, Neuropsychologia, 45, 7, 1400-1407). Here, studying a sample of 60 healthy participants we verified whether this correlation still holds when the endpoints of number intervals are defined verbally, i.e. with no visual-spatial cues suggesting their left-to-right arrangement. Participants bisected horizontal lines (2 cm, 10 cm and 20 cm), short number intervals (3-, 5-, 7- and 9-unit) and large number intervals (16-, 24-, 32-, 40-, 48-, 56-, and 64-unit). Pseudoneglect was observed both in line and number interval bisection, confirming the results of Longo and Lourenco (2007). Nonetheless, the study of correlations between bisection biases averaged across different line and number intervals lengths and between all possible pairings of line and number interval lengths revealed no significant or systematic pattern. During line bisection pseudoneglect increased as a function of line length while with short number intervals pseudoneglect decreased and turned into an opposite positive bias as a function of interval length. With large number intervals no linear relationship was present between bisection bias and interval length and, as in Longo and Lourenco (2007), the higher was the starting point of the number interval the larger was pseudoneglect. These results show that verbally defined number intervals are not mentally inspected with the same mechanisms that

  18. Synthetic CA II Triplet Lines

    NASA Astrophysics Data System (ADS)

    Erdelyi, M. M.; Barbuy, B.

    1990-11-01

    RESUMEN. Se hicieron calculos de sintesis del espectro en el ititervalo de longitud de onda - 8700 A, ara ? oder verificar el comporta- mien to de diferentes lineas moleculares y at5micas como funci5n de los parametros esteldres de temperatura, gravedad y metalicidad. El espec- tro sintetico ha sido generado para:(a) todas las , (b) solamente de CN, (c) solamente de TiO, y (d) solamente lineas at6micas. Abstract. Spectrum synthesis calculations are carried out in the wavelength interval X 8300 - 8700 A, in order to verify the behaviour of different molecular and atomic lines as a function of the stellar para meters temperature, gravity and metallicity. Synthetic spectra were ge nerated for: (a) all lines, (b) only CN lines, (c) only TiO lines, and (d) only atomic lines Key `td6: LINE-PROFILE - ST S-AThOSPHERES

  19. ALFA: Automated Line Fitting Algorithm

    NASA Astrophysics Data System (ADS)

    Wesson, R.

    2015-12-01

    ALFA fits emission line spectra of arbitrary wavelength coverage and resolution, fully automatically. It uses a catalog of lines which may be present to construct synthetic spectra, the parameters of which are then optimized by means of a genetic algorithm. Uncertainties are estimated using the noise structure of the residuals. An emission line spectrum containing several hundred lines can be fitted in a few seconds using a single processor of a typical contemporary desktop or laptop PC. Data cubes in FITS format can be analysed using multiple processors, and an analysis of tens of thousands of deep spectra obtained with instruments such as MUSE will take a few hours.

  20. A proposed neutral line signature

    NASA Technical Reports Server (NTRS)

    Doxas, I.; Speiser, T. W.; Dusenbery, P. B.; Horton, W.

    1992-01-01

    An identifying signature is proposed for the existence and location of the neutral line in the magnetotail. The signature, abrupt density, and temperature changes in the Earthtail direction, was first discovered in test particle simulations. Such temperature variations have been observed in ISEE data (Huang et. al. 1992), but their connection to the possible existence of a neutral line in the tail has not yet been established. The proposed signature develops earlier than the ion velocity space ridge of Martin and Speiser (1988), but can only be seen by spacecraft in the vicinity of the neutral line, while the latter can locate a neutral line remotely.

  1. LINE-ABOVE-GROUND ATTENUATOR

    DOEpatents

    Wilds, R.B.; Ames, J.R.

    1957-09-24

    The line-above-ground attenuator provides a continuously variable microwave attenuator for a coaxial line that is capable of high attenuation and low insertion loss. The device consists of a short section of the line-above- ground plane type transmission lime, a pair of identical rectangular slabs of lossy material like polytron, whose longitudinal axes are parallel to and indentically spaced away from either side of the line, and a geared mechanism to adjust amd maintain this spaced relationship. This device permits optimum fineness and accuracy of attenuator control which heretofore has been difficult to achieve.

  2. Recombinant human laminin isoforms can support the undifferentiated growth of human embryonic stem cells

    SciTech Connect

    Miyazaki, Takamichi; Futaki, Sugiko; Hasegawa, Kouichi; Kawasaki, Miwa; Sanzen, Noriko; Hayashi, Maria; Kawase, Eihachiro; Sekiguchi, Kiyotoshi Nakatsuji, Norio; Suemori, Hirofumi

    2008-10-10

    Human embryonic stem cells (hESCs) are thought to be a promising cell source for cell transplantation therapy. For such a clinical application, the hESCs should be manipulated using appropriate and qualified materials. In this study, we examined the efficacy of recombinant human laminin (rhLM) isoforms on the undifferentiated growth of hESCs. We first determined the major integrins expressed on the hESCs to reveal the preference of the hESCs for rhLMs, and found that the hESCs mainly expressed integrin {alpha}6{beta}1, which binds predominantly to laminin-111, -332 and -511/-521. When the hESCs were seeded onto rhLMs, the cells indeed adhered markedly to rhLM-332, and to rhLM-511 and rhLM-111 to a lesser extent. The hESCs proliferated on these three rhLMs for several passages while preserving their pluripotency. These results show that rhLM-111, -332, and -511 are good substrates to expand undifferentiated hESCs due to their high affinity to integrin {alpha}6{beta}1 expressed on hESCs.

  3. Ship-produced cloud line of 13 July 1991

    SciTech Connect

    Hindman, E.E.; Porch, W.M.; Hudson, J.G.; Durkee, P.A.

    1992-12-31

    Steaming ships can produce long linear cloud lines in regions of fog and broken stratus as well as in marine stratus layers. The lines are not always detected in 0.63 {mu}m satellite images, but are often detected in the corresponding 3.7 {mu}m images because the lines contain smaller and more numerous droplets than the stratus in which they are embedded as deduced by Coakley, et al. and measured by Radke, et al. They postulate cloud condensation nuclei (CCN) from steaming ships produced the more numerous and, hence, smaller cloud droplets. The ship-produced clouds are not always detected in 0.63 {mu}m images because this wavelength is not as sensitive to changes in droplet size as is 3.7 {mu}m. On 13 July 1991 a dramatic, ship-produced cloud line formed offshore of Baja California. The authors present satellite images of the line and corresponding photographs from the R/V EGABRAG III which passed under the line. The images and photos reveal the structure of the line. The EGABRAG was a source of CCN but did not produce a cloud line; they attempt to explain this important finding.

  4. Ship-produced cloud line of 13 July 1991

    SciTech Connect

    Hindman, E.E. ); Porch, W.M. ); Hudson, J.G. ); Durkee, P.A. )

    1992-01-01

    Steaming ships can produce long linear cloud lines in regions of fog and broken stratus as well as in marine stratus layers. The lines are not always detected in 0.63 [mu]m satellite images, but are often detected in the corresponding 3.7 [mu]m images because the lines contain smaller and more numerous droplets than the stratus in which they are embedded as deduced by Coakley, et al. and measured by Radke, et al. They postulate cloud condensation nuclei (CCN) from steaming ships produced the more numerous and, hence, smaller cloud droplets. The ship-produced clouds are not always detected in 0.63 [mu]m images because this wavelength is not as sensitive to changes in droplet size as is 3.7 [mu]m. On 13 July 1991 a dramatic, ship-produced cloud line formed offshore of Baja California. The authors present satellite images of the line and corresponding photographs from the R/V EGABRAG III which passed under the line. The images and photos reveal the structure of the line. The EGABRAG was a source of CCN but did not produce a cloud line; they attempt to explain this important finding.

  5. 132. FORKS DIVERSION, HIGH LINE AND LOW LINE CANALS, TWIN ...

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

    132. FORKS DIVERSION, HIGH LINE AND LOW LINE CANALS, TWIN FALLS COUNTY, SOUTH OF HANSEN, IDAHO; POWER GATES FOR HYDRO-ELECTRIC. - Milner Dam & Main Canal: Twin Falls Canal Company, On Snake River, 11 miles West of city of Burley, Idaho, Twin Falls, Twin Falls County, ID

  6. GERM-LINE SPECIFIC FACTORS IN CHEMICAL MUTAGENESIS

    EPA Science Inventory

    Chemical mutagenesis test results ave not revealed evidence of germ-line specific mutagens. owever, conventional assays have indicated that there are male-female differences in mutagenic response, as well as quantitative/qualitative differences in induced mutations which depend u...

  7. Assessment of an On-Line Educational Administration Course

    ERIC Educational Resources Information Center

    Kim, Lori

    2004-01-01

    An on-line education administration course was evaluated using Seven Lessons Learned by Graham, Cagiltay, Lim, Craner, and Duffy (2001 ) as the framework. The framework was found to be useful in revealing areas of strengths and weaknesses in offering virtual education administration courses. The evaluation provided information that clear…

  8. Establishment and characterization of unique human gallbladder cancer cell lines.

    PubMed

    Ghosh, Mila; Koike, Naoto; Yanagimoto, Go; Tsunoda, Shin-Ichi; Kaul, Sunil; Hirano, Takashi; Emura, Fabian; Kashiwagi, Hironobu; Kawamoto, Toru; Ohkohchi, Nobuhiro; Saijo, Kaoru; Ohno, Tadao; Miwa, Masanao; Todoroki, Takeshi

    2004-05-01

    Gallbladder cancer has a dismal prognosis. Understanding the disease at the biological, genetic, molecular, cellular, and clinical level is essential for effective diagnostics and therapeutics. However, the currently established gallbladder cell lines are insufficient for better understanding and further research. The aim of our present study was to establish and characterize human gallbladder cancer cell lines. We established 5 cell lines from resected specimens of gallbladder cancers. These cell lines revealed typical tumor histopathological characteristics. We examined growth characteristics and the colony-forming ability of established cell lines in terms of their cell cycle parameters, expression of tumor markers (carcinoembryonic antigen; CEA, carbohydrated antigen 19-9; CA19-9, MUC-1 and c-kit) and the oncogene c-erbB2 by flow cytometer. Comparative genomic hybridization (CGH) analysis with specific gene probes was performed to detect changes in the gene copy numbers. Human origin of cell lines was confirmed by chromosomal analysis. Cells maintained differentiation characteristics of the original tumors. The doubling time of different cell lines varied from 30 to 96 h. All 5 cell lines formed colonies in the colony forming assays and expressed CEA, CA19-9, MUC-1 and the oncogene c-erbB2 and showed chromosomal aneuploidy. CGH analysis demonstrated gain of chromosomal region bearing SRC, RAB1, and PAP in all cell lines and hTERT in 4 cell lines. These newly established cell lines might serve as a useful model for studying the molecular pathogenesis of gallbladder cancer. Furthermore, they may serve as a model for testing new therapeutics against gallbladder cancer. These chromosomal aberrations and imbalances provide a starting point for molecular analyses of genomic regions and genes in gallbladder carcinogenesis. PMID:15067341

  9. Genomic Heterogeneity and Structural Variation in Soybean Near Isogenic Lines

    PubMed Central

    Stec, Adrian O.; Bhaskar, Pudota B.; Bolon, Yung-Tsi; Nolan, Rebecca; Shoemaker, Randy C.; Vance, Carroll P.; Stupar, Robert M.

    2013-01-01

    Near isogenic lines (NILs) are a critical genetic resource for the soybean research community. The ability to identify and characterize the genes driving the phenotypic differences between NILs is limited by the degree to which differential genetic introgressions can be resolved. Furthermore, the genetic heterogeneity extant among NIL sub-lines is an unaddressed research topic that might have implications for how genomic and phenotypic data from NILs are utilized. In this study, a recently developed high-resolution comparative genomic hybridization (CGH) platform was used to investigate the structure and diversity of genetic introgressions in two classical soybean NIL populations, respectively varying in protein content and iron deficiency chlorosis (IDC) susceptibility. There were three objectives: assess the capacity for CGH to resolve genomic introgressions, identify introgressions that are heterogeneous among NIL sub-lines, and associate heterogeneous introgressions with susceptibility to IDC. Using the CGH approach, introgression boundaries were refined and previously unknown introgressions were revealed. Furthermore, heterogeneous introgressions were identified within seven sub-lines of the IDC NIL “IsoClark.” This included three distinct introgression haplotypes linked to the major iron susceptible locus on chromosome 03. A phenotypic assessment of the seven sub-lines did not reveal any differences in IDC susceptibility, indicating that the genetic heterogeneity among the lines does not have a significant impact on the primary NIL phenotype. PMID:23630538

  10. Dynamic Proteomic Analysis of Pancreatic Mesenchyme Reveals Novel Factors That Enhance Human Embryonic Stem Cell to Pancreatic Cell Differentiation

    PubMed Central

    Russ, Holger A.; Landsman, Limor; Moss, Christopher L.; Higdon, Roger; Greer, Renee L.; Kaihara, Kelly; Salamon, Randy; Kolker, Eugene; Hebrok, Matthias

    2016-01-01

    Current approaches in human embryonic stem cell (hESC) to pancreatic beta cell differentiation have largely been based on knowledge gained from developmental studies of the epithelial pancreas, while the potential roles of other supporting tissue compartments have not been fully explored. One such tissue is the pancreatic mesenchyme that supports epithelial organogenesis throughout embryogenesis. We hypothesized that detailed characterization of the pancreatic mesenchyme might result in the identification of novel factors not used in current differentiation protocols. Supplementing existing hESC differentiation conditions with such factors might create a more comprehensive simulation of normal development in cell culture. To validate our hypothesis, we took advantage of a novel transgenic mouse model to isolate the pancreatic mesenchyme at distinct embryonic and postnatal stages for subsequent proteomic analysis. Refined sample preparation and analysis conditions across four embryonic and prenatal time points resulted in the identification of 21,498 peptides with high-confidence mapping to 1,502 proteins. Expression analysis of pancreata confirmed the presence of three potentially important factors in cell differentiation: Galectin-1 (LGALS1), Neuroplastin (NPTN), and the Laminin α-2 subunit (LAMA2). Two of the three factors (LGALS1 and LAMA2) increased expression of pancreatic progenitor transcript levels in a published hESC to beta cell differentiation protocol. In addition, LAMA2 partially blocks cell culture induced beta cell dedifferentiation. Summarily, we provide evidence that proteomic analysis of supporting tissues such as the pancreatic mesenchyme allows for the identification of potentially important factors guiding hESC to pancreas differentiation. PMID:26681951

  11. Characteristic impedance of microstrip lines

    NASA Technical Reports Server (NTRS)

    Bailey, M. C.; Deshpande, M. D.

    1989-01-01

    The dyadic Green's function for a current embedded in a grounded dielectric slab is used to analyze microstrip lines at millimeter wave frequencies. The dyadic Green's function accounts accurately for fringing fields and dielectric cover over the microstrip line. Using Rumsey's reaction concept, an expression for the characteristic impedance is obtained. The numerical results are compared with other reported results.

  12. Off-Line Catalog Production.

    ERIC Educational Resources Information Center

    OCLC Online Computer Library Center, Inc., Dublin, OH.

    The Ohio College Library Center's off-line catalog system is a limited technique for production of card catalogs. Unlike the on-line system, it cannotmake the resources of a region available to users in an individual institution, and it does not have the potential for significantly reducing rate of rise of library per-unit costs. In short, it is…

  13. Wiswesser Line Notation: An Introduction.

    ERIC Educational Resources Information Center

    Vollmer, John J.

    1983-01-01

    Wiswesser Line Notation, a system for representing structural formulas by a single line of common symbols (numerals, capital letters, and three punctuation marks), is gaining acceptance as an effective indexing tool. The resulting notations, denoting specific structural fragments of a molecule, can be retrieved by humans or computer. (Author/JN)

  14. relline: Relativistic line profiles calculation

    NASA Astrophysics Data System (ADS)

    Dauser, Thomas

    2015-05-01

    relline calculates relativistic line profiles; it is compatible with the common X-ray data analysis software XSPEC (ascl:9910.005) and ISIS (ascl:1302.002). The two basic forms are an additive line model (RELLINE) and a convolution model to calculate relativistic smearing (RELCONV).

