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Sample records for cells support feeder-independent

  1. Feeder-cell-independent culture of the pig-embryonic-stem-cell-derived exocrine pancreatic cell line, PICM-31

    Science.gov (United States)

    The adaptation to feeder-independent growth of a pig embryonic stem cell-derived pancreatic cell line is described. The parental PICM-31 cell line, previously characterized as an exocrine pancreas cell line, was colony-cloned two times in succession resulting in the subclonal cell line, PICM-31A1. P...

  2. Identification of molecules derived from human fibroblast feeder cells that support the proliferation of human embryonic stem cells

    DEFF Research Database (Denmark)

    Anisimov, Sergey V.; Christophersen, Nicolaj S.; Correia, Ana S.

    2011-01-01

    The majority of human embryonic stem cell lines depend on a feeder cell layer for continuous growth in vitro, so that they can remain in an undifferentiated state. Limited knowledge is available concerning the molecular mechanisms that underlie the capacity of feeder cells to support both...... the proliferation and pluripotency of these cells. Importantly, feeder cells generally lose their capacity to support human embryonic stem cell proliferation in vitro following long-term culture. In this study, we performed large-scale gene expression profiles of human foreskin fibroblasts during early...... foreskin fibroblasts to serve as feeder cells for human embryonic stem cell cultures. Among these, the C-KIT, leptin and pigment epithelium-derived factor (PEDF) genes were the most interesting candidates....

  3. Feeder cells support the culture of induced pluripotent stem cells even after chemical fixation.

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    Xiao-Shan Yue

    Full Text Available Chemically fixed mouse embryonic fibroblasts (MEFs, instead of live feeder cells, were applied to the maintenance of mouse induced pluripotent stem (miPS cells. Formaldehyde and glutaraldehyde were used for chemical fixation. The chemically fixed MEF feeders maintained the pluripotency of miPS cells, as well as their undifferentiated state. Furthermore, the chemically fixed MEF feeders were reused several times without affecting their functions. These results indicate that chemical fixation can be applied to modify biological feeders chemically, without losing their original functions. Chemically fixed MEF feeders will be applicable to other stem cell cultures as a reusable extracellular matrix candidate that can be preserved on a long-term basis.

  4. Human induced pluripotent stem cells on autologous feeders.

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    Kazutoshi Takahashi

    Full Text Available BACKGROUND: For therapeutic usage of induced Pluripotent Stem (iPS cells, to accomplish xeno-free culture is critical. Previous reports have shown that human embryonic stem (ES cells can be maintained in feeder-free condition. However, absence of feeder cells can be a hostile environment for pluripotent cells and often results in karyotype abnormalities. Instead of animal feeders, human fibroblasts can be used as feeder cells of human ES cells. However, one still has to be concerned about the existence of unidentified pathogens, such as viruses and prions in these non-autologous feeders. METHODOLOGY/PRINCIPAL FINDINGS: This report demonstrates that human induced Pluripotent Stem (iPS cells can be established and maintained on isogenic parental feeder cells. We tested four independent human skin fibroblasts for the potential to maintain self-renewal of iPS cells. All the fibroblasts tested, as well as their conditioned medium, were capable of maintaining the undifferentiated state and normal karyotypes of iPS cells. Furthermore, human iPS cells can be generated on isogenic parental fibroblasts as feeders. These iPS cells carried on proliferation over 19 passages with undifferentiated morphologies. They expressed undifferentiated pluripotent cell markers, and could differentiate into all three germ layers via embryoid body and teratoma formation. CONCLUSIONS/SIGNIFICANCE: These results suggest that autologous fibroblasts can be not only a source for iPS cells but also be feeder layers. Our results provide a possibility to solve the dilemma by using isogenic fibroblasts as feeder layers of iPS cells. This is an important step toward the establishment of clinical grade iPS cells.

  5. Effects of Feeder Cells on Dopaminergic Differentiation of Human Embryonic Stem Cells

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    Zhenqiang Zhao

    2016-12-01

    Full Text Available Mouse embryonic fibroblasts (MEFs and human foreskin fibroblasts (HFFs are used for the culture of human embryonic stem cells (hESCs. MEFs and HFFs differed in their capacity to support the proliferation and pluripotency of hESCs and could affect cardiac differentiation potential of hESCs. The aim of this study was to evaluate the effect of MEFs and HFFs feeders on dopaminergic differentiation of hESCs lines. To minimize the impact of culture condition variation, two hESCs lines were cultured on mixed feeder cells (MFCs, MEFs: HFFs =1:1 and HFFs feeder respectively, and then were differentiated into DA neurons under the identical protocol. Dopaminergic differentiation was evaluated by immunocytochemistry, quantitative fluorescent real-time PCR (qRT-PCR, transmission and scanning electron microscopy, and patch clamp. Our results demonstrated that these hESCs-derived neurons were genuine and functional DA neurons. However, compared to hESCs line on MFCs feeder, hESCs line on HFFs feeder had a higher proportion of TH positive cells and expressed higher levels of FOXA2, PITX3, NURR1 and TH genes. In addition, the values of threshold intensity and threshold membrane potential of DA neurons from hESCs line on HFFs feeder were lower than those of DA neurons from hESCs line on the MFCs feeder. In conclusion, HFFs feeder not only facilitated the differentiation of hESCs cells into dopaminergic neurons, but also induced hESCs-derived DA neurons to express higher electrophysiological excitability. Therefore, feeder cells could affect not only dopaminergic differentiation potential of different hESCs lines, but also electrophysiological properties of hESCs-derived DA neurons.

  6. Neural stem cells achieve and maintain pluripotency without feeder cells.

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    Hyun Woo Choi

    Full Text Available BACKGROUND: Differentiated cells can be reprogrammed into pluripotency by transduction of four defined transcription factors. Induced pluripotent stem cells (iPS cells are expected to be useful for regenerative medicine as well as basic research. Recently, the report showed that mouse embryonic fibroblasts (MEF cells are not essential for reprogramming. However, in using fibroblasts as donor cells for reprogramming, individual fibroblasts that had failed to reprogram could function as feeder cells. METHODOLOGY/PRINCIPAL FINDING: Here, we show that adult mouse neural stem cells (NSCs, which are not functional feeder cells, can be reprogrammed into iPS cells using defined four factors (Oct4, Sox2, Klf4, and c-Myc under feeder-free conditions. The iPS cells, generated from NSCs expressing the Oct4-GFP reporter gene, could proliferate for more than two months (passage 20. Generated and maintained without feeder cells, these iPS cells expressed pluripotency markers (Oct4 and Nanog, the promoter regions of Oct4 and Nanog were hypomethylated, could differentiated into to all three germ layers in vitro, and formed a germline chimera. These data indicate that NSCs can achieve and maintain pluripotency under feeder-free conditions. CONCLUSION/SIGNIFICANCE: This study suggested that factors secreted by feeder cells are not essential in the initial/early stages of reprogramming and for pluripotency maintenance. This technology might be useful for a human system, as a feeder-free reprogramming system may help generate iPS cells of a clinical grade for tissue or organ regeneration.

  7. Utilization of human amniotic mesenchymal cells as feeder layers to sustain propagation of human embryonic stem cells in the undifferentiated state.

    Science.gov (United States)

    Zhang, Kehua; Cai, Zhe; Li, Yang; Shu, Jun; Pan, Lin; Wan, Fang; Li, Hong; Huang, Xiaojie; He, Chun; Liu, Yanqiu; Cui, Xiaohui; Xu, Yang; Gao, Yan; Wu, Liqun; Cao, Shanxia; Li, Lingsong

    2011-08-01

    Human embryonic stem (ES) cells are usually maintained in the undifferentiated state by culturing on feeder cells layers of mouse embryonic fibroblasts (MEFs). However, MEFs are not suitable to support human ES cells used for clinical purpose because of risk of zoonosis from animal cells. Therefore, human tissue-based feeder layers need to be developed for human ES cells for clinical purpose. Hereof we report that human amniotic mesenchymal cells (hAMCs) could act as feeder cells for human ES cells, because they are easily obtained and relatively exempt from ethical problem. Like MEFs, hAMCs could act as feeder cells for human ES cells to grow well on. The self-renewal rate of human ES cells cultured on hAMCs feeders was higher than that on MEFs and human amniotic epithelial cells determined by measurement of colonial diameters and growth curve as well as cell cycle analysis. Both immunofluorescence staining and immunoblotting showed that human ES cells cultured on hAMCs expressed stem cell markers such as Oct-3/4, Sox2, and NANOG. Verified by embryoid body formation in vitro and teratoma formation in vivo, we found out that after 20 passages of culture, human ES cells grown on hAMCs feeders could still retain the potency of differentiating into three germ layers. Taken together, our data suggested hAMCs may be safe feeder cells to sustain the propagation of human ES cells in undifferentiated state for future therapeutic use.

  8. Effects of different feeder layers on culture of bovine embryonic stem cell-like cells in vitro.

    Science.gov (United States)

    Cong, Shan; Cao, Guifang; Liu, Dongjun

    2014-12-01

    To find a suitable feeder layer is important for successful culture conditions of bovine embryonic stem cell-like cells. In this study, expression of pluripotency-related genes OCT4, SOX2 and NANOG in bovine embryonic stem cell-like cells on mouse embryonic fibroblast feeder layers at 1-5 passages were monitored in order to identify the possible reason that bovine embryonic stem cell-like cells could not continue growth and passage. Here, we developed two novel feeder layers, mixed embryonic fibroblast feeder layers of mouse and bovine embryonic fibroblast at different ratios and sources including mouse fibroblast cell lines. The bovine embryonic stem cell-like cells generated in our study displayed typical stem cell morphology and expressed specific markers such as OCT4, stage-specific embryonic antigen 1 and 4, alkaline phosphatase, SOX2, and NANOG mRNA levels. When feeder layers and cell growth factors were removed, the bovine embryonic stem cell-like cells formed embryoid bodies in a suspension culture. Furthermore, we compared the expression of the pluripotent markers during bovine embryonic stem cell-like cell in culture on mixed embryonic fibroblast feeder layers, including mouse fibroblast cell lines feeder layers and mouse embryonic fibroblast feeder layers by real-time quantitative polymerase chain reaction. Results suggested that mixed embryonic fibroblast and sources including mouse fibroblast cell lines feeder layers were more suitable for long-term culture and growth of bovine embryonic stem cell-like cells than mouse embryonic fibroblast feeder layers. The findings may provide useful experimental data for the establishment of an appropriate culture system for bovine embryonic stem cell lines.

  9. A feeder-free culture using autogeneic conditioned medium for undifferentiated growth of human embryonic stem cells: Comparative expression profiles of mRNAs, microRNAs and proteins among different feeders and conditioned media

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    Chou Chi-Hsien

    2010-10-01

    Full Text Available Abstract Background Human embryonic stem (hES cell lines were derived from the inner cell mass of human blastocysts, and were cultured on mouse embryonic fibroblast (MEF feeder to maintain undifferentiated growth, extensive renewal capacity, and pluripotency. The hES-T3 cell line with normal female karyotype was previously used to differentiate into autogeneic fibroblast-like cells (T3HDF as feeder to support the undifferentiated growth of hES-T3 cells (T3/HDF for 14 passages. Results A feeder-free culture on Matrigel in hES medium conditioned by the autogeneic feeder cells (T3HDF was established to maintain the undifferentiated growth of hES-T3 cells (T3/CMHDF for 8 passages in this investigation. The gene expression profiles of mRNAs, microRNAs and proteins between the undifferentiated T3/HDF and T3/CMHDF cells were shown to be very similar, and their expression profiles were also found to be similar to those of T3/MEF and T3/CMMEF cells grown on MEF feeder and feeder-free Matrigel in MEF-conditioned medium, respectively. The undifferentiated state of T3/HDF and T3/CMHDF as well as T3/MEF andT3/CMMEF cells was evidenced by the very high expression levels of "stemness" genes and low expression levels of differentiation markers of ectoderm, mesoderm and endoderm in addition to the strong staining of OCT4 and NANOG. Conclusion The T3HDF feeder and T3HDF-conditioned medium were able to support the undifferentiated growth of hES cells, and they would be useful for drug development and toxicity testing in addition to the reduced risks of xenogeneic pathogens when used for medical applications such as cell therapies.

  10. Fabrication of corneal epithelial cell sheets maintaining colony-forming cells without feeder cells by oxygen-controlled method.

    Science.gov (United States)

    Nakajima, Ryota; Takeda, Shizu

    2014-01-01

    The use of murine 3T3 feeder cells needs to be avoided when fabricating corneal epithelial cell sheets for use in treating ocular surface diseases. However, the expression level of the epithelial stem/progenitor cell marker, p63, is down-regulated in feeder-free culture systems. In this study, in order to fabricate corneal epithelial cell sheets that maintain colony-forming cells without using any feeder cells, we investigated the use of an oxygen-controlled method that was developed previously to fabricate cell sheets efficiently. Rabbit limbal epithelial cells were cultured under hypoxia (1-10% O2) and under normoxia during stratification after reaching confluence. Multilayered corneal epithelial cell sheets were fabricated using an oxygen-controlled method, and immunofluorescence analysis showed that cytokeratin 3 and p63 was expressed in appropriate localization in the cell sheets. The colony-forming efficiency of the cell sheets fabricated by the oxygen-controlled method without feeder cells was significantly higher than that of cell sheets fabricated under 20% O2 without feeder cells. These results indicate that the oxygen-controlled method has the potential to achieve a feeder-free culture system for fabricating corneal epithelial cell sheets for corneal regeneration. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Final report on the evolution of supporting conditions for the feeders of 500 MWe PHWR

    International Nuclear Information System (INIS)

    Mishra, Rajesh; Soni, R.S.; Kushawaha, H.S.; Mahajan, S.C.; Kakodkar, A.; Hariprasad, K.

    1994-01-01

    This report deals with the evolution of generic supporting conditions for the feeders of 500 MWe PHWR based on the analysis and qualification of a few representative feeders. There are 196 different feeder pipe configurations for a total of 748 feeders. The present analysis was aimed at evolving a generalised supporting criteria based on the analysis of some representative feeders. The analysis was carried out for various loadings viz. pressure, temperature, dead weight, operating basis earthquake (OBE), safe shutdown earthquake (SSE) and creep loadings. The analysis for OBE and SSE loadings were carried out using response spectrum method. The effect of spacers between various feeders was modelled using higher damping values than those prescribed in ASME code. Based on the above analyses, generic supporting arrangements for the feeders of various groups have been finalized. This report gives details about the mathematical modelling, the analysis approach, the optimised supporting criteria, finalization of grouping and fixing of boundaries between various groups of feeders. (author). 34 refs., 51 figs., 69 tabs

  12. Developmental Competence of Buffalo (Bubalus bubalis) Pluripotent Embryonic Stem Cells Over Different Homologous Feeder Layers and the Comparative Evaluation with Various Extracellular Matrices.

    Science.gov (United States)

    Sharma, Manjinder; Dubey, Pawan K; Kumar, Rajesh; Nath, Amar; Kumar, G Sai; Sharma, G Taru

    2013-05-01

    Use of somatic cells as a feeder layer to maintain the embryonic stem cells (ESCs) in undifferentiated state limits the stem cell research design, since experimental data may result from a combined ESCs and feeder cell response to various stimuli. Therefore, present study was designed to evaluate the developmental competence of the buffalo ESCs over different homogenous feeders and compare with various extracellular matrices using different concentrations of LIF. Inner cell masses (ICMs) of in vitro hatched blastocysts were cultured onto homologous feeders viz. fetal fibroblast, granulosa and oviductal cell feeder layers and synthetic matrices viz. fibronectin, collagen type I and matrigel in culture medium. Developmental efficiency was found higher for ESCs cultured on fetal fibroblast and granulosa layers (83.33%) followed by fibronectin (77.78%) at 30 ng LIF. Oviductal feeder was found to be the least efficient feeder showing only 11.11% undifferentiated primary ESC colonies at 30 ng LIF. However, neither feeder layer nor synthetic matrix could support the development of primary colonies at 10 ng LIF. Expression of SSEA- 4, TRA-1-60 and Oct-4 were found positive in ESC colonies from all the feeders and synthetic matrices with 20 ng and 30 ng LIF. Fetal fibroblast and granulosa cell while, amongst synthetic matrices, fibronectin were found to be equally efficient to support the growth and maintenance of ESCs pluripotency with 30 ng LIF. This well-defined culture conditions may provide an animal model for culturing human embryonic stem cells in the xeno-free or feeder-free conditions for future clinical applications.

  13. "The good into the pot, the bad into the crop!"--a new technology to free stem cells from feeder cells.

    Directory of Open Access Journals (Sweden)

    Annette Schneider

    Full Text Available A variety of embryonic and adult stem cell lines require an initial co-culturing with feeder cells for non-differentiated growth, self renewal and maintenance of pluripotency. However for many downstream ES cell applications the feeder cells have to be considered contaminations that might interfere not just with the analysis of experimental data but also with clinical application and tissue engineering approaches. Here we introduce a novel technique that allows for the selection of pure feeder-freed stem cells, following stem cell proliferation on feeder cell layers. Complete and reproducible separation of feeder and embryonic stem cells was accomplished by adaptation of an automated cell selection system that resulted in the aspiration of distinct cell colonies or fraction of colonies according to predefined physical parameters. Analyzing neuronal differentiation we demonstrated feeder-freed stem cells to exhibit differentiation potentials comparable to embryonic stem cells differentiated under standard conditions. However, embryoid body growth as well as differentiation of stem cells into cardiomyocytes was significantly enhanced in feeder-freed cells, indicating a feeder cell dependent modulation of lineage differentiation during early embryoid body development. These findings underline the necessity to separate stem and feeder cells before the initiation of in vitro differentiation. The complete separation of stem and feeder cells by this new technology results in pure stem cell populations for translational approaches. Furthermore, a more detailed analysis of the effect of feeder cells on stem cell differentiation is now possible, that might facilitate the identification and development of new optimized human or genetically modified feeder cell lines.

  14. Effects of different feeder layers on culture of bovine embryonic stem cell-like cells in vitro

    OpenAIRE

    Cong, Shan; Cao, Guifang; Liu, Dongjun

    2014-01-01

    To find a suitable feeder layer is important for successful culture conditions of bovine embryonic stem cell-like cells. In this study, expression of pluripotency-related genes OCT4, SOX2 and NANOG in bovine embryonic stem cell-like cells on mouse embryonic fibroblast feeder layers at 1–5 passages were monitored in order to identify the possible reason that bovine embryonic stem cell-like cells could not continue growth and passage. Here, we developed two novel feeder layers, mixed embryonic ...

  15. Feeder Cell Type Affects the Growth of In Vitro Cultured Bovine Trophoblast Cells

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    Islam M. Saadeldin

    2017-01-01

    Full Text Available Trophectoderm cells are the foremost embryonic cells to differentiate with prospective stem-cell properties. In the current study, we aimed at improving the current approach for trophoblast culture by using granulosa cells as feeders. Porcine granulosa cells (PGCs compared to the conventional mouse embryonic fibroblasts (MEFs were used to grow trophectoderm cells from hatched bovine blastocysts. Isolated trophectoderm cells were monitored and displayed characteristic epithelial/cuboidal morphology. The isolated trophectoderm cells expressed mRNA of homeobox protein (CDX2, cytokeratin-8 (KRT8, and interferon tau (IFNT. The expression level was higher on PGCs compared to MEFs throughout the study. In addition, primary trophectoderm cell colonies grew faster on PGCs, with a doubling time of approximately 48 hrs, compared to MEFs. PGCs feeders produced a fair amount of 17β-estradiol and progesterone. We speculated that the supplementation of sex steroids and still-unknown factors during the trophoblasts coculture on PGCs have helped to have better trophectoderm cell’s growth than on MEFs. This is the first time to use PGCs as feeders to culture trophectoderm cells and it proved superior to MEFs. We propose PGCs as alternative feeders for long-term culture of bovine trophectoderm cells. This model will potentially benefit studies on the early trophoblast and embryonic development in bovines.

  16. Tumorigenic risk of human induced pluripotent stem cell explants cultured on mouse SNL76/7 feeder cells

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    Kamada, Mizuna; Mitsui, Youji, E-mail: y-mitsui8310@hb.tp1.jp; Kumazaki, Tsutomu; Kawahara, Yuta; Matsuo, Taira; Takahashi, Tomoko, E-mail: t-takahashi@kph.bunri-u.ac.jp

    2014-10-24

    Highlights: • hiPS cell explants formed malignant tumors when SNL76/7 feeder cells were used. • Multi type tumors developed by interaction of SNL76/7 feeder cells with hiPS cells. • Tumorigenic risk occurs by co-culture of hiPS cells with SNL76/7 feeder cells. - Abstract: The potential for tumor formation from transplanted human induced pluripotent stem cell (hiPSC) derivatives represents a high risk in their application to regenerative medicine. We examined the genetic origin and characteristics of tumors, that were formed when 13 hiPSC lines, established by ourselves, and 201B7 hiPSC from Kyoto University were transplanted into severe combined immune-deficient (SCID) mice. Though teratomas formed in 58% of mice, five angiosarcomas, one malignant solitary fibrous tumor and one undifferentiated pleomorphic sarcoma formed in the remaining mice. Three malignant cell lines were established from the tumors, which were derived from mitomycin C (MMC)-treated SNL76/7 (MMC-SNL) feeder cells, as tumor development from fusion cells between MMC-SNL and hiPSCs was negative by genetic analysis. While parent SNL76/7 cells produced malignant tumors, neither MMC-SNL nor MMC-treated mouse embryo fibroblast (MEF) produced malignant tumors. When MMC-SNL feeder cells were co-cultured with hiPSCs, growing cell lines were generated, that expressed genes similar to the parent SNL76/7 cells. Thus, hiPSCs grown on MMC-SNL feeder cells have a high risk of generating feeder-derived malignant tumors. The possible mechanism(s) of growth restoration and the formation of multiple tumor types are discussed with respect of the interactions between MMC-SNL and hiPSC.

  17. Irradiated murine fibroblasts as feeder layer used in human cell culture

    International Nuclear Information System (INIS)

    Almeida, Tiago L.; Klingbeil, Fatima G.; Yoshito, Daniele; Caproni, Priscila; Mathor, Monica B.; Herson, Marisa R.

    2007-01-01

    In 1975, Rheinwald and Green published an in vitro model for keratinocyte cell cultures in which the use of murine fibroblasts, as a feeder layer was introduced. These cells are modified fibroblasts, which presence render keratinocyte cells to remain proliferative for longer periods of time. This optimization of culture outputs has allowed for several clinical applications of confluent keratinocyte cultures as skin substitutes or wound dressings in situations such as post burn extensive skin loss, loss of oral mucosa, and other skin disorders. Nevertheless, proliferation of fibroblast in co-culture with keratinocytes must be controlled by anti-proliferative measures such as irradiation; at the same time, keratinocytes require specific nutrients in the culture medium, which may interfere with the fibroblast feeder layer viability. Therefore, the thorough understanding of the impact of different issues such as culture media composition, irradiation dose and pre-plating storage conditions of irradiated fibroblast to be used as feeder layer in these co-culture systems is important. In this work, changes as far as viability and proliferative rates of irradiated fibroblasts in culture were evaluated in relation to the type of culture medium used, dose of gamma radiation exposure, storage and timing of cell plating post irradiation. Results indicate that the type of culture medium used and time-lag between irradiation, refrigeration and plating of irradiated cells do not have significant impact in culture outcomes. However, the dose of gamma radiation administered to the cells may influence the final quality of these cells if to be used as a feeder layer. (author)

  18. An evaluation of the choice of feeder cell growth arrest for the production of cultured epidermis.

    Science.gov (United States)

    Chugh, Rishi Man; Chaturvedi, Madhusudan; Yerneni, Lakshmana Kumar

    2015-12-01

    Growth arrested 3T3 cells have been used as feeder cells in human epidermal keratinocyte cultures to produce cultured epidermal autografts for the treatment of burns. The feeder cells were ideally growth-arrested by gamma-irradiation. Alternatively, growth arrest by mitomycin C treatment is a cost effective option. We compared the functional efficacy of these two approaches in keratinocyte cultures by colony forming efficiency, the net growth area of colonies, BrdU labeling and histological features of cultured epidermal sheets. The growth area estimation involved a semi-automated digital technique using the Adobe Photoshop and comprised of isolation and enumeration of red pixels in Rhodamine B-stained keratinocyte colonies. A further refinement of the technique led to the identification of critical steps to increasing the degree of accuracy and enabling its application as an extension of colony formation assay. The results on feeder cell functionality revealed that the gamma irradiated feeders influenced significantly higher colony forming efficiency and larger growth area than the mitomycin C treated feeders. The BrdU labeling study indicated significant stimulation of the overall keratinocyte proliferation by the gamma irradiated feeders. The cultured epidermal sheets produced by gamma feeders were relatively thicker than those produced by mitomycin C feeders. We discussed the clinical utility of mitomycin C feeders from the viewpoint of cost-effective burn care in developing countries. Copyright © 2015 Elsevier Ltd and ISBI. All rights reserved.

  19. Mesenchymal stem cell like (MSCl) cells generated from human embryonic stem cells support pluripotent cell growth

    International Nuclear Information System (INIS)

    Varga, Nóra; Veréb, Zoltán; Rajnavölgyi, Éva; Német, Katalin; Uher, Ferenc; Sarkadi, Balázs; Apáti, Ágota

    2011-01-01

    Highlights: ► MSC like cells were derived from hESC by a simple and reproducible method. ► Differentiation and immunosuppressive features of MSCl cells were similar to bmMSC. ► MSCl cells as feeder cells support the undifferentiated growth of hESC. -- Abstract: Mesenchymal stem cell like (MSCl) cells were generated from human embryonic stem cells (hESC) through embryoid body formation, and isolated by adherence to plastic surface. MSCl cell lines could be propagated without changes in morphological or functional characteristics for more than 15 passages. These cells, as well as their fluorescent protein expressing stable derivatives, efficiently supported the growth of undifferentiated human embryonic stem cells as feeder cells. The MSCl cells did not express the embryonic (Oct4, Nanog, ABCG2, PODXL, or SSEA4), or hematopoietic (CD34, CD45, CD14, CD133, HLA-DR) stem cell markers, while were positive for the characteristic cell surface markers of MSCs (CD44, CD73, CD90, CD105). MSCl cells could be differentiated toward osteogenic, chondrogenic or adipogenic directions and exhibited significant inhibition of mitogen-activated lymphocyte proliferation, and thus presented immunosuppressive features. We suggest that cultured MSCl cells can properly model human MSCs and be applied as efficient feeders in hESC cultures.

  20. Mesenchymal stem cell like (MSCl) cells generated from human embryonic stem cells support pluripotent cell growth

    Energy Technology Data Exchange (ETDEWEB)

    Varga, Nora [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary); Vereb, Zoltan; Rajnavoelgyi, Eva [Department of Immunology, Medical and Health Science Centre, University of Debrecen, Debrecen (Hungary); Nemet, Katalin; Uher, Ferenc; Sarkadi, Balazs [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary); Apati, Agota, E-mail: apati@kkk.org.hu [Membrane Research Group of the Hungarian Academy of Sciences, Semmelweis University, Budapest (Hungary)

    2011-10-28

    Highlights: Black-Right-Pointing-Pointer MSC like cells were derived from hESC by a simple and reproducible method. Black-Right-Pointing-Pointer Differentiation and immunosuppressive features of MSCl cells were similar to bmMSC. Black-Right-Pointing-Pointer MSCl cells as feeder cells support the undifferentiated growth of hESC. -- Abstract: Mesenchymal stem cell like (MSCl) cells were generated from human embryonic stem cells (hESC) through embryoid body formation, and isolated by adherence to plastic surface. MSCl cell lines could be propagated without changes in morphological or functional characteristics for more than 15 passages. These cells, as well as their fluorescent protein expressing stable derivatives, efficiently supported the growth of undifferentiated human embryonic stem cells as feeder cells. The MSCl cells did not express the embryonic (Oct4, Nanog, ABCG2, PODXL, or SSEA4), or hematopoietic (CD34, CD45, CD14, CD133, HLA-DR) stem cell markers, while were positive for the characteristic cell surface markers of MSCs (CD44, CD73, CD90, CD105). MSCl cells could be differentiated toward osteogenic, chondrogenic or adipogenic directions and exhibited significant inhibition of mitogen-activated lymphocyte proliferation, and thus presented immunosuppressive features. We suggest that cultured MSCl cells can properly model human MSCs and be applied as efficient feeders in hESC cultures.

  1. Establishment of feeder-free culture system for human induced pluripotent stem cell on DAS nanocrystalline graphene

    Science.gov (United States)

    Lee, Hyunah; Nam, Donggyu; Choi, Jae-Kyung; Araúzo-Bravo, Marcos J.; Kwon, Soon-Yong; Zaehres, Holm; Lee, Taehee; Park, Chan Young; Kang, Hyun-Wook; Schöler, Hans R.; Kim, Jeong Beom

    2016-02-01

    The maintenance of undifferentiated human pluripotent stem cells (hPSC) under xeno-free condition requires the use of human feeder cells or extracellular matrix (ECM) coating. However, human-derived sources may cause human pathogen contamination by viral or non-viral agents to the patients. Here we demonstrate feeder-free and xeno-free culture system for hPSC expansion using diffusion assisted synthesis-grown nanocrystalline graphene (DAS-NG), a synthetic non-biological nanomaterial which completely rule out the concern of human pathogen contamination. DAS-NG exhibited advanced biocompatibilities including surface nanoroughness, oxygen containing functional groups and hydrophilicity. hPSC cultured on DAS-NG could maintain pluripotency in vitro and in vivo, and especially cell adhesion-related gene expression profile was comparable to those of cultured on feeders, while hPSC cultured without DAS-NG differentiated spontaneously with high expression of somatic cell-enriched adhesion genes. This feeder-free and xeno-free culture method using DAS-NG will facilitate the generation of clinical-grade hPSC.

  2. Differentiation of oligodendrocyte progenitor cells from dissociated monolayer and feeder-free cultured pluripotent stem cells.

    Science.gov (United States)

    Yamashita, Tomoko; Miyamoto, Yuki; Bando, Yoshio; Ono, Takashi; Kobayashi, Sakurako; Doi, Ayano; Araki, Toshihiro; Kato, Yosuke; Shirakawa, Takayuki; Suzuki, Yutaka; Yamauchi, Junji; Yoshida, Shigetaka; Sato, Naoya

    2017-01-01

    Oligodendrocytes myelinate axons and form myelin sheaths in the central nervous system. The development of therapies for demyelinating diseases, including multiple sclerosis and leukodystrophies, is a challenge because the pathogenic mechanisms of disease remain poorly understood. Primate pluripotent stem cell-derived oligodendrocytes are expected to help elucidate the molecular pathogenesis of these diseases. Oligodendrocytes have been successfully differentiated from human pluripotent stem cells. However, it is challenging to prepare large amounts of oligodendrocytes over a short amount of time because of manipulation difficulties under conventional primate pluripotent stem cell culture methods. We developed a proprietary dissociated monolayer and feeder-free culture system to handle pluripotent stem cell cultures. Because the dissociated monolayer and feeder-free culture system improves the quality and growth of primate pluripotent stem cells, these cells could potentially be differentiated into any desired functional cells and consistently cultured in large-scale conditions. In the current study, oligodendrocyte progenitor cells and mature oligodendrocytes were generated within three months from monkey embryonic stem cells. The embryonic stem cell-derived oligodendrocytes exhibited in vitro myelinogenic potency with rat dorsal root ganglion neurons. Additionally, the transplanted oligodendrocyte progenitor cells differentiated into myelin basic protein-positive mature oligodendrocytes in the mouse corpus callosum. This preparative method was used for human induced pluripotent stem cells, which were also successfully differentiated into oligodendrocyte progenitor cells and mature oligodendrocytes that were capable of myelinating rat dorsal root ganglion neurons. Moreover, it was possible to freeze, thaw, and successfully re-culture the differentiating cells. These results showed that embryonic stem cells and human induced pluripotent stem cells maintained in a

  3. Evolution of generic supporting conditions for feeders of 500 M We PHWR

    International Nuclear Information System (INIS)

    Prasad, K.H.; Gupta, K.N.; Bapat, C.N.; Sharma, V.K.; Mishra, R.; Soni, R.S.; Kushwaha, H.S.; Mahajan, S.C.; Kakodkar, A.

    1995-01-01

    Feeder piping consists of 784 number of closely spaced, inter-connected flexible pipes catering for large thermal and creep movements besides seismic and other loadings. The analytical strategy for evolving generic supporting conditions is outlined in this paper. (author). 4 refs., 7 figs

  4. CATTLE FEEDER BEHAVIOR AND FEEDER CATTLE PLACEMENTS

    OpenAIRE

    Kastens, Terry L.; Schroeder, Ted C.

    1994-01-01

    Cattle feeders appear irrational when they place cattle on feed when projected profit is negative. Long futures positions appear to offer superior returns to cattle feeding investment. Cattle feeder behavior suggests that they believe a downward bias in live cattle futures persists and that cattle feeders use different expectations than the live cattle futures market price when making placement decisions. This study examines feeder cattle placement determinants, comparing performance of expec...

  5. Comparative Proteomic Analysis of Supportive and Unsupportive Extracellular Matrix Substrates for Human Embryonic Stem Cell Maintenance*

    Science.gov (United States)

    Soteriou, Despina; Iskender, Banu; Byron, Adam; Humphries, Jonathan D.; Borg-Bartolo, Simon; Haddock, Marie-Claire; Baxter, Melissa A.; Knight, David; Humphries, Martin J.; Kimber, Susan J.