  15. Cyclotron Line Measurements with INTEGRAL

    NASA Technical Reports Server (NTRS)

    Pottschmidt, K.; Kreykenbohm, I.; Caballero, I.; Fritz, S.; Schoenherr, G.; Kretschmar, P.; Wilms, J.; McBride, V. A.; Suchy, S.; Rothschild, R. E.

    2008-01-01

    Due to its broadband energy coverage, INTEGRAL has made important contributions to observing and interpreting cyclotron lines, which are present in the 10-100 keV range of a sample of accreting pulsars. In these systems photons with energies fulfilling the resonance condition inelastically Compton scatter off electrons quantized in the accretion column above the neutron star's magnetic pole(s). This process gives rise to the broad, absorption-like lines or 'cyclotron resonant scattering features' (CRSF). The observed lines allow to directly measure the B-fields of these sources, resulting in values of a few times 1E12G. In this overview I will present recent highlights regarding CRSF observations as well as discuss current ideas and models for the physical conditions in the accretion column. Among the former are the stability of the spectrum of Vela X-1 during giant flares in 2003, the observation of three cyclotron lines during the 2004 outburst of V0332+53, the confirmation of the fundamental line at approximately 45 keV during a 2005 normal outburst of A0535-26, and the simultaneous detection of the two lines in the dipping source 4U 1907+09 (for which also a torque reversal was detected for the first time). Through these and other observations it has become increasingly apparent that two types of observations can potentially be used to constrain the accretion column geometry: the determination of energy ratios for multiple harmonic lines (only two sources with greater than 2 lines are known), was well as the evolution of the fundamental line centroid, which, for different sources, may or may not be correlated with flux. Furthermore, first steps have been taken away from the usual phenomenological description of the lines, towards a physical approach based on self-consistent CRSF modeling. Initial applications are presented.

  16. Histone modification profiling reveals differential signatures associated with human embryonic stem cell self-renewal and differentiation.

    PubMed

    Bhanu, Natarajan V; Sidoli, Simone; Garcia, Benjamin A

    2016-02-01

    In this study, we trace developmental stages using epigenome changes in human embryonic stem cells (hESCs) treated with drugs modulating either self-renewal or differentiation. Based on microscopy, qPCR and flow cytometry, we classified the treatment outcome as inducing pluripotency (hESC, flurbiprofen and gatifloxacin), mesendoderm (sinomenine), differentiation (cyamarin, digoxin, digitoxin, selegeline and theanine) and lineage-commitment (RA). When we analyzed histone PTMs that imprinted these gene and protein expressions, the above classification was reassorted. Hyperacetylation at H3K4, 9, 14, 18, 56 and 122 as well as H4K5, 8, 12 and 16 emerged as the pluripotency signature of hESCs. Methylations especially of H3 at K9, K20, K27 and K36 characterized differentiation initiation as seen in no-drug control and fluribiprofen. Sinomenine-treated cells clustered close to "differentiation initiators", consistent with flow cytometry where it induced mesendoderm, along with cyamarin and possibly selegnine. Neurectoderm, induced by RA and theanine manifested methylations on H3 shifts to H3.3. By both flow cytometry and histone PTM clustering, it appears that cells treated with gatifloxacin, flurbiprofen, digitoxin and digoxin were not yet lineage-committed or mixed cell types. Taken together, our moderate-throughput histone PTM profiling approach highlighted subtle epigenetic signatures that permitted us to predict divergent lineage progression even in differentiating cells with similar phenotype and gene expression. PMID:26631989

  17. Histone modification profiling reveals differential signatures associated with human embryonic stem cell self-renewal and differentiation

    PubMed Central

    Bhanu, Natarajan V.; Sidoli, Simone; Garcia, Benjamin A.

    2016-01-01

    In this study, we trace developmental stages using epigenome changes in human embryonic stem cells (hESCs) treated with drugs modulating either self-renewal or differentiation. Based on microscopy, qPCR and flow cytometry, we classified the treatment outcome as inducing pluripotency (hESC, flurbiprofen and gatifloxacin), mesendoderm (sinomenine), differentiation (cyamarin, digoxin, digitoxin, selegeline and theanine) and lineage-commitment (RA). When we analyzed histone PTMs that imprinted these gene and protein expressions, the above classification was reassorted. Hyperacetylation at H3K4, 9, 14, 18, 56 and 122 as well as H4K5, 8, 12 and 16 emerged as the pluripotency signature of hESCs. Methylations especially of H3 at K9, K20, K27 and K36 characterized differentiation initiation as seen in no-drug control and fluribiprofen. Sinomenine-treated cells clustered close to “differentiation initiators”, consistent with flow cytometry where it induced mesendoderm, along with cyamarin and possibly selegnine. Neurectoderm, induced by RA and theanine manifested methylations on H3 shifts to H3.3. By both flow cytometry and histone PTM clustering, it appears that cells treated with gatifloxacin, flurbiprofen, digitoxin and digoxin were not yet lineage-committed or mixed cell types. Taken together, our moderate-throughput histone PTM profiling approach highlighted subtle epigenetic signatures that permitted us to predict divergent lineage progression even in differentiating cells with similar phenotype and gene expression. PMID:26631989

  18. Teaching Braille Line Tracking Using Stimulus Fading

    ERIC Educational Resources Information Center

    Scheithauer, Mindy C.; Tiger, Jeffrey H.

    2014-01-01

    Line tracking is a prerequisite skill for braille literacy that involves moving one's finger horizontally across a line of braille text and identifying when a line ends so the reader may reset his or her finger on the subsequent line. Current procedures for teaching line tracking are incomplete, because they focus on tracking lines with only…

  19. Characteristic Work Function Variations of Graphene Line Defects.

    PubMed

    Long, Fei; Yasaei, Poya; Sanoj, Raj; Yao, Wentao; Král, Petr; Salehi-Khojin, Amin; Shahbazian-Yassar, Reza

    2016-07-20

    Line defects, including grain boundaries and wrinkles, are commonly seen in graphene grown by chemical vapor deposition. These one-dimensional defects are believed to alter the electrical and mechanical properties of graphene. Unfortunately, it is very tedious to directly distinguish grain boundaries from wrinkles due to their similar morphologies. In this report, high-resolution Kelvin potential force microscopy (KPFM) is employed to measure the work function distribution of graphene line defects. The characteristic work function variations of grain boundaries, standing-collapsed wrinkles, and folded wrinkles could be clearly identified. Classical and quantum molecular dynamics simulations reveal that the unique work function distribution of each type of line defects is originated from the doping effect induced by the SiO2 substrate. Our results suggest that KPFM can be an easy-to-use and accurate method to detect graphene line defects, and also propose the possibility to tune the graphene work function by defect engineering. PMID:27356179

  20. Issues related to line-oriented flight training

    NASA Technical Reports Server (NTRS)

    Lauber, J. K.

    1981-01-01

    The use of a training simulator along with carefully structured, detailed, line trip scenarios was envisioned by NASA as a means of providing a controllable, repeatable way to observe line crews in a highly realistic simulation of their working environment and obtain better understanding operationally significant human factors problems and issues. Relevant research done by the agency and the results of full-mission simulation scenarios revealed potential implications for flight training. Aspects to be considered in creating training programs closely related to the actual line environment with a total crew application in real world incident experiences include: (1) operational, environmental, equipment, and crew problems in scenario design; (2) real time line oriented flight training operation; (3) performance assessment and debriefing; (4) instructor qualification and training; and (5) other issues such as ub un initial, transition, and upgrade training; procedures developent and evaluation, and equipment evaluation.

  1. Solar gamma-ray lines and interplanetary solar protons

    NASA Astrophysics Data System (ADS)

    Yoshimori, M.

    1985-03-01

    Solar gamma-ray lines and protons were simultaneously observed for six flares on April 1, 4, and 27, 1981, May 13, 1981, February 1, 1982, and June 6, 1982 by the Hinotori and Himawari satellites. The flare list is presented, and the time histories of gamma-rays and protons are shown. The relationship between the gamma-ray line fluences and peak proton fluxes for these flares does not reveal an apparent correlation between them. The present results imply that the protons producing gamma-ray lines in the flare region, and protons observed near the earth, do not always belong to the same population, and favor the downward streaming model for the gamma-ray line production.

  2. Radio Recombination Lines as Tools for Astronomers and Physicists

    NASA Astrophysics Data System (ADS)

    Gordon, M. A.

    2008-10-01

    Described by simple atomic theory published in 1913 by Niels Bohr, spectral lines in the radio range arising from transitions between large principal quantum numbers of atoms have proved to be useful tools for astronomers and physicists. Called ``radio recombination lines'' because of the wavelength range where most are observed, they are usually easy to detect, give unique information about astronomical objects, and facilitate the study of physical effects in environments that cannot be created in terrestrial laboratories. Observations have revealed unexpected results regarding thermodynamic populations of the principal quantum levels and about pressure broadening in astronomical environments. Detections of large-n lines, such as the n = 1006-->1010 absorption line of interstellar carbon, show the existence of atoms with classical diameters of about 0.1 mm, the thickness of a sheet of typing paper. This paper briefly discusses observations of Stark broadening reported by Bell et al. in 2002.

  3. Plan competitions reveal entrepreneurial talent

    SciTech Connect

    Madison, Alison L.

    2011-05-15

    Monthly economic diversity column for Tri-City Herald business section. Excerpt below: There’s something to be said for gaining valuable real-world experience in a structured, nurturing environment. Take for instance learning to scuba dive in the comfort of my resort pool rather than immediately hanging out with sharks while I figure out little things like oxygen tanks and avoiding underwater panic attacks. Likewise, graduate students are getting some excellent, supportive real-world training through university business plan competitions. These competitions are places where smart minds, new technologies, months of preparation and coaching, and some healthy pre-presentation jitters collide to reveal not only solid new business ideas, but also some promising entrepreneurial talent. In fact, professionals from around our region descend upon college campuses every spring to judge these events, which help to bridge the gap between academics and the real technology and business-driven economy.

  4. Revealing ontological commitments by magic.

    PubMed

    Griffiths, Thomas L

    2015-03-01

    Considering the appeal of different magical transformations exposes some systematic asymmetries. For example, it is more interesting to transform a vase into a rose than a rose into a vase. An experiment in which people judged how interesting they found different magic tricks showed that these asymmetries reflect the direction a transformation moves in an ontological hierarchy: transformations in the direction of animacy and intelligence are favored over the opposite. A second and third experiment demonstrated that judgments of the plausibility of machines that perform the same transformations do not show the same asymmetries, but judgments of the interestingness of such machines do. A formal argument relates this sense of interestingness to evidence for an alternative to our current physical theory, with magic tricks being a particularly pure source of such evidence. These results suggest that people's intuitions about magic tricks can reveal the ontological commitments that underlie human cognition. PMID:25490128

  5. CMB source apportionment during REVEAL

    SciTech Connect

    Lowenthal, D.H.; Gertler, A.W.; Wittorff, D.; Sakiyama, S.

    1997-01-01

    Source contributions to PM{sub 2.5} aerosol measured at Chilliwack and Pitt Meadows in the Lower Fraser Valley, B.C., Canada, during the REVEAL study, were estimated using chemical mass balance (CMB) receptor modeling. ON average, motor vehicles accounted for 34 and 43% of PM{sub 2.5} at Chilliwack and Pitt Meadows, respectively. Secondary sulfate and secondary nitrate were the next most significant PM{sub 2.5} components, accounting for 25 and 27%, respectively, at Chilliwack, and 27 and 12%, respectively, at Pitt Meadows. Geological material accounted for 3 and 5% of PM{sub 2.5} at Chilliwack and Pitt Meadows, respectively. A significant contribution of wood smoke was estimated for both sites: 8% at Chilliwack and 9% at Pitt Meadows.

  6. Archimedes: Accelerator Reveals Ancient Text

    SciTech Connect

    Bergmann, Uwe

    2004-02-24

    Archimedes (287-212 BC), who is famous for shouting 'Eureka' (I found it) is considered one of the most brilliant thinkers of all times. The 10th-century parchment document known as the 'Archimedes Palimpsest' is the unique source for two of the great Greek's treatises. Some of the writings, hidden under gold forgeries, have recently been revealed at the Stanford Synchrotron Radiation Laboratory at SLAC. An intense x-ray beam produced in a particle accelerator causes the iron in original ink, which has been partly erased and covered, to send out a fluorescence glow. A detector records the signal and a digital image showing the ancient writings is produced. Please join us in this fascinating journey of a 1,000-year-old parchment from its origin in the Mediterranean city of Constantinople to a particle accelerator in Menlo Park.

  7. Erosion and what it Reveals

    NASA Technical Reports Server (NTRS)

    2003-01-01

    [figure removed for brevity, see original site]

    Released 20 November 2003

    This image is located near the boundary between Syrtis Major and Isidis Planitia. The top of the image shows rough material that has eroded away from the lower portion of the image, revealing an underlying surface that has many small craters. It also reveals an ancient flow lobe that is barely discernable, crossing the southern part of the image (this flow lobe is much easier to see as a smooth region in the context image).

    Image information: VIS instrument. Latitude 16.4, Longitude 77.9 East (282.1 West). 19 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

  8. Dunes reveal Titan's recent history

    NASA Astrophysics Data System (ADS)

    Savage, Christopher J.; Radebaugh, Jani

    2010-04-01

    Large fields of linear dunes are abundant on Titan, covering nearly 20% of the surface. They are among the youngest features and represent interactions between near-surface winds and sediment. This interaction may vary from area to area creating unique populations of eolian features identified by dune field parameters such as crest-to-crest spacing, dune width and orientation. These parameters respond to changes in near-surface conditions over periods of time ranging from minutes to many thousands of years depending on dune size and the duration of the changes. While pattern analysis of dune field parameters on Earth and, in this study, Titan reveals much about current climatic conditions, such as wind regimes and wetter vs. drier areas, many inferences about past conditions can also be made. Initial pattern analysis of linear dunes on Titan reveals a single population of linear dunes representing a large percentage of all observed dunes. This single population is the result of two leading possibilities: Either there has been only one long period of dune building, leading to very old cores that have been built upon over long periods of time, perhaps punctuated with few or many intervals of non-deposition; or the current conditions of dune building have persisted long enough to completely erase any evidence of previous conditions. We have not yet worked through all the input parameters to adjust Earth's time scales to Titan's, and thus it is not yet possible to give a precise age for Titan's dunes. However, if these large linear dunes are similar to Earth's large linear dunes, they may represent at least several thousand years of dune building.

  9. Coordinating Numeric and Linear Units: Elementary Students' Strategies for Locating Whole Numbers on the Number Line

    ERIC Educational Resources Information Center

    Saxe, Geoffrey B.; Shaughnessy, Meghan M.; Gearhart, Maryl; Haldar, Lina Chopra

    2013-01-01

    Two investigations of fifth graders' strategies for locating whole numbers on number lines revealed patterns in students' coordination of numeric and linear units. In Study 1, we investigated the effects of context on students' placements of three numbers on an open number line. For one group ("n"?=?24), the line was…

  10. A Web-Based Synchronous Collaborative Review Tool: A Case Study of an On-Line Graduate Course

    ERIC Educational Resources Information Center

    Serce, Fatma Cemile; Yildirim, Soner

    2006-01-01

    On-line collaboration is an instructional method that facilitates collaboration in an on-line learning setting. To promote effective collaboration, it is vital to reveal both the student's and the instructor's point of view pertaining to effective on-line collaboration. In this study, the effectiveness of a learning management system in on-line…

  11. Pulse shaping with transmission lines

    DOEpatents

    Wilcox, R.B.

    1985-08-15

    A method and apparatus for forming shaped voltage pulses uses passive reflection from a transmission line with nonuniform impedance. The impedance of the reflecting line varies with length in accordance with the desired pulse shape. A high voltage input pulse is transmitted to the reflecting line. A reflected pulse is produced having the desired shape and is transmitted by pulse removal means to a load. Light activated photoconductive switches made of silicon can be utilized. The pulse shaper can be used to drive a Pockels cell to produce shaped optical pulses.

  12. Pulse shaping with transmission lines

    DOEpatents

    Wilcox, Russell B.

    1987-01-01

    A method and apparatus for forming shaped voltage pulses uses passive reflection from a transmission line with nonuniform impedance. The impedance of the reflecting line varies with length in accordance with the desired pulse shape. A high voltage input pulse is transmitted to the reflecting line. A reflected pulse is produced having the desired shape and is transmitted by pulse removal means to a load. Light activated photoconductive switches made of silicon can be utilized. The pulse shaper can be used to drive a Pockels cell to produce shaped optical pulses.

  13. Electrical transmission line diametrical retainer

    DOEpatents

    Hall, David R.; Hall, Jr., H. Tracy; Pixton, David; Dahlgren, Scott; Sneddon, Cameron; Briscoe, Michael; Fox, Joe

    2004-12-14

    The invention is a mechanism for retaining an electrical transmission line. In one embodiment of the invention it is a system for retaining an electrical transmission line within down hole components. In accordance with one aspect of the invention, the system includes a plurality of downhole components, such as sections of pipe in a drill string. The system also includes a coaxial cable running between the first and second end of a drill pipe, the coaxial cable having a conductive tube and a conductive core within it. The invention allows the electrical transmission line to with stand the tension and compression of drill pipe during routine drilling cycles.

  14. Spitzer Reveals Stellar 'Family Tree'

    NASA Technical Reports Server (NTRS)

    2008-01-01

    [figure removed for brevity, see original site] High resolution poster version

    Generations of stars can be seen in this new infrared portrait from NASA's Spitzer Space Telescope. In this wispy star-forming region, called W5, the oldest stars can be seen as blue dots in the centers of the two hollow cavities (other blue dots are background and foreground stars not associated with the region). Younger stars line the rims of the cavities, and some can be seen as pink dots at the tips of the elephant-trunk-like pillars. The white knotty areas are where the youngest stars are forming. Red shows heated dust that pervades the region's cavities, while green highlights dense clouds.

    W5 spans an area of sky equivalent to four full moons and is about 6,500 light-years away in the constellation Cassiopeia. The Spitzer picture was taken over a period of 24 hours.

    Like other massive star-forming regions, such as Orion and Carina, W5 contains large cavities that were carved out by radiation and winds from the region's most massive stars. According to the theory of triggered star-formation, the carving out of these cavities pushes gas together, causing it to ignite into successive generations of new stars.

    This image contains some of the best evidence yet for the triggered star-formation theory. Scientists analyzing the photo have been able to show that the ages of the stars become progressively and systematically younger with distance from the center of the cavities.

    This is a three-color composite showing infrared observations from two Spitzer instruments. Blue represents 3.6-micron light and green shows light of 8 microns, both captured by Spitzer's infrared array camera. Red is 24-micron light detected by Spitzer's multiband imaging photometer.

  15. Spitzer Reveals Stellar 'Family Tree'

    NASA Technical Reports Server (NTRS)

    2008-01-01

    [figure removed for brevity, see original site] High resolution poster version

    Generations of stars can be seen in this new infrared portrait from NASA's Spitzer Space Telescope. In this wispy star-forming region, called W5, the oldest stars can be seen as blue dots in the centers of the two hollow cavities (other blue dots are background and foreground stars not associated with the region). Younger stars line the rims of the cavities, and some can be seen as dots at the tips of the elephant-trunk-like pillars. The white knotty areas are where the youngest stars are forming.

    W5 spans an area of sky equivalent to four full moons and is about 6,500 light-years away in the constellation Cassiopeia. The Spitzer picture was taken over a period of 24 hours.

    Like other massive star-forming regions, such as Orion and Carina, W5 contains large cavities that were carved out by radiation and winds from the region's most massive stars. According to the theory of triggered star-formation, the carving out of these cavities pushes gas together, causing it to ignite into successive generations of new stars.

    This image contains some of the best evidence yet for the triggered star-formation theory. Scientists analyzing the photo have been able to show that the ages of the stars become progressively and systematically younger with distance from the center of the cavities.

    This picture was taken with Spitzer's infrared array camera. It is a four-color composite, in which light with a wavelength of 3.6 microns is blue; 4.5-micron light is green; 5.8-micron light is orange; and 8-micron light is red.

  16. Strike-slip movements and thrusting along a transpressive fault zone: The North Giudicarie line (Insubric line, northern Italy)

    NASA Astrophysics Data System (ADS)

    Prosser, Giacomo

    1998-12-01

    This paper analyzes the kinematic evolution and the deformation partitioning within an important transpressive fault zone located in the central part of the Alpine chain. The North Giudicarie line is a NNE trending fault which offsets the dextral Insubric line with an apparent left-lateral displacement of about 70 km. The main fault plane of the North Giudicarie line dips about 35°-45° to the NW. The footwall is characterized by N-S striking strike-slip faults, which reactivate extensional faults of Early Jurassic to Late Cretaceous age. The early deformation history of the North Giudicarie line is revealed by basement-and limestone-mylonites. Shear sense of mylonites indicates on average top-to-the-east thrusting. These movements took place during the late Oligocene-early Miocene, when the Insubric line was active as a right-lateral strike-slip fault. Therefore, in this time span the North Giudicarie line can be interpreted as a dextral transpressive bend of the Insubric line. Mylonites have later been overprinted by brittle faults related to top-to-the-SE thrusting of middle-late Miocene age. During this event the shape of the Insubric line was strongly modified by left-lateral transpression along the Giudicarie fault zone. Deformation was partitioned between prevailing compression along the Giudicarie line and left-lateral strike-slip movements along the N-S striking faults. These faults transferred the strike-slip component of the Giudicarie line into a wider area of the central southern Alps.

  17. APEX reveals glowing stellar nurseries

    NASA Astrophysics Data System (ADS)

    2008-11-01

    Illustrating the power of submillimetre-wavelength astronomy, an APEX image reveals how an expanding bubble of ionised gas about ten light-years across is causing the surrounding material to collapse into dense clumps that are the birthplaces of new stars. Submillimetre light is the key to revealing some of the coldest material in the Universe, such as these cold, dense clouds. Glowing Stellar Nurseries ESO PR Photo 40/08 Glowing Stellar Nurseries The region, called RCW120, is about 4200 light years from Earth, towards the constellation of Scorpius. A hot, massive star in its centre is emitting huge amounts of ultraviolet radiation, which ionises the surrounding gas, stripping the electrons from hydrogen atoms and producing the characteristic red glow of so-called H-alpha emission. As this ionised region expands into space, the associated shock wave sweeps up a layer of the surrounding cold interstellar gas and cosmic dust. This layer becomes unstable and collapses under its own gravity into dense clumps, forming cold, dense clouds of hydrogen where new stars are born. However, as the clouds are still very cold, with temperatures of around -250˚ Celsius, their faint heat glow can only be seen at submillimetre wavelengths. Submillimetre light is therefore vital in studying the earliest stages of the birth and life of stars. The submillimetre-wavelength data were taken with the LABOCA camera on the 12-m Atacama Pathfinder Experiment (APEX) telescope, located on the 5000 m high plateau of Chajnantor in the Chilean Atacama desert. Thanks to LABOCA's high sensitivity, astronomers were able to detect clumps of cold gas four times fainter than previously possible. Since the brightness of the clumps is a measure of their mass, this also means that astronomers can now study the formation of less massive stars than they could before. The plateau of Chajnantor is also where ESO, together with international partners, is building a next generation submillimetre telescope, ALMA

  18. In-line hydraulic dashpot

    NASA Astrophysics Data System (ADS)

    Moody, Paul E.

    1992-10-01

    An in-line hydraulic dashpot is disclosed that effectively decelerates the piston of a power cylinder by controllably choking off the oil which is providing pressure to the piston. The in-line hydraulic dashpot of the invention includes a valve spool member movable between an open and closed position along a fluid flow path that supplies oil to the power cylinder. An actuator rod is cooperative with the valve spool member and the piston shaft of the power cylinder to move tile valve spool member between its open and closed positions. The in-line hydraulic dashpot eliminates the clashing of mechanical parts and therewith eliminates the noise that would otherwise be generated thereby. The in-line hydraulic dashpot of the present invention makes possible the adaptation of a fixed stroke power cylinder to applications that call for a variable stroke length.

  19. Coplanar Waveguide Radial Line Stub

    NASA Technical Reports Server (NTRS)

    Simons, R. N.; Taub, S. R.

    1993-01-01

    A coplanar waveguide radial line stub resonator is experimentally characterized with respect to stub radius, sectoral angle, substrate thickness, and relative dielectric constant. A simple closed-form design equation which predicts the resonance radius of the stub is presented.

  20. Winslow Homer: "The Life Line."

    ERIC Educational Resources Information Center

    Brigham, Diane

    1986-01-01

    Using a color print of Winslow Homer's oil painting, "The Life Line," the goal of this senior high school art lesson is to have students debate the importance of dramatic effect in a work of art. (JDH)

  1. Umbilic Lines in Orientational Order

    NASA Astrophysics Data System (ADS)

    Machon, Thomas; Alexander, Gareth P.

    2016-01-01

    Three-dimensional orientational order in systems whose ground states possess nonzero gradients typically exhibits linelike structures or defects: λ lines in cholesterics or Skyrmion tubes in ferromagnets, for example. Here, we show that such lines can be identified as a set of natural geometric singularities in a unit vector field, the generalization of the umbilic points of a surface. We characterize these lines in terms of the natural vector bundles that the order defines and show that they give a way to localize and identify Skyrmion distortions in chiral materials—in particular, that they supply a natural representative of the Poincaré dual of the cocycle describing the topology. Their global structure leads to the definition of a self-linking number and helicity integral which relates the linking of umbilic lines to the Hopf invariant of the texture.

  2. Line competition in gas lasers

    SciTech Connect

    Pikulev, A A

    2000-08-31

    Simultaneous lasing on two lines having a common upper level is analysed. Within the framework of the two-level model, a formula for the gain in each line is obtained. Two possible types of line competition are found: symbiosis and quenching. It is shown that competition in the quenching regime can be used to study the rates of population of the lower levels. The results are illustrated by competition diagrams for the 2.65-{mu}m and 2.03-{mu}m of the Xe atom in the Ar-Xe mixture and the 703.2-nm and 724.5-nm lines of the Ne atom in the Ne-Ar mixture. (active media)

  3. Horizontal and Vertical Line Designs.

    ERIC Educational Resources Information Center

    Johns, Pat

    2003-01-01

    Presents an art lesson in which students learn about the artist Piet Mondrian and create their own abstract artworks. Focuses on geometric shapes using horizontal and vertical lines. Includes background information about the artist. (CMK)

  4. Plant transcriptomes reveal hidden guests.

    PubMed

    Zhu, Jiao; Wang, Guirong; Pelosi, Paolo

    2016-06-01

    With the wide adoption of transcriptome sequencing an ever increasing amount of information is becoming available, together with spurious data originating from contamination. We show that sometimes errors and inaccuracy can turn beneficial, revealing insect and arthropod pests when analysing plant transcriptomes. We have found a large number of soluble olfactory proteins, odorant-binding proteins (OBPs) and chemosensory proteins (CSPs), in plant databases, likely due to contamination by guest insects. In fact, both classes of proteins are only expressed in insects, with few CSPs also present in other arthropods. In addition, we found many sequences of the Niemann-Pick (Npc2) family, proteins dedicated to cholesterol transport in vertebrates and hypothesised to be involved in chemical communication in insects, but absent in plants. In several cases we were able to trace down members of the three classes of proteins to the insect or arthopod species responsible for contamination. Our work suggests that genes found in plants and recognised as contaminants can be turned into useful information to investigate plant-insect relationships or to identify new sequences from insects species not yet investigated. PMID:27130825

  5. Revealing ionic motion molecular solids

    NASA Astrophysics Data System (ADS)

    Thurzo, I.; Zahn, D. R. T.

    2006-01-01

    Thin films of mixed valence semiconductor copper-tetracyano-quinodimethane (Cu-TCNQ) and small molecule tris(8-hydroxyquinolinato)aluminum (Alq3) were investigated by current-voltage (I-V) and admittance (C-V,G-V) techniques in single-layer configuration with different combinations of electrodes. The predicted hysteresis of I-V curves and nearly constant loss (NCL) could be observed for both materials. When cycling devices between negative and positive biases, slowly decaying ionic transient currents interfere with steady-state currents and point to unidirectional motions of the ionic species with subsequent redox reaction at one of the electrodes. Plotting the frequency f dependence of the equivalent parallel capacitance at zero bias as log10 C(0) versus log10(f), the dielectric behavior of Cu-TCNQ and Alq3 complies with the effective-medium model for NCL in ionic conductors [J. R. Macdonald J. Appl. Phys. 94, 558 (2003)]. It also holds for a similar plot of the equivalent parallel zero-bias conductance G(0). The nature of the revealed mobile ions is discussed with emphasis on their sources.

  6. Challenges in LER/CDU metrology in DSA: placement error and cross-line correlations

    NASA Astrophysics Data System (ADS)

    Constantoudis, Vassilios; Kuppuswamy, Vijaya-Kumar M.; Gogolides, Evangelos; Pret, Alessandro V.; Pathangi, Hari; Gronheid, Roel

    2016-03-01

    DSA lithography poses new challenges in LER/LWR metrology due to its self-organized and pitch-based nature. To cope with these challenges, a novel characterization approach with new metrics and updating the older ones is required. To this end, we focus on two specific challenges of DSA line patterns: a) the large correlations between the left and right edges of a line (line wiggling, rms(LWR)line correlations, i.e. the resemblance of wiggling fluctuations of nearby lines. The first is quantified by the Line Center Roughness whose low-frequency part is related to the local placement errors of device structures. For the second, we propose the c-factor correlation function which quantifies the strength of the correlations between lines versus their horizontal distance in pitches. Also, we define roughness and uniformity parameters for the pitch changes along and across lines. The proposed characterization approach is applied to the analysis of line/space patterns obtained with the Liu-Nealey (LiNe) flow (post PMMA removal and pattern transfer) revealing the effects of pattern transfer on roughness and uniformity. Finally, we calculate the cfactor function of various Next-Generation Lithography techniques and reveal their distinct footprint on the extent of cross-line correlations.

  7. Fin-line horn antenna

    DOEpatents

    Reindel, John

    1990-01-01

    A fin line circuit card containing a fin line slot feeds a dipole antenna ich extends a quarterwave outside the waveguide and provides an energy beam focal point at or near the open end of the waveguide. The dipole antenna thus maintains a wide and nearly constant beamwidth, low VSWR and a circular symmetric radiation pattern for use in electronic warfare direction finding and surveillance applications.

  8. Diffusion from a line source

    NASA Technical Reports Server (NTRS)

    Burns, R. E.

    1973-01-01

    The problem with predicting pollutant diffusion from a line source of arbitrary geometry is treated. The concentration at the line source may be arbitrarily varied with time. Special attention is given to the meteorological inputs which act as boundary conditions for the problem, and a mixing layer of arbitrary depth is assumed. Numerical application of the derived theory indicates the combinations of meteorological parameters that may be expected to result in high pollution concentrations.

  9. Trace Metal Associations with Manganese-Rich Surface Coatings of Lead Service Lines

    EPA Science Inventory

    Analysis of lead service line samples from U. S. Environmental Protection Agency’s long-term research program to evaluate control and metal release from domestic drinking water service lines has revealed that Manganese-rich solids also contain Iron and sometimes Aluminum have fre...

  10. Linear rotary optical delay lines.

    PubMed

    Skorobogatiy, Maksim

    2014-05-19

    I present several classes of analytical and semi-analytical solutions for the design of high-speed rotary optical delay lines that use a combination of stationary and rotating curvilinear reflectors. Detailed analysis of four distinct classes of optical delay lines is presented. Particularly, I consider delay lines based on a single rotating reflector, a single rotating reflector and a single stationary reflector, two rotating reflectors, and two rotating reflectors and a single stationary reflector. I demonstrate that in each of these cases it is possible to design an infinite variety of the optical delay lines featuring linear dependence of the optical delay on the rotation angle. This is achieved via shape optimization of the rotating and stationary reflector surfaces. Moreover, in the case of two rotating reflectors a convenient spatial separation of the incoming and outgoing beams is possible. For the sake of example, all the blades presented in this paper are chosen to fit into a circle of 10 cm diameter and these delay lines feature in excess of 600 ps of optical delay. Finally, two prototypes of rotary delay lines were fabricated using CNC machining, and their optical properties are characterized. PMID:24921303

  11. A robust line extraction and matching algorithm

    NASA Technical Reports Server (NTRS)

    Hussien, B.; Sridhar, B.