    2013-01-01

    Human embryonic stem cells (hESCs) are pluripotent cells that have indefinite replicative potential and the ability to differentiate into derivatives of all three germ layers. hESCs are conventionally grown on mitotically inactivated mouse embryonic fibroblasts (MEFs) or feeder cells of human origin. In addition, feeder-free culture systems can be used to support hESCs, in which the adhesive substrate plays a key role in the regulation of stem cell self-renewal or differentiation. Extracellular matrix (ECM) components define the microenvironment of the niche for many types of stem cells, but their role in the maintenance of hESCs remains poorly understood. We used a proteomic approach to characterize in detail the composition and interaction networks of ECMs that support the growth of self-renewing hESCs. Whereas many ECM components were produced by supportive and unsupportive MEF and human placental stromal fibroblast feeder cells, some proteins were only expressed in supportive ECM, suggestive of a role in the maintenance of pluripotency. We show that identified candidate molecules can support attachment and self-renewal of hESCs alone (fibrillin-1) or in combination with fibronectin (perlecan, fibulin-2), in the absence of feeder cells. Together, these data highlight the importance of specific ECM interactions in the regulation of hESC phenotype and provide a resource for future studies of hESC self-renewal. PMID:23658023

  6. A single-cell and feeder-free culture system for monkey embryonic stem cells.

    Science.gov (United States)

    Ono, Takashi; Suzuki, Yutaka; Kato, Yosuke; Fujita, Risako; Araki, Toshihiro; Yamashita, Tomoko; Kato, Hidemasa; Torii, Ryuzo; Sato, Naoya

    2014-01-01

    Primate pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), hold great potential for research and application in regenerative medicine and drug discovery. To maximize primate PSC potential, a practical system is required for generating desired functional cells and reproducible differentiation techniques. Much progress regarding their culture systems has been reported to date; however, better methods would still be required for their practical use, particularly in industrial and clinical fields. Here we report a new single-cell and feeder-free culture system for primate PSCs, the key feature of which is an originally formulated serum-free medium containing FGF and activin. In this culture system, cynomolgus monkey ESCs can be passaged many times by single-cell dissociation with traditional trypsin treatment and can be propagated with a high proliferation rate as a monolayer without any feeder cells; further, typical PSC properties and genomic stability can be retained. In addition, it has been demonstrated that monkey ESCs maintained in the culture system can be used for various experiments such as in vitro differentiation and gene manipulation. Thus, compared with the conventional culture system, monkey ESCs grown in the aforementioned culture system can serve as a cell source with the following practical advantages: simple, stable, and easy cell maintenance; gene manipulation; cryopreservation; and desired differentiation. We propose that this culture system can serve as a reliable platform to prepare primate PSCs useful for future research and application.

  7. Decision Support for Optimal Repositioning of Containers in a Feeder System

    Directory of Open Access Journals (Sweden)

    Danijela Tuljak-Suban

    2008-03-01

    Full Text Available The transport of empty containers represents a seriousproblem in the fast growing sphere of maritime container transport.The most widespread type of container transport organizationin maritime transport is the hub and spoke mode, whichenables the transport of a great number of containers via largevessels between hub ports, from where feeder ships transport tosmaller ports that thus gravitate to the central hub port. The articlecontains a detailed analysis of the northern Adriatic portsand the feeder connections with the hub ports of the Mediterranean.A two-level VRPPD (Vehicle Routing Problem withPickup and Delivery problem is modelled on a graph, wherethe transport of full containers is privileged over the transport ofempty containers. This enables the simulation of the feeder systemin the n01them Adriatic, meaning that it shows the ship'soperator the movement programme with minimal transportcosts for the superfluous empty containers in the complex of theregular transports of full containers in the feeder system.

  8. Risk-informed prediction of feeder end of life

    International Nuclear Information System (INIS)

    Jyrkama, M.; Pandey, M.

    2011-01-01

    The operating life of feeder piping is negatively impacted by flow accelerated corrosion (FAC). In this study, an assessment of a large set of inspection data reveals that FAC in feeders is a relatively stationary process, with variability only at the local scale. Given the added uncertainty from inspection coverage, a new method for estimating the thinning rate and feeder EOL is developed using a probabilistic approach. The results of the study illustrate the benefits of the methodology in supporting risk-informed decision making at the station by quantifying the present and incremental risk in the feeder system over time. (author)

  9. Risk-informed prediction of feeder end of life

    Energy Technology Data Exchange (ETDEWEB)

    Jyrkama, M.; Pandey, M. [Univ. of Waterloo, Ontario (Canada)

    2011-07-01

    The operating life of feeder piping is negatively impacted by flow accelerated corrosion (FAC). In this study, an assessment of a large set of inspection data reveals that FAC in feeders is a relatively stationary process, with variability only at the local scale. Given the added uncertainty from inspection coverage, a new method for estimating the thinning rate and feeder EOL is developed using a probabilistic approach. The results of the study illustrate the benefits of the methodology in supporting risk-informed decision making at the station by quantifying the present and incremental risk in the feeder system over time. (author)

  10. Robust and highly-efficient differentiation of functional monocytic cells from human pluripotent stem cells under serum- and feeder cell-free conditions.

    Directory of Open Access Journals (Sweden)

    Masakatsu D Yanagimachi

    Full Text Available Monocytic lineage cells (monocytes, macrophages and dendritic cells play important roles in immune responses and are involved in various pathological conditions. The development of monocytic cells from human embryonic stem cells (ESCs and induced pluripotent stem cells (iPSCs is of particular interest because it provides an unlimited cell source for clinical application and basic research on disease pathology. Although the methods for monocytic cell differentiation from ESCs/iPSCs using embryonic body or feeder co-culture systems have already been established, these methods depend on the use of xenogeneic materials and, therefore, have a relatively poor-reproducibility. Here, we established a robust and highly-efficient method to differentiate functional monocytic cells from ESCs/iPSCs under serum- and feeder cell-free conditions. This method produced 1.3 × 10(6 ± 0.3 × 10(6 floating monocytes from approximately 30 clusters of ESCs/iPSCs 5-6 times per course of differentiation. Such monocytes could be differentiated into functional macrophages and dendritic cells. This method should be useful for regenerative medicine, disease-specific iPSC studies and drug discovery.

  11. Keratinocytes propagated in serum-free, feeder-free culture conditions fail to form stratified epidermis in a reconstituted skin model.

    Directory of Open Access Journals (Sweden)

    Rebecca Lamb

    Full Text Available Primary human epidermal stem cells isolated from skin tissues and subsequently expanded in tissue culture are used for human therapeutic use to reconstitute skin on patients and to generate artificial skin in culture for academic and commercial research. Classically, epidermal cells, known as keratinocytes, required fibroblast feeder support and serum-containing media for serial propagation. In alignment with global efforts to remove potential animal contaminants, many serum-free, feeder-free culture methods have been developed that support derivation and growth of these cells in 2-dimensional culture. Here we show that keratinocytes grown continually in serum-free and feeder-free conditions were unable to form into a stratified, mature epidermis in a skin equivalent model. This is not due to loss of cell potential as keratinocytes propagated in serum-free, feeder-free conditions retain their ability to form stratified epidermis when re-introduced to classic serum-containing media. Extracellular calcium supplementation failed to improve epidermis development. In contrast, the addition of serum to commercial, growth media developed for serum-free expansion of keratinocytes facilitated 3-dimensional stratification in our skin equivalent model. Moreover, the addition of heat-inactivated serum improved the epidermis structure and thickness, suggesting that serum contains factors that both aid and inhibit stratification.

  12. Human amniotic epithelial cell feeder layers maintain mouse embryonic stem cell pluripotency via epigenetic regulation of the c-Myc promoter.

    Science.gov (United States)

    Liu, Te; Cheng, Weiwei; Liu, Tianjin; Guo, Lihe; Huang, Qin; Jiang, Lizhen; Du, Xiling; Xu, Fuhui; Liu, Zhixue; Lai, Dongmei

    2010-02-01

    Mouse embryonic stem cells (ESCs) are typically cultured on a feeder layer of mouse embryonic fibroblasts (MEFs), with leukemia inhibitory factor (LIF) added to maintain them in an undifferentiated state. We have previously shown that human amniotic epithelial cells (hAECs) can be used as feeder cells to maintain mouse ESC pluripotency, but the mechanism for this is unknown. In the present study, we found that CpG islands 5' of the c-Myc gene remain hypomethylated in mouse ESCs cultured on hAECs. In addition, levels of acetylation of histone H3 and trimethylation of histone H3K4 in the c-Myc gene promoter were higher in ES cells cultured on hAECs than those in ES cells cultured on MEFs. These data suggested that hAECs can alter mouse ESC gene expression via epigenetic modification of c-Myc, providing a possible mechanism for the hAEC-induced maintenance of ESCs in an undifferentiated state.

  13. Human amniotic epithelial cell feeder layers maintain human iPS cell pluripotency via inhibited endogenous microRNA-145 and increased Sox2 expression

    International Nuclear Information System (INIS)

    Liu, Te; Cheng, Weiwei; Huang, Yongyi; Huang, Qin; Jiang, Lizhen; Guo, Lihe

    2012-01-01

    Currently, human induced pluripotent stem (iPS) cells were generated from patient or disease-specific sources and share the same key properties as embryonic stem cells. This makes them attractive for personalized medicine, drug screens or cellular therapy. Long-term cultivation and maintenance of normal iPS cells in an undifferentiated self-renewing state are a major challenge. Our previous studies have shown that human amniotic epithelial cells (HuAECs) could provide a good source of feeder cells for mouse and human embryonic stem cells, or spermatogonial stem cells, but the mechanism for this is unknown. Here, we examined the effect of endogenous microRNA-145 regulation on Sox2 expression in human iPS cells by HuAECs feeder cells regulation, and in turn on human iPS cells pluripotency. We found that human IPS cells transfected with a microRNA-145 mutant expressed Sox2 at high levels, allowing iPS to maintain a high level of AP activity in long-term culture and form teratomas in SCID mice. Expression of stem cell markers was increased in iPS transfected with the microRNA-145 mutant, compared with iPS was transfected with microRNA-145. Besides, the expression of Drosha proteins of the microRNA-processor complex, required for the generation of precursor pre-miRNA, was significantly increased in human iPS cells cultured on MEF but not on HuAECs. Taken together, these results suggest that endogenous Sox2 expression may be regulated by microRNA-145 in human iPS cells with HuAECs feeder cells, and Sox2 is a crucial component required for maintenance of them in an undifferentiated, proliferative state capable of self-renewal. Highlights: ► microRNA-145 inhibits Sox2 expression in human iPS cells. ► microRNA-145 suppresses the self-renewal and pluripotency of human iPS cells. ► HuAECs regulate expression of microRNA-145 and Sox2 in human iPS cells. ► HuAECs feeder layers maintain human iPS cells pluripotency. ► HuAECs negatively regulates the synthesis of

  14. Human amniotic epithelial cell feeder layers maintain human iPS cell pluripotency via inhibited endogenous microRNA-145 and increased Sox2 expression

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Te, E-mail: liute79@yahoo.com [School of Environmental Science and Engineering, Donghua University, Shanghai 201620 (China); Shanghai Geriatric Institute of Chinese Medicine, Shanghai 200031 (China); Cheng, Weiwei [International Peace Maternity and Child Health Hospital, Shanghai Jiaotong University, Shanghai 200030 (China); Huang, Yongyi [Laboratoire PROTEE, Batiment R, Universite du Sud Toulon-Var, 83957 LA GARDE Cedex (France); Huang, Qin; Jiang, Lizhen [Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China); Guo, Lihe, E-mail: liute79@yahoo.com [Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China)

    2012-02-15

    Currently, human induced pluripotent stem (iPS) cells were generated from patient or disease-specific sources and share the same key properties as embryonic stem cells. This makes them attractive for personalized medicine, drug screens or cellular therapy. Long-term cultivation and maintenance of normal iPS cells in an undifferentiated self-renewing state are a major challenge. Our previous studies have shown that human amniotic epithelial cells (HuAECs) could provide a good source of feeder cells for mouse and human embryonic stem cells, or spermatogonial stem cells, but the mechanism for this is unknown. Here, we examined the effect of endogenous microRNA-145 regulation on Sox2 expression in human iPS cells by HuAECs feeder cells regulation, and in turn on human iPS cells pluripotency. We found that human IPS cells transfected with a microRNA-145 mutant expressed Sox2 at high levels, allowing iPS to maintain a high level of AP activity in long-term culture and form teratomas in SCID mice. Expression of stem cell markers was increased in iPS transfected with the microRNA-145 mutant, compared with iPS was transfected with microRNA-145. Besides, the expression of Drosha proteins of the microRNA-processor complex, required for the generation of precursor pre-miRNA, was significantly increased in human iPS cells cultured on MEF but not on HuAECs. Taken together, these results suggest that endogenous Sox2 expression may be regulated by microRNA-145 in human iPS cells with HuAECs feeder cells, and Sox2 is a crucial component required for maintenance of them in an undifferentiated, proliferative state capable of self-renewal. Highlights: Black-Right-Pointing-Pointer microRNA-145 inhibits Sox2 expression in human iPS cells. Black-Right-Pointing-Pointer microRNA-145 suppresses the self-renewal and pluripotency of human iPS cells. Black-Right-Pointing-Pointer HuAECs regulate expression of microRNA-145 and Sox2 in human iPS cells. Black-Right-Pointing-Pointer HuAECs feeder

  15. Feeder replacement tooling and processes

    International Nuclear Information System (INIS)

    Mallozzi, R.; Goslin, R.; Pink, D.; Askari, A.

    2008-01-01

    Primary heat transport system feeder integrity has become a concern at some CANDU nuclear plants as a result of thinning caused by flow accelerated corrosion (FAC). Feeder inspections are indicating that life-limiting wall thinning can occur in the region between the Grayloc hub weld and second elbow of some outlet feeders. In some cases it has become necessary to replace thinned sections of affected feeders to restore feeder integrity to planned end of life. Atomic Energy of Canada Limited (AECL) and Babcock and Wilcox Canada Ltd. (B and W) have developed a new capability for replacement of single feeders at any location on the reactor face without impacting or interrupting operation of neighbouring feeders. This new capability consists of deploying trained crews with specialized tools and procedures for feeder replacements during planned outages. As may be expected, performing single feeder replacement in the congested working environment of an operational CANDU reactor face involves overcoming many challenges with respect to access to feeders, available clearances for tooling, and tooling operation and performance. This paper describes some of the challenges encountered during single feeder replacements and actions being taken by AECL and B and W to promote continuous improvement of feeder replacement tooling and processes and ensure well-executed outages. (author)

  16. Growth factor expression pattern of homologous feeder layer for culturing buffalo embryonic stem cell-like cells.

    Science.gov (United States)

    Sharma, Ruchi; George, Aman; Kamble, Nitin M; Chauhan, Manmohan S; Singla, Suresh; Manik, Radhey S; Palta, Prabhat

    2012-01-01

    The present study examined the expression profile of buffalo fetal fibroblasts (BFF) used as a feeder layer for embryonic stem (ES) cell-like cells. The expression of important growth factors was detected in cells at different passages. Mitomycin-C inactivation increased relative expression levels of ACTIVIN-A, TGF-β1, BMP-4 and GREMLIN but not of fibroblast growth factor-2 (FGF-2). The expression level of ACTIVIN-A, transforming growth factor-β1 (TGF-β1), bone morphogenetic protein-4 (BMP-4) and FGF-2 was similar in buffalo fetal fibroblast (BFF) cultured in stem cell medium (SCM), SCM+1000IU mL(-1) leukemia inhibitory factor (LIF), SCM+5 ngmL(-1) FGF-2 or SCM+LIF+FGF-2 for 24 h whereas GREMLIN expression was higher in FGF-2-supplemented groups. In spent medium, the concentration of ACTIVIN-A was higher in FGF-2-supplemented groups whereas that of TGF-β1 was similar in SCM and LIF+FGF-2, which was higher than when either LIF or FGF-2 was used alone. Following culture of ES cell-like cells on a feeder layer for 24 h, the TGF-β1 concentration was higher with LIF+FGF-2 than with LIF or FGF-2 alone which, in turn, was higher than that in SCM. In the LIF+FGF-2 group, the concentration of TGF-β1 was lower and that of ACTIVIN-A was higher in spent medium at 24 h than at 48 h of culture. These results suggest that BFF produce signalling molecules that may help in self-renewal of buffalo ES cell-like cells.

  17. Advanced feeder-free generation of induced pluripotent stem cells directly from blood cells.

    Science.gov (United States)

    Trokovic, Ras; Weltner, Jere; Nishimura, Ken; Ohtaka, Manami; Nakanishi, Mahito; Salomaa, Veikko; Jalanko, Anu; Otonkoski, Timo; Kyttälä, Aija

    2014-12-01

    Generation of validated human induced pluripotent stem cells (iPSCs) for biobanking is essential for exploring the full potential of iPSCs in disease modeling and drug discovery. Peripheral blood mononuclear cells (PBMCs) are attractive targets for reprogramming, because blood is collected by a routine clinical procedure and is a commonly stored material in biobanks. Generation of iPSCs from blood cells has previously been reported using integrative retroviruses, episomal Sendai viruses, and DNA plasmids. However, most of the published protocols require expansion and/or activation of a specific cell population from PBMCs. We have recently collected a PBMC cohort from the Finnish population containing more than 2,000 subjects. Here we report efficient generation of iPSCs directly from PBMCs in feeder-free conditions in approximately 2 weeks. The produced iPSC clones are pluripotent and transgene-free. Together, these properties make this novel method a powerful tool for large-scale reprogramming of PBMCs and for iPSC biobanking. ©AlphaMed Press.

  18. A Decentralized Storage Strategy for Residential Feeders with Photovoltaics

    DEFF Research Database (Denmark)

    Marra, Francesco; Yang, Guangya; Træholt, Chresten

    2014-01-01

    . The power sizing of the ESSs is performed with linear programming (LP) method, based on voltage sensitivity analysis. A Belgian residential LV feeder with private PV systems is used as a case study to demonstrate the effectiveness of the proposed strategy. Quantification of the required energy levels...... domestic energy storage systems (ESS). The traditional way of operating a domestic ESS to increase the local consumption rate does not take into account the need of voltage support in a feeder; the proposed storage concept improves the traditional one, by mitigating voltage rise due to PV in the feeder...

  19. SAFIRE - a robotic inspection system for CANDU feeders

    Energy Technology Data Exchange (ETDEWEB)

    Buckingham, R. [OC Robotics, Bristol (United Kingdom)

    2011-07-01

    The condition of primary circuit feeder pipes in CANDU reactors is relevant to the commercial viability and plant life. One known wear mechanism is external fretting between feeder pipes and adjacent services or support structures, particularly within the Upper Feeder Cabinet (UFC). Fretting leads to wall thinning which must not exceed certain agreed limits. Chafe shields have been added to protect the feeder pipes. Regular inspections are required of the chafe shields, feeder pipes and other structures that may cause feeder damage. Historically, the dose received by inspectors conducting this work has been significant. For this reason Ontario Power Generation has invested in a remotely operated robot system to conduct visual inspections within the UFC. This system, called SAFIRE for 'Snake-Arm Feeder Inspection Robot Equipment' has been deployed at Pickering during 2010 and 2011 and has been used to inspect areas that are extremely difficult to inspect with existing manual techniques. The 2011 scope of work included inspection of a total of 660 feeder pipes in three UFC quadrants, in two reactors. The full scope was completed over a one-month period in Autumn 2011 in which SAFIRE was used during 23, twelve hour shifts. This included two periods each of 72 hours of continuous operation using multiple teams of operators. SAFIRE is remote controlled delivery system for multiple cameras to record still images and video. The main system elements include a snake-arm robot mounted on a mobile vehicle. It can be controlled from up to 500m away using a fibre/copper connection. The snake-arm is 2.2m long, 25mm wide and has 18 degrees of freedom. It is designed to snake between the rows of feeder pipes to inspect feeder/hanger interfaces, both above and below the feeder cabinet catwalks. Future upgrades offer the potential to add additional tools to increase functionality. This paper describes the SAFIRE development process from inception to operational experience

  20. SAFIRE - a robotic inspection system for CANDU feeders

    International Nuclear Information System (INIS)

    Buckingham, R.

    2011-01-01

    The condition of primary circuit feeder pipes in CANDU reactors is relevant to the commercial viability and plant life. One known wear mechanism is external fretting between feeder pipes and adjacent services or support structures, particularly within the Upper Feeder Cabinet (UFC). Fretting leads to wall thinning which must not exceed certain agreed limits. Chafe shields have been added to protect the feeder pipes. Regular inspections are required of the chafe shields, feeder pipes and other structures that may cause feeder damage. Historically, the dose received by inspectors conducting this work has been significant. For this reason Ontario Power Generation has invested in a remotely operated robot system to conduct visual inspections within the UFC. This system, called SAFIRE for 'Snake-Arm Feeder Inspection Robot Equipment' has been deployed at Pickering during 2010 and 2011 and has been used to inspect areas that are extremely difficult to inspect with existing manual techniques. The 2011 scope of work included inspection of a total of 660 feeder pipes in three UFC quadrants, in two reactors. The full scope was completed over a one-month period in Autumn 2011 in which SAFIRE was used during 23, twelve hour shifts. This included two periods each of 72 hours of continuous operation using multiple teams of operators. SAFIRE is remote controlled delivery system for multiple cameras to record still images and video. The main system elements include a snake-arm robot mounted on a mobile vehicle. It can be controlled from up to 500m away using a fibre/copper connection. The snake-arm is 2.2m long, 25mm wide and has 18 degrees of freedom. It is designed to snake between the rows of feeder pipes to inspect feeder/hanger interfaces, both above and below the feeder cabinet catwalks. Future upgrades offer the potential to add additional tools to increase functionality. This paper describes the SAFIRE development process from inception to operational experience gained

  1. Power Hardware-in-the-Loop Evaluation of PV Inverter Grid Support on Hawaiian Electric Feeders: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Nelson, Austin; Prabakar, Kumaraguru; Nagarajan, Adarsh; Nepal, Shaili; Hoke, Anderson; Asano, Marc; Ueda, Reid; Ifuku, Earle

    2017-05-08

    As more grid-connected photovoltaic (PV) inverters become compliant with evolving interconnections requirements, there is increased interest from utilities in understanding how to best deploy advanced grid-support functions (GSF) in the field. One efficient and cost-effective method to examine such deployment options is to leverage power hardware-in-the-loop (PHIL) testing methods. Two Hawaiian Electric feeder models were converted to real-time models in the OPAL-RT real-time digital testing platform, and integrated with models of GSF capable PV inverters that were modeled from characterization test data. The integrated model was subsequently used in PHIL testing to evaluate the effects of different fixed power factor and volt-watt control settings on voltage regulation of the selected feeders. The results of this study were provided as inputs for field deployment and technical interconnection requirements for grid-connected PV inverters on the Hawaiian Islands.

  2. Urbanization affects neophilia and risk-taking at bird-feeders

    Science.gov (United States)

    Tryjanowski, Piotr; Møller, Anders Pape; Morelli, Federico; Biaduń, Waldemar; Brauze, Tomasz; Ciach, Michał; Czechowski, Paweł; Czyż, Stanisław; Dulisz, Beata; Goławski, Artur; Hetmański, Tomasz; Indykiewicz, Piotr; Mitrus, Cezary; Myczko, Łukasz; Nowakowski, Jacek J.; Polakowski, Michał; Takacs, Viktoria; Wysocki, Dariusz; Zduniak, Piotr

    2016-06-01

    Urban environments cover vast areas with a high density of humans and their dogs and cats causing problems for exploitation of new resources by wild animals. Such resources facilitate colonization by individuals with a high level of neophilia predicting that urban animals should show more neophilia than rural conspecifics. We provided bird-feeders across urban environments in 14 Polish cities and matched nearby rural habitats, testing whether the presence of a novel item (a brightly coloured green object made out of gum with a tuft of hair) differentially delayed arrival at feeders in rural compared to urban habitats. The presence of a novel object reduced the number of great tits Parus major, but also the total number of all species of birds although differentially so in urban compared to rural areas. That was the case independent of the potentially confounding effects of temperature, population density of birds, and the abundance of cats, dogs and pedestrians. The number of great tits and the total number of birds attending feeders increased in urban compared to rural areas independent of local population density of birds. This implies that urban birds have high levels of neophilia allowing them to readily exploit unpredictable resources in urban environments.

  3. Power Hardware-in-the-Loop Evaluation of PV Inverter Grid Support on Hawaiian Electric Feeders

    Energy Technology Data Exchange (ETDEWEB)

    Nelson, Austin A [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Prabakar, Kumaraguru [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Nagarajan, Adarsh [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Nepal, Shaili [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Hoke, Anderson F [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Asano, Marc [Hawaiian Electric Company; Ueda, Reid [Hawaiian Electric Company; Ifuku, Earle [Hawaiian Electric Company

    2017-10-03

    As more grid-connected photovoltaic (PV) inverters become compliant with evolving interconnections requirements, there is increased interest from utilities in understanding how to best deploy advanced grid-support functions (GSF) in the field. One efficient and cost-effective method to examine such deployment options is to leverage power hardware-in-the-loop (PHIL) testing methods, which combine the fidelity of hardware tests with the flexibility of computer simulation. This paper summarizes a study wherein two Hawaiian Electric feeder models were converted to real-time models using an OPAL-RT real-time digital testing platform, and integrated with models of GSF capable PV inverters based on characterization test data. The integrated model was subsequently used in PHIL testing to evaluate the effects of different fixed power factor and volt-watt control settings on voltage regulation of the selected feeders using physical inverters. Selected results are presented in this paper, and complete results of this study were provided as inputs for field deployment and technical interconnection requirements for grid-connected PV inverters on the Hawaiian Islands.

  4. CD34+ Testicular Stromal Cells Support Long-Term Expansion of Embryonic and Adult Stem and Progenitor Cells

    Science.gov (United States)

    Kim, Jiyeon; Seandel, Marco; Falciatori, Ilaria; Wen, Duancheng; Rafii, Shahin

    2010-01-01

    Stem cells reside in specialized microenvironments created by supporting stromal cells that orchestrate self-renewal and lineage-specific differentiation. However, the precise identity of the cellular and molecular pathways that support self-renewal of stem cells is not known. For example, long-term culture of prototypical stem cells, such as adult spermatogonial stem and progenitor cells (SPCs), in vitro has been impeded by the lack of an optimal stromal cell line that initiates and sustains proliferation of these cells. Indeed, current methods, including the use of mouse embryonic fibroblasts (MEFs), have not been efficient and have generally led to inconsistent results. Here, we report the establishment of a novel CD34-positive cell line, referred to as JK1, derived from mouse testicular stromal cells that not only facilitated long-term SPC culture but also allowed faithful generation of SPCs and multipotent stem cells. SPCs generated on JK1 maintained key features of germ line stem cells, including expression of PLZF, DAZL, and GCNA. Furthermore, these feeders also promoted the long-term cultivation of other types of primitive cells including multi-potent adult spermatogonial-derived stem cells, pluripotent murine embryonic stem cells, and embryonic germ cells derived from primordial germ cells. Stem cells could be passaged serially and still maintained expression of characteristic markers such as OCT4 and NANOG in vitro, as well as the ability to generate all three germ layers in vivo. These results indicate that the JK1 cell line is capable of promoting long-term culture of primitive cells. As such, this cell line allows for identification of stromal-derived factors that support long-term proliferation of various types of stem cells and constitutes a convenient alternative to other types of feeder layers. PMID:18669907

  5. Human fetal liver stromal cells that overexpress bFGF support growth and maintenance of human embryonic stem cells.

    Directory of Open Access Journals (Sweden)

    Jiafei Xi

    Full Text Available In guiding hES cell technology toward the clinic, one key issue to be addressed is to culture and maintain hES cells much more safely and economically in large scale. In order to avoid using mouse embryonic fibroblasts (MEFs we isolated human fetal liver stromal cells (hFLSCs from 14 weeks human fetal liver as new human feeder cells. hFLSCs feeders could maintain hES cells for 15 passages (about 100 days. Basic fibroblast growth factor (bFGF is known to play an important role in promoting self-renewal of human embryonic stem (hES cells. So, we established transgenic hFLSCs that stably express bFGF by lentiviral vectors. These transgenic human feeder cells--bFGF-hFLSCs maintained the properties of H9 hES cells without supplementing with any exogenous growth factors. H9 hES cells culturing under these conditions maintained all hES cell features after prolonged culture, including the developmental potential to differentiate into representative tissues of all three embryonic germ layers, unlimited and undifferentiated proliferative ability, and maintenance of normal karyotype. Our results demonstrated that bFGF-hFLSCs feeder cells were central to establishing the signaling network among bFGF, insulin-like growth factor 2 (IGF-2, and transforming growth factor β (TGF-β, thereby providing the framework in which hES cells were instructed to self-renew or to differentiate. We also found that the conditioned medium of bFGF-hFLSCs could maintain the H9 hES cells under feeder-free conditions without supplementing with bFGF. Taken together, bFGF-hFLSCs had great potential as feeders for maintaining pluripotent hES cell lines more safely and economically.

  6. Establishment of rat embryonic stem-like cells from the morula using a combination of feeder layers.

    Science.gov (United States)

    Sano, Chiaki; Matsumoto, Asako; Sato, Eimei; Fukui, Emiko; Yoshizawa, Midori; Matsumoto, Hiromichi

    2009-08-01

    Embryonic stem (ES) cells are characterized by pluripotency, in particular the ability to form a germline on injection into blastocysts. Despite numerous attempts, ES cell lines derived from rat embryos have not yet been established. The reason for this is unclear, although certain intrinsic biological differences among species and/or strains have been reported. Herein, using Wistar-Imamichi rats, specific characteristics of preimplantation embryos are described. At the blastocyst stage, Oct4 (also called Pou5f1) was expressed in both the inner cell mass (ICM) and the trophectoderm (TE), whereas expression of Cdx2 was localized to the TE. In contrast, at an earlier stage, expression of Oct4 was detected in all the nuclei in the morula. These stages were examined using a combination of feeder layers (rat embryonic fibroblast [REF] for primary outgrowth and SIM mouse embryo-derived thioguanine- and ouabain-resistant [STO] cells for passaging) to establish rat ES-like cell lines. The rat ES-like cell lines obtained from the morula maintained expression of Oct4 over long-term culture, whereas cell lines derived from blastocysts lost pluripotency during early passage. The morula-derived ES-like cell lines showed Oct4 expression in a long-term culture, even after cryogenic preservation, thawing and EGFP transfection. These results indicate that rat ES-like cell lines with long-term Oct4 expression can be established from the morula of Wistar-Imamichi rats using a combination of feeder layers.

  7. The ROCK inhibitor Y-27632 improves recovery of human embryonic stem cells after fluorescence-activated cell sorting with multiple cell surface markers.

    Directory of Open Access Journals (Sweden)

    Nil Emre

    Full Text Available BACKGROUND: Due to the inherent sensitivity of human embryonic stem cells (hESCs to manipulations, the recovery and survival of hESCs after fluorescence-activated cell sorting (FACS can be low. Additionally, a well characterized and robust methodology for performing FACS on hESCs using multiple-cell surface markers has not been described. The p160-Rho-associated coiled kinase (ROCK inhibitor, Y-27632, previously has been identified as enhancing survival of hESCs upon single-cell dissociation, as well as enhancing recovery from cryopreservation. Here we examined the application of Y-27632 to hESCs after FACS to improve survival in both feeder-dependent and feeder-independent growth conditions. METHODOLOGY/PRINCIPAL FINDINGS: HESCs were sorted using markers for SSEA-3, TRA-1-81, and SSEA-1. Cells were plated after sorting for 24 hours in either the presence or the absence of Y-27632. In both feeder-dependent and feeder-independent conditions, cell survival was greater when Y-27632 was applied to the hESCs after sort. Specifically, treatment of cells with Y-27632 improved post-sort recovery up to four fold. To determine the long-term effects of sorting with and without the application of Y-27632, hESCs were further analyzed. Specifically, hESCs sorted with and without the addition of Y-27632 retained normal morphology, expressed hESC-specific markers as measured by immunocytochemistry and flow cytometry, and maintained a stable karyotype. In addition, the hESCs could differentiate into three germ layers in vitro and in vivo in both feeder-dependent and feeder-independent growth conditions. CONCLUSIONS/SIGNIFICANCE: The application of Y-27632 to hESCs after cell sorting improves cell recovery with no observed effect on pluripotency, and enables the consistent recovery of hESCs by FACS using multiple surface markers. This improved methodology for cell sorting of hESCs will aid many applications such as removal of hESCs from secondary cell types

  8. Seismic analysis of ITER fourth PF (Poloidal Field Coil) feeder

    International Nuclear Information System (INIS)

    Liu, Sumei; Chen, Wei; Song, Yuntao; Ni, Xiaojun; Wang, Zhongwei; Chen, Yonghua; Gong, Chenyu

    2014-01-01

    The ITER feeder systems connect the ITER magnet systems located inside the main cryostat to the cryo-plant, power-supply and control system interfaces outside the cryostat. The main purpose of the feeders is to convey the cryogenic supply and electrical power to the coils as well as house the instrumentation wiring. The PF busbar which carries 52 kA current will suffer from high Lorentz force due to the background magnetic field inspired by the coils and the self-field between every pair of busbars. Except their mechanical strength and thermal insulation performance must be achieved, the dynamic mechanism on PF structure should be assessed. This paper presents the simulation and seismic analysis on ITER 4th PF feeder including the Coil Terminal Box and S-bend Box (CTB and SBB), the Cryostat Feed-through (CFT), the In-Cryostat-Feeder (ICF), especially for the ground supports and main outer-tube firstly. This analysis aims to study seismic resistance on system design under local seismograms with floor response spectrum, the structural response vibration mode and response duration results of displacement, membrane stress, and bending stress on structure under different directions actuating signals were obtained by using the single-seismic spectrum analysis and Dead Weight analysis respectively. Based on the simulative and analytical results, the system seismic resistance and the integrity of the support structure in the 4th PF feeder have been studied and the detail design confirmed

  9. Improvement of Local Voltage in Feeders with Photovoltaic using Electric Vehicles

    DEFF Research Database (Denmark)

    Marra, Francesco; Yang, Guangya; Fawzy, Y. T.

    2013-01-01

    In low-voltage (LV) feeders with high penetration of photovoltaic (PV), a major issue to be solved is voltage rise due to the active power injection. If no measures are taken, this may lead to generation’s interruptions and to the malfunctioning of domestic appliances due to non-standard voltage...... profiles. This paper proposes a storage strategy to alleviate voltage rise in feeders with PV, using coordinated electric vehicle (EV) load as the storage solution. The voltage support strategy is easy to implement practically and it is demonstrated on a test feeder emulating a household with roof...

  10. Feederism in a woman.

    Science.gov (United States)

    Terry, Lesley L; Vasey, Paul L

    2011-06-01

    Feederism is a fat fetish subculture in which individuals eroticize weight gain and feeding. Feeders are individuals who claim to become sexually aroused by feeding their partners and encouraging them to gain weight. Conversely, Feedees are individuals who claim to become sexually aroused by eating, being fed, and the idea or act of gaining weight. Very little is known about this population. This report describes a woman who self-identified as a Feedee. It is unclear, at present, whether female Feederism represents a unique paraphilia or a thematic variation of morphophilia or sexual masochism.