    1993-01-01

    This paper presents an algorithm for extracting straight lines from intensity mages and describes a line matching algorithm for solving the line correspondence problem. The line extraction process begins by detecting edges in the intensity image. Next, line support regions are formed where image points (pixels) have similar gradient orientation. A line fitting algorithm is then used to fit a line to the points in the line support region based on a least means square fitting algorithm. Finally, line segments are linked together to form the final lines by using an adaptive line linking method; this results in much stronger lines and a smaller set of lines to be considered. Once the lines are detected in a sequence of images, a line matching algorithm is used to match lines in one image to the lines in the other image. The images are either from a motion or stereo sequence. The matched lines may then be used with the sensor position and orientation data to estimate range to objects corresponding to the lines. We present results based on applying the line extraction and line matching algorithms to a synthetic image and an outdoor scene captured by a camera on a helicopter.

  12. Line-by-line spectroscopic simulations on graphics processing units

    NASA Astrophysics Data System (ADS)

    Collange, Sylvain; Daumas, Marc; Defour, David

    2008-01-01

    We report here on software that performs line-by-line spectroscopic simulations on gases. Elaborate models (such as narrow band and correlated-K) are accurate and efficient for bands where various components are not simultaneously and significantly active. Line-by-line is probably the most accurate model in the infrared for blends of gases that contain high proportions of H 2O and CO 2 as this was the case for our prototype simulation. Our implementation on graphics processing units sustains a speedup close to 330 on computation-intensive tasks and 12 on memory intensive tasks compared to implementations on one core of high-end processors. This speedup is due to data parallelism, efficient memory access for specific patterns and some dedicated hardware operators only available in graphics processing units. It is obtained leaving most of processor resources available and it would scale linearly with the number of graphics processing units in parallel machines. Line-by-line simulation coupled with simulation of fluid dynamics was long believed to be economically intractable but our work shows that it could be done with some affordable additional resources compared to what is necessary to perform simulations on fluid dynamics alone. Program summaryProgram title: GPU4RE Catalogue identifier: ADZY_v1_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/ADZY_v1_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.: 62 776 No. of bytes in distributed program, including test data, etc.: 1 513 247 Distribution format: tar.gz Programming language: C++ Computer: x86 PC Operating system: Linux, Microsoft Windows. Compilation requires either gcc/g++ under Linux or Visual C++ 2003/2005 and Cygwin under Windows. It has been tested using gcc 4.1.2 under Ubuntu Linux 7.04 and using Visual C

  13. Hepatocellular carcinoma cell lines retain the genomic and transcriptomic landscapes of primary human cancers

    PubMed Central

    Qiu, Zhixin; Zou, Keke; Zhuang, Liping; Qin, Jianjie; Li, Hong; Li, Chao; Zhang, Zhengtao; Chen, Xiaotao; Cen, Jin; Meng, Zhiqiang; Zhang, Haibin; Li, Yixue; Hui, Lijian

    2016-01-01

    Hepatocellular carcinoma (HCC) cell lines are useful in vitro models for the study of primary HCCs. Because cell lines acquire additional mutations in culture, it is important to understand to what extent HCC cell lines retain the genetic landscapes of primary HCCs. Most HCC cell lines were established during the last century, precluding comparison between cell lines and primary cancers. In this study, 9 Chinese HCC cell lines with matched patient-derived cells at low passages (PDCs) were established in the defined culture condition. Whole genome analyses of 4 HCC cell lines showed that genomic mutation landscapes, including mutations, copy number alterations (CNAs) and HBV integrations, were highly stable during cell line establishment. Importantly, genetic alterations in cancer drivers and druggable genes were reserved in cell lines. HCC cell lines also retained gene expression patterns of primary HCCs during in vitro culture. Finally, sequential analysis of HCC cell lines and PDCs at different passages revealed their comparable and stable genomic and transcriptomic levels if maintained within proper passages. These results show that HCC cell lines largely retain the genomic and transcriptomic landscapes of primary HCCs, thus laying the rationale for testing HCC cell lines as preclinical models in precision medicine. PMID:27273737

  14. Hepatocellular carcinoma cell lines retain the genomic and transcriptomic landscapes of primary human cancers.

    PubMed

    Qiu, Zhixin; Zou, Keke; Zhuang, Liping; Qin, Jianjie; Li, Hong; Li, Chao; Zhang, Zhengtao; Chen, Xiaotao; Cen, Jin; Meng, Zhiqiang; Zhang, Haibin; Li, Yixue; Hui, Lijian

    2016-01-01

    Hepatocellular carcinoma (HCC) cell lines are useful in vitro models for the study of primary HCCs. Because cell lines acquire additional mutations in culture, it is important to understand to what extent HCC cell lines retain the genetic landscapes of primary HCCs. Most HCC cell lines were established during the last century, precluding comparison between cell lines and primary cancers. In this study, 9 Chinese HCC cell lines with matched patient-derived cells at low passages (PDCs) were established in the defined culture condition. Whole genome analyses of 4 HCC cell lines showed that genomic mutation landscapes, including mutations, copy number alterations (CNAs) and HBV integrations, were highly stable during cell line establishment. Importantly, genetic alterations in cancer drivers and druggable genes were reserved in cell lines. HCC cell lines also retained gene expression patterns of primary HCCs during in vitro culture. Finally, sequential analysis of HCC cell lines and PDCs at different passages revealed their comparable and stable genomic and transcriptomic levels if maintained within proper passages. These results show that HCC cell lines largely retain the genomic and transcriptomic landscapes of primary HCCs, thus laying the rationale for testing HCC cell lines as preclinical models in precision medicine. PMID:27273737

  15. [Staple line presenting as a suspicious pulmonary nodule].

    PubMed

    Agrafiotis, A C; Prieto, M; Siat, J; Grosdidier, G

    2016-05-01

    The case reported herein concerns a 47-year-old female patient with a right apical pulmonary nodule, which was hypermetabolic on positron emission tomography scan. Eighteen years ago, she underwent a right axillary thoracotomy with apical blebectomy and pleurectomy for a recurrent primary spontaneous pneumothorax. Percutaneous transthoracic needle biopsy was unsuccessful. Surgical exploration revealed that the lesion was in fact the reinforcement material of the staple line that was used in the previous operation. Even though it is a rare presentation, previous use of surgical staples should alert the care team, especially when a new nodule is located near the ancient staple line, in order to avoid unnecessary invasive procedures. PMID:27113616

  16. Establishment and characterization of a cell line from the Chinese soft-shelled turtle Pelodiscus sinensis.

    PubMed

    Guo, Haijie; Xia, Zhaonan; Tang, Wei; Mao, Zhijuan; Qian, Guoying; Wang, Caisheng

    2016-06-01

    The establishment and partial characterization of Pelodiscus sinensis continuous cell line is described here. A novel P. sinensis fibroblast cell line, designated PSF, was established from heart tissue by the semi-digestion explant culture technique. Since its initiation in July 2013, the cell line has been subcultured at 30°C in minimal essential medium (MEM) containing 15% (v/v) fetal bovine serum for more than 50 passages. The growth curve of the cell line revealed the population doubling time was 51.1 h. Karyotyping analysis indicated the modal chromosome number was 66, and no microbial contamination was detected. The PSF cell line produced significant fluorescent signals after transfection with plasmid pEGFP-C3. Analysis of mitochondrial cytochrome D-loop sequences revealed 96% identity among other Chinese turtle subspecies. Several cell line characterizations included morphological analysis and immunocytochemistry, which revealed the origin of the PSF cell line was fibroblast-like cells. Measurement of the isoenzymes lactic dehydrogenase and malic dehydrogenase showed no cross-contamination of this cell line with other species. This newly established cell line will be a valuable tool for transgenic and genetic manipulation studies and will act as an efficient instrument for studies of the viral diseases of the soft-shelled turtle. PMID:27059326

  17. Building Bridges One Line at a Time

    ERIC Educational Resources Information Center

    Grigsby, Cathy Murray

    2012-01-01

    In this article, first-grade students were taught the different kinds of lines that were part of the construction of various bridges--the curved lines of the arches of stone bridges, straight lines connecting the cables of a suspension bridge, vertical lines, horizontal lines, and so on. They gained practice in drawing structures and in fine brush…

  18. 25 CFR 169.22 - Service lines.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... undertaken to construct a service line across such land. Such a service line shall be limited in the case of power lines to a voltage of 14.5 kv. or less except lines to serve irrigation pumps and commercial and industrial uses which shall be limited to a voltage not to exceed 34.5 kv. A service line shall be for...

  19. Towards Perfect Water Line Intensities

    NASA Astrophysics Data System (ADS)

    Lodi, L.; Tennyson, J.

    2012-06-01

    Over the last ten years the increased availability of computational resources and the steady refinement of theoretical methods have permitted more and more accurate first principle calculations of water-vapor spectra as exemplified, e.g., by the very successful BT2 line list both line positions and intensities, a reliable dipole moment surface (DMS), affecting line intensities. It is also very useful to several application to give reasonable uncertainty bars for computed quantities, an aspect which traditionally has received little attention. We report here recent progress leading to very accurate room-temperature linelists covering the range 0.05-20 000 cm-1, complete with uncertainty bars, for the H_218O and H_217O water isotopologues Line intensities were produced using a recent DMS produced by our group which is capable of giving line intensites accurate to 1% for most medium and strong transitions. Line positions are based if possible on the experimentally derived energy levels recently produced by a IUPAC task group and have a typical accuracy of 0.0002 cm-1; when experimentally derived energy levels are unavailable calculated line position are provided, with an accuracy of the order of 0.2 cm-1. An extension to the main isotopologue H_216O is currently underway. R. J. Barber, J. Tennyson, G. J. Harris and R. N. Tolchenov, Mon. Not. R. Astron. Soc. {368}, 1087-1094 (2006). L. Lodi and J. Tennyson, J. Quant. Spectrosc. Radiat. Trans. (2012), doi:10.1016/j.jqsrt.2012.02.023 L. Lodi, J. Tennyson and O. L. Polyansky, J. Chem. Phys. {135}, 034113 (2011). J. Tennyson at al., J. Quant. Spectrosc. Radiat. Trans. {110}, 573-96 (2009).

  20. Focus groups reveal consumer ambivalence.

    PubMed

    1983-01-01

    According to qualitative research, Salvadoreans are ambivalent about the use of contraceptives. Since complete responsibility for management of the CSM project was accepted by the Association Demografica Salvadorena (ADS), the agency which operates the contraceptive social marketing project in El Salvador, in November 1980, the need for decisions in such areas as product price increases, introduction of new condom brands, promotion of the vaginal foaming tablet, and assessment of product sales performance had arisen. The ICSMP funded market research, completed during 1983, was intended to provide the data on which such decisions by ADS could be based. The qualitative research involved 8 focus groups, comprised of men and women, aged 18-45, contraceptive users and nonusers, from the middle and lower socioeconomic strata of the city of San Salvador and other suburban areas. In each group a moderator led discussion of family planning and probed respondents for specific attitudes, knowledge, and behavior regarding the use of contraceptives. To assess attitudes at a more emotional level, moderators asked respondents to "draw" their ideas on certain issues. A marked discrepancy was revealed between respondents' intellectual responses to the issues raised in group discussion, as opposed to their feelings expressed in the drawings. Intellectually, participants responded very positively to family planning practice, but when they were asked to draw their perceptions, ambivalent feelings emerged. Drawings of both the user and the nonuser convey primarily negative aspects for either choice. The user is tense and moody toward her children; the nonuser loses her attractiveness and "dies." Figures also show drawings of some of the attitudes of single and married male participants. 1 drawing shows an incomplete and a complete circle, symbolizing a sterilized man (incomplete) and a nonsterilized man (complete). Another picture depicts a chained man who has lost his freedom

  1. Long-term survival and differentiation of retinal neurons derived from human embryonic stem cell lines in un-immunosuppressed mouse retina

    PubMed Central

    Hambright, Dustin; Park, Kye-Yoon; Brooks, Matthew; McKay, Ron; Swaroop, Anand

    2012-01-01

    Purpose To examine the potential of NIH-maintained human embryonic stem cell (hESC) lines TE03 and UC06 to differentiate into retinal progenitor cells (hESC-RPCs) using the noggin/Dkk-1/IGF-1/FGF9 protocol. An additional goal is to examine the in vivo dynamics of maturation and retinal integration of subretinal and epiretinal (vitreous space) hESC-RPC grafts without immunosuppression. Methods hESCs were neuralized in vitro with noggin for 2 weeks and expanded to derive neuroepithelial cells (hESC-neural precursors, NPs). Wnt (Integration 1 and wingless) blocking morphogens Dickkopf-1 (Dkk-1) and Insulin-like growth factor 1 (IGF-1) were used to direct NPs to a rostral neural fate, and fibroblast growth factor 9 (FGF9)/fibroblast growth factor-basic (bFGF) were added to bias the differentiation of developing anterior neuroectoderm cells to neural retina (NR) rather than retinal pigment epithelium (RPE). Cells were dissociated and grafted into the subretinal and epiretinal space of young adult (4–6-week-old) mice (C57BL/6J x129/Sv mixed background). Remaining cells were replated for (i) immunocytochemical analysis and (ii) used for quantitative reverse transcription polymerase chain reaction (qRT–PCR) analysis. Mice were sacrificed 3 weeks or 3 months after grafting, and the grafts were examined by histology and immunohistochemistry for survival of hESC-RPCs, presence of mature neuronal and retinal markers, and the dynamics of in vivo maturation and integration into the host retina. Results At the time of grafting, hESC-RPCs exhibited immature neural/neuronal immunophenotypes represented by nestin and neuronal class III β-tubulin, with about half of the cells positive for cell proliferation marker Kiel University -raised antibody number 67 (Ki67), and no recoverin-positive (recoverin [+]) cells. The grafted cells expressed eye field markers paired box 6 (PAX6), retina and anterior neural fold homeobox (RAX), sine oculis homeobox homolog 6 (SIX6), LIM homeobox 2

  2. Line-Shape Transition of Collision Broadened Lines

    NASA Astrophysics Data System (ADS)

    Harde, H.; Katzenellenbogen, N.; Grischkowsky, D.

    1995-02-01

    Using the newly developed technique of THz time-domain spectroscopy, we have measured the far-wing absorption line profile of the ensemble of collision broadened ground state rotational lines of methylchloride vapor out to more than 200 linewidths from resonance, corresponding to frequency offsets as much as 5× the resonant frequency. On these far wings the measured absorption is approximately an order of magnitude less than that predicted by the van Vleck-Weisskopf theory. Our observations show that at higher frequencies a transition occurs from the regime of the van Vleck-Weisskopf theory to the regime of the Lorentz theory.

  3. Auroral resonance line radiative transfer

    SciTech Connect

    Gladstone, G.R. )

    1992-02-01

    A model is developed for simulating the two-dimensional radiative transfer of resonance line emissions in auroras. The method of solution utilizes Fourier decomposition of the horizontal dependence in the intensity field so that the two-dimensional problem becomes a set of one-dimensional problems having different horizontal wavenumbers. The individual one-dimensional problems are solved for using a Feautrier-type solution of the differential-integral form of the radiative transfer equation. In the limit as the horizontal wavenumber becomes much larger than the local line-center extinction coefficient, the scattering integral becomes considerably simplified, and the final source function is evaluated in closed form. The two-dimensional aspects of the model are tested against results for nonresonance radiative transfer studies, and the resonance line part of the model is tested against results of existing plane-parallel resonance line radiative transfer codes. Finally, the model is used to simulate the intensity field of O{sub I} 1,304{angstrom} for hard and soft auroras of various Gaussian horizontal widths. The results demonstrate the importance of considering the effects of two-dimensional radiative transfer when analyzing auroral resonance line data.