  11. Network Reduction Algorithm for Developing Distribution Feeders for Real-Time Simulators: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Nagarajan, Adarsh; Nelson, Austin; Prabakar, Kumaraguru; Hoke, Andy; Asano, Marc; Ueda, Reid; Nepal, Shaili

    2017-06-15

    As advanced grid-support functions (AGF) become more widely used in grid-connected photovoltaic (PV) inverters, utilities are increasingly interested in their impacts when implemented in the field. These effects can be understood by modeling feeders in real-time systems and testing PV inverters using power hardware-in-the-loop (PHIL) techniques. This paper presents a novel feeder model reduction algorithm using a Monte Carlo method that enables large feeders to be solved and operated on real-time computing platforms. Two Hawaiian Electric feeder models in Synergi Electric's load flow software were converted to reduced order models in OpenDSS, and subsequently implemented in the OPAL-RT real-time digital testing platform. Smart PV inverters were added to the real-time model with AGF responses modeled after characterizing commercially available hardware inverters. Finally, hardware inverters were tested in conjunction with the real-time model using PHIL techniques so that the effects of AGFs on the choice feeders could be analyzed.

  12. Design of apron feeders

    Energy Technology Data Exchange (ETDEWEB)

    Ramos, C.M.

    1987-12-01

    This paper discusses practical aspects of apron feeder design, and includes information on how to make the first selection. Various aspects of design such as feeder speed, material depth, chain-pitch, roller loading, wheels/sprockets and chain roller friction are elaborated in detail. 8 refs., 14 figs., 12 tabs.

  13. The development and manufacture of size for size feeder pipe for feeder replacement

    International Nuclear Information System (INIS)

    Legate, G.; Schreiter, D.; Townley, N.

    2008-01-01

    The recently recognised problem of feeder pipe thinning created a unique sourcing problem. Operators require relatively small quantities of nuclear class 1 seamless feeder pipe for such replacement which prior to the introduction of this product in 2006 was not available. It was desired that the pipe be produced at the exact size of the pipe currently in use at the specific reactor site (feeder pipe size varies from site to site). Secondly the pipe had to be made in conformance to the original code year of issue and to conform to the intent of the original material specifications. Finally a supply strategy had to be implemented allowing for timely manufacture of replacement piping. This presentation will report upon how replacement size for size feeder tube was developed and is currently manufactured at Nu-Tech Precision Metals. The paper will also detail the current supply strategy to ensure timely manufacture of the product.

  14. Cultivation and irradiation of human fibroblasts in a medium enriched with platelet lysate for obtaining feeder layer in epidermal cell culture

    International Nuclear Information System (INIS)

    Yoshito, Daniele

    2011-01-01

    For over 30 years, the use of culture medium, enriched with bovine serum, and murines fibroblasts, with the rate of proliferation controlled by irradiation or by share anticarcinogenic drugs, has been playing successfully its role in assisting in the development of keratinocytes in culture, for clinical purposes. However, currently there is a growing concern about the possibility of transmitting prions and animals viruses to transplanted patients. Taking into account this concern, the present work aims to cultivate human fibroblasts in a medium enriched with human platelets lysate and determine the irradiation dose of these cells, for obtaining feeder layer in epidermal cell culture. For carrying out the proposed objective, platelets lysis has standardized, this lysate was used for human fibroblasts cultivation and the irradiation dose enough to inhibit its duplication was evaluated. Human keratinocytes were cultivated in these feeder layers, in culture medium enriched with the lysate. With these results we conclude that the 10% platelets lysate promoted a better adhesion and proliferation of human fibroblasts and in all dose levels tested (60 to 300 Gy), these had their mitotic activity inactivated by ionizing irradiation, being that the feeder layers obtained with doses from 70 to 150 Gy were those that provided the best development of keratinocytes in medium containing 2.5% of human platelet lysate. Therefore, it was possible to standardize both the cultivation of human fibroblasts as its inactivation for use as feeder layer in culture of keratinocytes, so as to eliminate xenobiotics components. (author)

  15. Immunoflourescence and mRNA analysis of human embryonic stem cells (hESCs) grown under feeder-free conditions

    DEFF Research Database (Denmark)

    Awan, Aashir; Oliveri, Roberto S; Jensen, Pernille L

    2010-01-01

    onto 16-well glass chambers, and continuing with the general IF and qPCR steps will be provided. The techniques will be illustrated with new results on cellular localization of transcriptional factors and components of the Hedgehog, Wnt, and PDGF signaling pathways to primary cilia in stem cell......This chapter describes the procedures in order to do immunofluorescence (IF) microscopy and quantitative PCR (qPCR) analysis of human embryonic stem cells (hESCs) grown specifically under feeder-free conditions. A detailed protocol outlining the steps from initially growing the cells, passaging...

  16. A bend thickness sensitivity study of Candu feeder piping

    International Nuclear Information System (INIS)

    Li, M.; Aggarwal, M.L.; Meysner, A.; Micelotta, C.

    2005-01-01

    In CANDU reactors, feeder bends close to the connection at the fuel channel may be subjected to the highest Flow Accelerated Corrosion (FAC) and stresses. Feeder pipe stress analysis is crucial in the life extension of aging CANDU plants. Typical feeder pipes are interconnected by upper link plates and spacers. It is well known that the stresses at the bends are sensitive to the local bend thicknesses. It is also known from the authors' study (Li and et al, 2005) that feeder inter linkage effect is significant and cannot be ignored. The field measurement of feeder bend thickness is difficult and may be subjected to uncertainty in accuracy. Hence, it is desirable to know how the stress on a subject feeder could be affected by the bend thickness variation of the neighboring feeders. This effect cannot be evaluated by the traditional 'single' feeder model approach. In this paper, the 'row' and 'combined' models developed in the previous study (Li and et al, 2005), which include the feeder interactions, are used to investigate the sensitivity of bend thickness. A series of random thickness bounded by maximum and minimum measured values were applied to feeders in the model. The results show that an individual feeder is not sensitive to the bend thickness variation of the remaining feeders in the model, but depends primarily on its own bend thickness. The highest stress at a feeder always occurs when the feeder has the smallest possible bend thickness. A minimum acceptable bend thickness for individual feeders can be computed by an iterative computing process. The dependency of field thickness measurement and the amount of required analysis work can be greatly reduced. (authors)

  17. A reliable and economical method for gaining mouse embryonic fibroblasts capable of preparing feeder layers.

    Science.gov (United States)

    Jiang, Guangming; Wan, Xiaoju; Wang, Ming; Zhou, Jianhua; Pan, Jian; Wang, Baolong

    2016-08-01

    Mouse embryonic fibroblasts (MEFs) are widely used to prepare feeder layers for culturing embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) in vitro. Transportation lesions and exorbitant prices make the commercially obtained MEFs unsuitable for long term research. The aim of present study is to establish a method, which enables researchers to gain MEFs from mice and establish feeder layers by themselves in ordinary laboratories. MEFs were isolated from ICR mouse embryos at 12.5-17.5 day post-coitum (DPC) and cultured in vitro. At P2-P7, the cells were inactivated with mitomycin C or by X-ray irradiation. Then they were used to prepare feeder layers. The key factors of the whole protocol were analyzed to determine the optimal conditions for the method. The results revealed MEFs isolated at 12.5-13.5 DPC, and cultured to P3 were the best choice for feeder preparation, those P2 and P4-P5 MEFs were also suitable for the purpose. The P3-P5 MEFs treated with 10 μg/ml of mitomycin C for 3 h, or irradiated with X-ray at 1.5 Gy/min for 25 Gy were the most suitable feeder cells. Treating MEFs with 10 μg/ml of mitomycin C for 2.5 h, 15 μg/ml for 2.0 h, or irradiating the cells with 20 Gy of X-ray at 2.0 Gy/min could all serve as alternative methods for P3-P4 cells. Our study provides a reliable and economical way to obtain large amount of qualified MEFs for long term research of ESCs or iPSCs.

  18. Differentiation of monkey embryonic stem cells to hepatocytes by feeder-free dispersion culture and expression analyses of cytochrome p450 enzymes responsible for drug metabolism.

    Science.gov (United States)

    Maruyama, Junya; Matsunaga, Tamihide; Yamaori, Satoshi; Sakamoto, Sakae; Kamada, Noboru; Nakamura, Katsunori; Kikuchi, Shinji; Ohmori, Shigeru

    2013-01-01

    We reported previously that monkey embryonic stem cells (ESCs) were differentiated into hepatocytes by formation of embryoid bodies (EBs). However, this EB formation method is not always efficient for assays using a large number of samples simultaneously. A dispersion culture system, one of the differentiation methods without EB formation, is able to more efficiently provide a large number of feeder-free undifferentiated cells. A previous study demonstrated the effectiveness of the Rho-associated kinase inhibitor Y-27632 for feeder-free dispersion culture and induction of differentiation of monkey ESCs into neural cells. In the present study, the induction of differentiation of cynomolgus monkey ESCs (cmESCs) into hepatocytes was performed by the dispersion culture method, and the expression and drug inducibility of cytochrome P450 (CYP) enzymes in these hepatocytes were examined. The cmESCs were successfully differentiated into hepatocytes under feeder-free dispersion culture conditions supplemented with Y-27632. The hepatocytes differentiated from cmESCs expressed the mRNAs for three hepatocyte marker genes (α-fetoprotein, albumin, CYP7A1) and several CYP enzymes, as measured by real-time polymerase chain reaction. In particular, the basal expression of cmCYP3A4 (3A8) in these hepatocytes was detected at mRNA and enzyme activity (testosterone 6β-hydroxylation) levels. Furthermore, the expression and activity of cmCYP3A4 (3A8) were significantly upregulated by rifampicin. These results indicated the effectiveness of Y-27632 supplementation for feeder-free dispersed culture and induction of differentiation into hepatocytes, and the expression of functional CYP enzyme(s) in cmESC-derived hepatic cells.

  19. A study of inter linkage effects on Candu feeder piping

    International Nuclear Information System (INIS)

    Li, M.; Aggarwal, M.L.; Meysner, A.

    2005-01-01

    A CANDU (Canadian Deuterium Uranium) reactor core consists of a large number of fuel channels where heat is generated. Two feeder pipes are connected to each fuel channel to transport D 2 O coolant into and out of the reactor core. The feeder piping is designed to the requirements of Class 1 piping of Section III NB of the ASME Boiler and Pressure Vessel and CSA Codes. Feeder piping stress analysis is being performed to demonstrate the code compliance check and the fitness for service of feeders. In the past, stress analyses were conducted for each individual feeder without including interaction effects among connected feeders. Interaction effects occur as a result of linkages that exist between feeders to prevent fretting and impacting damage during normal, abnormal and accident conditions. In this paper, a 'combined' approach is adopted to include all feeders connected by inter linkages into one feeder piping model. MSC/NASTRAN finite element software was used in the stress simulation, which contains up to 127 feeder pipes. The ASME Class 1 piping analysis was conducted to investigate the effects of the linkages between feeders. Both seismic time history and broadened response spectra methods were used in the seismic stress calculation. The results show that the effect of linkages is significant in dynamic stresses for all feeder configurations, as well as in static stresses for certain feeder configurations. The single feeder analysis could either underestimate or overestimate feeder stresses depending on the pipe geometry and bend wall thickness. (authors)

  20. Development of dry coal feeders

    Science.gov (United States)

    Bonin, J. H.; Cantey, D. E.; Daniel, A. D., Jr.; Meyer, J. W.

    1977-01-01

    Design and fabrication of equipment of feed coal into pressurized environments were investigated. Concepts were selected based on feeder system performance and economic projections. These systems include: two approaches using rotating components, a gas or steam driven ejector, and a modified standpipe feeder concept. Results of development testing of critical components, design procedures, and performance prediction techniques are reviewed.

  1. EV Charging Facilities and Their Application in LV Feeders with Photovoltaics

    DEFF Research Database (Denmark)

    Marra, Francesco; Yang, Guangya; Træholt, Chresten

    2013-01-01

    Low-voltage (LV) grid feeders with high penetration of photovoltaics (PVs) are often affected by voltage magnitude problems. To solve such issues, previous research has shown that reactive power methods, active power curtailment and grid reinforcement can be used for voltage support, yet showing...... several limits. We introduce the use of electric vehicle (EV) public charging stations with energy storage system (ESS) as a solution for voltage regulation in LV feeders with PV. A novel method is proposed to determine the ESS charging load required for voltage regulation and compare the results...... for the different locations in the feeder. With time-series simulations, we quantify the energy size required for a station ESS. A Belgian LV residential grid, modeled using real PV generation and load profiles, is used as case study. The method and simulation results show the effectiveness of using public EV...

  2. Economic feeder for recharging and ``topping off''

    Science.gov (United States)

    Fickett, Bryan; Mihalik, G.

    2000-04-01

    Increasing the size of the melt charge significantly increases yield and reduces costs. Siemens Solar Industries is optimizing a method to charge additional material after meltdown (top-off) using an external feeder system. A prototype feeder system was fabricated consisting of a hopper and feed delivery system. The low-cost feeder is designed for simple operation and maintenance. The system is capable of introducing up to 60 kg of granular silicon while under vacuum. An isolation valve permits refilling of the hopper while maintaining vacuum in the growth furnace. Using the feeder system in conjunction with Siemens Solar Industries' energy efficient hot zone dramatically reduces power and argon consumption. Throughput is also improved as faster pull speeds can be attained. The increased pull speeds have an even greater impact when the charge size is increased. Further cost reduction can be achieved by refilling the crucible after crystal growth and pulling a second ingot run. Siemens Solar Industries is presently testing the feeder in production.

  3. A feeder-free, human plasma-derived hydrogel for maintenance of a human embryonic stem cell phenotype in vitro

    Directory of Open Access Journals (Sweden)

    Lewis Fiona C

    2012-08-01

    Full Text Available Abstract Background Human embryonic stem cells (hESCs represent a tremendous resource for cell therapies and the study of human development; however to maintain their undifferentiated state in vitro they routinely require the use of mouse embryonic fibroblast (MEF feeder-layers and exogenous protein media supplementation. Results These well established requirements can be overcome and in this study, it will be demonstrated that phenotypic stability of hESCs can be maintained using a novel, human plasma protein-based hydrogel as an extracellular culture matrix without the use of feeder cell co-culture. hESCs were resuspended in human platelet poor plasma (PPP, which was gelled by the addition of calcium containing DMEM-based hESC culture medium. Phenotypic and genomic expression of the pluripotency markers OCT4, NANOG and SOX2 were measured using immunohistochemistry and qRT-PCR respectively. Typical hESC morphology was demonstrated throughout in vitro culture and both viability and phenotypic stability were maintained throughout extended culture, up to 25 passages. Conclusions PPP-derived hydrogel has demonstrated to be an efficacious alternative to MEF co-culture with its hydrophilicity allowing for this substrate to be delivered via minimally invasive procedures in a liquid phase with polymerization ensuing in situ. Together this provides a novel technique for the study of this unique group of stem cells in either 2D or 3D both in vitro and in vivo.

  4. Gut-associated lymphoid tissue contains the molecular machinery to support T-cell-dependent and T-cell-independent class switch recombination.

    Science.gov (United States)

    Barone, F; Patel, P; Sanderson, J D; Spencer, J

    2009-11-01

    A PRoliferation-Inducing Ligand (APRIL) is a secreted cytokine member of the tumor necrosis factor family. It is a B-cell survival factor that also induces class switch recombination (CSR) toward immunoglobulin A (IgA), independent of T cells. It is therefore an important contributor to the maintenance of the mucosal immunological barrier, which has been linked to a putative extrafollicular inductive phase of the IgA response in lamina propria. By immunohistochemistry (IHC) and quantitative real-time PCR (qRT-PCR) on microdissected tissue from normal human gut, we observed APRIL expression, together with TACI (transmembrane activator and CAML interactor) and BCMA (B-cell maturation antigen), in gut-associated lymphoid tissue (GALT), lamina propria, and in the epithelium of stomach, small and large intestine, and rectum. However, no activation-induced cytidine deaminase (AID) expression (an absolute requirement for class switching) was detected in lamina propria by IHC or qRT-PCR. APRIL and its receptors were only observed alongside AID in GALT, showing that GALT contains the apparatus to support both T-independent and T-dependent routes to IgA CSR.

  5. An RFID Based Smart Feeder for Hummingbirds.

    Science.gov (United States)

    Ibarra, Vicente; Araya-Salas, Marcelo; Tang, Yu-ping; Park, Charlie; Hyde, Anthony; Wright, Timothy F; Tang, Wei

    2015-12-16

    We present an interdisciplinary effort to record feeding behaviors and control the diet of a hummingbird species (Phaethornis longirostris, the long-billed hermit or LBH) by developing a Radio Frequency Identification (RFID) based smart feeder. The system contains an RFID reader, a microcontroller, and a servo-controlled hummingbird feeder opener; the system is presented as a tool for studying the cognitive ability of the LBH species. When equipped with glass capsule RFID tags (which are mounted on the hummingbird), the smart feeder can provide specific diets for predetermined sets of hummingbirds at the discretion of biologists. This is done by reading the unique RFID tag on the hummingbirds and comparing the ID number with the pre-programmed ID numbers stored in the smart feeder. The smart feeder records the time and ID of each hummingbird visit. The system data is stored in a readily available SD card and is powered by two 9 V batteries. The detection range of the system is approximately 9-11 cm. Using this system, biologists can assign the wild hummingbirds to different experimental groups and monitor their diets to determine if they develop a preference to any of the available nectars. During field testing, the smart feeder system has demonstrated consistent detection (when compared to detections observed by video-recordings) of RFID tags on hummingbirds and provides pre-designed nectars varying water and sugar concentrations to target individuals. The smart feeder can be applied to other biological and environmental studies in the future.

  6. An RFID Based Smart Feeder for Hummingbirds

    Directory of Open Access Journals (Sweden)

    Vicente Ibarra

    2015-12-01

    Full Text Available We present an interdisciplinary effort to record feeding behaviors and control the diet of a hummingbird species (Phaethornis longirostris, the long-billed hermit or LBH by developing a Radio Frequency Identification (RFID based smart feeder. The system contains an RFID reader, a microcontroller, and a servo-controlled hummingbird feeder opener; the system is presented as a tool for studying the cognitive ability of the LBH species. When equipped with glass capsule RFID tags (which are mounted on the hummingbird, the smart feeder can provide specific diets for predetermined sets of hummingbirds at the discretion of biologists. This is done by reading the unique RFID tag on the hummingbirds and comparing the ID number with the pre-programmed ID numbers stored in the smart feeder. The smart feeder records the time and ID of each hummingbird visit. The system data is stored in a readily available SD card and is powered by two 9 V batteries. The detection range of the system is approximately 9–11 cm. Using this system, biologists can assign the wild hummingbirds to different experimental groups and monitor their diets to determine if they develop a preference to any of the available nectars. During field testing, the smart feeder system has demonstrated consistent detection (when compared to detections observed by video-recordings of RFID tags on hummingbirds and provides pre-designed nectars varying water and sugar concentrations to target individuals. The smart feeder can be applied to other biological and environmental studies in the future.

  7. Comparing the engineering program feeders from SiF and convention models

    Science.gov (United States)

    Roongruangsri, Warawaran; Moonpa, Niwat; Vuthijumnonk, Janyawat; Sangsuwan, Kampanart

    2018-01-01

    This research aims to compare the relationship between two types of engineering program feeder models within the technical education systems of Rajamangala University of Technology Lanna (RMUTL), Chiangmai, Thailand. To illustrate, the paper refers to two typologies of feeder models, which are the convention and the school in factory (SiF) models. The new SiF model is developed through a collaborative educational process between the sectors of industry, government and academia, using work-integrated learning. The research methodology were use to compared features of the the SiF model with conventional models in terms of learning outcome, funding budget for the study, the advantages and disadvantages from the point of view of students, professors, the university, government and industrial partners. The results of this research indicate that the developed SiF feeder model is the most pertinent ones as it meet the requirements of the university, the government and the industry. The SiF feeder model showed the ability to yield positive learning outcomes with low expenditures per student for both the family and the university. In parallel, the sharing of knowledge between university and industry became increasingly important in the process, which resulted in the improvement of industrial skills for professors and an increase in industrial based research for the university. The SiF feeder model meets its demand of public policy in supporting a skilled workforce for the industry, which could be an effective tool for the triple helix educational model of Thailand.

  8. Expression and secretory profile of buffalo fetal fibroblasts and Wharton's jelly feeder layers.

    Science.gov (United States)

    Parmar, Mehtab S; Mishra, Smruti Ranjan; Somal, Anjali; Pandey, Sriti; Kumar, G Sai; Sarkar, Mihir; Chandra, Vikash; Sharma, G Taru

    2017-05-01

    The present study examined the comparative expression and secretory profile of vital signaling molecules in buffalo fetal fibroblasts (BFF) and Wharton's jelly (BWJ) feeder layers at different passages. Both feeder layers were expanded up to 8th passage. Signaling molecules viz. bone morphogenetic protein 4 (BMP4), fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF) and transforming growth factor beta 1 (TGFB1) and pluripotency-associated transcriptional factors (POU5F1, SOX2, NANOG, KLF4, MYC and FOXD3) were immunolocalized in the both feeder types. A clear variation in the expression pattern of key signaling molecules with passaging was registered in both feeders compared to primary culture (0 passage). The conditioned media (CM) was collected from different passages (2, 4, 6, 8) of both the feeder layers and was quantified using enzyme-linked immunosorbent assay (ELISA). Concomitant to expression profile, protein quantification also revealed differences in the concentration of signaling molecules at different time points. Conjointly, expression and secretory profile revealed that 2nd passage of BFF and 6th passage of BWJ exhibit optimal levels of key signaling molecules thus may be selected as best passages for embryonic stem cells (ESCs) propagation. Further, the effect of mitomycin-C (MMC) treatment on the expression profile of signaling molecules in the selected passages of BFF and BWJ revealed that MMC modulates the expression profile of these molecules. In conclusion, the results indicate that feeder layers vary in expression and secretory pattern of vital signaling molecules with passaging. Based on these findings, the appropriate feeder passages may be selected for the quality propagation of buffalo ESCs. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Expectations of Cattle Feeding Investors in Feeder Cattle Placements

    OpenAIRE

    Kastens, Terry L.; Schroeder, Ted C.

    1993-01-01

    Cattle feeders appear irrational when they place cattle on feed when projected profits are negative. Long futures positions appear to offer superior returns to cattle feeding investment. Cattle feeder behavior suggests that they believe a downward bias in live cattle futures persists and that cattle feeders use different information than the live cattle futures market price when making placement decisions. This paper examines feeder cattle placement determinants and compares performance of ex...

  10. Feeder density enhances house finch disease transmission in experimental epidemics.

    Science.gov (United States)

    Moyers, Sahnzi C; Adelman, James S; Farine, Damien R; Thomason, Courtney A; Hawley, Dana M

    2018-05-05

    Anthropogenic food provisioning of wildlife can alter the frequency of contacts among hosts and between hosts and environmental sources of pathogens. Despite the popularity of garden bird feeding, few studies have addressed how feeders influence host contact rates and disease dynamics. We experimentally manipulated feeder density in replicate aviaries containing captive, pathogen-naive, groups of house finches ( Haemorhous mexicanus ) and continuously tracked behaviours at feeders using radio-frequency identification devices. We then inoculated one bird per group with Mycoplasma gallisepticum (Mg), a common bacterial pathogen for which feeders are fomites of transmission, and assessed effects of feeder density on house finch behaviour and pathogen transmission. We found that pathogen transmission was significantly higher in groups with the highest density of bird feeders, despite a significantly lower rate of intraspecific aggressive interactions relative to the low feeder density groups. Conversely, among naive group members that never showed signs of disease, we saw significantly higher concentrations of Mg-specific antibodies in low feeder density groups, suggesting that birds in low feeder density treatments had exposure to subclinical doses of Mg. We discuss ways in which the density of garden bird feeders could play an important role in mediating the intensity of Mg epidemics.This article is part of the theme issue 'Anthropogenic resource subsidies and host-parasite dynamics in wildlife'. © 2018 The Author(s).

  11. Colour preferences of UK garden birds at supplementary seed feeders.

    Directory of Open Access Journals (Sweden)

    Luke Rothery

    Full Text Available Supplementary feeding of garden birds generally has benefits for both bird populations and human wellbeing. Birds have excellent colour vision, and show preferences for food items of particular colours, but research into colour preferences associated with artificial feeders is limited to hummingbirds. Here, we investigated the colour preferences of common UK garden birds foraging at seed-dispensing artificial feeders containing identical food. We presented birds simultaneously with an array of eight differently coloured feeders, and recorded the number of visits made to each colour over 370 30-minute observation periods in the winter of 2014/15. In addition, we surveyed visitors to a garden centre and science festival to determine the colour preferences of likely purchasers of seed feeders. Our results suggest that silver and green feeders were visited by higher numbers of individuals of several common garden bird species, while red and yellow feeders received fewer visits. In contrast, people preferred red, yellow, blue and green feeders. We suggest that green feeders may be simultaneously marketable and attractive to foraging birds.

  12. Round-bale feeder design affects hay waste and economics during horse feeding.

    Science.gov (United States)

    Martinson, K; Wilson, J; Cleary, K; Lazarus, W; Thomas, W; Hathaway, M

    2012-03-01

    Many horse owners find round bales convenient, less labor intensive, and more affordable than other hay types, but report an inability to control horse BW gain and excessive hay waste. The objectives were to compare hay waste, hay intake, and payback of 9 round-bale feeders and a no-feeder control when used during horse feeding. Nine round-bale feeders were tested: Cinch Net, Cone, Covered Cradle, Hayhut, Hay Sleigh, Ring, Tombstone, Tombstone Saver, and Waste Less. Each feeder design was placed on the ground in a dirt paddock. Five groups of 5 horses were fed in rotation for a 4-d period with each feeder. Every fourth day, groups were rotated among paddocks and a new round bale was placed in each feeder. In the 5 paddocks used, 5 feeders were installed for d 1 through 20, and the remaining 4 feeders and no-feeder control were installed for d 21 through 40. Groups of horses were sequentially assigned to feeders using two 5 × 5 Latin squares, the first for d 1 through 20, the second for d 21 through 40. Horse groups of similar age, BW, breed, and sex were formed from 25 Quarter Horse and Thoroughbred geldings and open mares (means: 11 yr; 541 kg of BW). Hay on the ground surrounding the feeder was collected daily, dried, and weighed. The total amount of hay removed around each feeder for a 4-d period was considered waste. Dry matter intake was estimated as the difference between hay disappearance and waste. Number of months for the reduction in waste to repay feeder cost (payback) were calculated using hay valued at $110/t, and improved feeder efficiency over the control. Feeder design did not affect hay intake (P > 0.05); all feeders resulted in an estimated hay intake of 2.0 to 2.4% BW; the no-feeder control resulted in a reduced intake of 1.3% BW (P = 0.001). Mean percentage of hay waste differed among feeders (P feeder control, 57%. Feeder design also affected payback (P feeder design affected hay waste and payback, but not estimated hay intake or BW change

  13. Impact of flow accelerated corrosion (FAC) on feeder refurbishment planning

    International Nuclear Information System (INIS)

    Jyrkama, M.; Pandey, M.

    2010-01-01

    Feeder wall thinning due to flow accelerated corrosion (FAC) may result in a large number of feeder replacements in the future. In this study, the process of FAC is modelled using a probabilistic approach and used to predict the expected number of degraded feeders and their replacements in the future. Because of the high cost associated with feeder replacements, it may be optimal to replace the entire feeder population during a single refurbishment outage when the unit cost of replacement is likely to be less. The results of this study demonstrate, however, that the unit cost of feeder replacement must be sufficiently lower than the standard replacement cost and the refurbishment performed at an optimal time to realize the economic benefits associated with the refurbishment. (author)

  14. Identification of pectenotoxins in plankton, filter feeders, and isolated cells of a Dinophysis acuminata with an atypical toxin profile, from Chile.

    Science.gov (United States)

    Blanco, Juan; Alvarez, Gonzalo; Uribe, Eduardo

    2007-04-01

    A bloom of Dinophysis acuminata produced, in autumn of 2005, a closure of the scallop harvesting in Bahía Inglesa, in the Chilean III region. Isolated cells of this Dinophysis species were shown to contain 180 pg cell(-1) of pectenotoxin 2 but neither okadaic acid nor any of its analogs or derivatives (at least at a detectable level). Examination of plankton and filter-feeder samples covering an area of ca. 350 km, from the location where the toxicity was recorded to Bahía Tongoy, showed that the unique toxin profile found in the first bloom was widespread over that part of Chile and persisted for months. The analysis were carried out by HPLC-ESI-MS using positive ionization mode, with a detection limit below 2 ng of OA mL(-1) of methanolic extract. This is the first report of the presence of pectenotoxins in the plankton of the Pacific coast of America and in the studied filter feeders. This is also the first report of a Dinophysis species containing pectenotoxins and not any toxin of the okadaic acid group.

  15. in meat production III. Feeder - breeder dimorphism

    African Journals Online (AJOL)

    Feeder- breeder dimorphism is advantageous when large offspring for slaughter is obtained from small breeding animals. The effect of feeder- breeder dimorphism on herd efficiency is evaluated for terminal crossbreeding and growth modification by biotechnological or dietary means. Selection criteria for breeds or lines in ...

  16. Development of ceramic-free antenna feeder

    International Nuclear Information System (INIS)

    Moriyama, S.; Kimura, H.; Fujii, T.; Saigusa, M.; Arai, H.

    1994-01-01

    We have proposed a ceramics-free antenna feeder line employing a ridged waveguide as a local support for IC antenna of next-generation tokamaks. One fourth mock-up model of the all metal waveguide designed for the ITER ICRF system is fabricated and electrical characteristics of the model including the coaxial line - waveguide converter are measured. Power reflection coefficient of the model including the coax-waveguide converter to the input coaxial line is estimated to be less than 15% below the cut-off frequency of 107 MHz and less than 3% above the cut-off frequency. It is found that this ceramics-free antenna support employing a ridged waveguide is quite available for IC antenna of next-generation tokamaks. (author)

  17. Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System.

    Science.gov (United States)

    Nandal, Anjali; Mallon, Barbara; Telugu, Bhanu P

    2017-11-08

    Embryonic and induced pluripotent stem cells can self-renew and differentiate into multiple cell types of the body. The pluripotent cells are thus coveted for research in regenerative medicine and are currently in clinical trials for eye diseases, diabetes, heart diseases, and other disorders. The potential to differentiate into specialized cell types coupled with the recent advances in genome editing technologies including the CRISPR/Cas system have provided additional opportunities for tailoring the genome of iPSC for varied applications including disease modeling, gene therapy, and biasing pathways of differentiation, to name a few. Among the available editing technologies, the CRISPR/Cas9 from Streptococcus pyogenes has emerged as a tool of choice for site-specific editing of the eukaryotic genome. The CRISPRs are easily accessible, inexpensive, and highly efficient in engineering targeted edits. The system requires a Cas9 nuclease and a guide sequence (20-mer) specific to the genomic target abutting a 3-nucleotide "NGG" protospacer-adjacent-motif (PAM) for targeting Cas9 to the desired genomic locus, alongside a universal Cas9 binding tracer RNA (together called single guide RNA or sgRNA). Here we present a step-by-step protocol for efficient generation of feeder-independent and footprint-free iPSC and describe methodologies for genome editing of iPSC using the Cas9 ribonucleoprotein (RNP) complexes. The genome editing protocol is effective and can be easily multiplexed by pre-complexing sgRNAs for more than one target with the Cas9 protein and simultaneously delivering into the cells. Finally, we describe a simplified approach for identification and characterization of iPSCs with desired edits. Taken together, the outlined strategies are expected to streamline generation and editing of iPSC for manifold applications.

  18. Leaky feeder: the communication backbone

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-01-01

    The need to communicate with all areas of the underground operation in monitoring the movement of men, materials and vehicles, as well as the optimum performance of conveyor systems, has been effectively met with the installation of a Flexcom leaky feeder cable network at a coliery in Mpumalanga. Installed by the South African subsidiary of Mine Radio Systems (MRS), based in Canada, Flexcom is an RF communications highway for underground mines. The system can provide up to 32 voice/data control channels and up to 16 video channels, all operating simultaneously. The system uses a series of bi-directional amplifiers (or signal boosters) spaced at 350 m intervals along the leaky feeder cable, with branching units and termination units added as required. Communication is possible within 50 m of the leaky feeder cable. MRS has 11 conveyors monitored via the SCADA program at the mine and the system produces reports as required which are accessible via cellphone from anywhere in the world. The wireless monitoring of miners and equipment contributes to mine safety. 3 figs.

  19. Determination of representative CANDU feeder dimensions for engineering simulator

    International Nuclear Information System (INIS)

    Cho, S.; Muzumdar, A.

    1996-01-01

    This paper describes a logic for selection of representative channel groups and a methodology for determination of representative CANDU feeder dimensions and the pressure drops between inlet/outlet header and fuel channel in the primary loop. A code, MEDOC, was developed based on this logic and methodology and helps perform a calculation of representative feeder dimensions for a selected channel group on the basis of feeder geometry data (fluid volume, mass flow rate, loss factor) and given property data (pressure, quality, density) at inlet/outlet header. The representative feeder dimensions calculated based on this methodology will be useful for the engineering simulator for the CANDU type reactor. (author)

  20. Assessment of CANDU feeders subject to flow accelerated corrosion

    Energy Technology Data Exchange (ETDEWEB)

    Iyer, S. [Atomic Energy of Canada Ltd., Mississauga, Ontario (Canada); Slade, J.P. [New Brunswick Power, Point Lepreau Generating Station, Lepreau, New Brunswick (Canada)

    2003-07-01

    'Full Text:' Inspections of CANDU feeders have indicated greater than expected wall thinning of outlet feeders. This wall thinning is attributed to Flow-Accelerated Corrosion (FAC). The rate of wall loss due to FAC is highest in the close radius bends near the end fitting. The minimum allowed thickness for a feeder pipes is based on design pressure during the design stage. Extended operation of the thinned feeders beyond their design basis, i.e., operation of feeders with thickness below design pressure based minimum thickness has economic benefits for the utilities. In such cases, it is important to establish the remaining life and evaluate the adequacy of the components for safe operation. ASME Code Case N-597 provides the guidelines for acceptance for continued service of Classes 2 and 3 piping components experiencing wall thinning during operation. However, for Class 1 systems, the Code Case recommends that the owner develop the methodology and criteria for the assessment of wall thinning. Therefore, under the CANDU Owner's Group's (COG) Feeder Integrity Joint Program (FIJP), the 'Fitness for Service Guidelines (FFSG) for Feeders Affected by Wall Thinning in Operating CANDU Reactors' was developed and subsequently conditionally approved by CNSC. This paper illustrates the underlying concepts in the FFSG methodology and its benefits to utilities. Specific examples of the application and benefits of the FFSG at Point Lepreau G.S. are described in this paper. The assessment of feeders is based on the requirements of the construction Code (Section III of the ASME Boiler and Pressure Vessel Code): The following points briefly describe the assessment methodology. Satisfying the requirements of NB-3650 for design and service loadings are sufficient for continued service and extended life if the predicted minimum wall thickness of the component is greater than or equal to 90% of the pressure based thickness calculated as per NB-3641. The B

  1. Feeding Against Gravity with Spot Feeders in High Silicon Ductile Iron

    DEFF Research Database (Denmark)

    Vedel-Smith, Nikolaj Kjelgaard; Tiedje, Niels Skat

    2014-01-01

    and classified using X-ray imaging and ultrasound analysis. The effect of the different feeder configurations were classified in reference to defect location, sleeve material, and feeder type, modulus, and location. The analysis showed that exothermal feeder sleeves with the right configurations can feed up......-hill against gravity. This effect may contribute to the thermal expansion created by the exothermal reaction. It was also found that the optimum feeder size does not scale linearly with the casting modulus but that larger casting modulus requires relatively smaller modulus feeders. The thermal gradient created...

  2. 26 CFR 1.502-1 - Feeder organizations.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Feeder organizations. 1.502-1 Section 1.502-1...) INCOME TAXES (CONTINUED) Exempt Organizations § 1.502-1 Feeder organizations. (a) In the case of an organization operated for the primary purpose of carrying on a trade or business for profit, exemption is not...

  3. Novel Methods to Determine Feeder Locational PV Hosting Capacity and PV Impact Signatures

    Energy Technology Data Exchange (ETDEWEB)

    Reno, Matthew J. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Coogan, Kyle [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Seuss, John [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Broderick, Robert Joseph [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2017-05-01

    Often PV hosting capacity analysis is performed for a limited number of distribution feeders. For medium - voltage distribution feeders, previous results generally analyze less than 20 feeders, and then the results are extrapolated out to similar types of feeders. Previous hosting capacity research has often focused on determining a single value for the hosting capacity for the entire feeder, whereas this research expands previous hosting capacity work to investigate all the regions of the feeder that may allow many different hosting capacity values wit h an idea called locational hosting capacity (LHC)to determine the largest PV size that can be interconnected at different locations (buses) on the study feeders. This report discusses novel methods for analyzing PV interconnections with advanced simulati on methods. The focus is feeder and location - specific impacts of PV that determine the locational PV hosting capacity. Feeder PV impact signature are used to more precisely determine the local maximum hosting capacity of individual areas of the feeder. T he feeder signature provides improved interconnection screening with certain zones that show the risk of impact to the distribution feeder from PV interconnections.

  4. A Dynamic and Heuristic Phase Balancing Method for LV Feeders

    Directory of Open Access Journals (Sweden)

    Samad Taghipour Boroujeni

    2016-01-01

    Full Text Available Due to the single-phase loads and their stochastic behavior, the current in the distribution feeders is not balanced. In addition, the single-phase loads are located in different positions along the LV feeders. So the amount of the unbalanced load and its location affect the feeder losses. An unbalanced load causes the feeder losses and the voltage drop. Because of time-varying behavior of the single-phase loads, phase balancing is a dynamic and combinatorial problem. In this research, a heuristic and dynamic solution for the phase balancing of the LV feeders is proposed. In this method, it is supposed that the loads’ tie could be connected to all phases through a three-phase switch. The aim of the proposed method is to make the feeder conditions as balanced as possible. The amount and the location of single-phase loads are considered in the proposed phase balancing method. Since the proposed method needs no communication interface or no remote controller, it is inexpensive, simple, practical, and robust. Applying this method provides a distributed and dynamic phase balancing control. In addition, the feasibility of reducing the used switches is investigated. The ability of the proposed method in the phase balancing of the LV feeders is approved by carrying out some simulations.

  5. Economic Contribution of Regional Feeder Ports to The Local Economy In Indonesia

    Directory of Open Access Journals (Sweden)

    Ibnu Syabri

    2017-07-01

    Full Text Available Indonesia regional feeder ports’ main function is to serve marine transport activities between regency/city in a province in limited quantities. The intention of the government to make marine transportation as the backbone of national logistic transportation system puts these ports to an important position, as suppliers for the main and/or collector ports and as goods distribution centers for the region from the bigger ports. Not only as a provider of accessibility between regions, regional feeder ports should be seen as an investation to boost regional economy. This study aims to explain the influences of regional feeder ports on regency/city economy, based on the port types: (1 cargo ports, and (2 cargo & passengers ports. Results showed that both types of ports have positive influences on the regency/city economy, although it is not related to region’s economic base sector. Cargo ports which are mainly located on west area of Indonesia, supports the region’s economy more than cargo & passenger ports which are mainly located on the east.

  6. Structural integrity investigations of feeder pipe ice plugging procedures

    International Nuclear Information System (INIS)

    Flaman, M.T.; Shah, N.N.

    1985-03-01

    A procedure involving the use of a liquid nitrogen cooled heat exchanger to form internal ice plugs in feeder pipes is routinely used in nuclear generating stations. The use of this procedure has caused concerns with regard to the safety of station maintenance personnel, and in regard to the integrity of the feeder pipes. This report describes the results of laboratory stress and pressure measurements which were performed on a feeder pipe section during ice plugging operations to investigate these concerns. From the results of this study, and from the results of previous studies of material behaviour at low temperatures, it has been determined that the ice plugging procedure can be performed on feeder pipes in a safe and effective manner

  7. Operation of a cell feeder for coal controlled by a frequency converter. [Czechoslovakia

    Energy Technology Data Exchange (ETDEWEB)

    Stonawski, J; Sikora, G

    1984-02-01

    Steam boiler of the Trinec coal-fired power plant with a capacity of 125 t/h, 545 C, 9.5 MPa was equipped with a screw coal feeder with the KT26A-6 Winter-Eichberg motor with a transformer. The screw fee

  8. Derivation and characterization of the NIH registry human stem cell line NYSCF100 line under defined feeder-free conditions

    Directory of Open Access Journals (Sweden)

    Ana Sevilla

    2018-05-01

    Full Text Available The human embryonic stem cell line NYSCFe001-A was derived from a day 6 blastocyst in feeder-free and antibiotic free conditions. The blastocyst was voluntarily donated for research as surplus after in vitro fertilization treatment following informed consent. The NYSCFe001-A line, registered as NYSCF100 on the NIH registry, presents normal karyotype, is mycoplasma free, expresses all the pluripotency markers and has the potential to differentiate into all three germ layers in vitro.

  9. Vertical-Screw-Auger Conveyer Feeder

    Science.gov (United States)

    Walton, Otis (Inventor); Vollmer, Hubert J. (Inventor)

    2016-01-01

    A conical feeder is attached to a vertically conveying screw auger. The feeder is equipped with scoops and rotated from the surface to force-feed regolith the auger. Additional scoops are possible by adding a cylindrical section above the conical funnel section. Such then allows the unit to collect material from swaths larger in diameter than the enclosing casing pipe of the screw auger. A third element includes a flexible screw auger. All three can be used in combination in microgravity and zero atmosphere environments to drill and recover a wide area of subsurface regolith and entrained volatiles through a single access point on the surface.

  10. Indirect effects of Wnt3a/β-catenin signalling support mouse spermatogonial stem cells in vitro.

    Directory of Open Access Journals (Sweden)

    Jonathan R Yeh

    Full Text Available Proper regulation of spermatogonial stem cells (SSCs is crucial for sustaining steady-state spermatogenesis. Previous work has identified several paracrine factors involved in this regulation, in particular, glial cell line-derived neurotrophic factor and fibroblast growth factor 2, which promote long-term SSC self-renewal. Using a SSC culture system, we have recently reported that Wnt5a promotes SSC self-renewal through a β-catenin-independent Wnt mechanism whereas the β-catenin-dependent Wnt pathway is not active in SSCs. In contrast, another study has reported that Wnt3a promotes SSC self-renewal through the β-catenin-dependent pathway, as it can stimulate the proliferation of a spermatogonia cell line. To reconcile these two contradictory reports, we assessed Wnt3a effects on SSCs and progenitor cells, rather than a cell line, in vitro. We observed that Wnt3a induced β-catenin-dependent signalling in a large subset of germ cells and increased SSC numbers. However, further investigation revealed that cell populations with greater β-catenin-signalling activity contained fewer SSCs. The increased maintenance of SSCs by Wnt3a coincided with more active cell cycling and the formation of germ cell aggregates, or communities, under feeder-free conditions. Therefore, the results of this study suggest that Wnt3a selectively stimulates proliferation of progenitors that are committed to differentiation or are in the process of exiting the SSC state, leading to enhanced formation of germ cell communities, which indirectly support SSCs and act as an in vitro niche.

  11. Photovoltaic Hosting Capacity of Feeders with Reactive Power Control and Tap Changers

    Energy Technology Data Exchange (ETDEWEB)

    Ceylan, Oğuzhan; Paudyal, Sumit; Bhattarai, Bishnu P.; Myers, Kurt S.

    2017-06-01

    This paper proposes an algorithm to determine photovoltaic (PV) hosting capacity of power distribution networks as a function of number of PV injection nodes, reactive power support from the PVs, and the sub-station load tap changers (LTCs). In the proposed method, several minute by minute simulations are run based on randomly chosen PV injection nodes, daily PV output profiles, and daily load profiles from a pool of high-resolution realistic data set. The simulation setup is built using OpenDSS and MATLAB. The performance of the proposed method is investigated in the IEEE 123-node distribution feeder for multiple scenarios. The case studies are performed particularly for one, two, five and ten PV injection nodes, and looking at the maximum voltage deviations. Case studies show that the PV hosting capacity of the 123-node feeder greatly differs with the number of PV injection nodes. We have also observed that the PV hosting capacity increases with reactive power support and higher tap position of sub-station LTC.

  12. Damping considerations in CANDU feeder pipe design and analysis

    International Nuclear Information System (INIS)

    Usmani, S.A.; Saleem, M.A.; So, G.

    1990-01-01

    Recent developments in pipe damping indicate a trend towards more realistic and less conservative values, which result in less rigid and safer pipe designs. The CANDU-PHW (Canada deuterium uranium, pressurized heavy water) reactor feeder pipe designs have applied similar approaches which permit seismic qualifications without overly restraining these compact arrays of pipes to cater for the large creep and thermal anchor movement. This paper reviews the feeder design aspects, especially pertaining to the design provisions, experimental verification and analytical modelling for seismic qualification in the light of recent pipe dynamic developments. Using illustrative examples, comparison of seismic analysis results is provided for the ASME Code Case N-411 dampings, and those traditionally used in the feeder seismic qualification. The results confirm acceptability of the traditional approach which permit simplified analysis to demonstrate seismic qualificationqualification of CANDU feeder pipes

  13. Automation Selection and Sequencing of Traps for Vibratory Feeders

    DEFF Research Database (Denmark)

    Mathiesen, Simon; Ellekilde, Lars-Peter

    2017-01-01

    Vibratory parts feeders with mechanical orienting devices are used extensively in the assembly automation industry. Even so, the design process is based on trial-and-error approaches and is largely manual. In this paper, a methodology is presented for automatic design of this type of feeder....... The approach uses dynamic simulation for generating the necessary data for configuring a feeder with a sequence of mechanical orienting devices called traps, with the goal of reorienting all parts from a random to fixed orientation. Then, a fast algorithm for facilitating this configuration task automatically...

  14. Urban Bird Feeders Dominated by a Few Species and Individuals

    Directory of Open Access Journals (Sweden)

    Josie A. Galbraith

    2017-08-01

    Full Text Available The practice of garden bird feeding is a global phenomenon, involving millions of people and vast quantities of food annually. Many people engage in the practice of feeding assuming that birds gain some benefit from the food they provide, yet recent studies have revealed the potential for detrimental impacts as well. However, there is still a paucity of information on the impacts of feeding, including the ubiquity of these impacts among and within feeder-visiting species. Consistency in feeder use among birds is likely an important determinant of this. Individual birds and species that make frequent use of feeders are more likely to experience both the benefits and detrimental impacts of supplementary food. We investigated patterns of feeder use by garden birds visiting experimental feeding stations in Auckland, New Zealand, with the specific aim of determining whether use of supplementary food was consistent or variable among individuals and species. We used camera traps as well as Radio Frequency Identification (RFID technology to examine intra- and interspecific feeder visitation patterns and to discern species associations. Eleven bird species were detected using feeding stations, however, two introduced species (house sparrow Passer domesticus and spotted dove Streptopelia chinensis dominated visitation events. These species were present at feeders most frequently, with the largest conspecific group sizes. Significant associations were detected among a number of species, suggesting interspecific interactions are important in determining feeder use. We also found within-species differences in feeder use for all focal species, with individual variation greatest in house sparrows. Furthermore, season had an important influence on most visitation parameters. The observed individual and species-specific differences in supplementary food resource use imply that the impacts of garden bird feeding are not universal. Crucially, particularly given

  15. Development of a feeder device for logging residue for a drum chipper; Hakkurin syoetoen kehittaeminen energiapuulle

    Energy Technology Data Exchange (ETDEWEB)

    Leinonen, T.; Kemppainen, J P [Oulu Univ. (Finland). Dept. of Mechanical Engineering

    1997-12-01

    The aim of this project is to increase the productivity of a large drum chipper by developing a feeder device to be used especially when chipping logging residue. Logging residue from regeneration areas is the most important reserve of forest biomass for the production of renewable energy in Finland. It is possible ecologically and economically to use about 5 million m{sup 3} of logging residue yearly for energy purposes. Currently both large lorry based chippers and smaller forwarder based chippers are used for chipping logging residue. However even the latest versions of these chippers are not well equipped for chipping logging residue. The ball like shape of residue piles causes blockades, if too large amounts of residue are fed into the chipper. Keeping the material flow even is difficult. In this project a prototype of a feeder device will be designed and manufactured to exactly find out the problems, that occur when feeding logging residue to a chipper. The designed prototype is based on a tubular chassis, which supports a conveyor made of steel plates. Prototype also has a 2,5 m wide feeding table and a feeder roll, which is located above the conveyor just before the chipper opening. Both the conveyor and the feeder roll are powered by hydraulic motors. The maximum feeding force is 20 kN. For research purposes both hydraulic pressure and flow can be adjusted individually for both motors. During 1996 activities included designing and building the prototype feeder device. In 1997 the prototype will be tested to find out possible problems, which need to be cured. After problems are solved and other improvements are made, the feeder device will be installed to an existing chipper and then tested again to determine the productivity of the new feeder device compared to the old unit. (orig.)

  16. Easy xeno-free and feeder-free method for isolating and growing limbal stromal and epithelial stem cells of the human cornea.

    Directory of Open Access Journals (Sweden)

    Djida Ghoubay-Benallaoua

    Full Text Available Epithelial and stromal stem cells are required to maintain corneal transparency. The aim of the study was to develop a new method to isolate and grow both corneal stromal (SSC and epithelial limbal (LSC stem cells from small human limbal biopsies under culture conditions in accordance with safety requirements mandatory for clinical use in humans. Superficial limbal explants were retrieved from human donor corneo-scleral rims. Human limbal cells were dissociated by digestion with collagenase A, either after epithelial scraping or with no scraping. Isolated cells were cultured with Essential 8 medium (E8, E8 supplemented with EGF (E8+ or Green's medium with 3T3 feeder-layers. Cells were characterized by immunostaining, RT-qPCR, colony forming efficiency, sphere formation, population doubling, second harmonic generation microscopy and differentiation potentials. LSC were obtained from unscraped explants in E8, E8+ and Green's media and were characterized by colony formation and expression of PAX6, ΔNP63α, Bmi1, ABCG2, SOX9, CK14, CK15 and vimentin, with a few cells positive for CK3. LSC underwent 28 population doublings still forming colonies. SSC were obtained from both scraped and unscraped explants in E8 and E8+ media and were characterized by sphere formation, expression of PAX6, SOX2, BMI1, NESTIN, ABCG2, KERATOCAN, VIMENTIN, SOX9, SOX10 and HNK1, production of collagen fibrils and differentiation into keratocytes, fibroblasts, myofibroblasts, neurons, adipocytes, chondrocytes and osteocytes. SSC underwent 48 population doublings still forming spheres, Thus, this new method allows both SSC and LSC to be isolated from small superficial limbal biopsies and to be primary cultured in feeder-free and xeno-free conditions, which will be useful for clinical purposes.

  17. SATB2 expression increased anchorage-independent growth and cell migration in human bronchial epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Feng; Jordan, Ashley; Kluz, Thomas [Department of Environmental Medicine, New York University School of Medicine, 57 Old Forge Road, Tuxedo, NY 10987 (United States); Shen, Steven [Center for Health Informatics and Bioinformatics, New York University Langone Medical Center, New York, NY 10016 (United States); Sun, Hong; Cartularo, Laura A. [Department of Environmental Medicine, New York University School of Medicine, 57 Old Forge Road, Tuxedo, NY 10987 (United States); Costa, Max, E-mail: Max.Costa@nyumc.org [Department of Environmental Medicine, New York University School of Medicine, 57 Old Forge Road, Tuxedo, NY 10987 (United States)

    2016-02-15

    The special AT-rich sequence-binding protein 2 (SATB2) is a protein that binds to the nuclear matrix attachment region of the cell and regulates gene expression by altering chromatin structure. In our previous study, we reported that SATB2 gene expression was induced in human bronchial epithelial BEAS-2B cells transformed by arsenic, chromium, nickel and vanadium. In this study, we show that ectopic expression of SATB2 in the normal human bronchial epithelial cell-line BEAS-2B increased anchorage-independent growth and cell migration, meanwhile, shRNA-mediated knockdown of SATB2 significantly decreased anchorage-independent growth in Ni transformed BEAS-2B cells. RNA sequencing analyses of SATB2 regulated genes revealed the enrichment of those involved in cytoskeleton, cell adhesion and cell-movement pathways. Our evidence supports the hypothesis that SATB2 plays an important role in BEAS-2B cell transformation. - Highlights: • We performed SATB2 overexpression in the BEAS-2B cell line. • We performed SATB2 knockdown in a Ni transformed BEAS-2B cell line. • SATB2 induced anchorage-independent growth and increased cell migration. • SATB2 knockdown significantly decreased anchorage-independent growth. • We identified alterations in gene involved in cytoskeleton, cell adhesion.

  18. SATB2 expression increased anchorage-independent growth and cell migration in human bronchial epithelial cells

    International Nuclear Information System (INIS)

    Wu, Feng; Jordan, Ashley; Kluz, Thomas; Shen, Steven; Sun, Hong; Cartularo, Laura A.; Costa, Max

    2016-01-01

    The special AT-rich sequence-binding protein 2 (SATB2) is a protein that binds to the nuclear matrix attachment region of the cell and regulates gene expression by altering chromatin structure. In our previous study, we reported that SATB2 gene expression was induced in human bronchial epithelial BEAS-2B cells transformed by arsenic, chromium, nickel and vanadium. In this study, we show that ectopic expression of SATB2 in the normal human bronchial epithelial cell-line BEAS-2B increased anchorage-independent growth and cell migration, meanwhile, shRNA-mediated knockdown of SATB2 significantly decreased anchorage-independent growth in Ni transformed BEAS-2B cells. RNA sequencing analyses of SATB2 regulated genes revealed the enrichment of those involved in cytoskeleton, cell adhesion and cell-movement pathways. Our evidence supports the hypothesis that SATB2 plays an important role in BEAS-2B cell transformation. - Highlights: • We performed SATB2 overexpression in the BEAS-2B cell line. • We performed SATB2 knockdown in a Ni transformed BEAS-2B cell line. • SATB2 induced anchorage-independent growth and increased cell migration. • SATB2 knockdown significantly decreased anchorage-independent growth. • We identified alterations in gene involved in cytoskeleton, cell adhesion.

  19. Advances in technologies for feeder pipe inspections

    Energy Technology Data Exchange (ETDEWEB)

    Ten Grotenhuis, R.; Verma, Y.; Hitchcox, T.; Sakuta, A. [Ontario Power Generation, Inspection and Maintenance Div., Toronto, Ontario (Canada)

    2014-07-15

    The successful development of the Matrix Inspection Technique (MIT) for feeder weld area applications has been followed up with a project to leverage the technology to address other aspects of feeder inspection. The goal of the project is to adapt the technology to provide full circumferential inspection of feeder pipes for FAC thinning and, potentially, for axial cracking. The project necessitated evolving a new generation of high speed, high element count data acquisition instruments. It also required the development of custom inspection arrays, innovative approaches to sealing the water column, use of inertial motion sensors to synthesize encoder inputs, real-time visual feedback for the operator, and enhanced automated analysis software capable of plotting the inspected configuration in 3D. The individual components of the system are currently being integrated into a whole. The results obtained to date demonstrate the approach to be fundamentally sound. (author)

  20. Object categorization by wild ranging birds-Winter feeder experiments.

    Science.gov (United States)

    Nováková, Nela; Veselý, Petr; Fuchs, Roman

    2017-10-01

    The object categorization is only scarcely studied using untrained wild ranging animals and relevant stimuli. We tested the importance of the spatial position of features salient for categorization of a predator using wild ranging birds (titmice) visiting a winter feeder. As a relevant stimulus we used a dummy of a raptor, the European sparrowhawk (Accipiter nisus), placed at the feeding location. This dummy was designed to be dismantled into three parts and rearranged with the head in the correct position, in the middle or at the bottom of the dummy. When the birds had the option of visiting an alternative feeder with a dummy pigeon, they preferred this option to visiting the feeder with the dummy sparrowhawk with the head in any of the three positions. When the birds had the option of visiting an alternative feeder with an un-rearranged dummy sparrowhawk, they visited both feeders equally often, and very scarcely. This suggests that the titmice considered all of the sparrowhawk modifications as being dangerous, and equally dangerous as the un-rearranged sparrowhawk. The position of the head was not the most important cue for categorization. The presence of the key features was probably sufficient for categorization, and their mutual spatial position was of lower importance. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Stress analysis of feeder bends using neutrons: new results and cumulative impacts

    Energy Technology Data Exchange (ETDEWEB)

    Banks, D.; Donaberger, R. [Canadian Neutron Beam Centre, Chalk River, ON (Canada); Leitch, B. [Atomic Energy of Canada Limited, Chalk River, ON (Canada); Rogge, R.B. [Canadian Neutron Beam Centre, Chalk River, ON (Canada)

    2014-07-01

    Neutron diffraction has played a vital role in stress analysis of bends in carbon steel pipes, known as feeder pipes, in CANDU reactors. Due to incidents of cracking of feeders, extensive R&D programs to manage feeder cracking have been implemented over about ten years. We review the cumulative impacts of this research from the view point of the stress analysis using neutrons, and present new results by examining a feeder bend with a partial crack both experimentally using neutron diffraction and theoretically using a finite element model. (author)

  2. Impacting effects of seismic loading in feeder pipes of PHWR power plants

    International Nuclear Information System (INIS)

    Kumar, R.

    1996-01-01

    The core of a pressurized heavy water reactor (PHWR) consists of a large number of fuel channels. These fuel channels are connected to the feeder pipes through which the heavy water flows and transports heat from the reactor core to the steam generators. The feeder pipes are several hundreds in number. They run close to each other with small gaps and have several bends. Thus they represent a complex piping system. Under seismic loading, the adjacent feeder pipes may impact each other. In this paper a simplified procedure has been established to assess such impacting effects. The results of the proposed analysis include bending moment and impact force, which provide the stresses due to impacting effects. These results are plotted in nondimensional form so that they could be utilized for any set of feeder pipes. The procedure used for studying the impacting effects includes seismic analysis of individual feeder pipes without impacting effects, selection of pipes for impact analysis, and estimating their maximum impact velocity. Based on the static and dynamic characteristics of the selected feeder pipes, the maximum bending moment, impact force, and stresses are obtained. The results of this study are useful for quick evaluation of the impacting effects in feeder pipes

  3. S08: investigation and repair of a cracked feeder at Point Lepreau GS

    International Nuclear Information System (INIS)

    A Celovsky, A.; Wright, M.D.; Gendron, T.S.

    1997-01-01

    Early in 1997 investigation of a low level leak in the Point Lepreau GS (PLGS) PHTS revealed that an outlet feeder, S08, was leaking. Ultrasonic inspection, and subsequent failure analysis, revealed that the leak was a consequence of a crack. Given the unusual nature of this event, and current concerns over feeder thinning, a detailed and careful removal and examination procedure was developed. The S08 outlet feeder was removed and shipped to Chalk River Laboratories for examination. The examination confirmed that the failure was a through-wall crack, most likely the consequence of stress corrosion cracking. A critical point of the analysis was to determine how the crack initiated, and subsequently propagated. High residual stresses and possible abnormal loading in conjunction with chemistry environments resulted in the Stress Corrosion Cracking (SCC) of the S08 outlet feeder bend. It is recognized that some of the causative factors implicated in the S08 failure apply to other outlet feeders. In particular, residual stresses in the non-stress-relieved, short-radius cold bent pipes will remain relatively high over the future life of the feeders. However, the risk of CANDU feeder failure by SCC is judged to be extremely low based on the evidence of the inspections carried out to date and the good performance record of feeder pipe in the CANDU industry. The channel was restored to its locked configuration, and the failed section of feeder replaced. (author)

  4. Hybrid RSOA and fibre raman amplified long reach feeder link for WiMAX-on-fibre

    DEFF Research Database (Denmark)

    Amaya Fernández, Ferney Orlando; Martinez, Javier; Yu, Xianbin

    2009-01-01

    A distributed fibre Raman amplified long reach optical access feeder link using a reflective semiconductor optical amplifier in the remote base station is experimentally demonstrated for supporting WiMAXover- fibre transmission. The measured values for the error vector magnitude for quadrature...

  5. Dry piston coal feeder

    Science.gov (United States)

    Hathaway, Thomas J.; Bell, Jr., Harold S.

    1979-01-01

    This invention provides a solids feeder for feeding dry coal to a pressurized gasifier at elevated temperatures substantially without losing gas from the gasifier by providing a lock having a double-acting piston that feeds the coals into the gasifier, traps the gas from escaping, and expels the trapped gas back into the gasifier.

  6. Induced pluripotent stem cells generated from human adipose-derived stem cells using a non-viral polycistronic plasmid in feeder-free conditions.

    Directory of Open Access Journals (Sweden)

    Xinjian Qu

    Full Text Available Induced pluripotent stem cells (iPSCs can be generated from somatic cells by ectopic expression of defined transcription factors (TFs. However, the optimal cell type and the easy reprogramming approaches that minimize genetic aberrations of parent cells must be considered before generating the iPSCs. This paper reports a method to generate iPSCs from adult human adipose-derived stem cells (hADSCs without the use of a feeder layer, by ectopic expression of the defined transcription factors OCT4, SOX2, KLF4 and C-MYC using a polycistronic plasmid. The results, based on the expression of pluripotent marker, demonstrated that the iPSCs have the characteristics similar to those of embryonic stem cells (ESCs. The iPSCs differentiated into three embryonic germ layers both in vitro by embryoid body generation and in vivo by teratoma formation after being injected into immunodeficient mice. More importantly, the plasmid DNA does not integrate into the genome of human iPSCs as revealed by Southern blotting experiments. Karyotypic analysis also demonstrated that the reprogramming of hADSCs by the defined factors did not induce chromosomal abnormalities. Therefore, this technology provides a platform for studying the biology of iPSCs without viral vectors, and can hopefully overcome immune rejection and ethical concerns, which are the two important barriers of ESC applications.

  7. Improvement of automatic fish feeder machine design

    Science.gov (United States)

    Chui Wei, How; Salleh, S. M.; Ezree, Abdullah Mohd; Zaman, I.; Hatta, M. H.; Zain, B. A. Md; Mahzan, S.; Rahman, M. N. A.; Mahmud, W. A. W.

    2017-10-01

    Nation Plan of action for management of fishing is target to achieve an efficient, equitable and transparent management of fishing capacity in marine capture fisheries by 2018. However, several factors influence the fishery production and efficiency of marine system such as automatic fish feeder machine could be taken in consideration. Two latest fish feeder machines have been chosen as the reference for this study. Based on the observation, it has found that the both machine was made with heavy structure, low water and temperature resistance materials. This research’s objective is to develop the automatic feeder machine to increase the efficiency of fish feeding. The experiment has conducted to testing the new design of machine. The new machine with maximum storage of 5 kg and functioning with two DC motors. This machine able to distribute 500 grams of pellets within 90 seconds and longest distance of 4.7 meter. The higher speed could reduce time needed and increase the distance as well. The minimum speed range for both motor is 110 and 120 with same full speed range of 255.

  8. Pressurized feeder for biofuels; Paineistettu syoetin biopolttoaineille

    Energy Technology Data Exchange (ETDEWEB)

    Lundqvist, R.; Makkonen, P. [Foster Wheeler Energia Oy, Karhula (Finland). Karhula R and D Center

    1995-12-31

    A new type of pressurized piston feeder has been tested for further development of the IGCC concept. The project is funded by the Finnish Ministry of Trade and Industry. The objective has been to develop a substitute for lock-hopper systems, which are commercial, but far too expensive from an operational point of view, due to high inert gas consumption for pressurization. The new feeder system, which is of dual piston type, uses little inert gas, because the pressurization sequence is performed partially by the piston movement. The project comprised further development of the concept, mechanical testing under atmospheric and pressurized conditions and testing with wood chips, peat and coal at elevated pressures. The main concern has been the wear of seals and gaskets during long term operation and for this reason over 600 hours of testing has been carried out. The tests were carried out between May 1994 and January 1995 with wood chips, peat and coal mainly at 17 bar(a) pressure. During the operation, the system was fine-tuned, and generally, the testing was successful. At the early stage of the tests, the ring seal design was modified and several alternative seal constructions were tested. During the tests, the seal was developed further and improved. The present ring seal design has been proven reliable and it meets the commercial requirements. After the tests, the feeder system was inspected thoroughly and found to be in good condition. A commercially available concept of the feeder has been developed on the basis of the knowledge obtained during the test runs

  9. Pressurized feeder for biofuels; Paineistettu syoetin biopolttoaineille

    Energy Technology Data Exchange (ETDEWEB)

    Lundqvist, R; Makkonen, P [Foster Wheeler Energia Oy, Karhula (Finland). Karhula R and D Center

    1996-12-31

    A new type of pressurized piston feeder has been tested for further development of the IGCC concept. The project is funded by the Finnish Ministry of Trade and Industry. The objective has been to develop a substitute for lock-hopper systems, which are commercial, but far too expensive from an operational point of view, due to high inert gas consumption for pressurization. The new feeder system, which is of dual piston type, uses little inert gas, because the pressurization sequence is performed partially by the piston movement. The project comprised further development of the concept, mechanical testing under atmospheric and pressurized conditions and testing with wood chips, peat and coal at elevated pressures. The main concern has been the wear of seals and gaskets during long term operation and for this reason over 600 hours of testing has been carried out. The tests were carried out between May 1994 and January 1995 with wood chips, peat and coal mainly at 17 bar(a) pressure. During the operation, the system was fine-tuned, and generally, the testing was successful. At the early stage of the tests, the ring seal design was modified and several alternative seal constructions were tested. During the tests, the seal was developed further and improved. The present ring seal design has been proven reliable and it meets the commercial requirements. After the tests, the feeder system was inspected thoroughly and found to be in good condition. A commercially available concept of the feeder has been developed on the basis of the knowledge obtained during the test runs

  10. A Feeder-Bus Dispatch Planning Model for Emergency Evacuation in Urban Rail Transit Corridors

    Science.gov (United States)

    Wang, Yun; Yan, Xuedong; Zhou, Yu; Zhang, Wenyi

    2016-01-01

    The mobility of modern metropolises strongly relies on urban rail transit (URT) systems, and such a heavy dependence causes that even minor service interruptions would make the URT systems unsustainable. This study aims at optimally dispatching the ground feeder-bus to coordinate with the urban rails’ operation for eliminating the effect of unexpected service interruptions in URT corridors. A feeder-bus dispatch planning model was proposed for the collaborative optimization of URT and feeder-bus cooperation under emergency situations and minimizing the total evacuation cost of the feeder-buses. To solve the model, a concept of dummy feeder-bus system is proposed to transform the non-linear model into traditional linear programming (ILP) model, i.e., traditional transportation problem. The case study of Line #2 of Nanjing URT in China was adopted to illustrate the model application and sensitivity analyses of the key variables. The modeling results show that as the evacuation time window increases, the total evacuation cost as well as the number of dispatched feeder-buses decrease, and the dispatched feeder-buses need operate for more times along the feeder-bus line. The number of dispatched feeder-buses does not show an obvious change with the increase of parking spot capacity and time window, indicating that simply increasing the parking spot capacity would cause huge waste for the emergent bus utilization. When the unbalanced evacuation demand exists between stations, the more feeder-buses are needed. The method of this study will contribute to improving transportation emergency management and resource allocation for URT systems. PMID:27676179

  11. Human neural progenitor cells decrease photoreceptor degeneration, normalize opsin distribution and support synapse structure in cultured porcine retina.

    Science.gov (United States)

    Mollick, Tanzina; Mohlin, Camilla; Johansson, Kjell

    2016-09-01

    Retinal neurodegenerative disorders like retinitis pigmentosa, age-related macular degeneration, diabetic retinopathy and retinal detachment decrease retinal functionality leading to visual impairment. The pathological events are characterized by photoreceptor degeneration, synaptic disassembly, remodeling of postsynaptic neurons and activation of glial cells. Despite intense research, no effective treatment has been found for these disorders. The current study explores the potential of human neural progenitor cell (hNPC) derived factors to slow the degenerative processes in adult porcine retinal explants. Retinas were cultured for 3 days with or without hNPCs as a feeder layer and investigated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), immunohistochemical, western blot and quantitative real time-polymerase chain reaction (qRT-PCR) techniques. TUNEL showed that hNPCs had the capacity to limit photoreceptor cell death. Among cone photoreceptors, hNPC coculture resulted in better maintenance of cone outer segments and reduced opsin mislocalization. Additionally, maintained synaptic structural integrity and preservation of second order calbindin positive horizontal cells was also observed. However, Müller cell gliosis only seemed to be alleviated in terms of reduced Müller cell density. Our observations indicate that at 3 days of coculture, hNPC derived factors had the capacity to protect photoreceptors, maintain synaptic integrity and support horizontal cell survival. Human neural progenitor cell applied treatment modalities may be an effective strategy to help maintain retinal functionality in neurodegenerative pathologies. Whether hNPCs can independently hinder Müller cell gliosis by utilizing higher concentrations or by combination with other pharmacological agents still needs to be determined. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. limit loads for wall-thinning feeder pipes under combined bending and internal pressure

    International Nuclear Information System (INIS)

    Je, Jin Ho; Lee, Kuk Hee; Chung, Ha Joo; Kim, Ju Hee; Han, Jae Jun; Kim, Yun Jae

    2009-01-01

    Flow Accelerated Corrosion (FAC) during inservice conditions produces local wall-thinning in the feeder pipes of CANDU. The Wall-thinning in the feeder pipes is main degradation mechanisms affecting the integrity of piping systems. This paper discusses the integrity assessment of wall-thinned feeder pipes using limit load analysis. Based on finite element limit analyses, this paper compare limit loads for wall-thinning feeder pipes under combined bending and internal pressure with proposed limit loads. The limit loads are determined from limit analyses based on rectangular wall-thinning and elastic-perfectly-plastic materials using the large geometry change.