  4. Cryopreservation of Xenopus transgenic lines.

    PubMed

    Buchholz, Daniel R; Fu, Liezhen; Shi, Yun-Bo

    2004-01-01

    Xenopus laevis has been widely used for molecular, cellular, and developmental studies. With the development of the sperm-mediated transgenic method, it is now possible to study gene function during vertebrate development by using this popular model. On the other hand, like other animal species, it is labor intensive, and the maintenance of transgenic lines is expensive. In this article, we investigated the possibility of using sperm-cryopreservation as a means to preserve transgenic frog lines. We demonstrated that cryopreserved sperms are viable but not fertile under our in vitro fertilization (IVF) conditions. However, by microinjecting cryopreserved sperm nuclei, we successfully regenerated a transgenic line carrying a double promoter transgene construct, where the marker gene encoding the green fluorescent protein (GFP) is driven by the gamma-crystallin gene promoter and a gene of interest, encoding a fusion protein of GFP with the matrix metalloproteinase stromelysin-3 (ST3-GFP), is driven by a heat shock-inducible promoter. We demonstrated the functional transmission of the ST3-GFP transgene by analyzing the phenotype of the F1 animals after heat-shock to induce its expression. Our method thus provides an inexpensive means to preserve transgenic frog lines and a convenient way for distribution of transgenic lines. Furthermore, the ease with which to microinject nuclei compared to the technically demanding transgenesis procedure with variable outcome should facilitate more laboratories to use transgenic Xenopus laevis for functional studies in vivo. Mol. Reprod. Dev. 67: 65-69, 2004. PMID:14648875

  5. Magnetic field line lengths inside interplanetary magnetic flux ropes

    NASA Astrophysics Data System (ADS)

    Hu, Qiang; Qiu, Jiong; Krucker, Sam

    2015-07-01

    We report on the detailed and systematic study of field line twist and length distributions within magnetic flux ropes embedded in interplanetary coronal mass ejections (ICMEs). The Grad-Shafranov reconstruction method is utilized together with a constant-twist nonlinear force-free (Gold-Hoyle) flux rope model to reveal the close relation between the field line twist and length in cylindrical flux ropes, based on in situ Wind spacecraft measurements. We show that the field line twist distributions within interplanetary flux ropes are inconsistent with the Lundquist model. In particular, we utilize the unique measurements of magnetic field line lengths within selected ICME events as provided by Kahler et al. () based on energetic electron burst observations at 1 AU and the associated type III radio emissions detected by the Wind spacecraft. These direct measurements are compared with our model calculations to help assess the flux rope interpretation of the embedded magnetic structures. By using the different flux rope models, we show that the in situ direct measurements of field line lengths are consistent with a flux rope structure with spiral field lines of constant and low twist, largely different from that of the Lundquist model, especially for relatively large-scale flux ropes.

  6. Analysis of spontaneous chromosomal rearrangements in neuroblasts of genetically unstable mutant lines of Drosophila melanogaster

    SciTech Connect

    Derzhavets, E.M.; Kim, A.I.; Aslanyan, M.M.

    1988-11-01

    The spectrum and frequency of chromosomal aberrations in the somatic cells of III instar larvae of Drosophila melanogaster mutator line were studied using three of its derivatives (sbt, if, and w/sup a/) and line w as control. It has been demonstrated that the frequency of anaphases with bridges and acentric fragments increases in the neuroblast of flies of the mutator line as well as in the neuroblasts of the larvae of the lines sbt, if, and w/sup a/. The metaphase analysis revealed that the mutator line and its derivatives are characterized by higher frequencies of chromosomal aberrations as compared to the control. Chromatid breaks are predominant type of rearrangements. These results, suggest probably presence of the specific mutator factor or factors in the line studied, affecting chromosomal structure and, possibly, activating migration of the mobile genetic elements in the mutator line.

  7. 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

  8. The line profile variable B stars

    NASA Technical Reports Server (NTRS)

    Smith, M. A.

    1980-01-01

    The observational status of the line profile variable B stars is summarized. The following areas are discussed: recent history; evolutionary status; line profile variations; line strength variations; and photometric variations.

  9. Standardization of beam line representations

    NASA Astrophysics Data System (ADS)

    Carey, David C.

    1999-05-01

    Standardization of beam line representations means that a single set of data can be used in many situations to represent a beam line. This set of data should be the same no matter what the program to be run or the calculation to be made. We have concerned ourselves with three types of standardization: (1) The same set of data should be usable by different programs. (2) The inclusion of other items in the data, such as calculations to be done, units to be used, or preliminary specifications, should be in a notation similar to the lattice specification. (3) A single set of data should be used to represent a given beam line, no matter what is being modified or calculated. The specifics of what is to be modified or calculated can be edited into the data as part of the calculation. These three requirements all have aspects not previously discussed in a public forum. Implementations into TRANSPORT [1] will be discussed.

  10. Identification of interstellar methanol lines

    NASA Astrophysics Data System (ADS)

    Sutton, E. C.; Herbst, Eric

    1988-10-01

    The extended internal axis method Hamiltonian of Herbst et al. has been employed to study the rotational spectrum of methanol out to high values of the rotational quantum number J. For 12CH3OH the available laboratory data, consisting of 783 lines out to J = 22, have been fitted with a Hamiltonian containing 32 free parameters. For 13CH3OH a Hamiltonian with 23 free parameters is sufficient for fitting 455 lines, also out to J = 22. Frequency predictions based on these fits have permitted the identification of a number of previously unidentified interstellar lines from OMC-1. The majority of these are b-type R-branch transitions of 12CH3OH.

  11. Balmer line shifts in quasars

    NASA Astrophysics Data System (ADS)

    Sulentic, J. W.; Marziani, P.; Del Olmo, A.; Zamfir, S.

    2016-02-01

    We offer a broad review of Balmer line phenomenology in type 1 active galactic nuclei, briefly summarising luminosity and radio loudness effects, and discussing interpretation in terms of nebular physics along the 4D eigenvector 1 sequence of quasars. We stress that relatively rare, peculiar Balmer line profiles (i.e., with large shifts with respect to the rest frame or double and multiple peaked) that start attracted attentions since the 1970s are still passable of multiple dynamical interpretation. More mainstream objects are still not fully understood as well, since competing dynamical models and geometries are possible. Further progress may come from inter-line comparison across the 4D Eigenvector 1 sequence.

  12. Cryopreservation of transgenic mouse lines.

    PubMed

    Pomeroy, K O

    1993-01-01

    A transgenic animal represents an enormous investment in time and money. Animals can be destroyed through disease, fire, malfuncnons in the control of the environment, negligence, sabotage, or accidental disposal. Researchers can protect valuable transgenic lines from accrdental destruction by "banking" them in liquid nitrogen. Cryopreservation can also reduce animal costs by decreasing the number of live animals investigators must maintain. Often, when one is trying to produce a transgenic animal, some lines will be derived that may not initially appear interesting. These animals can be stored in liquid nitrogen for future recovery and study. The maintenance of just one line of mice, say 25 mice at 15 cents/d, can cost over $1000 (US) in a single year. PMID:21390665

  13. Cutting of bent vortex lines

    SciTech Connect

    Wagenleithner, P.

    1982-07-01

    One of the major problems in the application of type II superconductors is the appearance of resistivity in case where a current-carrying specimen is in a longitudinal magnetic field. This is explained by the onset of flux-line cutting events, followed by cross-joining of the line parts. The calculation given here shows the amount of repulsive force and energy between two curved vortex lines and examines the general stability of the vortex-vortex system. First, the actual interaction potential between curved vortices is computed. It includes all electromagnetic and core overlap terms of interactions and self-interaction, and allows computation of the system energy under all curved vortex-line configurations. A computer program is used to find the form of lowest free energy. To do this, special trial functions are established to describe the three-dimensional form of the vortex-vortex system. In these functions parameters determine the qualitative and quantitative form. The asymptotic boundary conditions are built into the nature of the trial functions. The computer program now minimizes the free energy with respect to these parameters. The resulting repulsive energy and force are more than ten times less than the known results for straight flux lines, especially for small asymptotic cutting angles. There is no sharp maximum in the plot of repulsive force versus flux-line separation. A remarkable results is the loss of general stability below a separation distance of several London penetration depths, depending on the cutting angle and the Ginzburg-Landau parameter. The explanation lies in the local attraction of central sections of the vortices as a result of configurational adaption. This explains the onset of resistance at small currents and small magnetic fields.

  14. Stable line defects in silicene

    NASA Astrophysics Data System (ADS)

    Ghosh, Dibyajyoti; Parida, Prakash; Pati, Swapan K.

    2015-11-01

    Line defects in two-dimensional (2D) materials greatly modulate various properties of their pristine form. Using ab initio molecular dynamics (AIMD) simulations, we investigate the structural reconstructions of different kinds of grain boundaries in the silicene sheets. It is evident that depending upon the presence of silicon adatoms and edge shape of grain boundaries (i.e., armchair or zigzag), stable extended line defects (ELDs) can be introduced in a controlled way. Further studies show the stability of these line-defects in silicene, grown on Ag(111) surface at room-temperature. Importantly, unlike most of the 2D sheet materials such as graphene and hexagonal boron nitride, 5-5-8 line defects modify the nonmagnetic semimetallic pristine silicene sheet to spin-polarized metal. As ferromagnetically ordered magnetic moments remain strongly localized at the line defect, a one-dimensional spin channel gets created in silicene. Interestingly, these spin channels are quite stable because, unlike the edge of nanoribbons, structural reconstruction or contamination cannot destroy the ordering of magnetic moments here. Zigzag silicene nanoribbons with a 5-5-8 line defect also exhibit various interesting electronic and magnetic properties depending upon their width as well as the nature of the magnetic coupling between edge and defect spin states. Upon incorporation of other ELDs, such as 4-4-4 and 4-8 defects, 2D sheets and nanoribbons of silicene show a nonmagnetic metallic or semiconducting ground state. Highlighting the controlled formation of ELDs and consequent emergence of technologically important properties in silicene, we propose new routes to realize silicene-based nanoelectronic and spintronic devices.

  15. Assembly-line Simulation Program

    NASA Technical Reports Server (NTRS)

    Chamberlain, Robert G.; Zendejas, Silvino; Malhotra, Shan

    1987-01-01

    Costs and profits estimated for models based on user inputs. Standard Assembly-line Manufacturing Industry Simulation (SAMIS) program generalized so useful for production-line manufacturing companies. Provides accurate and reliable means of comparing alternative manufacturing processes. Used to assess impact of changes in financial parameters as cost of resources and services, inflation rates, interest rates, tax policies, and required rate of return of equity. Most important capability is ability to estimate prices manufacturer would have to receive for its products to recover all of costs of production and make specified profit. Written in TURBO PASCAL.

  16. Two terminal line voltage thermostat

    SciTech Connect

    Stalsberg, K.J.; Ingalls, J.E.; Hoglund, S.R.

    1995-10-10

    A two terminal line voltage thermostat includes a switch which effectively connects line voltage to a heater load. The entire process is controlled by an integrated circuit microcontroller which is powered by a rectified voltage from a transformer secondary connected to a primary which is in series with the heater load. Backup battery power is provided to maintain limited functions of the microcontroller in the event of overall power loss. The microcontroller is programmed to meet the temperature sensing and control requirements specific to a two terminal electric baseboard heating installation. 7 figs.

  17. Linearization algorithms for line transfer

    SciTech Connect

    Scott, H.A.

    1990-11-06

    Complete linearization is a very powerful technique for solving multi-line transfer problems that can be used efficiently with a variety of transfer formalisms. The linearization algorithm we describe is computationally very similar to ETLA, but allows an effective treatment of strongly-interacting lines. This algorithm has been implemented (in several codes) with two different transfer formalisms in all three one-dimensional geometries. We also describe a variation of the algorithm that handles saturable laser transport. Finally, we present a combination of linearization with a local approximate operator formalism, which has been implemented in two dimensions and is being developed in three dimensions. 11 refs.

  18. Recombination Line vs. Forbidden Line Abundances in Planetary Nebulae

    NASA Astrophysics Data System (ADS)

    Robertson-Tessi, M.; Garnett, D. R.

    2004-05-01

    Recombination lines (RLs) of C II, N II, and O II in planetary nebulae (PNe) have been found to give abundances that are much larger in some cases than abundances from collisionally-excited forbidden lines (CELs). The origins of this abundance discrepancy are highly debated. We present new spectroscopic observations of O II and C II recombination lines for six planetary nebulae. With these data we compare the abundances derived from the optical recombination lines with those determined from collisionally-excited lines. Combining our new data with published results on RLs in other PNe, we examine the discrepancy in abundances derived from RLs and CELs. We find that there is a wide range in the measured abundance discrepancy Δ (O+2) = log O+2(RL) - log O+2(CEL), ranging from approximately 0.1 dex (within the 1σ measurement errors) up to 1.3 dex. This tends to rule out errors in the recombination coefficients as a source of the discrepancy. Most RLs yield similar abundances, with the notable exception of O II multiplet V15, known to arise primarily from dielectronic recombination, which gives abundances averaging 0.6 dex higher than other O II RLs. We compare Δ (O+2) against a variety of physical properties of the PNe to look for clues as to the mechanism responsible for the abundance discrepancy. The strongest correlation is found with Balmer surface brightness; high surface brightness, compact PNe show small values of Δ (O+2), while large low surface brightness PNe show the largest discrepancies. Rougher correlations of Δ (O+2) are found with He+2/He+ and with the expansion velocity. No correlations are seen with electron temperature, electron density, central star effective temperature and luminosity, stellar mass loss rate, or nebular morphology. Similar results are found for carbon, comparing C II RL abundances with ultraviolet measurements of C III]. This work is supported by NSF grant AST-0203905.

  19. Label-free separation of human embryonic stem cells and their differentiating progenies by phasor fluorescence lifetime microscopy

    PubMed Central

    Stringari, Chiara; Sierra, Robert; Donovan, Peter J.

    2012-01-01

    Abstract. We develop a label-free optical technique to image and discriminate undifferentiated human embryonic stem cells (hESCs) from their differentiating progenies in vitro. Using intrinsic cellular fluorophores, we perform fluorescence lifetime microscopy (FLIM) and phasor analysis to obtain hESC metabolic signatures. We identify two optical biomarkers to define the differentiation status of hESCs: Nicotinamide adenine dinucleotide (NADH) and lipid droplet-associated granules (LDAGs). These granules have a unique lifetime signature and could be formed by the interaction of reactive oxygen species and unsaturated metabolic precursor that are known to be abundant in hESC. Changes in the relative concentrations of these two intrinsic biomarkers allow for the discrimination of undifferentiated hESCs from differentiating hESCs. During early hESC differentiation we show that NADH concentrations increase, while the concentration of LDAGs decrease. These results are in agreement with a decrease in oxidative phosphorylation rate. Single-cell phasor FLIM signatures reveal an increased heterogeneity in the metabolic states of differentiating H9 and H1 hESC colonies. This technique is a promising noninvasive tool to monitor hESC metabolism during differentiation, which can have applications in high throughput analysis, drug screening, functional metabolomics and induced pluripotent stem cell generation. PMID:22559690

  20. Label-free separation of human embryonic stem cells and their differentiating progenies by phasor fluorescence lifetime microscopy

    NASA Astrophysics Data System (ADS)

    Stringari, Chiara; Sierra, Robert; Donovan, Peter J.; Gratton, Enrico

    2012-04-01

    We develop a label-free optical technique to image and discriminate undifferentiated human embryonic stem cells (hESCs) from their differentiating progenies in vitro. Using intrinsic cellular fluorophores, we perform fluorescence lifetime microscopy (FLIM) and phasor analysis to obtain hESC metabolic signatures. We identify two optical biomarkers to define the differentiation status of hESCs: Nicotinamide adenine dinucleotide (NADH) and lipid droplet-associated granules (LDAGs). These granules have a unique lifetime signature and could be formed by the interaction of reactive oxygen species and unsaturated metabolic precursor that are known to be abundant in hESC. Changes in the relative concentrations of these two intrinsic biomarkers allow for the discrimination of undifferentiated hESCs from differentiating hESCs. During early hESC differentiation we show that NADH concentrations increase, while the concentration of LDAGs decrease. These results are in agreement with a decrease in oxidative phosphorylation rate. Single-cell phasor FLIM signatures reveal an increased heterogeneity in the metabolic states of differentiating H9 and H1 hESC colonies. This technique is a promising noninvasive tool to monitor hESC metabolism during differentiation, which can have applications in high throughput analysis, drug screening, functional metabolomics and induced pluripotent stem cell generation.

  1. The missing UV absorption lines of NGC 4151

    NASA Technical Reports Server (NTRS)

    Leech, K. J.; Penston, M. V.; Snijders, M. A. J.; Ward, M. J.; Gull, T. R.

    1990-01-01

    Near simultaneous high dispersion long and short wavelength International Ultraviolet Explorer (IUE) observations of the Seyfert galaxy NGC 4151 are discussed. Previous observations revealed a narrow absorption system in Mg II not present in Ly alpha or C IV. The new observations confirm the presence of this system in Mg II and its absence in the other lines. Possible reasons for this are discussed. Future Hubble Space Telescope studies of NGC 4151 are discussed.

  2. Reconnections of Wave Vortex Lines

    ERIC Educational Resources Information Center

    Berry, M. V.; Dennis, M. R.

    2012-01-01

    When wave vortices, that is nodal lines of a complex scalar wavefunction in space, approach transversely, their typical crossing and reconnection is a two-stage process incorporating two well-understood elementary events in which locally coplanar hyperbolas switch branches. The explicit description of this reconnection is a pedagogically useful…

  3. Checking the Lines for Communication

    ERIC Educational Resources Information Center

    Friedman, Rachel

    2005-01-01

    Lines of communication are often blurred in early childhood settings and relationships need to be reestablished regularly as children move through a program. In addition, the parent-teacher-child relationship is a triad that is built upon trust and need and not a clear hierarchy. This article discusses the following issues brought up during a…

  4. Teachers' Conceptions of Tangent Line

    ERIC Educational Resources Information Center

    Paez Murillo, Rosa Elvira; Vivier, Laurent

    2013-01-01

    In order to study the conceptions, and their evolutions, of the tangent line to a curve an updating workshop which took place in Mexico was designed for upper secondary school teachers. This workshop was planned using the methodology of cooperative learning, scientific debate and auto reflection (ACODESA) and the conception-knowing-concept model…

  5. Nonceramic insulators for transmission lines

    SciTech Connect

    Schneider, H.M. . High Voltage Transmission Research Center); Hall, J.F. ); Karady, G. ); Rendowden, J. )

    1989-10-01

    Progress in development of nonceramic transmission lines insulator technology is reviewed. The results of a survey to utilities in the United States are presented to indicate the range of applications, distribution by voltage class, appraisal of performance, and failure modes. Concerns expressed by utility representatives at a recent workshop are described and future research and standardization needs are presented.

  6. Collaboration across the Power Line.

    ERIC Educational Resources Information Center

    Romer, Karen T.; Whipple, William R.

    1991-01-01

    The college student who participates in genuine collaboration with a faculty member in an intellectual endeavor transcends the barrier of power. Once it has been breached, other power lines will be less constraining. The experience transforms the nature of learning and constructs a lasting authority in the student. (MSE)

  7. Calibrating the Mental Number Line

    ERIC Educational Resources Information Center

    Izard, Veronique; Dehaene, Stanislas

    2008-01-01

    Human adults are thought to possess two dissociable systems to represent numbers: an approximate quantity system akin to a mental number line, and a verbal system capable of representing numbers exactly. Here, we study the interface between these two systems using an estimation task. Observers were asked to estimate the approximate numerosity of…

  8. Gamma-ray line astrophysics

    NASA Technical Reports Server (NTRS)

    Lingenfelter, R. E.; Ramaty, R.

    1986-01-01

    Recent observations of gamma-ray line emission from solar flares, gamma-ray bursts, the galactic center, the interstellar medium and the jets of SS433 are reviewed. The implications of these observations on high energy processes in these sources are discussed.

  9. Priming the Mental Time Line

    ERIC Educational Resources Information Center

    Di Bono, Maria Grazia; Casarotti, Marco; Priftis, Konstantinos; Gava, Lucia; Umilta, Carlo; Zorzi, Marco

    2012-01-01

    Growing experimental evidence suggests that temporal events are represented on a mental time line, spatially oriented from left to right. Support for the spatial representation of time comes mostly from studies that have used spatially organized responses. Moreover, many of these studies did not avoid possible confounds attributable to target…

  10. MAINE CONTOUR LINES 500 FEET

    EPA Science Inventory

    MECON500 contains 500 foot contour intervals for Maine, generated from USGS 1:250,000 DEMs. Arcs are coded by elevation. Due to the nature of the source data, the positional accuracy of these contour lines varies from good to poor. Use of these data at scales of greater then 1:2...

  11. MAINE CONTOUR LINES 60 FEET

    EPA Science Inventory

    MECON60 contains contours at 60 foot intervals for the entire state of Maine as generated from USGS 1:250,000 scale digital elevation models using ARC/INFO software. Arcs are coded by elevation. Due to the nature of the source data, the positional accuracy of these contour line...

  12. MODELING MOLECULAR HYPERFINE LINE EMISSION

    SciTech Connect

    Keto, Eric; Rybicki, George

    2010-06-20

    In this paper, we discuss two approximate methods previously suggested for modeling hyperfine spectral line emission for molecules whose collisional transition rates between hyperfine levels are unknown. Hyperfine structure is seen in the rotational spectra of many commonly observed molecules such as HCN, HNC, NH{sub 3}, N{sub 2}H{sup +}, and C{sup 17}O. The intensities of these spectral lines can be modeled by numerical techniques such as {Lambda}-iteration that alternately solve the equations of statistical equilibrium and the equation of radiative transfer. However, these calculations require knowledge of both the radiative and collisional rates for all transitions. For most commonly observed radio frequency spectral lines, only the net collisional rates between rotational levels are known. For such cases, two approximate methods have been suggested. The first method, hyperfine statistical equilibrium, distributes the hyperfine level populations according to their statistical weight, but allows the population of the rotational states to depart from local thermal equilibrium (LTE). The second method, the proportional method, approximates the collision rates between the hyperfine levels as fractions of the net rotational rates apportioned according to the statistical degeneracy of the final hyperfine levels. The second method is able to model non-LTE hyperfine emission. We compare simulations of N{sub 2}H{sup +} hyperfine lines made with approximate and more exact rates and find that satisfactory results are obtained.

  13. Coiled transmission line pulse generators

    DOEpatents

    McDonald, Kenneth Fox

    2010-11-09

    Methods and apparatus are provided for fabricating and constructing solid dielectric "Coiled Transmission Line" pulse generators in radial or axial coiled geometries. The pour and cure fabrication process enables a wide variety of geometries and form factors. The volume between the conductors is filled with liquid blends of monomers, polymers, oligomers, and/or cross-linkers and dielectric powders; and then cured to form high field strength and high dielectric constant solid dielectric transmission lines that intrinsically produce ideal rectangular high voltage pulses when charged and switched into matched impedance loads. Voltage levels may be increased by Marx and/or Blumlein principles incorporating spark gap or, preferentially, solid state switches (such as optically triggered thyristors) which produce reliable, high repetition rate operation. Moreover, these Marxed pulse generators can be DC charged and do not require additional pulse forming circuitry, pulse forming lines, transformers, or an a high voltage spark gap output switch. The apparatus accommodates a wide range of voltages, impedances, pulse durations, pulse repetition rates, and duty cycles. The resulting mobile or flight platform friendly cylindrical geometric configuration is much more compact, light-weight, and robust than conventional linear geometries, or pulse generators constructed from conventional components. Installing additional circuitry may accommodate optional pulse shape improvements. The Coiled Transmission Lines can also be connected in parallel to decrease the impedance, or in series to increase the pulse length.

  14. On-Line Communications Devices.

    ERIC Educational Resources Information Center

    Sternick, Barbara R.

    These summaries have been compiled to assist users in selecting terminals for use with the National Library of Medicine (NLM) on-line systems. The summaries describe the salient characteristics and approximate prices of a large number of hard copy and display devices. Many of the terminals listed may be modified by the addition of various options…

  15. Characterization of a Novel Radiation-Induced Sarcoma Cell Line

    PubMed Central

    Lang, J.E.; Zhu, W.; Nokes, B.T.; Sheth, G.R.; Novak, P.; Fuchs, L.; Watts, G.S.; Futscher, B.W.; Mineyev, N.; Ring, A.; LeBeau, L.; Nagle, R.; Cranmer, L.D.

    2014-01-01

    Background Radiation-induced sarcoma (RIS) is a potential complication of cancer treatment. No widely available cell line models exist to facilitate studies of RIS. Methods We derived a spontaneously immortalized primary human cell line, UACC-SARC1, from a RIS. Results Short tandem repeat (STR) profiling of UACC-SARC1 was virtually identical to its parental tumor. Immunohistochemistry (IHC) analysis of the tumor and immunocytochemistry (ICC) analysis of UACC-SARC1 revealed shared expression of vimentin, osteonectin, CD68, Ki67 and PTEN but tumor-restricted expression of the histiocyte markers α1-antitrypsin and α1-antichymotrypsin. Karyotyping of the tumor demonstrated aneuploidy. Comparative genomic hybridization (CGH) provided direct genetic comparison between the tumor and UACC-SARC1. Sequencing of 740 mutation hotspots revealed no mutations in UACC-SARC1 nor in the tumor. NOD/SCID gamma mouse xenografts demonstrated tumor formation and metastasis. Clonogenicity assays demonstrated that 90% of single cells produced viable colonies. NOD/SCID gamma mice produced useful patient-derived xenografts for orthotopic or metastatic models. Conclusion Our novel RIS strain constitutes a useful tool for pre-clinical studies of this rare, aggressive disease. UACC-SARC1 is an aneuploid cell line with complex genomics lacking common oncogenes or tumor suppressor genes as drivers of its biology. The UACC-SARC1 cell line will enable further studies of the drivers of RIS. Synopsis We derived a spontaneously immortalized primary human cell line, UACC-SARC1, from a radiation-induced sarcoma (RIS). Our novel RIS cell line constitutes a useful tool for pre-clinical studies of this rare, aggressive disease. PMID:25644184

  16. X-ray Emission and Absorption Lines During the SSS Phase of RS Ophiuchi

    NASA Astrophysics Data System (ADS)

    Schönrich, R. A.; Ness, J.-U.

    2008-12-01

    The high-resolution X-ray spectra of the sixth outburst of RS Ophiuchi revealed P Cygni-like line profiles. We use the column densities of selected isolated absorption lines to derive the nitrogen-to-oxygen abundance ratio. We next discuss the origin of the emission lines, which may originate from the shock, and the absorption and emission lines may thus have a different formation history. Finally, we discuss the correlation of high-amplitude variability detected during the early SSS phase with variability in the hardness ratio that follows the same pattern but is shifted by 1000~sec.

  17. 76 FR 51367 - China Shipping Container Lines Co., Ltd.; COSCO Container Lines Company Limited; Evergreen Line A...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-18

    ... From the Federal Register Online via the Government Publishing Office FEDERAL MARITIME COMMISSION China Shipping Container Lines Co., Ltd.; COSCO Container Lines Company Limited; Evergreen Line A Joint Service Agreement; Hanjin Shipping Co., Ltd.; Horizon Lines, LLC; Kawasaki Kisen Kaisha, Ltd.; Nippon Yusen Kaisha; United Arab Shipping Company...

  18. Line Coupling in Atmospheric Spectra

    NASA Technical Reports Server (NTRS)

    Tipping, R. H.

    1996-01-01

    The theoretical modeling of atmospheric spectra is important for a number of different applications: for instance, in the determination of minor atmospheric constituents such as ozone, carbon dioxide, CFC's etc.; in monitoring the temperature profile for climate studies; and in measuring the incoming and outgoing radiation to input into global climate models. In order to accomplish the above mentioned goal, one needs to know the spectral parameters characterizing the individual spectral lines (frequency, width, strength, and shape) as well as the physical parameters of the atmosphere (temperature, abundances, and pressure). When all these parameters are known, it is usually assumed that the resultant spectra and concomitant absorption coefficient can then be calculated by a superposition of individual profiles of appropriate frequency, strength and shape. However, this is not true if the lines are 'coupled'. Line coupling is a subtle effect that takes place when lines of a particular molecule overlap in frequency. In this case when the initial states and the final states of two transitions are connected by collisions, there is a quantum interference resulting in perturbed shapes. In general, this results in the narrowing of Q-branches (those in which the rotational quantum number does not change), and vibration-rotational R- and P branches (those in which the rotational quantum number changes by +/- 1), and in the spectral region beyond band heads (regions where the spectral lines pile up due to centrifugal distortion). Because these features and spectral regions are often those of interest in the determination of the abundances and pressure-temperature profiles, one must take this effect into account in atmospheric models.

  19. 49 CFR 178.255-10 - Lining.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Specifications for Portable Tanks § 178.255-10 Lining. (a) If a lining is required, the material used for lining... metallic, and not less than 1/16 inch thick if nonmetallic, and shall be directly bonded or attached by... in the lining shall be made by fusing the material together or by other equally satisfactory...

  20. 19 CFR 142.41 - Line Release.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 19 Customs Duties 2 2013-04-01 2013-04-01 false Line Release. 142.41 Section 142.41 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY (CONTINUED) ENTRY PROCESS Line Release § 142.41 Line Release. Line Release is an automated system designed...

  1. 19 CFR 142.41 - Line Release.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 19 Customs Duties 2 2014-04-01 2014-04-01 false Line Release. 142.41 Section 142.41 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY (CONTINUED) ENTRY PROCESS Line Release § 142.41 Line Release. Line Release is an automated system designed...

  2. 19 CFR 142.41 - Line Release.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 2 2012-04-01 2012-04-01 false Line Release. 142.41 Section 142.41 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY (CONTINUED) ENTRY PROCESS Line Release § 142.41 Line Release. Line Release is an automated system designed...

  3. 19 CFR 142.41 - Line Release.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 2 2010-04-01 2010-04-01 false Line Release. 142.41 Section 142.41 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY (CONTINUED) ENTRY PROCESS Line Release § 142.41 Line Release. Line Release is an automated system designed...

  4. 18 CFR 2.2 - Transmission lines.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Transmission lines. 2.2... Under the Federal Power Act § 2.2 Transmission lines. In a public statement dated March 7, 1941, the Commission announced its determination that transmission lines which are not primary lines transmitting...

  5. 18 CFR 2.2 - Transmission lines.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 18 Conservation of Power and Water Resources 1 2014-04-01 2014-04-01 false Transmission lines. 2.2... Under the Federal Power Act § 2.2 Transmission lines. In a public statement dated March 7, 1941, the Commission announced its determination that transmission lines which are not primary lines transmitting...

  6. 18 CFR 2.2 - Transmission lines.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 18 Conservation of Power and Water Resources 1 2012-04-01 2012-04-01 false Transmission lines. 2.2... Under the Federal Power Act § 2.2 Transmission lines. In a public statement dated March 7, 1941, the Commission announced its determination that transmission lines which are not primary lines transmitting...

  7. 18 CFR 2.2 - Transmission lines.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 18 Conservation of Power and Water Resources 1 2013-04-01 2013-04-01 false Transmission lines. 2.2... Under the Federal Power Act § 2.2 Transmission lines. In a public statement dated March 7, 1941, the Commission announced its determination that transmission lines which are not primary lines transmitting...

  8. 18 CFR 2.2 - Transmission lines.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 18 Conservation of Power and Water Resources 1 2011-04-01 2011-04-01 false Transmission lines. 2.2... Under the Federal Power Act § 2.2 Transmission lines. In a public statement dated March 7, 1941, the Commission announced its determination that transmission lines which are not primary lines transmitting...

  9. 30 CFR 56.6803 - Blasting lines.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Blasting lines. 56.6803 Section 56.6803 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR METAL AND NONMETAL MINE SAFETY AND... Blasting lines. Permanent blasting lines shall be properly supported. All blasting lines shall be...