  13. Glial cell line-derived neurotrophic factor and endothelial cells promote self-renewal of rabbit germ cells with spermatogonial stem cell properties.

    Science.gov (United States)

    Kubota, Hiroshi; Wu, Xin; Goodyear, Shaun M; Avarbock, Mary R; Brinster, Ralph L

    2011-08-01

    Previous studies suggest that exogenous factors crucial for spermatogonial stem cell (SSC) self-renewal are conserved among several mammalian species. Since glial cell line-derived neurotrophic factor (GDNF) and fibroblast growth factor 2 (FGF2) are critical for rodent SSC self-renewal, we hypothesized that they might promote self-renewal of nonrodent SSCs. Therefore, we cultured testicular germ cells from prepubertal rabbits in the presence of GDNF and FGF2 and found they proliferated indefinitely as cellular clumps that displayed characteristics previously identified for rodent SSCs. The rabbit germ cells could not be maintained on mouse embryonic fibroblast (STO) feeders that support rodent SSC self-renewal in vitro but were rather supported on mouse yolk sac-derived endothelial cell (C166) feeder layers. Proliferation of rabbit germ cells was dependent on GDNF. Of critical importance was that clump-forming rabbit germ cells colonized seminiferous tubules of immunodeficient mice, proliferated for at least 6 mo, while retaining an SSC phenotype in the testes of recipient mice, indicating that they were rabbit SSCs. This study demonstrates that GDNF is a mitogenic factor promoting self-renewal that is conserved between rodent and rabbit SSCs; with an evolutionary separation of ∼ 60 million years. These findings provide a foundation to study the mechanisms governing SSC self-renewal in nonrodent species.

  14. A defined, feeder-free, serum-free system to generate in vitro hematopoietic progenitors and differentiated blood cells from hESCs and hiPSCs.

    Directory of Open Access Journals (Sweden)

    Giorgia Salvagiotto

    2011-03-01

    Full Text Available Human ESC and iPSC are an attractive source of cells of high quantity and purity to be used to elucidate early human development processes, for drug discovery, and in clinical cell therapy applications. To efficiently differentiate pluripotent cells into a pure population of hematopoietic progenitors we have developed a new 2-dimensional, defined and highly efficient protocol that avoids the use of feeder cells, serum or embryoid body formation. Here we showed that a single matrix protein in combination with growth factors and a hypoxic environment is sufficient to generate from pluripotent cells hematopoietic progenitors capable of differentiating further in mature cell types of different lineages of the blood system. We tested the differentiation method using hESCs and 9 iPSC lines generated from different tissues. These data indicate the robustness of the protocol providing a valuable tool for the generation of clinical-grade hematopoietic cells from pluripotent cells.

  15. Real-Time Distribution Feeder Performance Monitoring, Advisory Control, and Health Management System

    Energy Technology Data Exchange (ETDEWEB)

    Stoupis, James; Mousavi, Mirrasoul

    2014-09-30

    provides this time and helps accelerate service restoration ultimately. To demonstrate the effectiveness of this feature, a demo system using substation data alone was set up to compare the minutes saved over a period of 22 months for two feeders where the real-time notification system has been deployed. The metric used for performance assessment is the time difference between the actual outage time from the OMS versus the time the notification email was received on the operators desk. Over the period of 22 months, we have accumulated over 7600 minutes (32 hours) ahead of actual outage time compared to the OMS timestamps. The significance of this analysis is that it shows the potential to reduce the SAIDI minutes and directly impact utility performance in terms of outage duration. If deployed at scale, it would have a significant impact on system reliability. To put this number in perspective, it would be helpful to assign a dollar figure to the potential savings that could be realized. According to the host utility, the average cost for each customer-minute-out (CMO) is approximately $0.30 across the operating company. This includes both direct and indirect costs such as bad press. The outage data over the previous 4 years show that the average customer count on primary/tap level outages is about 56. Accordingly, the total minutes saved amounts to 425,600 CMOs on the average. This would in turn result in a potential cost savings figure of $127,680 for two feeders alone over the period of performance. This empirical evidence validates the strong value proposition of the project that was contemplated at the onset and its potential impact to reduce outage duration in support of DOE’s goal of 20%

  16. Urban and rural habitats differ in number and type of bird feeders and in bird species consuming supplementary food.

    Science.gov (United States)

    Tryjanowski, Piotr; Skórka, Piotr; Sparks, Tim H; Biaduń, Waldemar; Brauze, Tomasz; Hetmański, Tomasz; Martyka, Rafał; Indykiewicz, Piotr; Myczko, Łukasz; Kunysz, Przemysław; Kawa, Piotr; Czyż, Stanisław; Czechowski, Paweł; Polakowski, Michał; Zduniak, Piotr; Jerzak, Leszek; Janiszewski, Tomasz; Goławski, Artur; Duduś, Leszek; Nowakowski, Jacek J; Wuczyński, Andrzej; Wysocki, Dariusz

    2015-10-01

    Bird feeding is one of the most widespread direct interactions between man and nature, and this has important social and environmental consequences. However, this activity can differ between rural and urban habitats, due to inter alia habitat structure, human behaviour and the composition of wintering bird communities. We counted birds in 156 squares (0.25 km(2) each) in December 2012 and again in January 2013 in locations in and around 26 towns and cities across Poland (in each urban area, we surveyed 3 squares and also 3 squares in nearby rural areas). At each count, we noted the number of bird feeders, the number of bird feeders with food, the type of feeders, additional food supplies potentially available for birds (bread offered by people, bins) and finally the birds themselves. In winter, urban and rural areas differ in the availability of food offered intentionally and unintentionally to birds by humans. Both types of food availability are higher in urban areas. Our findings suggest that different types of bird feeder support only those species specialized for that particular food type and this relationship is similar in urban and rural areas.

  17. Comparison of the Ex Vivo Expansion of UCB-Derived CD34+ in 3D DBM/MBA Scaffolds with USSC as a Feeder Layer.

    Science.gov (United States)

    Sadat Hashemi, Zahra; Forouzandeh Moghadam, Mahdi; Soleimani, Masoud

    2013-10-01

    Ex vivo expansion of hematopoitic stem cells is an alternative way to increase umbilical cord blood (UCB)-CD34+ cells for bone marrow transplantation. For this purpose demineralized bone matrix (DBM) and mineralized bone allograft (MBA) as two scaffolds based on bone matrix and stem cell niche, were simultaneously used to enhance the effect of human mesenchymal progenitor cells (MPCs) - unrestricted somatic stem cells (USSCs) - as a feeder layer. USSCs were isolated and characterized by morphological and immunological analysis then seeded on both scaffolds as a feeder layer. UCB-CD34(+) were isolated by MACS method and were co-culture expanded by USSC in 3D and 2D environments. After 3 weeks expansion, cells were counted and were assessed by karyotype, flow cytometry, clonogenic activity, and long-term culture-initiating cells (LTC-IC). Co-culture expansion in DBM and MBA was 29.22-fold and 27.77-fold, no significant differences in colony and LTC-IC were obtained. Maximum number of colonies belonged to the day 14 with the 73% CFU-GM (Colony Forming Unit- Granulocyte/Macrophage) in contrast to the day 0 which was BFU-E/CFU-E (Burst/Colony Forming Unit-Erythroid). Flow cytometry indicated that the percentage of CD34+ marker was decreased in USSC co-culture and the highest percentage was observed in simple 2D culture. Because of acid extraction in the DBM production process, mineral materials were removed and the protein background that was more flexible was presented. Therefore these results suggest that USSC-DBM can be a suitable ex vivo mimicry niche by intensifying of surface/volume ratio and supporting the stem cell differentiation and expansion.

  18. A Markov chain model for CANDU feeder pipe degradation

    International Nuclear Information System (INIS)

    Datla, S.; Dinnie, K.; Usmani, A.; Yuan, X.-X.

    2008-01-01

    There is need for risk based approach to manage feeder pipe degradation to ensure safe operation by minimizing the nuclear safety risk. The current lack of understanding of some fundamental degradation mechanisms will result in uncertainty in predicting the rupture frequency. There are still concerns caused by uncertainties in the inspection techniques and engineering evaluations which should be addressed in the current procedures. A probabilistic approach is therefore useful in quantifying the risk and also it provides a tool for risk based decision making. This paper discusses the application of Markov chain model for feeder pipes in order to predict and manage the risks associated with the existing and future aging-related feeder degradation mechanisms. The major challenge in the approach is the lack of service data in characterizing the transition probabilities of the Markov model. The paper also discusses various approaches in estimating plant specific degradation rates. (author)

  19. A novel method for decomposing electricity feeder load into elementary profiles from customer information

    International Nuclear Information System (INIS)

    Gerossier, Alexis; Barbier, Thibaut; Girard, Robin

    2017-01-01

    Highlights: •Use of aggregated electricity load profiles and customer description at feeder level. •Statistical recovery of elementary load profiles with customer categorization. •Generation of load demand profiles for unknown feeders and new local areas. •Relevancy of the different categorizations. -- Abstract: To plan a distribution grid involves making a long-term forecast of sub-hourly demand, which requires modeling the demand and its dynamics with aggregated measurement data. Distribution system operators (DSOs) have been recording electricity sub-hourly demand delivered by their medium-voltage feeders (around 1000—10,000 customers) for several years. Demand profiles differ widely among the various considered feeders. This is partly due to the varying mix of customer categories from one feeder to another. To overcome this issue, elementary demand profiles are often associated with customer categories and then combined according to a mix description. This paper presents a novel method to estimate elementary profiles that only requires several feeder demand curves and a description of customers. The method relies on a statistical blind source model and a new estimation procedure based on the augmented Lagrangian method. The use of feeders to estimate elementary profiles means that measurements are fully representative and continuously updated. We illustrate the proposed method through a case study comprising around 1000 feeder demand curves operated by the main French DSO Enedis. We propose an application o that uses the obtained profiles to evaluate the contribution of any set of new customers to a feeder peak load. We show that profiles enable a simulation of new unmeasured areas with errors of around 20%. We also show how our method can be used to evaluate the relevancy of different customer categorizations.

  20. Stepwise Differentiation of Pluripotent Stem Cells into Osteoblasts Using Four Small Molecules under Serum-free and Feeder-free Conditions

    Directory of Open Access Journals (Sweden)

    Kosuke Kanke

    2014-06-01

    Full Text Available Pluripotent stem cells are a promising tool for mechanistic studies of tissue development, drug screening, and cell-based therapies. Here, we report an effective and mass-producing strategy for the stepwise differentiation of mouse embryonic stem cells (mESCs and mouse and human induced pluripotent stem cells (miPSCs and hiPSCs, respectively into osteoblasts using four small molecules (CHIR99021 [CHIR], cyclopamine [Cyc], smoothened agonist [SAG], and a helioxanthin-derivative 4-(4-methoxyphenylpyrido[4′,3′:4,5]thieno[2,3-b]pyridine-2-carboxamide [TH] under serum-free and feeder-free conditions. The strategy, which consists of mesoderm induction, osteoblast induction, and osteoblast maturation phases, significantly induced expressions of osteoblast-related genes and proteins in mESCs, miPSCs, and hiPSCs. In addition, when mESCs defective in runt-related transcription factor 2 (Runx2, a master regulator of osteogenesis, were cultured by the strategy, they molecularly recapitulated osteoblast phenotypes of Runx2 null mice. The present strategy will be a platform for biological and pathological studies of osteoblast development, screening of bone-augmentation drugs, and skeletal regeneration.

  1. Busbar arcs at large fusion magnets: Conductor to feeder tube arcing model experiments with the LONGARC device

    Energy Technology Data Exchange (ETDEWEB)

    Klimenko, Dmitry, E-mail: dmitry.klimenko@kit.edu; Pasler, Volker

    2014-10-15

    Highlights: •The LONGARC device was successfully implemented for busbar to feeder tubes arcing model experiments. •Arcing at an ITER busbar inside its feeder tube was simulated in scaled model experiments. •The narrower half tubes imply a slight increase of the arc propagation speed in compare to full tube experiments. •All simulated half tubes experiments show severe damage indicating that the ITER inner feeder tube will not withstand a busbar arc. -- Abstract: Electric arcs moving along the power cables (the so-called busbars) of the toroidal field (TF) coils of ITER may reach and penetrate the cryostat wall. Model experiments with the new LONGARC device continue the VACARC (VACuum ARC) experiments that were initiated to investigate the propagation and destruction mechanisms of busbar arcs in small scale [1]. The experiments are intended to support the development and validation of a numerical model. LONGARC overcomes the space limitations inside VACARC and allows also for advanced 1:3 (vs. ITER full scale) model setups. The LONGARC device and first results are presented below.

  2. EPICS: operating system independent device/driver support

    International Nuclear Information System (INIS)

    Kraimer, M.R.

    2003-01-01

    Originally EPICS input/output controllers (IOCs) were only supported on VME-based systems running the vxWorks operating system. Now IOCs are supported on many systems: vxWorks, RTEMS, Solaris, HPUX, Linux, WIN32, and Darwin. A challenge is to provide operating-system-independent device and driver support. This paper presents some techniques for providing such support. EPICS (Experimental Physics and Industrial Control System) is a set of software tools, libraries, and applications developed collaboratively and used worldwide to create distributed, real-time control systems for scientific instruments such as particle accelerators, telescopes, and other large scientific experiments. An important component of all EPICS-based control systems is a collection of input/output controllers (IOCs). An IOC has three primary components: (1) a real-time database; (2) channel access, which provides network access to the database; and (3) device/driver support for interfacing to equipment. This paper describes some projects related to providing device/driver support on non-vxWorks systems. In order to support IOCs on platforms other than vxWorks, operating-system-independent (OSI) application program interfaces (APIs) were defined for threads, semaphores, timers, etc. Providing support for a new platform consists of providing an operating-system-dependent implementation of the OSI APIs.

  3. BC Hydro's integrated approach on replacing underground feeder cables

    Energy Technology Data Exchange (ETDEWEB)

    Tarampi, D. [BC Hydro, Burnaby, BC (Canada)

    2008-07-01

    BC Hydro's integrated approach on replacing underground feeder cables was discussed. The feeder cables were all in ducts encased in concrete and splices were identified in concrete manholes. The feeder cables averaged 35 to 40 failures per year, with a large majority of failures occurring in manholes or near terminations. The age of the cables was not known. A graphical representation of a condition assessment was presented with reference to an electrical test, metallurgical test, hardness test, and dye penetrating test. Prioritization steps were also listed, including a condition index and replacement threshold. Steps in prioritizing the condition index included determining the insulation index based on electrical test results; determining the manhole condition based on metallurgical test results; determining the customer importance factor and feeder load factor; and calculating the overall condition index based on the indices. A process flowchart was also illustrated. A replacement strategy was then discussed and a cost comparison of two options was described. Option 1 was to replace only the unjacketed cables while option two involved selective replacement. tabs., figs.

  4. Efficient generation of transgene- and feeder-free induced pluripotent stem cells from human dental mesenchymal stem cells and their chemically defined differentiation into cardiomyocytes.

    Science.gov (United States)

    Tan, Xiaobing; Dai, Qingli; Guo, Tao; Xu, Jingshu; Dai, Qingyuan

    2018-01-22

    Advance in stem cell research resulted in several processes to generate induced pluripotent stem cells (iPSCs) from adult somatic cells. In our previous study, the reprogramming of iPSCs from human dental mesenchymal stem cells (MSCs) including SCAP and DPSCs, has been reported. Herein, safe iPSCs were reprogrammed from SCAP and DPSCs using non-integrating RNA virus vector, which is an RNA virus carrying no risk of altering host genome. DPSCs- and SCAP-derived iPSCs exhibited the characteristics of the classical morphology with human embryonic stem cells (hESCs) without integration of foreign genes, indicating the potential of their clinical application. Moreover, induced PSCs showed the capacity of self-renewal and differentiation into cardiac myocytes. We have achieved the differentiation of hiPSCs to cardiomyocytes lineage under serum and feeder-free conditions, using a chemically defined medium CDM3. In CDM3, hiPSCs differentiation is highly generating cardiomyocytes. The results showed this protocol produced contractile sheets of up to 97.2% TNNT2 cardiomyocytes after purification. Furthermore, derived hiPSCs differentiated to mature cells of the three embryonic germ layers in vivo and in vitro of beating cardiomyocytes. The above whole protocol enables the generation of large scale of highly pure cardiomyocytes as needed for cellular therapy. Copyright © 2017. Published by Elsevier Inc.

  5. Effect of artificial feeders on pollen loads of the hummingbirds of Cerro de La Muerte, Costa Rica

    Directory of Open Access Journals (Sweden)

    Gerardo Avalos

    2012-03-01

    Full Text Available Although sugar-water feeders are commonly used by enthusiasts to attract hummingbirds, little is known about how they affect hummingbird behavior and flower use. We studied the highland hummingbird assemblage of Cerro de La Muerte, Costa Rica, both at a site with permanent feeders (La Georgina Restaurant and further from it. We examined how feeder use and monopolization affected seasonal changes in pollen loads during four sampling periods, including dry and wet seasons, from 2003-2005. We expected that species monopolizing the feeders would carry little or no pollen whatsoever, and would have pollen loads characterized by low floral diversity, in contrast with species less dependent on feeders. We obtained pollen samples from 183 individuals of four hummingbird species captured around the feeders using mist nets, which were compared with a pollen reference collection of plants with a pollination syndrome by hummingbirds. The same methods were implemented at a site 3km away from the feeders. Feeder usage was quantified by counting the number of times hummingbirds drank from the feeders in periods of 4min separated by 1min. The effects of hummingbird species and season on pollen load categories were assessed using a nominal logistic regression. The alpha species at the site, the Fiery-throated Hummingbird (Panterpe insignis, dominated the feeders during the dry season. Meanwhile, in the wet season, feeder usage was more evenly distributed across species, with the exception of the Volcano Hummingbird, Selasphorus flammula, which occupies the last place in the dominance hierarchy. Pollen loads of hummingbirds captured near feeders were low in abundance (more than 50% of captured individuals had zero or low pollen loads, and low in species richness (96% of the hummingbirds with pollen from only one plant genus, Centropogon. Overall pollen loads increased during the dry season coinciding with peaks in flower availability, although the majority of

  6. Effect of artificial feeders on pollen loads of the hummingbirds of Cerro de la Muerte, Costa Rica.

    Science.gov (United States)

    Avalos, Gerardo; Soto, Alejandra; Alfaro, Willy

    2012-03-01

    Although sugar-water feeders are commonly used by enthusiasts to attract hummingbirds, little is known about how they affect hummingbird behavior and flower use. We studied the highland hummingbird assemblage of Cerro de La Muerte, Costa Rica, both at a site with permanent feeders (La Georgina Restaurant) and further from it. We examined how feeder use and monopolization affected seasonal changes in pollen loads during four sampling periods, including dry and wet seasons, from 2003-2005. We expected that species monopolizing the feeders would carry little or no pollen whatsoever, and would have pollen loads characterized by low floral diversity, in contrast with species less dependent on feeders. We obtained pollen samples from 183 individuals of four hummingbird species captured around the feeders using mist nets, which were compared with a pollen reference collection of plants with a pollination syndrome by hummingbirds. The same methods were implemented at a site 3km away from the feeders. Feeder usage was quantified by counting the number of times hummingbirds drank from the feeders in periods of 4min separated by 1min. The effects of hummingbird species and season on pollen load categories were assessed using a nominal logistic regression. The alpha species at the site, the Fiery-throated Hummingbird (Panterpe insignis), dominated the feeders during the dry season. Meanwhile, in the wet season, feeder usage was more evenly distributed across species, with the exception of the Volcano Hummingbird, Selasphorus flammula, which occupies the last place in the dominance hierarchy. Pollen loads of hummingbirds captured near feeders were low in abundance (more than 50% of captured individuals had zero or low pollen loads), and low in species richness (96% of the hummingbirds with pollen from only one plant genus, Centropogon). Overall pollen loads increased during the dry season coinciding with peaks in flower availability, although the majority of captured

  7. Optimal Design of the Feeder-Bus Network Based on the Transfer System

    Directory of Open Access Journals (Sweden)

    Lianbo Deng

    2013-01-01

    Full Text Available This paper studied the classic feeder-bus network design problem (FBNDP, which can be described as follows: for the passenger travel demand between rail stations and bus stops on a given urban transit network, it designs the optimal feeder bus routes and frequencies so as to minimize the passengers’ travel expense and the operator’s cost. We extended the demand pattern of M-to-1 in most existing researches to M-to-M. We comprehensively considered the passenger travel cost, which includes the waiting and riding cost on the bus, riding cost on rail, and transfer cost between these two transportation modes, and presented a new genetic algorithm that determines the optimal feeder-bus operating frequencies under strict constraint conditions. The numerical examples under different demand patterns have been experienced and analysed, which showed the robustness and efficiency of the presented algorithm. We also found that the distribution pattern of the travel demand has a significant influence on the feeder-bus network construction.

  8. Generation of megakaryocytic progenitors from human embryonic stem cells in a feeder- and serum-free medium.

    Directory of Open Access Journals (Sweden)

    Marjorie Pick

    Full Text Available BACKGROUND: The production of human platelets from embryonic stem cells in a defined culture system is a prerequisite for the generation of platelets for therapeutic use. As an important step towards this goal, we report the differentiation of human embryonic stem cells (hESCs towards the megakaryocyte (Mk lineage using a 'spin embryoid body' method in serum-free differentiation medium. METHODOLOGY AND PRINCIPAL FINDINGS: Immunophenotypic analyses of differentiating hESC identified a subpopulation of cells expressing high levels of CD41a that expressed other markers associated with the Mk lineage, including CD110, CD42b and CD61. Differentiated cells were sorted on the basis of their expression of CD41a, CD34 and CD45 and assessed for Mk colony formation, expression of myeloid and Mk genes and ability to endoreplicate DNA. In a collagen-based colony assay, the CD41a⁺ cells sorted from these differentiation cultures produced 100-800 Mk progenitors at day 13 and 25-160 Mk progenitors at day 20 of differentiation per 100,000 cells assayed. Differentiated Mk cells produced platelet-like particles which expressed CD42b and were activated by ADP, similar to platelets generated from precursors in cord blood. These studies were complemented by real time PCR analyses showing that subsets of cells enriched for CD41a⁺ Mk precursors expressed high levels of Mk associated genes such as PF4 and MPL. Conversely, high levels of myeloid and erythroid related transcripts, such as GATA1, TAL1/SCL and PU.1, were detected in sorted fractions containing CD34⁺ and CD45⁺ cells. CONCLUSIONS: We describe a serum- and feeder-free culture system that enabled the generation of Mk progenitors from human embryonic stem cells. These cells formed colonies that included differentiated Mks that fragmented to form platelet-like particles. This protocol represents an important step towards the generation of human platelets for therapeutic use.

  9. A feeder- and xeno-free human induced pluripotent stem cell line obtained from primary human dermal fibroblasts with epigenetic repression of reprogramming factors expression: GPCCi001-A

    Directory of Open Access Journals (Sweden)

    Michał Stefan Lach

    2017-04-01

    Full Text Available The primary human dermal fibroblasts (PHDFs from breast cancer patient were obtained to generate the human induced pluripotent stem cell line GPCCi001-A via lentiviral transfection. Thus, a modified EF1a-hSTEMCCA-loxP with tetO operator which regulates transgene expression was used. This method takes advantage of epigenetic regulation of transcription and allows for stable silencing of the reprogramming factors in obtained hiPS cells. To increase the potential utility of hiPSCs for clinical applications, they were adapted to feeder- and xeno-free conditions. The pluripotency of GPCCi001-A cell line and ability to differentiate into three germ layers was confirmed.

  10. Mass transfer effects in feeder flow-accelerated corrosion wall thinning

    International Nuclear Information System (INIS)

    Pietralik, J.

    2008-01-01

    Flow conditions play a dominant role in Flow-Accelerated Corrosion (FAC) under certain conditions, e.g., in CANDU feeders. While chemistry and materials set the overall potential for FAC, flow conditions determine the local distribution of wall thinning. Recent plant data of feeders and laboratory tests confirms that there is a close relationship between local flow conditions, expressed by mass transfer coefficient, and FAC rate in CANDU feeder bends. The knowledge of local effects can be useful for minimizing the number of inspected components, predicting the location of the highest FAC rate for a given piping component, and determining what components or feeders should be replaced. A similar evaluation applies also to FAC in heat transfer equipment such as heat exchangers and steam generators. The objective of this paper is to examine the relationship between FAC rate and local mass transfer parameters. For FAC where the flow is dominant, the FAC rate is proportional to mass flux of ferrous ions. The mass flux is the product of the mass transfer coefficient and the concentration difference, or degree of saturation. The mass transfer coefficient describes the intensity of the transport of corrosion products (ferrous ions) from the oxide-water interface into the bulk water. Therefore, this parameter can be used for predicting the local distribution of FAC rate in the mass-transfer controlled FAC. The degree of saturation reduces the mass flux, thus reducing the FAC rate. This effect can be significant in long piping, e.g., in outlet feeders. The paper presents plant and laboratory evidence for the relationship between local mass transfer conditions and the FAC rate. It shows correlations for mass transfer coefficient in components that are highly susceptible to FAC and most important flow parameters that affect mass transfer coefficient. The role of surface roughness, wall shear stress, and local turbulence is also discussed. (author)

  11. Evaluation of options for life cycle management of feeder cracking at the Point Lepreau Generating Station

    International Nuclear Information System (INIS)

    Gendron, T.S.; Slade, J.P.

    2003-01-01

    The CANDU industry has a predictive capability for most Heat Transport System (HTS) degradation issues that allows utilities to apply cost-effective maintenance programs. The standard approach for maintenance programs is focussed inspection and planned replacement. Some examples of degradation issues with deterministic failure rates are feeder thinning, and pressure tube elongation and deuterium ingress. However, the cracking observed in Point Lepreau Generating Station (PLGS) outlet feeder first bends is one notable exception to this behaviour. A predictive capability for feeder cracking does not currently exist for several reasons. First, the mechanism of feeder cracking, stress corrosion cracking (SCC), has to some degree a random nature. Second, although a probable environment causing cracking has been identified, the precise stress and environmental conditions for feeder crack initiation and propagation have not been defined. Finally, the very low incidence of feeder cracking observed to-date (four, all at PLGS) precludes a probabilistic or statistical prediction of failure rate. Generally, utilities select a Life Cycle Management Plan that ensures safe operation and has the lowest Net Present Value cost. In preparing a Feeder Life Cycle Management Plan, New Brunswick Power (NBP) has recognized that the Net Present Value cost is very sensitive to failure rate. Since the failure rate for feeder cracking is not well defined, the following three scenarios were considered to bound the probability of future failures at PLGS. (author)

  12. Computational fluid dynamics analysis for flow accelerated corrosion in CANDU6 feeder pipes

    International Nuclear Information System (INIS)

    Catana, A.; Pauna, E.; Ioan, M.

    2013-01-01

    CANDU6 plant management over a long time period includes various ageing and degradation mechanisms like FAC manifested mainly at first and second elbow of CANDU6 outlet feeders. FAC take place at all CANDU6 built before 2000 year with feeders made from SA106 grade B low alloy carbon-steel (with chromium at 0.02%). CFD method is used in this paper to investigate the feeder's wall thinning process taking place mainly due local flow conditions in complex 3D geometrical configurations. The 380 outlet feeders grouped in 2.5'' (320) and 2.0'' feeders (60). The objective of this paper is to help, as much as possible, to focus investigation on most probable maximum thinning rate locations through 3D distribution of some TH parameters. Application of CFD methods in CANDU6 nuclear reactors implies the knowledge of real plant operating data like: long term time averaged channel power and mass flow as well as temperature, pressure, pHa etc allowing the optimization and cost reduction of wall thinning monitoring process at CANDU6 nuclear power plants. (authors)

  13. Counseling in the Elementary Feeder Schools.

    Science.gov (United States)

    Dunham, Virginia

    This brief paper presents the concept of transition counseling between a junior high school and its feeder school(s), designed to make the change from elementary into junior high less traumatic. Aside from routine sixth grade counseling, the counselors expanded their base of counseling to include all types of problems as well as all grade levels.…

  14. Configuration System for Simulation Based Design of Vibratory Bowl Feeders

    DEFF Research Database (Denmark)

    Hansson, Michael Natapon; Mathiesen, Simon; Ellekilde, Lars-Peter

    2017-01-01

    Vibratory bowl feeders are still among the most commonly used production equipment for automated part feeding, where parts are correctly oriented for further manipulation by being conveyed through a set of orienting devices. Designing vibratory bowl feeders involves selecting and sequencing...... a number of these devices that either reorients or rejects the part until a desirable orientation is achieved. To aid the designer in this task, this work presents a configuration system where knowledge of the behaviour for each device is acquired through dynamic simulation, and used to solve...

  15. Caspase-independent cell death mediated by apoptosis-inducing factor (AIF) nuclear translocation is involved in ionizing radiation induced HepG2 cell death

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Hengwen [Department of Radiation, Cancer Center of Guangdong General Hospital (Guangdong Academy of Medical Science), Guangzhou, 510080, Guangdong (China); Yang, Shana; Li, Jianhua [Department of Physiology, Guangzhou Medical University, Guangzhou, 510182, Guangdong (China); Zhang, Yajie [Department of Pathology, Guangzhou Medical University, Guangzhou, 510182, Guangdong (China); Gao, Dongsheng [Department of Oncology, Guangdong Medical College Affiliated Pengpai Memorial Hospital, Hai Feng, 516400, Gungdong (China); Zhao, Shenting, E-mail: zhaoshenting@126.com [Department of Physiology, Guangzhou Medical University, Guangzhou, 510182, Guangdong (China)

    2016-03-25

    Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world. The aim of radiotherapy is to eradicate cancer cells with ionizing radiation. Except for the caspase-dependent mechanism, several lines of evidence demonstrated that caspase-independent mechanism is directly involved in the cell death responding to irradiation. For this reason, defining the contribution of caspase-independent molecular mechanisms represents the main goal in radiotherapy. In this study, we focused on the role of apoptosis-inducing factor (AIF), the caspase-independent molecular, in ionizing radiation induced hepatocellular carcinoma cell line (HepG2) cell death. We found that ionizing radiation has no function on AIF expression in HepG2 cells, but could induce AIF release from the mitochondria and translocate into nuclei. Inhibition of AIF could reduce ionizing radiation induced HepG2 cell death. These studies strongly support a direct relationship between AIF nuclear translocation and radiation induced cell death. What's more, AIF nuclear translocation is caspase-independent manner, but not caspase-dependent manner, in this process. These new findings add a further attractive point of investigation to better define the complex interplay between caspase-independent cell death and radiation therapy. - Highlights: • AIF nuclear translocation is involved in ionizing radiation induced hepatocellular carcinoma cell line HepG2 cell death. • AIF mediated cell death induced by ionizing radiation is caspase-independent. • Caspase-independent pathway is involved in ionzing radiation induced HepG2 cell death.

  16. Caspase-independent cell death mediated by apoptosis-inducing factor (AIF) nuclear translocation is involved in ionizing radiation induced HepG2 cell death

    International Nuclear Information System (INIS)

    Sun, Hengwen; Yang, Shana; Li, Jianhua; Zhang, Yajie; Gao, Dongsheng; Zhao, Shenting

    2016-01-01

    Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world. The aim of radiotherapy is to eradicate cancer cells with ionizing radiation. Except for the caspase-dependent mechanism, several lines of evidence demonstrated that caspase-independent mechanism is directly involved in the cell death responding to irradiation. For this reason, defining the contribution of caspase-independent molecular mechanisms represents the main goal in radiotherapy. In this study, we focused on the role of apoptosis-inducing factor (AIF), the caspase-independent molecular, in ionizing radiation induced hepatocellular carcinoma cell line (HepG2) cell death. We found that ionizing radiation has no function on AIF expression in HepG2 cells, but could induce AIF release from the mitochondria and translocate into nuclei. Inhibition of AIF could reduce ionizing radiation induced HepG2 cell death. These studies strongly support a direct relationship between AIF nuclear translocation and radiation induced cell death. What's more, AIF nuclear translocation is caspase-independent manner, but not caspase-dependent manner, in this process. These new findings add a further attractive point of investigation to better define the complex interplay between caspase-independent cell death and radiation therapy. - Highlights: • AIF nuclear translocation is involved in ionizing radiation induced hepatocellular carcinoma cell line HepG2 cell death. • AIF mediated cell death induced by ionizing radiation is caspase-independent. • Caspase-independent pathway is involved in ionzing radiation induced HepG2 cell death.

  17. Co-ordination of directional overcurrent protection with load current for parallel feeders

    Energy Technology Data Exchange (ETDEWEB)

    Wright, J.W.; Lloyd, G.; Hindle, P.J. [Alstom, Inc., Stafford (United Kingdom). T and D Protection and Control

    1999-11-01

    Directional phase overcurrent relays are commonly applied at the receiving ends of parallel feeders or transformer feeders. Their purpose is to ensure full discrimination of main or back-up power system overcurrent protection for a fault near the receiving end of one feeder. This paper reviews this type of relay application and highlights load current setting constraints for directional protection. Such constraints have not previously been publicized in well-known text books. A directional relay current setting constraint that is suggested in some text books is based purely on thermal rating considerations for older technology relays. This constraint may not exist with modern numerical relays. In the absence of any apparent constraint, there is a temptation to adopt lower current settings with modern directional relays in relation to reverse load current at the receiving ends of parallel feeders. This paper identifies the danger of adopting very low current settings without any special relay feature to ensure protection security with load current during power system faults. A system incident recorded by numerical relays is also offered to highlight this danger. In cases where there is a need to infringe the identified constraints an implemented and testing relaying technique is proposed.