  10. 30 CFR 57.6803 - Blasting lines.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Blasting lines. 57.6803 Section 57.6803 Mineral... and Underground § 57.6803 Blasting lines. Permanent blasting lines shall be properly supported. All blasting lines shall be insulated and kept in good repair. General Requirements—Surface and Underground...

  11. Recombination Line versus Forbidden Line Abundances in Planetary Nebulae

    NASA Astrophysics Data System (ADS)

    Robertson-Tessi, Mark; Garnett, Donald R.

    2005-04-01

    Recombination lines (RLs) of C II, N II, and O II in planetary nebulae (PNs) have been found to give abundances that are much larger in some cases than abundances from collisionally excited forbidden lines (CELs). The origins of this abundance discrepancy are highly debated. We present new spectroscopic observations of O II and C II recombination lines for six planetary nebulae. With these data we compare the abundances derived from the optical recombination lines with those determined from collisionally excited lines. Combining our new data with published results on RLs in other PNs, we examine the discrepancy in abundances derived from RLs and CELs. We find that there is a wide range in the measured abundance discrepancy Δ(O+2)=logO+2(RL)-logO+2(CEL), ranging from approximately 0.1 dex (within the 1 σ measurement errors) up to 1.4 dex. This tends to rule out errors in the recombination coefficients as a source of the discrepancy. Most RLs yield similar abundances, with the notable exception of O II multiplet V15, known to arise primarily from dielectronic recombination, which gives abundances averaging 0.6 dex higher than other O II RLs. We compare Δ(O+2) against a variety of physical properties of the PNs to look for clues as to the mechanism responsible for the abundance discrepancy. The strongest correlations are found with the nebula diameter and the Balmer surface brightness; high surface brightness, compact PNs show small values of Δ(O+2), while large low surface brightness PNs show the largest discrepancies. An inverse correlation of Δ(O+2) with nebular density is also seen. A marginal correlation of Δ(O+2) is found with expansion velocity. No correlations are seen with electron temperature, He+2/He+, central star effective temperature and luminosity, stellar mass-loss rate, or nebular morphology. Similar results are found for carbon in comparing C II RL abundances with ultraviolet measurements of C III].

  12. HERSCHEL OBSERVATIONS REVEAL ANOMALOUS MOLECULAR ABUNDANCES TOWARD THE GALACTIC CENTER

    SciTech Connect

    Sonnentrucker, P.; Neufeld, D. A.; Indriolo, N.; Gerin, M.; De Luca, M.; Lis, D. C.; Goicoechea, J. R.

    2013-01-20

    We report the Herschel detections of hydrogen fluoride (HF) and para-water (p-H{sub 2}O) in gas intercepting the sight lines to two well-studied molecular clouds in the vicinity of the Sgr A complex: G-0.02-0.07 (the {sup +}50 km s{sup -1} cloud{sup )} and G-0.13-0.08 (the {sup +}20 km s{sup -1} cloud{sup )}. Toward both sight lines, HF and water absorption components are detected over a wide range of velocities covering {approx}250 km s{sup -1}. For all velocity components with V{sub LSR} > -85 km s{sup -1}, we find that the HF and water abundances are consistent with those measured toward other sight lines probing the Galactic disk gas. The velocity components with V{sub LSR} {<=} -85 km s{sup -1}, which are known to trace gas residing within {approx}200 pc of the Galactic center, however, exhibit water vapor abundances with respect to HF at least a factor three higher than those found in the Galactic disk gas. Comparison with CH data indicates that our observations are consistent with a picture where HF and a fraction of the H{sub 2}O absorption arise in diffuse molecular clouds showing Galactic disk-like abundances while the bulk of the water absorption arises in warmer (T {>=} 400 K) diffuse molecular gas for V{sub LSR} {<=} -85 km s{sup -1}. This diffuse Interstellar Medium (ISM) phase has also been recently revealed through observations of CO, HF, H{sup +}{sub 3}, and H{sub 3}O{sup +} absorption toward other sight lines probing the Galactic center inner region.

  13. Behavioral idiosyncrasy reveals genetic control of phenotypic variability

    PubMed Central

    Ayroles, Julien F.; Buchanan, Sean M.; O’Leary, Chelsea; Skutt-Kakaria, Kyobi; Grenier, Jennifer K.; Clark, Andrew G.; Hartl, Daniel L.; de Bivort, Benjamin L.

    2015-01-01

    Quantitative genetics has primarily focused on describing genetic effects on trait means and largely ignored the effect of alternative alleles on trait variability, potentially missing an important axis of genetic variation contributing to phenotypic differences among individuals. To study the genetic effects on individual-to-individual phenotypic variability (or intragenotypic variability), we used Drosophila inbred lines and measured the spontaneous locomotor behavior of flies walking individually in Y-shaped mazes, focusing on variability in locomotor handedness, an assay optimized to measure variability. We discovered that some lines had consistently high levels of intragenotypic variability among individuals, whereas lines with low variability behaved as although they tossed a coin at each left/right turn decision. We demonstrate that the degree of variability is itself heritable. Using a genome-wide association study (GWAS) for the degree of intragenotypic variability as the phenotype across lines, we identified several genes expressed in the brain that affect variability in handedness without affecting the mean. One of these genes, Ten-a, implicates a neuropil in the central complex of the fly brain as influencing the magnitude of behavioral variability, a brain region involved in sensory integration and locomotor coordination. We validated these results using genetic deficiencies, null alleles, and inducible RNAi transgenes. Our study reveals the constellation of phenotypes that can arise from a single genotype and shows that different genetic backgrounds differ dramatically in their propensity for phenotypic variabililty. Because traditional mean-focused GWASs ignore the contribution of variability to overall phenotypic variation, current methods may miss important links between genotype and phenotype. PMID:25953335

  14. BAC transgenesis in human embryonic stem cells as a novel tool to define the human neural lineage.

    PubMed

    Placantonakis, Dimitris G; Tomishima, Mark J; Lafaille, Fabien; Desbordes, Sabrina C; Jia, Fan; Socci, Nicholas D; Viale, Agnes; Lee, Hyojin; Harrison, Neil; Tabar, Viviane; Studer, Lorenz

    2009-03-01

    Human embryonic stem cells (hESCs) have enormous potential for applications in basic biology and regenerative medicine. However, harnessing the potential of hESCs toward generating homogeneous populations of specialized cells remains challenging. Here we describe a novel technology for the genetic identification of defined hESC-derived neural cell types using bacterial artificial chromosome (BAC) transgenesis. We generated hESC lines stably expressing Hes5::GFP, Dll1::GFP, and HB9::GFP BACs that yield green fluorescent protein (GFP)(+) neural stem cells, neuroblasts, and motor neurons, respectively. Faithful reporter expression was confirmed by cell fate analysis and appropriate transgene regulation. Prospective isolation of HB9::GFP(+) cells yielded purified human motor neurons with proper marker expression and electrophysiological activity. Global mRNA and microRNA analyses of Hes5::GFP(+) and HB9::GFP(+) populations revealed highly specific expression signatures, suggesting that BAC transgenesis will be a powerful tool for establishing expression libraries that define the human neural lineage and for accessing defined cell types in applications of human disease. PMID:19074416

  15. 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. PMID:1726925

  16. LINE-1 retrotransposons: from 'parasite' sequences to functional elements.

    PubMed

    Paço, Ana; Adega, Filomena; Chaves, Raquel

    2015-02-01

    Long interspersed nuclear elements-1 (LINE-1) are the most abundant and active retrotransposons in the mammalian genomes. Traditionally, the occurrence of LINE-1 sequences in the genome of mammals has been explained by the selfish DNA hypothesis. Nevertheless, recently, it has also been argued that these sequences could play important roles in these genomes, as in the regulation of gene expression, genome modelling and X-chromosome inactivation. The non-random chromosomal distribution is a striking feature of these retroelements that somehow reflects its functionality. In the present study, we have isolated and analysed a fraction of the open reading frame 2 (ORF2) LINE-1 sequence from three rodent species, Cricetus cricetus, Peromyscus eremicus and Praomys tullbergi. Physical mapping of the isolated sequences revealed an interspersed longitudinal AT pattern of distribution along all the chromosomes of the complement in the three genomes. A detailed analysis shows that these sequences are preferentially located in the euchromatic regions, although some signals could be detected in the heterochromatin. In addition, a coincidence between the location of imprinted gene regions (as Xist and Tsix gene regions) and the LINE-1 retroelements was also observed. According to these results, we propose an involvement of LINE-1 sequences in different genomic events as gene imprinting, X-chromosome inactivation and evolution of repetitive sequences located at the heterochromatic regions (e.g. satellite DNA sequences) of the rodents' genomes analysed. PMID:25106509

  17. Heterozygous Embryonic Stem Cell Lines Derived from Nonhuman Primate Parthenotes

    PubMed Central

    Dighe, Vikas; Clepper, Lisa; Pedersen, Darlene; Byrne, James; Ferguson, Betsy; Gokhale, Sumita; Penedo, M. Cecilia T.; Wolf, Don; Mitalipov, Shoukhrat

    2009-01-01

    Monoparental parthenotes represent a potential source of histocompatible stem cells that should be isogenic with the oocyte donor and therefore suitable for use in cell or tissue replacement therapy. We generated five rhesus monkey parthenogenetic embryonic stem cell (PESC) lines with stable, diploid female karyotypes that were morphologically indistinguishable from biparental controls, expressed key pluripotent markers, and generated cell derivatives representative of all three germ layers following in vivo and in vitro differentiation. Interestingly, high levels of heterozygosity were observed at the majority of loci that were polymorphic in the oocyte donors. Some PESC lines were also heterozygous in the major histocompatibility complex region, carrying haplotypes identical to those of the egg donor females. Expression analysis revealed transcripts from some imprinted genes that are normally expressed from only the paternal allele. These results indicate that limitations accompanying the potential use of PESC-derived phenotypes in regenerative medicine, including aberrant genomic imprinting and high levels of homozygosity, are cell line-dependent and not always present. PESC lines were derived in high enough yields to be practicable, and their derivatives are suitable for autologous transplantation into oocyte donors or could be used to establish a bank of histocompatible cell lines for a broad spectrum of patients. PMID:18192229

  18. A Simulator of Periodically Switching Channels for Power Line Communications

    NASA Astrophysics Data System (ADS)

    Hayasaki, Taro; Umehara, Daisuke; Denno, Satoshi; Morikura, Masahiro

    An indoor power line is one of the most attractive media for in-home networks. However, there are many technical problems for achieving in-home power line communication (PLC) with high rate and high reliability. One of such problem is the degradation in the performance of the in-home PLC caused by periodically time-varying channel responses, particularly when connecting the switching power supply equipment. We present a measurement method for power line channel responses and reveal the switching of the channel responses synchronized with power-frequency voltage when connecting switching power supply equipment in sending or receiving outlets. In this paper, we term them periodically switching channel responses. The performance of PLC adapters is seriously affected by the periodically switching channel responses. Therefore, we provide a modeling of the periodically switching channel responses by using finite impulse response (FIR) filters with a shared channel memory and construct a simulator for in-home power line channels including the periodically switching channel responses in order to evaluate the various communication systems through the power line. We present the validity of the proposed simulator through the performance evaluation of OFDM/64QAM over periodically switching channels with additive white Gaussian noise. Furthermore, we evaluate the influence of the periodically switching channel responses on the communication quality of a time-invariant modulation scheme by using the proposed simulator.

  19. Cable and Line Inspection Mechanism

    NASA Technical Reports Server (NTRS)

    Ross, Terence J. (Inventor)

    2003-01-01

    An automated cable and line inspection mechanism visually scans the entire surface of a cable as the mechanism travels along the cable=s length. The mechanism includes a drive system, a video camera, a mirror assembly for providing the camera with a 360 degree view of the cable, and a laser micrometer for measuring the cable=s diameter. The drive system includes an electric motor and a plurality of drive wheels and tension wheels for engaging the cable or line to be inspected, and driving the mechanism along the cable. The mirror assembly includes mirrors that are positioned to project multiple images of the cable on the camera lens, each of which is of a different portion of the cable. A data transceiver and a video transmitter are preferably employed for transmission of video images, data and commands between the mechanism and a remote control station.

  20. Cable and line inspection mechanism

    NASA Technical Reports Server (NTRS)

    Ross, Terence J. (Inventor)

    2003-01-01

    An automated cable and line inspection mechanism visually scans the entire surface of a cable as the mechanism travels along the cable=s length. The mechanism includes a drive system, a video camera, a mirror assembly for providing the camera with a 360 degree view of the cable, and a laser micrometer for measuring the cable=s diameter. The drive system includes an electric motor and a plurality of drive wheels and tension wheels for engaging the cable or line to be inspected, and driving the mechanism along the cable. The mirror assembly includes mirrors that are positioned to project multiple images of the cable on the camera lens, each of which is of a different portion of the cable. A data transceiver and a video transmitter are preferably employed for transmission of video images, data and commands between the mechanism and a remote control station.

  1. The MICE Muon Beam Line

    SciTech Connect

    Apollonio, Marco

    2011-10-06

    In the Muon Ionization Cooling Experiment (MICE) at RAL, muons are produced and transported in a dedicated beam line connecting the production point (target) to the cooling channel. We discuss the main features of the beamline, meant to provide muons with momenta between 140 MeV/c and 240 MeV/c and emittances up to 10 mm rad, which is accomplished by means of a diffuser. Matching procedures to the MICE cooling channel are also described. In summer 2010 we performed an intense data taking campaign to finalize the calibration of the MICE Particle Identification (PID) detectors and the understanding of the beam line, which completes the STEPI phase of MICE. We highlight the main results from these data.

  2. Cohomology of line bundles: Applications

    NASA Astrophysics Data System (ADS)

    Blumenhagen, Ralph; Jurke, Benjamin; Rahn, Thorsten; Roschy, Helmut

    2012-01-01

    Massless modes of both heterotic and Type II string compactifications on compact manifolds are determined by vector bundle valued cohomology classes. Various applications of our recent algorithm for the computation of line bundle valued cohomology classes over toric varieties are presented. For the heterotic string, the prime examples are so-called monad constructions on Calabi-Yau manifolds. In the context of Type II orientifolds, one often needs to compute cohomology for line bundles on finite group action coset spaces, necessitating us to generalize our algorithm to this case. Moreover, we exemplify that the different terms in Batyrev's formula and its generalizations can be given a one-to-one cohomological interpretation. Furthermore, we derive a combinatorial closed form expression for two Hodge numbers of a codimension two Calabi-Yau fourfold.

  3. Second line of defense program

    SciTech Connect

    Cantut, L; Thomas, L L

    1999-07-15

    Since the collapse of the Soviet Union, the prospect of nuclear materials entering the world market has become an ever-increasing threat. The Second Line of Defense (SLD) program was developed by the U.S. Department of Energy's (DOE) Nuclear Transfer and Supplier Policy Division (NN-43) to assist the Russian Federation State Customs Committee (RFSCC) in strengthening its capability to prevent illicit trafficking of nuclear materials across Russia's borders. The SLD program is a natural complement to the Material Protection Control and Accounting (MPC and A) program, which represents a first line of defense against the theft and diversion of nuclear materials. The SLD program is the first U.S.-Russian cooperative program to combat the illicit trafficking of nuclear and nuclear-related materials to would-be proliferators across Russia's borders.

  4. Clinoptilolite and heulandite structural differences as revealed by multinuclear nuclear magnetic resonance spectroscopy

    SciTech Connect

    Ward, R.L.; McKague, H.L. )

    1994-01-27

    Wide-line proton NMR has revealed differences in the water environment in two zeolitic isomorphs: clinoptilolite and heulandite. The proton spectrum of clinoptilolite is Gaussian in shape, while that of heulandite is a (reduced splitting) Pake doublet. Studies of clinoptilolite from five different locations and heulandite from nine different sites agree with this observation. These results suggest a simple nondestructive NMR method for distinguishing heulandite from clinoptilolite. Dehydration experiments are particularly informative. Proton line widths of clinoptilolite as a function of the dehydration temperature reveal three types of water binding, none being absorbed water. Similar studies with heulandite reveal a change from the Pake doublet to a Gaussian, which is probably associated with the heulandite to heulandite B transformation. [sup 27]Al and [sup 29]Si NMR studies of clinoptilolite indicate a change in framework structure and/or cation binding with dehydration. [sup 27]Al NMR of heulandite exhibits an increase in line width with dehydration temperature to 175[degrees]C. At this temperature the increase stops and the line width remains constant to 215[degrees]C, the maximum temperature studied. This agrees with the proton studies and is attributed to the heulandite to heulandite B transformation. [sup 29]Si NMR of heulandite reveals a framework structural change and/or cation binding with dehydration. All of these observations are reversible upon rehydration. 21 refs., 9 figs.

  5. Emission lines in the optical spectrum of 3 Cen A

    NASA Astrophysics Data System (ADS)

    Wahlgren, G. M.; Hubrig, S.

    2004-05-01

    Previously, weak emission lines had been detected at red wavelengths in the spectra of a limited sample of mid to late B type main sequence stars. A fuller description of the occurrence and origins of these lines has yet to be forwarded, in part due to the lack of observations detailing the spectral transitions involved. To address this deficiency, we present a line list of weak emission features found in the optical and near infrared spectral region of the chemically peculiar He-weak star 3 Cen A (HD 120709). Nearly 350 features, mostly associated with allowed transitions from high-excitation states of first ions, are catalogued along with identifications. Prominent among the emission lines are the spectra P II, Mn II, Fe II, Ni II and Cu II. Emission lines from Ca II, Si II and Hg II are also evident. Abundances are determined for several elements from synthetic spectrum fitting, with anomalies detected for the ions O I/II, P II/III and Si II/III. The LTE synthetic spectrum fitting also revealed that the low excitation 4s-4p transitions of Fe II predict an abundance that is greater than that determined from higher excitation 4d-4f transitions. Several of these latter transitions have upper energy levels that are found to be associated with emission lines. We also present empirical considerations for the excitation processes leading to the weak emission lines. Based on observations obtained at the European Southern Observatory, La Silla, Chile, No. 65.L-0316 and Paranal, Chile No. 266.D-5655. Tables 2 and 3 are only available in electronic form at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130.79.128.5) or via http://cdsweb.u-strasbg.fr/cgi-bin/qcat?J/A+A/418/1073

  6. Gamma-ray line astronomy

    NASA Technical Reports Server (NTRS)

    Ramaty, R.; Lingenfelter, R. E.

    1979-01-01

    Gamma-ray astronomy is a valuable source of information on solar activity, supernovae, and nucleosynthesis. Cosmic gamma-ray lines were first observed from solar flares and more recently from the galactic center and a transient event. The latter may give an important insight into nuclear reactions taking place near neutron stars and black holes and a measure of the gravitational redshifts of such objects.

  7. Off line ion source terminal

    NASA Astrophysics Data System (ADS)

    Jayamanna, K.

    2014-01-01

    The off-line ion source (OLIS) terminal provides beams from stable isotopes to ISAC (see Fig. 1) experiments as well as for accelerator commissioning and for pilot beams for radioactive beam experiments. The OLIS terminal (see Fig. 2) is equipped with a microwave driven cusp source for single and double charge ions, a surface ion source for low energy spread alkali beams, and a multi-charge ion source.

  8. Firing up the front line.

    PubMed

    Katzenbach, J R; Santamaria, J A

    1999-01-01

    For many organizations, achieving competitive advantage means eliciting superior performance from employees on the front line--the burger flippers, hotel room cleaners, and baggage handlers whose work has an enormous effect on customers. That's no easy task. Front line workers are paid low wages, have scant hope of advancement, and--not surprisingly--often care little about the company's performance. But then how do some companies succeed in engaging the emotional energy of rank-and-file workers? A team of researchers at McKinsey & Company and the Conference Board recently explored that question and discovered that one highly effective route is demonstrated by the U.S. Marine Corps. The Marines' approach to motivation follows the "mission, values, and pride" path, which researchers say is practical and relevant for the business world. More specifically, the authors say the Marines follow five practices: they over-invest in cultivating core value; prepare every person to lead, including front line supervisors; learn when to create teams and when to create single-leader work groups; attend to all employees, not just the top half; and encourage self-discipline as a way of building pride. The authors admit there are critical differences between the Marines and most businesses. But using vivid examples from companies such as KFC and Marriott International, the authors illustrate how the Marines' approach can be translated for corporate use. Sometimes, the authors maintain, minor changes in a company's standard operating procedure can have a powerful effect on front line pride and can result in substantial payoffs in company performance. PMID:10387573

  9. Line-Editor Computer Program

    NASA Technical Reports Server (NTRS)

    Scott, Peter J.

    1989-01-01

    ZED editing program for DEC VAX computer simple, powerful line editor for text, program source code, and nonbinary data. Excels in processing of text by use of procedure files. Also features versatile search qualifiers, global changes, conditionals, online help, hexadecimal mode, space compression, looping, logical combinations of search strings, journaling, visible control characters, and automatic detabbing. Users of Cambridge implementation devised such ZED procedures as chess games, calculators, and programs for evaluating pi. Written entirely in C.

  10. Line formation in turbulent media

    NASA Astrophysics Data System (ADS)

    Stahlberg, J.

    The methods of mean field electrodynamics are used to investigate the influence of stochastic velocity fields on line formation. Due to numerical difficulties in such concepts, only two point correlations are taken into account. But there are some difficulties: Different models can yield different profiles - for the same parameter - or vice versa, a different interpretation of the same profile can yield different correlation lengths and also different velocities. Care is required in interpreting these parameters as real hydrodynamics ones.

  11. On real quadric line complexes

    NASA Astrophysics Data System (ADS)

    Krasnov, Vyacheslav A.

    2010-12-01

    We describe the topological types of the real parts of the Kummer surfaces associated with real three-dimensional quadric line complexes. The topological type of the real part of such a surface is shown to depend on the number of real singular points: it is determined by the number of such points if any exist, and otherwise the real part of the Kummer surface is either empty or consists of one or two tori.

  12. 46 CFR 42.13-30 - Lines to be used with the load line mark.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 2 2010-10-01 2010-10-01 false Lines to be used with the load line mark. 42.13-30 Section 42.13-30 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) LOAD LINES DOMESTIC AND FOREIGN VOYAGES BY SEA General Rules for Determining Load Lines § 42.13-30 Lines to be used with the load line mark. (a) The lines which...

  13. A new mathematical formulation of the line-by-line method in case of weak line overlapping

    NASA Technical Reports Server (NTRS)

    Ishov, Alexander G.; Krymova, Natalie V.

    1994-01-01

    A rigorous mathematical proof is presented for multiline representation on the equivalent width of a molecular band which consists in the general case of n overlapping spectral lines. The multiline representation includes a principal term and terms of minor significance. The principal term is the equivalent width of the molecular band consisting of the same n nonoverlapping spectral lines. The terms of minor significance take into consideration the overlapping of two, three and more spectral lines. They are small in case of the weak overlapping of spectral lines in the molecular band. The multiline representation can be easily generalized for optically inhomogeneous gas media and holds true for combinations of molecular bands. If the band lines overlap weakly the standard formulation of line-by-line method becomes too labor-consuming. In this case the multiline representation permits line-by-line calculations to be performed more effectively. Other useful properties of the multiline representation are pointed out.

  14. Cavity lining in primary teeth.

    PubMed

    Chauhan, Ravi

    2016-03-01

    Data sourcesEmbase, Medline, Cochrane Central, Biomed Central and Open Grey databases and bibliographies of identified studies.Study selectionRandomised controlled trials investigating humans with primary caries lesions receiving operative treatment involving caries removal and restoration, with minimum two treatment groups comparing different cavity treatments before restoration (no lining versus lining) were included.Data extraction and synthesisData were extracted independently by two reviewers and study quality assessed using the Cochrane risk of bias tool. Random effect meta-analysis was carried out.ResultsThree studies involving a total of 89 patients were included. All the studies involved primary teeth and were conducted in Brazil. Follow-up periods ranged from 26-53 months. All the studies were considered to be at high risk of bias. Restoring the cavity without lining did not significantly affect the risk of failure. The quality of the evidence was low.ConclusionsCurrent evidence does not support strong recommendations to use or not to use liners after caries removal and before restoring cavities. Our findings are restricted to primary teeth after selective excavation, with only one liner (calcium hydroxide) being used for comparison. PMID:27012571

  15. In-line thermoelectric module

    DOEpatents

    Pento, Robert; Marks, James E.; Staffanson, Clifford D.

    2000-01-01

    A thermoelectric module with a plurality of electricity generating units each having a first end and a second end, the units being arranged first end to second end along an in-line axis. Each unit includes first and second elements each made of a thermoelectric material, an electrically conductive hot member arranged to heat one side of the first element, and an electrically conductive cold member arranged to cool another side of the first element and to cool one side of the second element. The hot member, the first element, the cold member and the second element are supported in a fixture, are electrically connected respectively to provide an electricity generating unit, and are arranged respectively in positions along the in-line axis. The individual components of each generating unit and the respective generating units are clamped in their in-line positions by a loading bolt at one end of the fixture and a stop wall at the other end of the fixture. The hot members may have a T-shape and the cold members an hourglass shape to facilitate heat transfer. The direction of heat transfer through the hot members may be perpendicular to the direction of heat transfer through the cold members, and both of these heat transfer directions may be perpendicular to the direction of current flow through the module.

  16. Line-scanning laser ophthalmoscope

    NASA Astrophysics Data System (ADS)

    Hammer, Daniel X.; Ferguson, R. Daniel; Ustun, Teoman E.; Bigelow, Chad E.; Iftimia, Nicusor V.; Webb, Robert H.

    2006-07-01

    Scanning laser ophthalmoscopy (SLO) is a powerful imaging tool with specialized applications limited to research and ophthalmology clinics due in part to instrument size, cost, and complexity. Conversely, low-cost retinal imaging devices have limited capabilities in screening, detection, and diagnosis of diseases. To fill the niche between these two, a hand-held, nonmydriatic line-scanning laser ophthalmoscope (LSLO) is designed, constructed, and tested on normal human subjects. The LSLO has only one moving part and uses a novel optical approach to produce wide-field confocal fundus images. Imaging modes include multiwavelength illumination and live stereoscopic imaging with a split aperture. Image processing and display functions are controlled with two stacked prototype compact printed circuit boards. With near shot-noise limited performance, the digital LSLO camera requires low illumination power (<500 µW) at near-infrared wavelengths. The line-scanning principle of operation is examined in comparison to SLO and other imaging modes. The line-scanning approach produces high-contrast confocal images with nearly the same performance as a flying-spot SLO. The LSLO may significantly enhance SLO utility for routine use by ophthalmologists, optometrists, general practitioners, and also emergency medical personnel and technicians in the field for retinal disease detection and other diverse applications.

  17. Gemini spectroscopy of Galactic Bulge Sources: a population of hidden accreting binaries revealed?

    NASA Astrophysics Data System (ADS)

    Wu, Jianfeng; Jonker, P. G.; Torres, M. A. P.; Britt, C. T.; Johnson, C. B.; Hynes, R. I.; Greiss, S.; Steeghs, D. T. H.; Maccarone, T. J.; Heinke, C. O.; Wevers, T.

    2015-04-01

    We present Gemini spectroscopy for 21 candidate optical counterparts to X-ray sources discovered in the Galactic Bulge Survey (GBS). For the majority of the 21 sources, the optical spectroscopy establishes that they are indeed the likely counterparts. One of the criteria we used for the identification was the presence of an Hα emission line. The spectra of several sources revealed an Hα emission line only after careful subtraction of the F or G stellar spectral absorption lines. In a subclass of three of these sources, the residual Hα emission line is broad (≳ 400 km s-1) which suggests that it is formed in an accretion disc, whereas in other cases the line width is such that we currently cannot determine whether the line emission is formed in an active star/binary or in an accretion disc. GBS source CX377 shows this hidden-accretion behaviour most dramatically. The previously identified broad Hα emission of this source is not present in its Gemini spectra taken ˜1 yr later. However, broad emission is revealed after subtracting an F6 template star spectrum. The Gemini spectra of three sources (CX446, CX1004, and CXB2) as well as the presence of possible eclipses in light curves of these sources suggest that these sources are accreting binaries viewed under a high inclination.

  18. Unidentified line in x-ray spectra of the Andromeda galaxy and Perseus galaxy cluster.

    PubMed

    Boyarsky, A; Ruchayskiy, O; Iakubovskyi, D; Franse, J

    2014-12-19

    We report a weak line at 3.52±0.02  keV in x-ray spectra of the Andromeda galaxy and the Perseus galaxy cluster observed by the metal-oxide-silicon (MOS) and p-n (PN) CCD cameras of the XMM-Newton telescope. This line is not known as an atomic line in the spectra of galaxies or clusters. It becomes stronger towards the centers of the objects; is stronger for Perseus than for M31; is absent in the spectrum of a deep "blank sky" data set. Although for each object it is hard to exclude that the feature is due to an instrumental effect or an atomic line, it is consistent with the behavior of a dark matter decay line. Future (non-)detections of this line in multiple objects may help to reveal its nature. PMID:25554871

  19. Human amniotic epithelial cells as feeder layer to derive and maintain human embryonic stem cells from poor-quality embryos.

    PubMed

    Ávila-González, Daniela; Vega-Hernández, Eva; Regalado-Hernández, Juan Carlos; De la Jara-Díaz, Julio Francisco; García-Castro, Irma Lydia; Molina-Hernández, Anayansi; Moreno-Verduzco, Elsa Romelia; Razo-Aguilera, Guadalupe; Flores-Herrera, Héctor; Portillo, Wendy; Díaz-Martínez, Néstor Emmanuel; García-López, Guadalupe; Díaz, Néstor Fabián

    2015-09-01

    Data from the literature suggest that human embryonic stem cell (hESC) lines used in research do not genetically represent all human populations. The derivation of hESC through conventional methods involve the destruction of viable human embryos, as well the use of mouse embryonic fibroblasts as a feeder layer, which has several drawbacks. We obtained the hESC line (Amicqui-1) from poor-quality (PQ) embryos derived and maintained on human amniotic epithelial cells (hAEC). This line displays a battery of markers of pluripotency and we demonstrated the capacity of these cells to produce derivates of the three germ layers. PMID:26246271

  20. VIP contribution to the decidualization program: regulatory T cell recruitment.

    PubMed

    Grasso, Esteban; Paparini, Daniel; Agüero, Mariana; Mor, Gil; Pérez Leirós, Claudia; Ramhorst, Rosanna

    2014-04-01

    During early pregnancy, the human uterus undergoes profound tissue remodeling characterized by leukocyte invasion and production of proinflammatory cytokines, followed by tissue repair and tolerance maintenance induction. Vasoactive intestinal peptide (VIP) is produced by trophoblast cells and modulates the maternal immune response toward a tolerogenic profile. Here, we evaluated the contribution of the VIP/VPAC to endometrial renewal, inducing decidualization and the recruitment of induced regulatory T cells (iTregs) that accompany the implantation period. For that purpose, we used an in vitro model of decidualization with a human endometrial stromal cell line (HESC) stimulated with progesterone (P4) and lipopolysaccharide (LPS) simulating the inflammatory response during implantation and human iTregs (CD4(+)CD25(+)FOXP3(+)) differentiated from naïve T cells obtained from peripheral blood mononuclear cells of fertile women. We observed that VIP and its receptor VPAC1 are constitutively expressed in HESCs and that P4 increased VIP expression. Moreover, in HESC VIP induced expression of RANTES (CCL5), one of the main chemokines involved in T cell recruitment, and this effect is enhanced by the presence of P4 and LPS. Finally, assays of the migration of iTregs toward conditioned media from HESCs revealed that endogenous VIP production induced by P4 and LPS and RANTES production were involved, as anti-RANTES neutralizing Ab or VIP antagonist prevented their migration. We conclude that VIP may have an active role in the decidualization process, thus contributing to recruitment of iTregs toward endometrial stromal cells by increasing RANTES expression in a P4-dependent manner. PMID:24492467