  18. Response of deposit-feeders to exclusion of epibenthic predators in a Mediterranean intertidal flat

    NARCIS (Netherlands)

    Como, S.; Rossi, F.; Lardicci, C.

    2004-01-01

    Deposit-feeders are common components of macrofaunal assemblages in intertidal soft sediments. Predation has been considered to have a central role in affecting their distribution and population dynamics. This study investigates the effect of epibenthic predators on deposit-feeders, inhabiting the

  19. wnt3a but not wnt11 supports self-renewal of embryonic stem cells

    International Nuclear Information System (INIS)

    Singla, Dinender K.; Schneider, David J.; LeWinter, Martin M.; Sobel, Burton E.

    2006-01-01

    wnt proteins (wnts) promote both differentiation of midbrain dopaminergic cells and self-renewal of haematopoietic stem cells. Mouse embryonic stem (ES) cells can be maintained and self-renew on mouse feeder cell layers or in media containing leukemia inhibitory factor (LIF). However, the effects of wnts on ES cells self-renewal and differentiation are not clearly understood. In the present study, we found that conditioned medium prepared from L cells expressing wnt3a can replace feeder cell layers and medium containing LIF in maintaining ES cells in the proliferation without differentiation (self-renewal) state. By contrast, conditioned medium from NIH3T3 cells expressing wnt11 did not. Alkaline phosphatase staining and compact colony formation were used as criteria of cells being in the undifferentiated state. ES cells maintained in medium conditioned by Wnt3a expressing cells underwent freezing and thawing while maintaining properties seen with LIF maintained ES cells. Purified wnt3a did not maintain self-renewal of ES cells for prolonged intervals. Thus, other factors in the medium conditioned by wnt3a expressing cells may have contributed to maintenance of ES cells in a self-renewal state. Pluripotency of ES cells was determined with the use of embryoid bodies in vitro. PD98059, a MEK specific inhibitor, promoted the growth of undifferentiated ES cells maintained in conditioned medium from wnt3a expressing cells. By contrast, the P38 MAPK inhibitor SB230580 did not, suggesting a role for the MEK pathway in self-renewal and differentiation of ES cells maintained in the wnt3a cell conditioned medium. Thus, our results show that conditioned medium from wnt3a but not wnt11 expressing cells can maintain ES cells in self-renewal and in a pluripotent state

  20. harmonic load modeling: a case study of 33 kv abuja steel mill feeder

    African Journals Online (AJOL)

    HOD

    techniques are adopted. This paper studied the harmonic orders of the 33 kV Abuja Steel Feeder modeled as a ... (ETAP) software package was deployed to perform Discrete Fast Transform (DFT) while the input ... and documented in research and development articles ... network with 33kV Abuja Steel feeder as case study.

  1. Variations in Humanized and Defined Culture Conditions Supporting Derivation of New Human Embryonic Stem Cell Lines

    DEFF Research Database (Denmark)

    Fletcher, Judy M; Ferrier, Patricia M; Gardner, John O

    2006-01-01

    matrix substrate of purified human laminin (Ln) with transitional reliance on mitotically inactivated human fibroblast (HDF) feeder cells. With this integrated system hESC lines were isolated using either HDF conditioned medium supplemented with a bovine-sourced serum replacement (bSRM), or a defined...

  2. Diagnostic approaches for viruses and prions in stem cell banks

    International Nuclear Information System (INIS)

    Cobo, Fernando; Talavera, Paloma; Concha, Angel

    2006-01-01

    Some stem cell lines may contain an endogenous virus or can be contaminated with exogenous viruses (even of animal origin) and may secrete viral particles or express viral antigens on their surface. Moreover, certain biotechnological products (e.g. bovine fetal serum, murine feeder cells) may contain prion particles. Viral and prion contamination of cell cultures and 'feeder' cells, which is a common risk in all biotechnological products derived from the cell lines, is the most challenging and potentially serious outcome to address, due to the difficulty involved in virus and prion detection and the potential to cause serious disease in recipients of these cell products. Stem cell banks should introduce adequate quality assurance programs like the microbiological control program and can provide researchers with valuable support in the standardization and safety of procedures and protocols used for the viral and prion testing and in validation programs to assure the quality and safety of the cells

  3. Impact of Distribution Feeders that do not have Voltage Regulators on the number of Charged Electric Vehicles using IEEE 34 Bus Test Feeder

    Energy Technology Data Exchange (ETDEWEB)

    Allehyani, Ahmed [University of Southern California, Department of Electrical Engineering; Beshir, Mohammed [University of Southern California, Department of Electrical Engineering

    2015-02-01

    Voltage regulators help maintain an acceptable voltage profile for the system. This paper discusses the effect of installing voltage regulators to the system to fix the voltage drop resulting from the electrical vehicles loading increase when they are being charged. The effect will be studied in the afternoon, when the peak load occurs, using the IEEE 34 bus test feeder. First, only one spot node is used to charge the electric vehicles while a voltage regulator is present. Second, five spot nodes are loaded at the same time to charge the electric vehicles while voltage regulators are installed at each node. After that, the impact of electric vehicles on distribution feeders that do not have voltage regulators will appear.

  4. The effect of isolation and culture methods on epithelial stem cell populations and their progeny-toward an improved cell expansion protocol for clinical application.

    Science.gov (United States)

    Lenihan, Catherine; Rogers, Caroline; Metcalfe, Anthony D; Martin, Yella H

    2014-12-01

    The use of cultured epithelial keratinocytes in the treatment of burns and skin graft donor sites is well established in clinical practice. The most widely used culture method for clinical use was originally developed by Rheinwald and Green 40 years ago. This system uses irradiated mouse dermal fibroblasts as a feeder cell layer to promote keratinocyte growth, a process that is costly and labor-intensive for health care providers. The medium formulation contains several components of animal origin, which pose further safety risks for patients. Improvements and simplification in the culturing process would lead to clear advantages: improved safety through reduction of xenobiotic components and reduction in cost for health care providers by dispensing with feeder cells. We compared the Rheinwald and Green method to culture in three commercially available, feeder-free media systems with defined/absent components of animal origin. During the isolation process, short incubation times in high-strength trypsin resulted in increased numbers of liberated keratinocyte stem cells compared with longer incubation times. All three commercially available media tested in this study could support the expansion of keratinocytes, with phenotypes comparable to cells expanded using the established Rheinwald and Green method. Growth rates varied, with two of the media displaying comparable growth rates, whereas the third was significantly slower. Our study demonstrates the suitability of such feeder-free media systems in clinical use. It further outlines a range of techniques to evaluate keratinocyte phenotype when assessing the suitability of cells for clinical application. Copyright © 2014 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  5. Optimization of Human NK Cell Manufacturing: Fully Automated Separation, Improved Ex Vivo Expansion Using IL-21 with Autologous Feeder Cells, and Generation of Anti-CD123-CAR-Expressing Effector Cells.

    Science.gov (United States)

    Klöß, Stephan; Oberschmidt, Olaf; Morgan, Michael; Dahlke, Julia; Arseniev, Lubomir; Huppert, Volker; Granzin, Markus; Gardlowski, Tanja; Matthies, Nadine; Soltenborn, Stephanie; Schambach, Axel; Koehl, Ulrike

    2017-10-01

    The administration of ex vivo expanded natural killer (NK) cells as potential antitumor effector cells appears to be suitable for effector cell-based immunotherapies in high-risk cancer patients. However, good manufacturing practice (GMP)-compliant manufacturing of clinical-grade NK cells at sufficiently high numbers represents a great challenge. Therefore, previous expansion protocols for those effector cells were improved and optimized by using newly developed culture medium, interleukin (IL)-21, and autologous feeder cells (FCs). Separation of primary human NK cells (CD56 + CD3 - ) was carried out with the CliniMACS Prodigy ® in a single process, starting with approximately 1.2 × 10 9 leukocytes collected by small-scale lymphapheresis or from buffy coats. Enriched NK cells were adjusted to starting cell concentrations within approximately 1 × 10 6 effector cells/mL and cultured in comparative expansion experiments for 14 days with IL-2 (1,000 IU/mL) in different GMP-compliant media (X-VIVO ™ 10, CellGro ® , TexMACS ™ , and NK MACS ® ). After medium optimization, beneficial effects for functionality and phenotype were investigated at the beginning of cell expansion with irradiated (25 Gy) autologous FCs at a ratio of 20:1 (feeder: NK) in the presence or absence of IL-21 (100 ng/mL). Additionally, expanded NK cells were gene modified to express chimeric antigen receptors (CARs) against CD123, a common marker for acute myeloid leukemia (AML). Cytotoxicity, degranulation, and cytokine release of transduced NK cells were determined against KG1a cells in flow cytometric analysis and fluorescent imaging. The Prodigy manufacturing process revealed high target cell viabilities (median 95.4%), adequate NK cell recovery (median 60.4%), and purity of 95.4% in regard to CD56 + CD3 - target cells. The process in its early phase of development led to a median T-cell depletion of log 3.5 after CD3 depletion and log 3.6 after the whole process, including CD3

  6. Culture conditions for bovine embryonic stem cell-like cells isolated from blastocysts after external fertilization

    OpenAIRE

    Jin, Muzi; Wu, Asga; Dorzhin, Sergei; Yue, Qunhua; Ma, Yuzhen; Liu, Dongjun

    2012-01-01

    Although isolation and characterization of embryonic stem cells have been successful in cattle, maintenance of bovine embryonic stem cells in culture remains difficult. In this study, we compared different methods of cell passaging, feeder cell layers and medium conditions for bovine embryonic stem cell-like cells. We found that a murine embryonic fibroblast feeder layer is more suitable for embryonic stem cell-like cells than bovine embryonic fibroblasts. When murine embryonic fibroblasts we...

  7. Research, Development and Demonstration of Peak Load Reduction on Distribution Feeders Using Distributed Energy Resources for the City of Fort Collins

    Energy Technology Data Exchange (ETDEWEB)

    Sumner, Dennis [City of Fort Collins Utilities, CO (United States); Vosburg, Tom [City of Fort Collins Utilities, CO (United States); Brunner, Steve [Brendle Group, Fort Collins, CO (United States); Gates, Judy [Woodward, Inc., Fort Collins, CO (United States); Howard, Nathan [Spirae, Inc., Fort Collins, CO (United States); Merton, Andrew [Spirae, Inc., Fort Collins, CO (United States); Wright, Don [Spirae, Inc., Fort Collins, CO (United States); Birlingmair, Doug [Spirae, Inc., Fort Collins, CO (United States)

    2015-10-01

    This project titled “Research, Development and Demonstration of Peak Load Reduction on Distribution Feeders Using Distributed Energy Resources for the City of Fort Collins” evolved in response to the Department of Energy’s (DOE) Funding Opportunity Announcement (FOA) Number DE-PS26-07NT43119. Also referred to as the Fort Collins Renewable and Distributed System Integration (RDSI) Project, the effort was undertaken by a diverse group of local government, higher education and business organizations; and was driven by three overarching goals: I. Fulfill the requirements of the DOE FOA’s Area of Interest 2: Renewable and Distributed System Integration; most notably, to demonstrate the ability to reduce electric system distribution feeder peak load by 15% or more through the coordinated use of Distributed Energy Resources (DER). II. Advance the expertise, technologies and infrastructure necessary to support the long term vision of the Fort Collins Zero Energy District (FortZED) and move towards creating a zero energy district in the Fort Collins “Old Town” area. III. Further the goals of the City of Fort Collins Energy Policy, including the development of a Smart Grid-enabled distribution system in Fort Collins, expanded use of renewable energy, increased energy conservation, and peak load reduction. Through the collaborative efforts of the partner organizations, the Fort Collins RDSI project was successful in achieving all three of these goals. This report is organized into two distinct sections corresponding to the two phases of the project: • Part 1: Feeder Peak Load Reduction and the FortZED Initiative. • Part 2: Forming and Operating Utility Microgrids and Managing Load and Production Variability The original project scope addressed the Part 1 feeder peak load reduction. That work took place from 2009 through 2011 and was largely complete when the project scope was amended to include a demonstration of microgrid operations. While leveraging the

  8. Vertical linear feeder to elliptical igneous saucer-shaped sills: evidences from structural observations, geochemistry and experimental modeling

    Science.gov (United States)

    Galerne, C. Y.; Galland, O.; Neumann, E. R.; Planke, S.

    2009-12-01

    The structural relationships between sills and their feeders are poorly documented because they are rarely observed in the field and difficult to image on seismic data. For instance, it is unclear whether sills are fed by pipes, dikes or other sills. Nevertheless, the geometrical relationships between sills and their feeders provide first-order constraints on magma emplacement mechanisms. Here, we investigate the structural and geochemical relationships between sills and potential feeder dikes in a remarkably well-preserved and exposed sill complex, the Golden Valley Sill Complex (GVSC), Karoo Basin, South Africa. The GVSC consists of five major saucer-shaped sills and six dikes. The Golden Valley sill itself is an elliptical saucer, with a N-S trend. A one meter thick dike (D4) crops out underneath the southern tip of the Golden Valley sill. The strike of this dike is parallel to the long axis of the Golden Valley sill. Detailed sampling and geochemical analyses of the GVSC show that each sill and dike exhibits a specific geochemical signature. The Golden Valley sill and its underlying dike D4 have identical signatures. Although there is no clear structural evidence, the consistent geometrical and geochemical relationships between the Golden Valley sill and the D4 dike suggest that this vertical linear structure is the feeder of the overlying saucer-shaped sill. In order to investigate the relationships between sills and feeders, we resorted to scaled laboratory experiments. The experiments consisted of a low-viscosity vegetable oil representing magma and a cohesive fine-grained silica flour representing brittle rocks. We placed a horizontal weak layer into the silica flour, just above the top of the inlet, to simulate strata. Such a weak layer controlled the formation of horizontal sill that subsequently turned into a transgressive sheet leading to the formation of a saucer geometry. We ran experiments with varying inlet shapes: 1) a point inlet representing a

  9. Network integration modelling of feeder and BRT(bus rapid transit) to reduce the usage of private vehicles in Palembang’s suburban area

    Science.gov (United States)

    Nur'afalia, D.; Afifa, F.; Rubianto, L.; Handayeni, K. D. M. E.

    2018-01-01

    The aim of this research is to determine the optimal feeder network route that integrates with BRT (Bus Rapid Transit). Palembang, a high growing population city with unresolved transportation demand sector. BRT as main public transportation could not fulfill people’s demand in transportation, especially in Alang-Alang Lebar sub-district. As an impact, the usage of private vehicles increases along the movement toward the city center. The concept of Network Integration that integrates feeder network with BRT is expected to be a solution to suppress the rate of private vehicles’ usage and to improve public transportation service, so that the use of BRT will be increased in the suburban area of Palembang. The method used to identifying the optimal route using Route Analysis method is route analysis using Tranetsim 0.4. The best route is obtained based on 156 movement samples. The result is 58,7% from 199 mobility’s potency of private vehicle usage’s can be reduced if there is a feeder network’s route in Alang-Alang Lebar’s sub-district. From the result, the existance of integration between feeder network and BRT is potential enough to reduce the usage of private vehicles and supports the sustainability of transportation mobility in Palembang City.

  10. Immune suppressor factor confers stromal cell line with enhanced supporting activity for hematopoietic stem cells

    International Nuclear Information System (INIS)

    Nakajima, Hideaki; Shibata, Fumi; Fukuchi, Yumi; Goto-Koshino, Yuko; Ito, Miyuki; Urano, Atsushi; Nakahata, Tatsutoshi; Aburatani, Hiroyuki; Kitamura, Toshio

    2006-01-01

    Immune suppressor factor (ISF) is a subunit of the vacuolar ATPase proton pump. We earlier identified a short form of ISF (ShIF) as a stroma-derived factor that supports cytokine-independent growth of mutant Ba/F3 cells. Here, we report that ISF/ShIF supports self-renewal and expansion of primary hematopoietic stem cells (HSCs). Co-culture of murine bone marrow cells with a stromal cell line overexpressing ISF or ShIF (MS10/ISF or MS10/ShIF) not only enhanced their colony-forming activity and the numbers of long-term culture initiating cells, but also maintained the competitive repopulating activity of HSC. This stem cell supporting activity depended on the proton-transfer function of ISF/ShIF. Gene expression analysis of ISF/ShIF-transfected cell lines revealed down-regulation of secreted frizzled-related protein-1 and tissue inhibitor of metalloproteinase-3, and the restoration of their expressions in MS10/ISF cells partially reversed its enhanced LTC-IC supporting activity to a normal level. These results suggest that ISF/ShIF confers stromal cells with enhanced supporting activities for HSCs by modulating Wnt-activity and the extracellular matrix

  11. Evaluation of the effect of radiation levels and dose rates in irradiation of murine fibroblasts used as a feeder layer in the culture of human keratinocytes

    International Nuclear Information System (INIS)

    Yoshito, Daniele; Almeida, Tiago L.; Santin, Stefany Plumeri; Somessari, Elizabeth S.R.; Silveira, Carlos G. da; Mathor, Monica B.; Altran, Silvana C.; Isaac, Cesar

    2009-01-01

    In 1975, Rheinwald and Green published an effective methodology for obtaining and cultivating human keratinocytes. This methodology consisted of seeding keratinocytes onto a feeder layer composed of lineage 3T3 murine fibroblasts, the proliferation rate of which is then controlled through the action of ionizing radiation. The presence of the feeder layer encourages the development of keratinocyte colonies and their propagation in similar cultures, becoming possible several clinical applications as skin substitutes or wound dressings in situations such as post burn extensive skin loss and other skin disorders. However, good development of these keratinocytes depends on a high quality feeder layer among other factors. In the present work, we evaluated the relationship between radiation levels and dose rates applied to fibroblasts used in construction of feeder layers and the radiation effect on keratinocytes colonies forming efficiency. Results indicate 3T3 lineage murine fibroblasts irradiated with doses varying between 60 and 100 Gy can be used as a feeder layer immediately after irradiation or storage of the irradiated cells in suspension at 4 g C for 24 hours with similar results. The exception is when the irradiation dose rate is 2.75 Gyh -1 ; in this case, results suggested that the fibroblasts should be used immediately after irradiation. (author)

  12. Development of I and C system for the coal feeder of coal firing plant

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Teak Soo; Park, Chan Ho [Korea Electric Power Corp. (KEPCO), Taejon (Korea, Republic of). Research Center

    1996-12-31

    KECC(Kepco Coal Feeder Control System) receives coal weight, conveyor speed and boiler demand signals. It controls coal flow by generating speed signal of feeder which conveys coal in hopper to pulverizer, displaying measured coal quantity and providing local auto and manual manipulator (author). 33 figs.

  13. The Use of Filter-feeders to Manage Disease in a Changing World.

    Science.gov (United States)

    Burge, Colleen A; Closek, Collin J; Friedman, Carolyn S; Groner, Maya L; Jenkins, Cody M; Shore-Maggio, Amanda; Welsh, Jennifer E

    2016-10-01

    Rapid environmental change is linked to increases in aquatic disease heightening the need to develop strategies to manage disease. Filter-feeding species are effective biofilters and can naturally mitigate disease risk to humans and wildlife. We review the role of filter-feeders, with an emphasis on bivalves, in altering disease outcomes via augmentation and reduction. Filtration can reduce transmission by removing pathogens from the water column via degradation and release of pathogens in pseudofeces. In other cases, filtration can increase pathogen transmission and disease risk. The effect of filtration on pathogen transmission depends on the selectivity of the filter-feeder, the degree of infectivity by the pathogen, the mechanism(s) of pathogen transmission and the ability of the pathogen to resist degradation. For example, some bacteria and viruses can resist degradation and accumulate within a filter-feeder leading to disease transmission to humans and other wildlife upon ingestion. Since bivalves can concentrate microorganisms, they are also useful as sentinels for the presence of pathogenic microorganisms. While somewhat less studied, other invertebrates, including ascidians and sponges may also provide ecosystem services by altering pathogen transmission. In all scenarios, climate change may affect the potential for filter-feeders to mitigate disease risk. We conclude that an assessment including empirical data and modeling of system-wide impacts should be conducted before selection of filter-feeders to mitigate disease. Such studies should consider physiology of the host and microbe and risk factors for negative impacts including augmentation of other pathogens. © The Author 2016. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.

  14. High-speed optical feeder-link system using adaptive optics

    Science.gov (United States)

    Arimoto, Yoshinori; Hayano, Yutaka; Klaus, Werner

    1997-05-01

    We propose a satellite laser communication system between a ground station and a geostationary satellite, named high- speed optical feeder link system. It is based on the application of (a) high-speed optical devices, which have been developed for ground-based high-speed fiber-optic communications, and (b) the adaptive optics which compensates wavefront distortions due to atmospheric turbulences using a real time feedback control. A link budget study shows that a system with 10-Gbps bit-rate are available assuming the state-of-the-art device performance of the Er-doped fiber amplifier. We further discuss preliminary measurement results of the atmospheric turbulence at the telescope site in Tokyo, and present current study on the design of the key components for the feeder-link laser transceiver.

  15. A hybrid evolutionary algorithm for distribution feeder reconfiguration

    Indian Academy of Sciences (India)

    the reconfiguration of distribution networks has been proposed by .... An effective strategy to increase the loading margin of heavily loaded feeders is to ... social animals such as a flock of birds, a school of fish or a group of people that pursue.

  16. Digital optical feeder links system for broadband geostationary satellite

    Science.gov (United States)

    Poulenard, Sylvain; Mège, Alexandre; Fuchs, Christian; Perlot, Nicolas; Riedi, Jerome; Perdigues, Josep

    2017-02-01

    An optical link based on a multiplex of wavelengths at 1.55μm is foreseen to be a valuable solution for the feeder link of the next generation of high-throughput geostationary satellite. The main satellite operator specifications for such link are an availability of 99.9% over the year, a capacity around 500Gbit/s and to be bent-pipe. Optical ground station networks connected to Terabit/s terrestrial fibers are proposed. The availability of the optical feeder link is simulated over 5 years based on a state-of-the-art cloud mask data bank and an atmospheric turbulence strength model. Yearly and seasonal optical feeder link availabilities are derived and discussed. On-ground and on-board terminals are designed to be compliant with 10Gbit/s per optical channel data rate taking into account adaptive optic systems to mitigate the impact of atmospheric turbulences on single-mode optical fiber receivers. The forward and return transmission chains, concept and implementation, are described. These are based on a digital transparent on-off keying optical link with digitalization of the DVB-S2 and DVB-RCS signals prior to the transmission, and a forward error correcting code. In addition, the satellite architecture is described taking into account optical and radiofrequency payloads as well as their interfaces.

  17. Mesenchymal stem cells from the Wharton's jelly of umbilical cord segments provide stromal support for the maintenance of cord blood hematopoietic stem cells during long-term ex vivo culture.

    Science.gov (United States)

    Bakhshi, Tiki; Zabriskie, Ryan C; Bodie, Shamanique; Kidd, Shannon; Ramin, Susan; Paganessi, Laura A; Gregory, Stephanie A; Fung, Henry C; Christopherson, Kent W

    2008-12-01

    Hematopoietic stem cells (HSCs) are routinely obtained from marrow, mobilized peripheral blood, and umbilical cord blood. Mesenchymal stem cells (MSCs) are traditionally isolated from marrow. Bone marrow-derived MSCs (BM-MSCs) have previously demonstrated their ability to act as a feeder layer in support of ex vivo cord blood expansion. However, the use of BM-MSCs to support the growth, differentiation, and engraftment of cord blood may not be ideal for transplant purposes. Therefore, the potential of MSCs from a novel source, the Wharton's jelly of umbilical cords, to act as stromal support for the long-term culture of cord blood HSC was evaluated. Umbilical cord-derived MSCs (UC-MSCs) were cultured from the Wharton's jelly of umbilical cord segments. The UC-MSCs were then profiled for expression of 12 cell surface receptors and tested for their ability to support cord blood HSCs in a long-term culture-initiating cell (LTC-IC) assay. Upon culture, UC-MSCs express a defined set of cell surface markers (CD29, CD44, CD73, CD90, CD105, CD166, and HLA-A) and lack other markers (CD45, CD34, CD38, CD117, and HLA-DR) similar to BM-MSCs. Like BM-MSCs, UC-MSCs effectively support the growth of CD34+ cord blood cells in LTC-IC assays. These data suggest the potential therapeutic application of Wharton's jelly-derived UC-MSCs to provide stromal support structure for the long-term culture of cord blood HSCs as well as the possibility of cotransplantation of genetically identical, HLA-matched, or unmatched cord blood HSCs and UC-MSCs in the setting of HSC transplantation.

  18. Supported Housing and Supported Independent Living in the Netherlands, with a Comparison with England

    NARCIS (Netherlands)

    de Heer-Wunderink, Charlotte; Visser, Ellen; Caro-Nienhuis, Annemarie; Sytema, Sjoerd; Wiersma, Durk

    Research into community housing programs for people with severe mental illness is underexposed. The Dutch UTOPIA study describes characteristics of their service users, which may predict their allocation to either supported housing or supported independent living programs. Additionally, a comparison

  19. Impact of feeder space on laying hen feeding behavior and production performance in enriched colony housing.

    Science.gov (United States)

    Oliveira, J L; Xin, H; Wu, H

    2018-05-30

    Current feeder space recommendations in laying hen welfare guidelines are inconsistent among and within countries. One determining criterion forming the recommendations (e.g. 12.0 cm/hen for the EU guideline) is that all birds can feed simultaneously. However, if there are other resources in the environment, as in enriched colony housing (ECH), it is unknown whether group-housed hens will choose to feed simultaneously. This study assesses the impact of feeder space on feeding behavior of 60 laying hens (W-36) in ECH using a ultra-high frequency radio-frequency identification-based tracking system. The feeder spaces investigated were 12.0, 9.5, 8.5 and 6.5 cm/hen, achieved by blocking portions of the overall feeder access to keep hens at the same stocking density. Each feeder space treatment, randomly assigned over the course of the experiment, lasted for 7 consecutive days. Feeding behaviors were characterized as daily time spent at the feeder (TS, min/hen-day), daily frequency of visits to the feeder (FV, #/hen-day), and maximum or average percentage of hens feeding simultaneously (MPB, APB, %). Group-average daily feed intake (FI, g/hen-day), water use (WU, g/hen-day), and hen-day egg production (HDEP, %) were also measured. The results revealed that at 12.0 cm/hen, where unoccupied feeder space was present, a maximum of 59.0±1.4% (average of 31.7±0.3%) hens fed simultaneously. No significant differences were detected among 12.0, 9.5 and 8.5 cm/hen in TS (293±10, 286±10 and 281±10 min/hen-day) and MPB (59.0±1.4, 57.3±1.4 and 53.3±1.4%) (P>0.05). The outcome of no significant differences also held true between 12.0 and 9.5 cm/hen in APB (31.7±0.3 v. 30.8±0.3%) and between 9.5 and 8.5 cm/hen in all response variables measured (P>0.05). However, there were significant differences in APB between 6.5 cm/hen and all other treatments; in TS and FV between 6.5 and 9.5 cm/hen; and in MPB between 6.5 and 12 cm/hen (P0.05). The results revealed that synchronous

  20. Demonstration of Market-Based Real-Time Electricity Pricing on a Congested Feeder

    DEFF Research Database (Denmark)

    Larsen, Emil Mahler; Pinson, Pierre; le Ray, Guillaume

    2015-01-01

    Congestion management can delay grid reinforcements needed due to the growth of distributed technologies like photovoltaics and electric vehicles. This paper presents a method of congestion management for low voltage feeders using indirect control from the smart grid demonstration EcoGrid EU, where...... prices to 1900 houses, with a virtual feeder of 28 houses receiving congestion pricing. Simulations are used to calculate the cost from using this congestion management method, while demonstration results indicate that congestion can be managed successfully....

  1. Optimisation of Trap Design for Vibratory Bowl Feeders

    DEFF Research Database (Denmark)

    Mathiesen, Simon; Sørensen, Lars Carøe; Kraft, Dirk

    2018-01-01

    Vibratory bowl feeders (VBFs) are a widely used option for industrial part feeding, but their design is still largely manual. A subtask of VBF design is determining an optimal parameter set for the passive devices, called traps, which the VBF uses to ensure correct part orientation. This paper...

  2. A simple and efficient feeder-free culture system to up-scale iPSCs on polymeric material surface for use in 3D bioprinting.

    Science.gov (United States)

    Wong, Chui-Wei; Chen, You-Tzung; Chien, Chung-Liang; Yu, Tien-Yu; Rwei, Syang-Peng; Hsu, Shan-Hui

    2018-01-01

    The 3D bioprinting and cell/tissue printing techniques open new possibilities for future applications. To facilitate the 3D bioprinting process, a large amount of living cells are required. Induced pluripotent stem cells (iPSCs) represent a promising cell source for bioprinting. However, the maintenance and expansion of undifferentiated iPSCs are expensive and time consuming. Therefore, in this study a culture method to obtain a sufficient amount of healthy and undifferentiated iPSCs in a short-term period was established. The iPSCs could be passaged for twice on tissue culture polystyrene (TCPS) dish with the conditional medium and could adapt to the feeder-free environment. Feeder-free dishes were further prepared from chitosan, chitosan-hyaluronan, silk fibroin, and polyurethane (PU1 and PU2) two-dimensional substrates. The iPSCs cultured on the chitosan substrates showed a higher proliferation rate without losing the stemness feature. Among the different materials, PU2 could be prepared as a thermoresponsive hydrogel, which was a potential ink for 3D bioprinting. The iPSCs cultured on PU2 substrates well survived when further embedded in PU2 hydrogel. Moreover, PU2 hydrogel printed with iPSCs remained structural integrity. The use of PU2 hydrogel to embed iPSCs reduced the injury to iPSCs by shear stress. These results indicate that iPSCs could be expanded on chitosan or PU2 membranes without the feeder layer and then printed in PU2 hydrogel. The combination of these steps could offer a new possibility for future applications of iPSC-based 3D bioprinting in tissue engineering. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Convex relaxation of Optimal Power Flow in Distribution Feeders with embedded solar power

    DEFF Research Database (Denmark)

    Hermann, Alexander Niels August; Wu, Qiuwei; Huang, Shaojun

    2016-01-01

    There is an increasing interest in using Distributed Energy Resources (DER) directly coupled to end user distribution feeders. This poses an array of challenges because most of today’s distribution feeders are designed for unidirectional power flow. Therefore when installing DERs such as solar...... panels with uncontrolled inverters, the upper limit of installable capacity is quickly reached in many of today’s distribution feeders. This problem can often be mitigated by optimally controlling the voltage angles of inverters. However, the optimal power flow problem in its standard form is a large...... scale non-convex optimization problem, and thus can’t be solved precisely and also is computationally heavy and intractable for large systems. This paper examines the use of a convex relaxation using Semi-definite programming to optimally control solar power inverters in a distribution grid in order...

  4. Cultivation and irradiation of human fibroblasts in a medium enriched with platelet lysate for obtaining feeder layer in epidermal cell culture; Cultivo e irradiacao de fibroblastos humanos em meio enriquecido com lisado de plaquetas para obtencao de camada de sustentacao em culturas de celulas da epiderme

    Energy Technology Data Exchange (ETDEWEB)

    Yoshito, Daniele

    2011-07-01

    For over 30 years, the use of culture medium, enriched with bovine serum, and murines fibroblasts, with the rate of proliferation controlled by irradiation or by share anticarcinogenic drugs, has been playing successfully its role in assisting in the development of keratinocytes in culture, for clinical purposes. However, currently there is a growing concern about the possibility of transmitting prions and animals viruses to transplanted patients. Taking into account this concern, the present work aims to cultivate human fibroblasts in a medium enriched with human platelets lysate and determine the irradiation dose of these cells, for obtaining feeder layer in epidermal cell culture. For carrying out the proposed objective, platelets lysis has standardized, this lysate was used for human fibroblasts cultivation and the irradiation dose enough to inhibit its duplication was evaluated. Human keratinocytes were cultivated in these feeder layers, in culture medium enriched with the lysate. With these results we conclude that the 10% platelets lysate promoted a better adhesion and proliferation of human fibroblasts and in all dose levels tested (60 to 300 Gy), these had their mitotic activity inactivated by ionizing irradiation, being that the feeder layers obtained with doses from 70 to 150 Gy were those that provided the best development of keratinocytes in medium containing 2.5% of human platelet lysate. Therefore, it was possible to standardize both the cultivation of human fibroblasts as its inactivation for use as feeder layer in culture of keratinocytes, so as to eliminate xenobiotics components. (author)

  5. A feeder protection method against the phase-phase fault using symmetrical components

    DEFF Research Database (Denmark)

    Ciontea, Catalin-Iosif; Bak, Claus Leth; Blaabjerg, Frede

    2017-01-01

    generation and relatively reduced short-circuit currents, thus resembling the electric network on a ship. The simulation results demonstrate that the proposed method of protection provides an improved performance compared to the conventional OverCurrent relays in a radial feeder with variable short......The method of symmetrical components simplifies analysis of an electric circuit during the fault and represents an important tool for the protection engineers. In this paper, the symmetrical components of the fault current are used in a new feeder protection method for the maritime applications...

  6. Supervisory control and data acquisition system development for superconducting current feeder system of SST-1

    International Nuclear Information System (INIS)

    Patel, R.; Mahesuria, G.; Gupta, N.C.; Sonara, D.; Panchal, R.; Panchal, P.; Tanna, V.L.; Pradhan, S.

    2014-01-01

    The Current Feeders System (CFS) is essentially an optimized bridge between the power supply at room temperature and Super Conducting Magnet System (SCMS) of the SST-1 machine at 4.5 K.CFS is a complex electrical and cryogenic network which consists of ten pairs of 10 KA rating helium Vapor cooled Conventional Current Leads (VCCLs), superconducting (SC) current feeder and associated components. For the safe and reliable operation of CFS, it is equipped with different physical process parameters measuring instruments like flow, pressure, temperature, level, vacuum, voltage taps and final control element like control valves, heaters, vacuum pumps etc. PLC program is developed in ladder language for acquiring and controlling the process parameters. Independent SCADA applications developed in WonderwareIntouch software for data communication from PLC, front-end Graphical User Interface (GUI), auto-manual interface, real time trends, history trends, events and alarm pages. Time synchronized communication established between CFS control system and Industrial SQL server (InSQL) Historian for centralized storage of CFS process parameters which intern provides the CFS process data to SST-1 central control room. SCADA based data acquisition and data retrieval system is found to be satisfactory during the recent SST-1 cool down experiment. This paper describes the SCADA and PLC application development and their communication to InSQL server. (author)

  7. Hyperthermic survival of Chinese hamster ovary cells as a function of cellular population density at the time of plating

    International Nuclear Information System (INIS)

    Highfield, D.P.; Holahan, E.V.; Holahan, P.K.; Dewey, W.C.

    1984-01-01

    The survival of synchronous G 1 or asynchronous Chinese hamster ovary cells in vitro to heat treatment may depend on the cellular population density at the time of heating and/or as the cells are cultured after heating. The addition of lethally irradiated feeder cells may increase survival at 10 -3 by as much as 10- to 100-fold for a variety of conditions when cells are heated either in suspension culture or as monolayers with or without trypsinization. The protective effect associated with feeder cells appears to be associated with close cell-to-cell proximity. However, when cells are heated without trypsinization about 24 hr or later after plating, when adaptation to monolayer has occurred, the protective effect is reduced; i.e., addition of feeder cells enhances survival much less, for example, about 2- to 3-fold at 10 -2 -10 -3 survival. Also, the survival of a cell to heat is independent of whether the neighboring cell in a microcolony is destined to live or die. Finally, if protective effects associated with cell density do occur and are not controlled, serious artifacts can result as the interaction of heat and radiation is studied; for example, survival curves can be moved upward, and thus changed in shape as the number of cells plated is increased with an increase in the hyperthermic treatment or radiation dose following hyperthermia. Therefore, to understand mechanisms and to obtain information relevant to populations of cells in close proximity, such as those in vivo, these cellular population density effects should be considered and understood

  8. Evaluation of the cryogenic mechanical properties of the insulation material for ITER Feeder superconducting joint

    Science.gov (United States)

    Wu, Zhixiong; Huang, Rongjin; Huang, ChuanJun; Yang, Yanfang; Huang, Xiongyi; Li, Laifeng

    2017-12-01

    The Glass-fiber reinforced plastic (GFRP) fabricated by the vacuum bag process was selected as the high voltage electrical insulation and mechanical support for the superconducting joints and the current leads for the ITER Feeder system. To evaluate the cryogenic mechanical properties of the GFRP, the mechanical properties such as the short beam strength (SBS), the tensile strength and the fatigue fracture strength after 30,000 cycles, were measured at 77K in this study. The results demonstrated that the GFRP met the design requirements of ITER.

  9. Tumor Volume Decrease via Feeder Occlusion for Treating a Large, Firm Trigone Meningioma.

    Science.gov (United States)

    Nakashima, Takuma; Hatano, Norikazu; Kanamori, Fumiaki; Muraoka, Shinsuke; Kawabata, Teppei; Takasu, Syuntaro; Watanabe, Tadashi; Kojima, Takao; Nagatani, Tetsuya; Seki, Yukio

    2018-01-01

    Trigone meningiomas are considered a surgical challenge, as they tend to be considerably large and hypervascularized at the time of presentation. We experienced a case of a large and very hard trigone meningioma that was effectively treated using initial microsurgical feeder occlusion followed by surgery in stages. A 19-year-old woman who presented with loss of consciousness was referred to our hospital for surgical treatment of a brain tumor. Radiological findings were compatible with a left ventricular trigone meningioma extending laterally in proximity to the Sylvian fissure. At initial surgery using the transsylvian approach, main feeders originating from the anterior and lateral posterior choroidal arteries were occluded at the inferior horn; however, only a small section of the tumor could initially be removed because of its firmness. Over time, feeder occlusion resulted in tumor necrosis and a 20% decrease in its diameter; the mass effect was alleviated within 1 year. The residual meningioma was then totally excised in staged surgical procedures after resection became more feasible owing to ischemia-induced partial softening of the tumor. When a trigone meningioma is large and very hard, initial microsurgical feeder occlusion in the inferior horn can be a safe and effective option, and can lead to necrosis, volume decrease, and partial softening of the residual tumor to allow for its staged surgical excision.

  10. Diatom feeding across trophic guilds in tidal flat nematodes, and the importance of diatom cell size

    Science.gov (United States)

    Moens, Tom; Vafeiadou, Anna-Maria; De Geyter, Ellen; Vanormelingen, Pieter; Sabbe, Koen; De Troch, Marleen

    2014-09-01

    We examine the capacity of nematodes from three feeding types (deposit feeder, epistrate feeder, predator) to utilize microphytobenthos (MPB), and assess whether diatom cell size and consumer body size are important drivers of their feeding. We analyzed natural stable isotope ratios of carbon and nitrogen in abundant nematode genera and a variety of carbon sources at an estuarine intertidal flat. All nematodes had δ13C indicating that MPB is their major carbon source. δ15N, however, demonstrated that only one deposit and one epistrate feeder genus obtained most of their carbon from direct grazing on MPB, whereas other deposit feeders and predators obtained at least part of their carbon by predation on MPB grazers. We then performed a microcosm experiment in which equal cell numbers of each of three differently sized strains of the pennate diatom Seminavis were offered as food to four, one and one genera of deposit feeders, epistrate feeders and predators, respectively. Previous studies have shown that all but the epistrate feeder ingest whole diatoms, whereas the epistrate feeder pierces cells and sucks out their contents. Most genera showed markedly higher carbon absorption from medium and large cells than from small ones. When considering the number of cells consumed, however, none of the nematodes which ingest whole cells exhibited a clear preference for any specific diatom size. The epistrate feeder was the smallest nematode taxon considered here, yet it showed a marked preference for large cells. These results highlight that the feeding mechanism is much more important than consumer size as a driver of particle size selection in nematodes grazing MPB.

  11. Mesenchymal stem cells from the Wharton’s jelly of umbilical cord segments provide stromal support for the maintenance of cord blood hematopoietic stem cells during long-term ex vivo culture

    Science.gov (United States)

    Bakhshi, Tiki; Zabriskie, Ryan C.; Bodie, Shamanique; Kidd, Shannon; Ramin, Susan; Paganessi, Laura A.; Gregory, Stephanie A.; Fung, Henry C.; Christopherson, Kent W.

    2012-01-01

    BACKGROUND Hematopoietic stem cells (HSCs) are routinely obtained from marrow, mobilized peripheral blood, and umbilical cord blood. Mesenchymal stem cells (MSCs) are traditionally isolated from marrow. Bone marrow–derived MSCs (BM-MSCs) have previously demonstrated their ability to act as a feeder layer in support of ex vivo cord blood expansion. However, the use of BM-MSCs to support the growth, differentiation, and engraftment of cord blood may not be ideal for transplant purposes. Therefore, the potential of MSCs from a novel source, the Wharton’s jelly of umbilical cords, to act as stromal support for the long-term culture of cord blood HSC was evaluated. STUDY DESIGN AND METHODS Umbilical cord–derived MSCs (UC-MSCs) were cultured from the Wharton’s jelly of umbilical cord segments. The UC-MSCs were then profiled for expression of 12 cell surface receptors and tested for their ability to support cord blood HSCs in a long-term culture-initiating cell (LTC-IC) assay. RESULTS Upon culture, UC-MSCs express a defined set of cell surface markers (CD29, CD44, CD73, CD90, CD105, CD166, and HLA-A) and lack other markers (CD45, CD34, CD38, CD117, and HLA-DR) similar to BM-MSCs. Like BM-MSCs, UC-MSCs effectively support the growth of CD34+ cord blood cells in LTC-IC assays. CONCLUSION These data suggest the potential therapeutic application of Wharton’s jelly–derived UC-MSCs to provide stromal support structure for the long-term culture of cord blood HSCs as well as the possibility of cotransplantation of genetically identical, HLA-matched, or unmatched cord blood HSCs and UC-MSCs in the setting of HSC transplantation. PMID:18798803

  12. The effect of space allowance and cage size on laying hens housed in furnished cages, Part II: Behavior at the feeder.

    Science.gov (United States)

    Widowski, T M; Caston, L J; Casey-Trott, T M; Hunniford, M E

    2017-09-01

    Standards for feeder (a.k.a. feed trough) space allowance (SA) are based primarily on studies in conventional cages where laying hens tend to eat simultaneously, limiting feeder space. Large furnished cages (FC) offer more total space and opportunities to perform a greater variety of behaviors, which may affect feeding behavior and feeder space requirements. Our objective was to determine the effects of floor/feeder SA on behavior at the feeder. LSL-Lite hens were housed in FC equipped with a nest, perches, and a scratch mat. Hens with SA of either 520 cm2 (Low; 8.9 cm feeder space/hen) or 748 cm2 (High; 12.8 cm feeder space/hen) per bird resulted in groups of 40 vs. 28 birds in small FC (SFC) and 80 vs. 55 in large FC (LFC). Chain feeders ran at 0500, 0800, 1100, 1400, and 1700 with lights on at 0500 and off at 1900 hours. Digital recordings of FC were scanned at chain feeder onset and every 15 min for one h after (5 scans × 5 feeding times × 2 d) to count the number of birds with their head in the feeder. All occurrences of aggressive pecks and displacements during 2 continuous 30-minute observations at 0800 h and 1700 h also were counted. Mixed model repeated analyses tested the effects of SA, cage size, and time on the percent of hens feeding, and the frequency of aggressive pecks and displacements. Surprisingly, the percent of birds feeding simultaneously was similar regardless of cage size (LFC: 23.0 ± 0.9%; SFC: 24.0 ± 1.0%; P = 0.44) or SA (Low: 23.8 ± 0.9%; High: 23.3 ± 1.0%; P = 0.62). More birds were observed feeding at 1700 h (35.3 ± 0.1%) than any at other time (P Feeder use differed by cage area (nest, middle, or scratch) over the d (P feeder competition at the Low SA in this study. © The Author 2017. Published by Oxford University Press on behalf of Poultry Science Association.

  13. Downregulation of proapoptotic Bim augments IL-2-independent T-cell transformation by human T-cell leukemia virus type-1 Tax

    International Nuclear Information System (INIS)

    Higuchi, Masaya; Takahashi, Masahiko; Tanaka, Yuetsu; Fujii, Masahiro

    2014-01-01

    Human T-cell leukemia virus type 1 (HTLV-1), an etiological agent of adult T-cell leukemia, immortalizes and transforms primary human T cells in vitro in both an interleukin (IL)-2-dependent and IL-2-independent manner. Expression of the HTLV-1 oncoprotein Tax transforms the growth of the mouse T-cell line CTLL-2 from being IL-2-dependent to IL-2-independent. Withdrawal of IL-2 from normal activated T cells induces apoptosis, which is mediated through the inducible expression of several proapoptotic proteins, including Bim. In this study, we found that Tax protects IL-2-depleted T cells against Bim-induced apoptosis. Withdrawal of IL-2 from CTLL-2 cells induced a prominent increase in the level of Bim protein in CTLL-2 cells, but not in Tax-transformed CTLL-2 cells. This inhibition of Bim in Tax-transformed CTLL-2 cells was mediated by two mechanisms: downregulation of Bim mRNA and posttranscriptional reduction of Bim protein. Transient expression of Tax in CTLL-2 cells also inhibited IL-2 depletion–induced expression of Bim, however, this decrease in Bim protein expression was not due to downregulation of Bim mRNA, thus indicating that Bim mRNA downregulation in Tax-transformed CTLL-2 occurs only after long-term expression of Tax. Transient expression of Tax in CTLL-2 cells also induced Erk activation, however, this was not involved in the reduction of Bim protein. Knockdown of Bim expression in CTLL-2 cells augmented Tax-induced IL-2-independent transformation. HTLV-1 infection of human T cells also reduced their levels of Bim protein, and restoring Bim expression in HTLV-1-infected cells reduced their proliferation by inducing apoptosis. Taken together, these results indicate that Tax-induced downregulation of Bim in HTLV-1-infected T cells promotes their IL-2-independent growth, thereby supporting the persistence of HTLV-1 infection in vivo

  14. Downregulation of proapoptotic Bim augments IL-2-independent T-cell transformation by human T-cell leukemia virus type-1 Tax.

    Science.gov (United States)

    Higuchi, Masaya; Takahashi, Masahiko; Tanaka, Yuetsu; Fujii, Masahiro

    2014-12-01

    Human T-cell leukemia virus type 1 (HTLV-1), an etiological agent of adult T-cell leukemia, immortalizes and transforms primary human T cells in vitro in both an interleukin (IL)-2-dependent and IL-2-independent manner. Expression of the HTLV-1 oncoprotein Tax transforms the growth of the mouse T-cell line CTLL-2 from being IL-2-dependent to IL-2-independent. Withdrawal of IL-2 from normal activated T cells induces apoptosis, which is mediated through the inducible expression of several proapoptotic proteins, including Bim. In this study, we found that Tax protects IL-2-depleted T cells against Bim-induced apoptosis. Withdrawal of IL-2 from CTLL-2 cells induced a prominent increase in the level of Bim protein in CTLL-2 cells, but not in Tax-transformed CTLL-2 cells. This inhibition of Bim in Tax-transformed CTLL-2 cells was mediated by two mechanisms: downregulation of Bim mRNA and posttranscriptional reduction of Bim protein. Transient expression of Tax in CTLL-2 cells also inhibited IL-2 depletion-induced expression of Bim, however, this decrease in Bim protein expression was not due to downregulation of Bim mRNA, thus indicating that Bim mRNA downregulation in Tax-transformed CTLL-2 occurs only after long-term expression of Tax. Transient expression of Tax in CTLL-2 cells also induced Erk activation, however, this was not involved in the reduction of Bim protein. Knockdown of Bim expression in CTLL-2 cells augmented Tax-induced IL-2-independent transformation. HTLV-1 infection of human T cells also reduced their levels of Bim protein, and restoring Bim expression in HTLV-1-infected cells reduced their proliferation by inducing apoptosis. Taken together, these results indicate that Tax-induced downregulation of Bim in HTLV-1-infected T cells promotes their IL-2-independent growth, thereby supporting the persistence of HTLV-1 infection in vivo. © 2014 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

  15. Culture conditions affecting the survival response of Chinese hamster ovary cells treated by hyperthermia

    International Nuclear Information System (INIS)

    Highfield, D.P.; Holahan, E.V.; Dewey, W.C.

    1982-01-01

    Using lethally irradiated feeder cells to control cell population densities, researchers investigated the survival of Chinese hamster ovary cells heated between 42.2 and 45.5 degrees C. Test cells were plated into T25 flasks with or without feeder cells, incubated 2 hours at 37 degrees C, and then given various heat treatments. Under all heating conditions, survival increased in those flasks containing feeder cells. Increased survival (by as much as a factor of 100 for cells heated at 42.4 degrees C for 6-10 hr) was most apparent when cells were heated to thermotolerance. By adjustment of test and feeder cell numbers, survival increased as density increased; however, maximum survival followed a transition period that occurred between the plating of 1 X 10(4) and 6 X 10(4) cells. Experimental artifacts due to improper control of cell density was demonstrated

  16. Faecal nitrogen of browser and mixed feeder game species during ...

    African Journals Online (AJOL)

    Faecal nitrogen of browser and mixed feeder game species during different seasons. ... A practical measure of assessing periods of potential nutritional stress of game species is needed in the management of ... AJOL African Journals Online.

  17. A probabilistic degradation model for the estimation of the remaining life distribution of feeders

    International Nuclear Information System (INIS)

    Yuan, X.-X.; Pandey, M.D.; Bickel, G.A.

    2006-01-01

    Wall thinning due to flow accelerated corrosion (FAC) is a pervasive form of degradation in the outlet feeder pipes of the primary heat transport system of CANDU reactors. The prediction of the end-of-life of a feeder from wall thickness measurement data is confounded by the sampling and temporal uncertainties associated with the FAC degradation phenomenon. Traditional regression-based statistical methods deal with only the sampling uncertainties, leaving the temporal uncertainties unresolved. This paper presents an advanced probabilistic model, which is able to integrate the temporal uncertainties into the prediction of lifetime. In particular, a random gamma process model is proposed to model the FAC process and it is calibrated with a set of wall thickness measurements using the method of maximum likelihood. This information can be used to establish an optimum strategy for inspection and replacement of feeders. (author)

  18. CXCL10 can inhibit endothelial cell proliferation independently of CXCR3.

    Directory of Open Access Journals (Sweden)

    Gabriele S V Campanella

    2010-09-01

    Full Text Available CXCL10 (or Interferon-inducible protein of 10 kDa, IP-10 is an interferon-inducible chemokine with potent chemotactic activity on activated effector T cells and other leukocytes expressing its high affinity G protein-coupled receptor CXCR3. CXCL10 is also active on other cell types, including endothelial cells and fibroblasts. The mechanisms through which CXCL10 mediates its effects on non-leukocytes is not fully understood. In this study, we focus on the anti-proliferative effect of CXCL10 on endothelial cells, and demonstrate that CXCL10 can inhibit endothelial cell proliferation in vitro independently of CXCR3. Four main findings support this conclusion. First, primary mouse endothelial cells isolated from CXCR3-deficient mice were inhibited by CXCL10 as efficiently as wildtype endothelial cells. We also note that the proposed alternative splice form CXCR3-B, which is thought to mediate CXCL10's angiostatic activity, does not exist in mice based on published mouse CXCR3 genomic sequences as an in-frame stop codon would terminate the proposed CXCR3-B splice variant in mice. Second, we demonstrate that human umbilical vein endothelial cells and human lung microvascular endothelial cells that were inhibited by CXL10 did not express CXCR3 by FACS analysis. Third, two different neutralizing CXCR3 antibodies did not inhibit the anti-proliferative effect of CXCL10. Finally, fourth, utilizing a panel of CXCL10 mutants, we show that the ability to inhibit endothelial cell proliferation correlates with CXCL10's glycosaminoglycan binding affinity and not with its CXCR3 binding and signaling. Thus, using a very defined system, we show that CXCL10 can inhibit endothelial cell proliferation through a CXCR3-independent mechanism.

  19. Feederism: an exaggeration of a normative mate selection preference?

    Science.gov (United States)

    Terry, Lesley L; Suschinsky, Kelly D; Lalumière, Martin L; Vasey, Paul L

    2012-02-01

    Quinsey and Lalumière (1995) suggested that some, if not most, paraphilias are exaggerated manifestations of more normative and functional mate selection preferences. The present study tested whether Feederism, a fat fetish focused on erotic eating, feeding, and gaining weight, is an exaggeration of a sexual arousal pattern commonly seen in the general population. Thirty participants (15 men and 15 women) recruited from the general population were assessed using penile plethysmography and vaginal photoplethysmography, respectively. None of the participants were self-identified Feeders or Feedees. Participants were shown sexual, neutral, and feeding still images while listening to audio recordings of sexual, neutral, and feeding stories. Participants did not genitally respond to feeding stimuli. However, both men and women subjectively rated feeding stimuli as more sexually arousing than neutral stimuli. We discuss the discordance between physiological and self-reported sexual arousal in the context of sex differences in sexual concordance and implications for future research.

  20. Experience of superconducting current feeders system of SST-1

    International Nuclear Information System (INIS)

    Gupta, N.C.; Garg, A.; Sonara, D.

    2014-01-01

    The superconducting current feeder system for SST-1 which has been installed and commissioned recently along with SST-1, felicitates to energize the SST-1. The CFS consists of ten pairs of 10,000 Ampere (A) rating helium vapor cooled conventional current leads, interconnecting Cu-SC joints, three numbers of cryo-compatible SC feeders ducts, current leads assembly chamber, hydraulic network and three numbers of joint boxes operated at different current rating to charge Toroidal Field and Poloidal Field coils separately. During the last three campaigns, it was possible to achieve a controlled cool down up to 4 K and showed its rated operational performance. Actively cooled liquid nitrogen shield showed temperature profile in the temperature range of 80-85K and the whole system was evacuated up to 6x10 -6 mbar. The measured LHe consumption rates from TF VCCL were 0.3 g/s and 0.35 g/s at zero current and 1 kA respectively. (author)

  1. Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture.

    Science.gov (United States)

    Dahlmann, Julia; Awad, George; Dolny, Carsten; Weinert, Sönke; Richter, Karin; Fischer, Klaus-Dieter; Munsch, Thomas; Leßmann, Volkmar; Volleth, Marianne; Zenker, Martin; Chen, Yaoyao; Merkl, Claudia; Schnieke, Angelika; Baraki, Hassina; Kutschka, Ingo; Kensah, George

    2018-01-01

    The possibility to generate cardiomyocytes from pluripotent stem cells in vitro has enormous significance for basic research, disease modeling, drug development and heart repair. The concept of heart muscle reconstruction has been studied and optimized in the rat model using rat primary cardiovascular cells or xenogeneic pluripotent stem cell derived-cardiomyocytes for years. However, the lack of rat pluripotent stem cells (rPSCs) and their cardiovascular derivatives prevented the establishment of an authentic clinically relevant syngeneic or allogeneic rat heart regeneration model. In this study, we comparatively explored the potential of recently available rat embryonic stem cells (rESCs) and induced pluripotent stem cells (riPSCs) as a source for cardiomyocytes (CMs). We developed feeder cell-free culture conditions facilitating the expansion of undifferentiated rPSCs and initiated cardiac differentiation by embryoid body (EB)-formation in agarose microwell arrays, which substituted the robust but labor-intensive hanging drop (HD) method. Ascorbic acid was identified as an efficient enhancer of cardiac differentiation in both rPSC types by significantly increasing the number of beating EBs (3.6 ± 1.6-fold for rESCs and 17.6 ± 3.2-fold for riPSCs). These optimizations resulted in a differentiation efficiency of up to 20% cTnTpos rPSC-derived CMs. CMs showed spontaneous contractions, expressed cardiac markers and had typical morphological features. Electrophysiology of riPSC-CMs revealed different cardiac subtypes and physiological responses to cardio-active drugs. In conclusion, we describe rPSCs as a robust source of CMs, which is a prerequisite for detailed preclinical studies of myocardial reconstruction in a physiologically and immunologically relevant small animal model.

  2. The effect of space allowance and cage size on laying hens housed in furnished cages, Part II: Behavior at the feeder

    Science.gov (United States)

    Widowski, T. M; Caston, L. J; Casey-Trott, T. M; Hunniford, M. E

    2017-01-01

    Abstract Standards for feeder (a.k.a. feed trough) space allowance (SA) are based primarily on studies in conventional cages where laying hens tend to eat simultaneously, limiting feeder space. Large furnished cages (FC) offer more total space and opportunities to perform a greater variety of behaviors, which may affect feeding behavior and feeder space requirements. Our objective was to determine the effects of floor/feeder SA on behavior at the feeder. LSL-Lite hens were housed in FC equipped with a nest, perches, and a scratch mat. Hens with SA of either 520 cm2 (Low; 8.9 cm feeder space/hen) or 748 cm2 (High; 12.8 cm feeder space/hen) per bird resulted in groups of 40 vs. 28 birds in small FC (SFC) and 80 vs. 55 in large FC (LFC). Chain feeders ran at 0500, 0800, 1100, 1400, and 1700 with lights on at 0500 and off at 1900 hours. Digital recordings of FC were scanned at chain feeder onset and every 15 min for one h after (5 scans × 5 feeding times × 2 d) to count the number of birds with their head in the feeder. All occurrences of aggressive pecks and displacements during 2 continuous 30-minute observations at 0800 h and 1700 h also were counted. Mixed model repeated analyses tested the effects of SA, cage size, and time on the percent of hens feeding, and the frequency of aggressive pecks and displacements. Surprisingly, the percent of birds feeding simultaneously was similar regardless of cage size (LFC: 23.0 ± 0.9%; SFC: 24.0 ± 1.0%; P = 0.44) or SA (Low: 23.8 ± 0.9%; High: 23.3 ± 1.0%; P = 0.62). More birds were observed feeding at 1700 h (35.3 ± 0.1%) than any at other time (P < 0.001). Feeder use differed by cage area (nest, middle, or scratch) over the d (P < 0.001). The frequency of aggressive pecks was low overall and not affected by SA or cage size. Frequency of displacements was also low but greater at Low SA (P = 0.001). There was little evidence of feeder competition at the Low SA in this study. PMID:29050409

  3. The impact of small benthic passive suspension feeders in shallow marine ecosystems: the hydroids as an example

    NARCIS (Netherlands)

    Gili, J.-M.; Alvà, V.; Coma, R.; Orejas, C.; Pagès, F.; Ribes, M.; Zabala, M.; Arntz, W.; Bouillon, J.; Boero, F.; Hughes, R.G.

    1998-01-01

    Benthic suspension feeders are abundant in littoral and shallow sub-littoral ecosystems, where they feed on the plankton and on organic matter suspended in the water column. Recent studies indicate that active suspension feeders with powerful water filtration mechanisms (e.g., bivalve molluscs) may

  4. Feeders of Free-Roaming Cats: Personal Characteristics, Feeding Practices, and Data on Cat Health and Welfare in an Urban Setting of Israel.

    Science.gov (United States)

    Gunther, Idit; Raz, Tal; Even Zor, Yehonatan; Bachowski, Yuval; Klement, Eyal

    2016-01-01

    Cat feeders serve as an important source of available food for free-roaming cats (FRCs) and can play a central role in providing data on FRC distribution, welfare, and health. Data on cat feeder personalities as well as a better understanding of their feeding practices offer relevance for decision making concerning FRC population control strategies. The current study surveyed 222 FRC feeders who responded to a municipal trap-neuter-return (TNR) campaign in an Israeli central urban setting. The aim of the study was to describe their personal characteristics, feeding practices, and the FRC populations they feed. Feeders were divided into four groups according to the number of cats they claimed to feed per day (group 1: fed up to 5 cats, group 2: fed 6-10 cats, group 3: fed 11-20 cats, and group 4: fed ≥21 cats). Most feeders were women (81%), with a median age of 58 years (range 18-81). The feeders reported an overall feeding of 3337 cats in 342 different feeding locations. Feeders of group 4 comprised 15.31% (n = 34) of all feeders but fed 56% (n = 1869) of the FRC in 37.42% (n = 128) of the feeding locations. "Heavy" feeders (groups 3 and 4) reported that they traveled significantly longer distances in order to feed the cats. Commercial dry food consisted of 90% of the food they provided, with 66% of them feeding once a day, with less food per cat per day than the other feeder groups. Interestingly, "heavy" feeders were usually singles, had on average fewer siblings, a clear preference for owning cats as pets, and lived in lower income neighborhoods. According to the feeders' reports on the FRC populations they fed, 69.7% (2325/3337) cats were neutered and 11.8% (395/3337) were kittens. In addition, they reported that 1.6% (54/3337) of the cats were limping, 2% (67/3337) suffered from a systemic disease, 4% (135/3337) had skin lesions, and 3.9% (130/3337) were suffering from a chronic disability. Abundance of kittens and morbidity rate were

  5. Feeders of free-roaming cats: personal characteristics, feeding practices and data on cat health and welfare in an urban setting of Israel

    Directory of Open Access Journals (Sweden)

    Idit eGunther

    2016-03-01

    Full Text Available Cat feeders serve as an important source of available food for free-roaming cats (FRC and can play a central role in providing data on FRC distribution, welfare, and health. Data on cat feeder personalities as well as a better understanding of their feeding practices offer relevance for decision making concerning FRC population control strategies. The current study surveyed 222 FRC feeders who responded to a municipal trap-neuter-return (TNR campaign in an Israeli central urban setting. The aim of the study was to describe their personal characteristics, feeding practices and the FRC populations they feed. Feeders were divided into four groups according to the number of cats they claimed to feed per day (group 1: fed up to five cats; group 2: fed six to 10 cats; group 3: fed 11-20 cats; group 4: fed ≥21 cats. Most feeders were women (81%, with a median age of 58 years (range 18-81. The feeders reported an overall feeding of 3,337 cats in 342 different feeding locations. Feeders of group 4 comprised of 15.31% (n=34 of all feeders, but fed 56% (n=1869 of the FRC in 37.42% (n=128 of the feeding locations. 'Heavy' feeders (groups 3 and 4 reported that they traveled significantly longer distances in order to feed the cats. Commercial dry food consisted of 90% of the food they provided, with 66% of them feeding once a day, with less food per cat per day than the other feeder groups. Interestingly, 'heavy' feeders were usually singles, had on average fewer siblings, a clear preference for owning cats as pets and lived in lower income neighborhoods. According to the feeders' reports on the FRC populations they fed, 69.7% (2325/3337 cats were neutered and 11.8% (395/3337 were kittens. In addition, they reported that 1.6% (54/3337 of the cats were limping, 2% (67/3337 suffered from a systemic disease, 4% (135/3337 had skin lesions, and 3.9% (130/3337 were suffering from a chronic disability. Abundance of kittens and morbidity rate were significantly and

  6. Meta-analysis on the effects of the physical environment, animal traits, feeder and feed characteristics on the feeding behaviour and performance of growing-finishing pigs.

    Science.gov (United States)

    Averós, X; Brossard, L; Dourmad, J Y; de Greef, K H; Edwards, S A; Meunier-Salaün, M C

    2012-08-01

    A meta-analysis, using information from 45 experiments on growing-finishing pigs published in 39 manuscripts, was carried out to determine the simultaneous effects of the physical environment (space allowance, group size, flooring conditions, temperature, presence of enrichment), pig traits (initial body weight (BW) for each studied time interval, sex, genetics), feeder characteristics (water provision within the feeder, feeder design (individual/collective), feeder places/pig, presence of feeder protection) and feed characteristics (feed allowance (ad libitum/restricted), net energy content, crude protein (CP) content), as well as their potential interactions, on the feeding behaviour and performance of growing-finishing pigs. The detrimental effect of low temperature on performance was particularly evident for restricted-fed pigs (P < 0.05). At reduced feeder space allowance, a reduction in the percentage of time spent eating was predicted when increasing initial BW, whereas the opposite was predicted for larger feeder space allowances (P < 0.001). The reduction in visit duration to the feeder in higher BW groups became gradually more important with increasing feeder space allowance (P < 0.01), whereas the increase in the ingestion rate and average daily feed intake (ADFI) with increasing initial BW became smaller with increasing feeder space (P < 0.05). The model predicted a reduction in feed conversion ratio (FCR) with increasing group size (P < 0.05) and floor space allowance (P < 0.01) and on solid floors with or without bedding (P < 0.05). In comparison with other feeders, wet/dry feeders were associated with more frequent but shorter feeder visits (P < 0.05), higher ingestion rates (P < 0.001) and higher ADFI (P < 0.10). The use of protection within individual feeders increased the time spent feeding (P < 0.001), reduced the number of visits per day (P < 0.01), the ingestion rate (P < 0.001) and FCR (P < 0.01) in comparison with other feeder types. Sex

  7. Effect of artificial feeders on pollen loads of the hummingbirds of Cerro de La Muerte, Costa Rica

    Directory of Open Access Journals (Sweden)

    Gerardo Avalos

    2012-03-01

    Full Text Available Although sugar-water feeders are commonly used by enthusiasts to attract hummingbirds, little is known about how they affect hummingbird behavior and flower use. We studied the highland hummingbird assemblage of Cerro de La Muerte, Costa Rica, both at a site with permanent feeders (La Georgina Restaurant and further from it. We examined how feeder use and monopolization affected seasonal changes in pollen loads during four sampling periods, including dry and wet seasons, from 2003-2005. We expected that species monopolizing the feeders would carry little or no pollen whatsoever, and would have pollen loads characterized by low floral diversity, in contrast with species less dependent on feeders. We obtained pollen samples from 183 individuals of four hummingbird species captured around the feeders using mist nets, which were compared with a pollen reference collection of plants with a pollination syndrome by hummingbirds. The same methods were implemented at a site 3km away from the feeders. Feeder usage was quantified by counting the number of times hummingbirds drank from the feeders in periods of 4min separated by 1min. The effects of hummingbird species and season on pollen load categories were assessed using a nominal logistic regression. The alpha species at the site, the Fiery-throated Hummingbird (Panterpe insignis, dominated the feeders during the dry season. Meanwhile, in the wet season, feeder usage was more evenly distributed across species, with the exception of the Volcano Hummingbird, Selasphorus flammula, which occupies the last place in the dominance hierarchy. Pollen loads of hummingbirds captured near feeders were low in abundance (more than 50% of captured individuals had zero or low pollen loads, and low in species richness (96% of the hummingbirds with pollen from only one plant genus, Centropogon. Overall pollen loads increased during the dry season coinciding with peaks in flower availability, although the majority of

  8. Arthropods associated with fungal galls: do large galls support more abundant and diverse inhabitants?

    Science.gov (United States)

    Funamoto, Daichi; Sugiura, Shinji

    2017-02-01

    Fungus-induced galls can attract spore-feeding arthropods as well as gall-feeding ones, resulting in diverse communities. Do large fungal galls support more abundant and diverse arthropod communities than small fungal galls? To address this question, we investigated the structure of the arthropod community associated with bud galls induced by the fungus Melanopsichium onumae on the tree species Cinnamomum yabunikkei (Lauraceae) in central Japan. Thirteen species of arthropods were associated with M. onumae galls. Dominant arthropod species were represented by the larvae of a salpingid beetle (a spore feeder), a nitidulid beetle (a spore feeder), a cosmopterigid moth (a spore feeder), an unidentified moth (a gall tissue feeder), and a drosophilid species (a gall tissue feeder). Arthropod abundance and species richness were positively correlated with gall diameter. The majority of the most abundant species were more frequently found in large galls than in small ones, indicating that large fungal galls, which have more food and/or space for arthropods, could support a more abundant and diverse arthropod community.

  9. Expansion and differentiation of germline-derived pluripotent stem cells on biomaterials.

    Science.gov (United States)

    Hoss, Mareike; Šarić, Tomo; Denecke, Bernd; Peinkofer, Gabriel; Bovi, Manfred; Groll, Jürgen; Ko, Kinarm; Salber, Jochen; Halbach, Marcel; Schöler, Hans R; Zenke, Martin; Neuss, Sabine

    2013-05-01

    Stem cells with broad differentiation potential, such as the recently described germline-derived pluripotent stem cells (gPS cells), are an appealing source for tissue engineering strategies. Biomaterials can inhibit, support, or induce proliferation and differentiation of stem cells. Here we identified (1) polymers that maintain self-renewal and differentiation potential of gPS cells for feeder-free expansion and (2) polymers supporting the cardiomyogenic fate of gPS cells by analyzing a panel of polymers of an established biomaterial bank previously used to assess growth of diverse stem cell types. Identification of cytocompatible gPS cell/biomaterial combinations required analysis of several parameters, including morphology, viability, cytotoxicity, apoptosis, proliferation, and differentiation potential. Pluripotency of gPS cells was visualized by the endogenous Oct4-promoter-driven GFP and by Sox2 and Nanog immunofluorescence. Viability assay, proliferation assay, and flow cytometry showed that gPS cells efficiently adhere and are viable on synthetic polymers, such as Resomer(®) LR704 (poly(L-lactic-D,L-lactic acid), poly(tetrafluor ethylene) (PTFE), poly(vinylidene fluoride) (PVDF), and on gelatine-coated tissue culture polystyrene. Expansion experiments showed that Resomer LR704 is an alternative substrate for feeder-free gPS cell maintenance. Resomer LR704, PTFE, and PVDF were found to be suitable for gPS cell differentiation. Spontaneous beating in embryoid bodies cultured on Resomer LR704 occurred already on day 8 of differentiation, much earlier compared to the other surfaces. This indicates that Resomer LR704 supports spontaneous cardiomyogenic differentiation of gPS cells, which was also confirmed on molecular, protein and functional level.

  10. ATP-independent DNA synthesis in Vaccinia-infected L cells

    International Nuclear Information System (INIS)

    Berger, N.A.; Kauff, R.A.; Sikorski, G.W.

    1978-01-01

    Mouse L cells can be made permeable to exogenous nucleotides by a cold shock in 0.01 M Tris . HCl pH 7.8, 0.25 M sucrose, 1 mM EDTA, 30 mM 2-mercaptoethanol and 4 mM MgCl 2 . DNA synthesis in permeabilized L cells requires ATP whereas DNA synthesis in permeabilized L cells that are infected with Vaccinia virus is ATP-independent. Permeabilized L cells that are infected with ultraviolet-irradiated virus show a marked suppression of DNA synthesis which is not corrected by an excess of deoxynucleoside triphosphates and ATP. The ATP-dependent and ATP-independent processes of DNA synthesis are inhibited to the same extent by Mal-Net, pHMB, ara CTP and phosphonoacetate. Concentrations of daunorubicin and cytembena, which cause marked inhibition of the ATP-dependent enzymes, only cause partial inhibition of the ATP-independent enzymes. (Auth.)

  11. Two-photon laser-generated microtracks in 3D collagen lattices: principles of MMP-dependent and -independent collective cancer cell invasion

    Science.gov (United States)

    Ilina, Olga; Bakker, Gert-Jan; Vasaturo, Angela; Hoffman, Robert M.; Friedl, Peter

    2011-02-01

    Cancer invasion into an extracellular matrix (ECM) results from a biophysical reciprocal interplay between the expanding cancer lesion and tissue barriers imposed by the adjacent microenvironment. In vivo, connective tissue provides both densely packed ECM barriers adjacent to channel/track-like spaces and loosely organized zones, both of which may impact cancer invasion mode and efficiency; however little is known about how three-dimensional (3D) spaces and aligned tracks present in interstitial tissue guide cell invasion. We here describe a two-photon laser ablation procedure to generate 3D microtracks in dense 3D collagen matrices that support and guide collective cancer cell invasion. Whereas collective invasion of mammary tumor (MMT) breast cancer cells into randomly organized collagen networks required matrix metalloproteinase (MMP) activity for cell-derived collagen breakdown, re-alignment and track generation, preformed tracks supported MMP-independent collective invasion down to a track caliber of 3 µm. Besides contact guidance along the track of least resistance and initial cell deformation (squeezing), MMP-independent collective cell strands led to secondary track expansion by a pushing mechanism. Thus, two-photon laser ablation is useful to generate barrier-free microtracks in a 3D ECM which guide collective invasion independently of pericellular proteolysis.

  12. Two-photon laser-generated microtracks in 3D collagen lattices: principles of MMP-dependent and -independent collective cancer cell invasion

    International Nuclear Information System (INIS)

    Ilina, Olga; Bakker, Gert-Jan; Hoffman, Robert M; Friedl, Peter; Vasaturo, Angela

    2011-01-01

    Cancer invasion into an extracellular matrix (ECM) results from a biophysical reciprocal interplay between the expanding cancer lesion and tissue barriers imposed by the adjacent microenvironment. In vivo, connective tissue provides both densely packed ECM barriers adjacent to channel/track-like spaces and loosely organized zones, both of which may impact cancer invasion mode and efficiency; however little is known about how three-dimensional (3D) spaces and aligned tracks present in interstitial tissue guide cell invasion. We here describe a two-photon laser ablation procedure to generate 3D microtracks in dense 3D collagen matrices that support and guide collective cancer cell invasion. Whereas collective invasion of mammary tumor (MMT) breast cancer cells into randomly organized collagen networks required matrix metalloproteinase (MMP) activity for cell-derived collagen breakdown, re-alignment and track generation, preformed tracks supported MMP-independent collective invasion down to a track caliber of 3 µm. Besides contact guidance along the track of least resistance and initial cell deformation (squeezing), MMP-independent collective cell strands led to secondary track expansion by a pushing mechanism. Thus, two-photon laser ablation is useful to generate barrier-free microtracks in a 3D ECM which guide collective invasion independently of pericellular proteolysis

  13. [Effects of prefrontal ablations on the reaction of the active choice of feeder under different probability and value of the reinforcement on dog].

    Science.gov (United States)

    Preobrazhenskaia, L A; Ioffe, M E; Mats, V N

    2004-01-01

    The role of the prefrontal cortex was investigated on the reaction of the active choice of the two feeders under changes value and probability reinforcement. The experiments were performed on 2 dogs with prefrontal ablation (g. proreus). Before the lesions the dogs were taught to receive food in two different feeders to conditioned stimuli with equally probable alimentary reinforcement. After ablation in the inter-trial intervals the dogs were running from the one feeder to another. In the answer to conditioned stimuli for many times the dogs choose the same feeder. The disturbance of the behavior after some times completely restored. In the experiments with competition of probability events and values of reinforcement the dogs chose the feeder with low-probability but better quality of reinforcement. In the experiments with equal value but different probability the intact dogs chose the feeder with higher probability. In our experiments the dogs with prefrontal lesions chose the each feeder equiprobably. Thus in condition of free behavior one of different functions of the prefrontal cortex is the reactions choose with more probability of reinforcement.

  14. Identification of distinct topographical surface microstructures favoring either undifferentiated expansion or differentiation of murine embryonic stem cells.

    Science.gov (United States)

    Markert, Lotte D'Andrea; Lovmand, Jette; Foss, Morten; Lauridsen, Rune Hoff; Lovmand, Michael; Füchtbauer, Ernst-Martin; Füchtbauer, Annette; Wertz, Karin; Besenbacher, Flemming; Pedersen, Finn Skou; Duch, Mogens

    2009-11-01

    The potential of embryonic stem (ES) cells for both self-renewal and differentiation into cells of all three germ layers has generated immense interest in utilizing these cells for tissue engineering or cell-based therapies. However, the ability to culture undifferentiated ES cells without the use of feeder cells as well as means to obtain homogeneous, differentiated cell populations devoid of residual pluripotent ES cells still remain major challenges. Here we have applied murine ES cells to topographically microstructured surface libraries, BioSurface Structure Arrays (BSSA), and investigated whether these could be used to (i) identify topographically microstructured growth supports alleviating the need for feeder cells for expansion of undifferentiated ES cells and (ii) identify specific types of microstructures enforcing differentiation of ES cells. The BSSA surfaces arrays consisted of 504 different topographical microstructures each located in a tester field of 3 x 3 mm. The murine ES cell lines CJ7 and KH2 were seeded upon the BSSA libraries and specific topographical structures facilitating either undifferentiated ES cell growth or enhancing spreading indicative of differentiation of the ES cells were identified. Secondly serial passage of undifferentiated CJ7 ES cells on selected microstructures, identified in the screening of these BSSA libraries, showed that these cells had retained germ-line potential. These results indicate that one specific type of topographical surface microstructures, identified by the BSSA technology, can substitute for feeder cells and that another subset may be used to eliminate undifferentiated ES cells from a population of differentiated ES cells.

  15. Replacement of murine fibroblasts by human fibroblasts irradiated in obtaining feeder layer for the culture of human keratinocytes

    International Nuclear Information System (INIS)

    Yoshito, Daniele; Sufi, Bianca S.; Santin, Stefany P.; Mathor, Monica B.; Altran, Silvana C.; Isaac, Cesar

    2011-01-01

    Human autologous epithelia cultivated in vitro, have been used successfully in treating damage to skin integrity. The methodology allowed the cultivation of these epithelia was described by Rheinwald and Green in 1975, this methodology consisted in seeding keratinocytes onto a feeder layer composed of lineage 3T3 murine fibroblasts, the proliferation rate is controlled through the action of ionizing radiation. However, currently there is a growing concern about the possibility of transmitting prions and murine viruses to transplanted patients. Taking into account this concern, in this present work, we replaced the feeder layer originally composed of murine fibroblasts by human fibroblasts. To obtain this new feeder layer was necessary to standardize the enough irradiation dose to inhibit the replication of human fibroblasts and the verification of effectiveness of the development of keratinocytes culture on a feeder layer thus obtained. According to the obtained results we can verify that the human fibroblasts irradiated at various tested doses (60, 70, 100, 200, 250 and 300 Gy) had their mitotic activity inactivated by irradiation, allowing the use of any of these doses to confection of the feeder layer, since these fibroblasts irradiated still showed viable until fourteen days of cultivation. In the test of colony formation efficiency was observed that keratinocytes seeded on irradiated human fibroblasts were able to develop satisfactorily, preserving their clonogenic potential. Therefore it was possible the replacement of murine fibroblasts by human fibroblasts in confection of the feeder layer, in order to eliminate this xenobiotic component of the keratinocytes culture. (author)

  16. Replacement of murine fibroblasts by human fibroblasts irradiated in obtaining feeder layer for the culture of human keratinocytes

    Energy Technology Data Exchange (ETDEWEB)

    Yoshito, Daniele; Sufi, Bianca S.; Santin, Stefany P.; Mathor, Monica B. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Altran, Silvana C.; Isaac, Cesar [Universidade Sao Paulo (USP), Sao Paulo, SP (Brazil). Fac. de Medicina. Lab. de Microcirurgia Plastica; Esteves-Pedro, Natalia M. [Universidade Sao Paulo (USP), Sao Paulo, SP (Brazil). Fac. de Ciencias Farmaceuticas. Lab. de Controle Biologico; Herson, Marisa R. [DonorTissue Bank of Victoria (Australia)

    2011-07-01

    Human autologous epithelia cultivated in vitro, have been used successfully in treating damage to skin integrity. The methodology allowed the cultivation of these epithelia was described by Rheinwald and Green in 1975, this methodology consisted in seeding keratinocytes onto a feeder layer composed of lineage 3T3 murine fibroblasts, the proliferation rate is controlled through the action of ionizing radiation. However, currently there is a growing concern about the possibility of transmitting prions and murine viruses to transplanted patients. Taking into account this concern, in this present work, we replaced the feeder layer originally composed of murine fibroblasts by human fibroblasts. To obtain this new feeder layer was necessary to standardize the enough irradiation dose to inhibit the replication of human fibroblasts and the verification of effectiveness of the development of keratinocytes culture on a feeder layer thus obtained. According to the obtained results we can verify that the human fibroblasts irradiated at various tested doses (60, 70, 100, 200, 250 and 300 Gy) had their mitotic activity inactivated by irradiation, allowing the use of any of these doses to confection of the feeder layer, since these fibroblasts irradiated still showed viable until fourteen days of cultivation. In the test of colony formation efficiency was observed that keratinocytes seeded on irradiated human fibroblasts were able to develop satisfactorily, preserving their clonogenic potential. Therefore it was possible the replacement of murine fibroblasts by human fibroblasts in confection of the feeder layer, in order to eliminate this xenobiotic component of the keratinocytes culture. (author)

  17. Cell vertices as independent actors during cell intercalation in epithelial morphogenesis

    Science.gov (United States)

    Loerke, Dinah

    Epithelial sheets form the lining of organ surfaces and body cavities, and it is now appreciated that these sheets are dynamic structures that can undergo significant reorganizing events, e.g. during wound healing or morphogenesis. One of the key morphogenetic mechanisms that is utilized during development is tissue elongation, which is driven by oriented cell intercalation. In the Drosophila embryonic epithelium, this occurs through the contraction of vertical T1 interfaces and the subsequent resolution of horizontal T3 interfaces (analogous to so-called T1 transitions in soap foams), where the symmetry breaking behaviors are created by a system of planar polarity of actomyosin and adhesion complexes within the cell layer. The dominant physical model for this process posits that the anisotropy of line tension directs T1 contraction. However, this model is inconsistent with the in vivo observation that cell vertices of T1 interfaces lack physical coupling, and instead show independent movements. Thus, we propose that a more useful explanation of intercalary behaviors will be possible through a description of the radially-directed and adhesion-coupled force events that lead to vertex movements and produce subsequent dependent changes in interface lengths. This work is supported by NIH R15 GM117463-01 and by a Research Corporation for Science Advancement (RCSA) Cottrell Scholar Award.

  18. Differential effects of the extracellular microenvironment on human embryonic stem cell differentiation into keratinocytes and their subsequent replicative life span.

    Science.gov (United States)

    Movahednia, Mohammad Mehdi; Kidwai, Fahad Karim; Zou, Yu; Tong, Huei Jinn; Liu, Xiaochen; Islam, Intekhab; Toh, Wei Seong; Raghunath, Michael; Cao, Tong

    2015-04-01

    Culture microenvironment plays a critical role in the propagation and differentiation of human embryonic stem cells (hESCs) and their differentiated progenies. Although high efficiency of hESC differentiation to keratinocytes (hESC-Kert) has been achieved, little is known regarding the effects of early culture microenvironment and pertinent extracellular matrix (ECM) interactions during epidermal commitment on subsequent proliferative capacity of hESC-Kert. The aim of this study is to evaluate the effects of the different ECM microenvironments during hESC differentiation on subsequent replicative life span of hESC-Kert. In doing so, H1-hESCs were differentiated to keratinocytes (H1-Kert) in two differentiation systems. The first system employed autologous fibroblast feeder support, in which keratinocytes (H1-Kert(ACC)) were derived by coculture of hESCs with hESC-derived fibroblasts (H1-ebFs). The second system employed a novel decellularized matrix from H1-ebFs to create a dermoepidermal junction-like (DEJ) matrix. H1-Kert(AFF) were derived by differentiation of hESCs on the feeder-free system employing the DEJ matrix. Our study indicated that the feeder-free system with the use of DEJ matrix was more efficient in differentiation of hESCs toward epidermal progenitors. However, the feeder-free system was not sufficient to support the subsequent replicative capacity of differentiated keratinocytes. Of note, H1-Kert(AFF) showed limited replicative capacity with reduced telomere length and early cellular senescence. We further showed that the lack of cell-cell interactions during epidermal commitment led to heightened production of TGF-β1 by hESC-Kert during extended culture, which in turn was responsible for resulting in the limited replicative life span with cellular senescence of hESC-Kert derived under the feeder-free culture system. This study highlights for the first time the importance of the culture microenvironment and cell-ECM interactions during

  19. Microplastic in a macro filter feeder: humpback whale Megaptera novaeangliae

    NARCIS (Netherlands)

    Besseling, E.; Foekema, E.M.; Franeker, van J.A.; Leopold, M.F.; Bravo Rebolledo, E.; Kuehn, S.; Mielke, L.; Heberle-Bors, E.; Ijzer, J.; Kamminga, P.; Koelmans, A.A.

    2015-01-01

    Marine filter feeders are exposed to microplastic because of their selection of small particles as food source. Baleen whales feed by filtering small particles from large water volumes. Macroplastic was found in baleen whales before. This study is the first to show the presence of microplastic in

  20. Microplastic in a macro filter feeder: Humpback whale Megaptera novaeangliae.

    NARCIS (Netherlands)

    E., Besseling,; E.M., Foekema,; J.A. van, Franeker; Leopold, Mardik F; Kuhn, S.; Bravo Rebolledo, E.L.; Hese, E.; Mielke, L.; IJzer, J.|info:eu-repo/dai/nl/304839663; Kamminga, P.; Koelmans, A.A.

    2015-01-01

    Marine filter feeders are exposed to microplastic because of their selection of small particles as food source. Baleen whales feed by filtering small particles from large water volumes. Macroplastic was found in baleen whales before. This study is the first to show the presence of microplastic in

  1. The influence of interleukin-2, feeder cells, and timing of irradiation on the radiosensitivity of human T lymphocytes assessed by the colony-forming assay

    International Nuclear Information System (INIS)

    Gerber, M.; Guichard, M.; Pioch, Y.; Dubois, J.B.

    1989-01-01

    The radiosensitivity of human lymphocytes was investigated by the method of colony formation in the absence of interleukin-2 (IL2) and feeder cells, both of which enhance growth of T-cell colonies. The shape of the survival curve and the radiosensitivity was shown to depend upon the ability of lymphocytes to produce IL2: the survival curve for lymphocytes that were the most competent producers of IL2 is the closest to linearity; the lymphocytes that were poor producers show biphasic survival curves. The radiosensitivity of the lymphocytes from the first group is less than that of the latter, when the comparison is based on the first part of the biphasic survival curve. This is more easily seen when cultures are irradiated 24 h after stimulation by phytohemagglutinin (the time of the peak IL2 production) than when cultures are irradiated 2 h before stimulation. This study demonstrates that growth conditions influence the response of lymphocytes to irradiation and that optimal growth conditions result in a linear survival curve

  2. The synergistic effect of beta-boswellic acid and Nurr1 overexpression on dopaminergic programming of antioxidant glutathione peroxidase-1-expressing murine embryonic stem cells.

    Science.gov (United States)

    Abasi, M; Massumi, M; Riazi, G; Amini, H

    2012-10-11

    Parkinson's disease (PD) is a neurodegenerative disorder in which the nigro-striatal dopaminergic (DAergic) neurons have been selectively lost. Due to side effects of levodopa, a dopamine precursor drug, recently cell replacement therapy for PD has been considered. Lack of sufficient amounts of, embryos and ethical problems regarding the use of dopamine-rich embryonic neural cells have limited the application of these cells for PD cell therapy. Therefore, many investigators have focused on using the pluripotent stem cells to generate DAergic neurons. This study is aimed first to establish a mouse embryonic stem (mES) cell line that can stably co-express Nurr1 (Nuclear receptor subfamily 4, group A, member 2) transcription factor in order to efficiently generate DAergic neurons, and glutathione peroxidase-1 (GPX-1) to protect the differentiated DAergic-like cells against oxidative stress. In addition to genetic engineering of ES cells, the effect of Beta-boswellic acid (BBA) on DAergic differentiation course of mES cells was sought in the present study. To that end, the feeder-independent CGR8 mouse embryonic stem cells were transduced by Nurr1- and GPX-1-harboring Lentiviruses and the generated Nurr1/GPX-1-expresssing ES clones were characterized and verified. Gene expression analyses demonstrated that BBA treatment and overexpression of Nurr1 has a synergistic effect on derivation of DAergic neurons from Nurr1/GPX-1-expressing ES cells. The differentiated cells could exclusively synthesize and secrete dopamine in response to stimuli. Overexpression of GPX-1 in genetically engineered Nurr1/GPX-1-ES cells increased the viability of these cells during their differentiation into CNS stem cells. In conclusion, the results demonstrated that Nurr1-overexpressing feeder-independent ES cells like the feeder-dependent ES cells, can be efficiently programmed into functional DAergic neurons and additional treatment of cells by BBA can even augment this efficiency. GPX-1

  3. Transitioning to employment with a rheumatic disease: the role of independence, overprotection, and social support.

    Science.gov (United States)

    Jetha, Arif; Badley, Elizabeth; Beaton, Dorcas; Fortin, Paul R; Shiff, Natalie J; Rosenberg, Alan M; Tucker, Lori B; Mosher, Dianne P; Gignac, Monique A M

    2014-12-01

    To examine perceived independence, overprotection, and support, and their association with the employment participation of young adults with rheumatic disease. One hundred and forty-three young adults, ages 18 to 30 years, with systemic lupus erythematosus (54.5%) and juvenile arthritis (45.5%) completed a 30-min online questionnaire of their work and education experiences. Information collected was demographic, health (e.g., pain, fatigue, disease activity), work context (e.g., career satisfaction, helpfulness of job accommodation/benefits, and workplace activity limitations), and psychosocial (e.g., independence, social support, and overprotection). Log-Poisson regression analysis examined factors associated with employment status. Over half of respondents were employed (59%) and 26% were enrolled in school. Respondents reported moderate to high perceptions of independence and social support. However, 27% reported that "quite a bit" to "a great deal" of overprotection characterized their relationships with those closest to them. At the bivariate level, employed participants and those indicating greater perceived independence reported greater social support and less overprotection. Multivariable analysis revealed that being employed was associated with older age, more job accommodations/benefits perceived as being helpful, and greater perceived independence. This is one of the first studies examining the employment of young adults with rheumatic diseases. Findings highlight the importance of psychosocial perceptions such as independence and overprotection, in addition to support related to working. Additional research is needed to better understand the role of those close to young adults with rheumatic diseases in supporting independence and encouraging employment.

  4. Myogel supports the ex-vivo amplification of corneal epithelial cells.

    Science.gov (United States)

    Francis, D; Abberton, K; Thompson, E; Daniell, M

    2009-03-01

    Limbal stem cell deficiency leads to conjunctivalisation of the cornea and subsequent loss of vision. The recent development of transplantation of ex-vivo amplified corneal epithelium, derived from limbal stem cells, has shown promise in treating this challenging condition. The purpose of this research was to compare a variety of cell sheet carriers for their suitability in creating a confluent corneal epithelium from amplified limbal stem cells. Cadaveric donor limbal cells were cultured using an explant technique, free of 3T3 feeder cells, on a variety of cell sheet carriers, including denuded amniotic membrane, Matrigel, Myogel and stromal extract. Comparisons in rate of growth and degree of differentiation were made, using immunocytochemistry (CK3, CK19 and ABCG2). The most rapid growth was observed on Myogel and denuded amniotic membrane, these two cell carriers also provided the most reliable substrata for achieving confluence. The putative limbal stem cell marker, ABCG2, stained positively on cells grown over Myogel and Matrigel but not for those propagated on denuded amniotic membrane. In the clinical setting amniotic membrane has been demonstrated to provide a suitable carrier for limbal stem cells and the resultant epithelium has been shown to be successful in treating limbal stem cell deficiency. Myogel may provide an alternative cell carrier with a further reduction in risk as it is has the potential to be derived from an autologous muscle biopsy in the clinical setting.

  5. Assessing the role of benthic filter feeders on phytoplankton production in a shellfish farming site: Mont Saint Michel Bay, France

    Science.gov (United States)

    Cugier, Philippe; Struski, Caroline; Blanchard, Michel; Mazurié, Joseph; Pouvreau, Stéphane; Olivier, Frédéric; Trigui, Jihane R.; Thiébaut, Eric

    2010-07-01

    The macrobenthic community of Mont Saint Michel Bay (English Channel, France) is mainly dominated by filter feeders, including cultivated species (oysters and mussels). An ecological model of the bay was developed, coupling a 2D hydro-sedimentary model and two biological models for primary production and filter-feeder filtration. The filter-feeder model includes three cultivated species ( Mytilus edulis, Crassostrea gigas and Ostrea edulis), one invasive species ( Crepidula fornicata) and eight wild native species ( Abra alba, Cerastoderma edule, Glycymeris glycymeris, Lanice conchilega, Macoma balthica, Paphia rhomboides, Sabellaria alveolata, andSpisula ovalis). For cultivated and invasive species, the production of biodeposits was computed to assess their role in restimulating primary production. Chlorophyll a concentrations appeared to be strongly controlled by the filter feeders. When the pressure of each benthic compartment on phytoplankton was estimated separately wild species and the invasive slipper limpet C.fornicata were shown to be key elements in the control of primary production. Conversely, the role of cultivated species, particularly oysters, was weaker. Feedback due to the mineralization of biodeposits also appears to be crucial to fully evaluate the role of filter feeders in primary production.

  6. GROα regulates human embryonic stem cell self-renewal or adoption of a neuronal fate

    Science.gov (United States)

    Krtolica, Ana; Larocque, Nick; Genbacev, Olga; Ilic, Dusko; Coppe, Jean-Philippe; Patil, Christopher K.; Zdravkovic, Tamara; McMaster, Michael; Campisi, Judith; Fisher, Susan J.

    2012-01-01

    Previously we reported that feeders formed from human placental fibroblasts (hPFs) support derivation and long-term self-renewal of human embryonic stem cells (hESCs) under serum-free conditions. Here, we show, using antibody array and ELISA platforms, that hPFs secrete ~6-fold higher amounts of the CXC-type chemokine, GROα, than IMR 90, a human lung fibroblast line, which does not support hESC growth. Furthermore, immunocytochemistry and immunoblot approaches revealed that hESCs express CXCR, a GROα receptor. We used this information to develop defined culture medium for feeder-free propagation of hESCs in an undifferentiated state. Cells passaged as small aggregates and maintained in the GROα-containing medium had a normal karyotype, expressed pluripotency markers, and exhibited apical–basal polarity, i.e., had the defining features of pluripotent hESCs. They also differentiated into the three primary (embryonic) germ layers and formed teratomas in immunocompromised mice. hESCs cultured as single cells in the GROα-containing medium also had a normal karyotype, but they downregulated markers of pluripotency, lost apical–basal polarity, and expressed markers that are indicative of the early stages of neuronal differentiation—βIII tubulin, vimentin, radial glial protein, and nestin. These data support our hypothesis that establishing and maintaining cell polarity is essential for the long-term propagation of hESCs in an undifferentiated state and that disruption of cell–cell contacts can trigger adoption of a neuronal fate. PMID:21396766

  7. The Important Role of FLT3-L in Ex Vivo Expansion of Hematopoietic Stem Cells following Co-Culture with Mesenchymal Stem Cells.

    Science.gov (United States)

    Oubari, Farhad; Amirizade, Naser; Mohammadpour, Hemn; Nakhlestani, Mozhdeh; Zarif, Mahin Nikougoftar

    2015-01-01

    Hematopoietic stem cells (HSCs) transplantation using umbilical cord blood (UCB) has improved during the last decade. Because of cell limitations, several studies focused on the ex vivo expansion of HSCs. Numerous investigations were performed to introduce the best cytokine cocktails for HSC expansion The majority used the Fms-related tyrosine kinase 3 ligand (FLT3-L) as a critical component. According to FLT3-L biology, in this study we have investigated the hypothesis that FLT3-L only effectively induces HSCs expansion in the presence of a mesenchymal stem cell (MSC) feeder. In this experimental study, HSCs and MSCs were isolated from UCB and placenta, respectively. HSCs were cultured in different culture conditions in the presence and absence of MSC feeder and cytokines. After ten days of culture, total nucleated cell count (TNC), cluster of differentiation 34+(CD34(+)) cell count, colony forming unit assay (CFU), long-term culture initiating cell (LTC-IC), homeobox protein B4 (HoxB4) mRNA and surface CD49d expression were evaluated. The fold increase for some culture conditions was compared by the t test. HSCs expanded in the presence of cytokines and MSCs feeder. The rate of expansion in the co-culture condition was two-fold more than culture with cytokines (Pculture condition at a level of 20-fold equal to the presence of stem cell factor (SCF), thrombopoietin (TPO) and FLT3-L without feeder cells. The number of extracted colonies from LTC-IC and CD49d expression compared with a cytokine cocktail condition meaningfully increased (Pculture with MSCs can induce high yield expansion of HSCs and be a substitute for the universal cocktail of SCF, TPO and FLT3-L in feeder-free culture.

  8. Bacterial Feeders, the Nematode Caenorhabditis elegans and the Flagellate Cercomonas longicauda, have different Effects on Outcome of Competition among the Pseudomonas Biocontrol Strains CHA0 and DSS73

    DEFF Research Database (Denmark)

    Pedersen, Annette; Nybroe, Ole; Winding, Anne

    2009-01-01

    How bacterial feeding fauna affects colonization and survival of bacteria in soil is not well understood, which constrains the applicability of bacterial inoculants in agriculture. This study aimed to unravel how food quality of bacteria and bacterial feeders with different feeding habits (the......50090 or one of two biocontrol strains P. fluorescens CHA0 or Pseudomonas sp. DSS73) or combinations of two bacterial strains. DSM50090 is a suitable food bacterium, DSS73 is of intermediate food quality, and CHA0 is inedible to the bacterial feeders. Bacterial and protozoan cell numbers were measured...... predation pressure. Hence, the results suggested that the outcome of competition among bacteria depended on their ability to cope with the prevailing bacterial predator....

  9. Effect of portion size and milk flow on the use of a milk feeder and the development of cross-sucking in dairy calves

    DEFF Research Database (Denmark)

    Nielsen, Per Peetz; Jensen, Margit Bak; Halekoh, Ulrich

    2018-01-01

    milk feeder in each pen, and exposed to four milk-feeding treatments in a cross-over design with four periods of 1 week. All four treatments involved a daily allowance of 8. L of whole milk, with variation in the maximum portion size and the rate at which milk entered the teat of the milk feeder......This study aimed to investigate whether reducing the milk flow and increasing the milk portion size of a computer-controlled milk feeder would lead to less cross-sucking and fewer unrewarded feeder visits in dairy calves. Five groups, each with 9 or 10 calves (n = 48), were housed in pens with one...

  10. Collagen-IV supported embryoid bodies formation and differentiation from buffalo (Bubalus bubalis) embryonic stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Taru Sharma, G., E-mail: gts553@gmail.com [Reproductive Physiology Laboratory, Division of Physiology and Climatology, Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, U.P. (India); Dubey, Pawan K.; Verma, Om Prakash; Pratheesh, M.D.; Nath, Amar; Sai Kumar, G. [Reproductive Physiology Laboratory, Division of Physiology and Climatology, Indian Veterinary Research Institute, Izatnagar-243 122, Bareilly, U.P. (India)

    2012-08-03

    Graphical abstract: EBs formation, characterization and expression of germinal layers marker genes of in vivo developed teratoma using four different types of extracellular matrices. Highlights: Black-Right-Pointing-Pointer Collagen-IV matrix is found cytocompatible for EBs formation and differentiation. Black-Right-Pointing-Pointer Established 3D microenvironment for ES cells development and differentiation into three germ layers. Black-Right-Pointing-Pointer Collagen-IV may be useful as promising candidate for ES cells based therapeutic applications. -- Abstract: Embryoid bodies (EBs) are used as in vitro model to study early extraembryonic tissue formation and differentiation. In this study, a novel method using three dimensional extracellular matrices for in vitro generation of EBs from buffalo embryonic stem (ES) cells and its differentiation potential by teratoma formation was successfully established. In vitro derived inner cell masses (ICMs) of hatched buffalo blastocyst were cultured on buffalo fetal fibroblast feeder layer for primary cell colony formation. For generation of EBs, pluripotent ES cells were seeded onto four different types of extracellular matrices viz; collagen-IV, laminin, fibronectin and matrigel using undifferentiating ES cell culture medium. After 5 days of culture, ESCs gradually grew into aggregates and formed simple EBs having circular structures. Twenty-six days later, they formed cystic EBs over collagen matrix with higher EBs formation and greater proliferation rate as compared to other extracellular matrices. Studies involving histological observations, fluorescence microscopy and RT-PCR analysis of the in vivo developed teratoma revealed that presence of all the three germ layer derivatives viz. ectoderm (NCAM), mesoderm (Flk-1) and endoderm (AFP). In conclusion, the method described here demonstrates a simple and cost-effective way of generating EBs from buffalo ES cells. Collagen-IV matrix was found cytocompatible as it

  11. Collagen-IV supported embryoid bodies formation and differentiation from buffalo (Bubalus bubalis) embryonic stem cells

    International Nuclear Information System (INIS)

    Taru Sharma, G.; Dubey, Pawan K.; Verma, Om Prakash; Pratheesh, M.D.; Nath, Amar; Sai Kumar, G.

    2012-01-01

    Graphical abstract: EBs formation, characterization and expression of germinal layers marker genes of in vivo developed teratoma using four different types of extracellular matrices. Highlights: ► Collagen-IV matrix is found cytocompatible for EBs formation and differentiation. ► Established 3D microenvironment for ES cells development and differentiation into three germ layers. ► Collagen-IV may be useful as promising candidate for ES cells based therapeutic applications. -- Abstract: Embryoid bodies (EBs) are used as in vitro model to study early extraembryonic tissue formation and differentiation. In this study, a novel method using three dimensional extracellular matrices for in vitro generation of EBs from buffalo embryonic stem (ES) cells and its differentiation potential by teratoma formation was successfully established. In vitro derived inner cell masses (ICMs) of hatched buffalo blastocyst were cultured on buffalo fetal fibroblast feeder layer for primary cell colony formation. For generation of EBs, pluripotent ES cells were seeded onto four different types of extracellular matrices viz; collagen-IV, laminin, fibronectin and matrigel using undifferentiating ES cell culture medium. After 5 days of culture, ESCs gradually grew into aggregates and formed simple EBs having circular structures. Twenty-six days later, they formed cystic EBs over collagen matrix with higher EBs formation and greater proliferation rate as compared to other extracellular matrices. Studies involving histological observations, fluorescence microscopy and RT-PCR analysis of the in vivo developed teratoma revealed that presence of all the three germ layer derivatives viz. ectoderm (NCAM), mesoderm (Flk-1) and endoderm (AFP). In conclusion, the method described here demonstrates a simple and cost-effective way of generating EBs from buffalo ES cells. Collagen-IV matrix was found cytocompatible as it supported buffalo EBs formation, their subsequent differentiation could prove to

  12. The effect of stocking density and bin feeder space on performance ...

    African Journals Online (AJOL)

    The effect of stocking density and bin feeder space on performance in pigs. G.A. Lavers, N.S. Ferguson. Abstract. (South African J of Animal Science, 2000, 30, Supplement 1: 70-71). Full Text: EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT.

  13. A Branch-and-Price Approach to the Feeder Network Design Problem

    DEFF Research Database (Denmark)

    Santini, Alberto; Plum, Christian Edinger Munk; Røpke, Stefan

    2017-01-01

    In this paper we consider the problem of designing a container liner shipping feeder network. The designer has to choose which port to serve during many rotations that start and end at a central hub. Many operational characteristics are considered, such as variable leg-by-leg speeds and cargo...

  14. Mitochondrial calcium uniporter silencing potentiates caspase-independent cell death in MDA-MB-231 breast cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Curry, Merril C.; Peters, Amelia A. [School of Pharmacy, The University of Queensland, Brisbane, Queensland 4072 (Australia); Kenny, Paraic A. [Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, New York 10461 (United States); Roberts-Thomson, Sarah J. [School of Pharmacy, The University of Queensland, Brisbane, Queensland 4072 (Australia); Monteith, Gregory R., E-mail: gregm@uq.edu.au [School of Pharmacy, The University of Queensland, Brisbane, Queensland 4072 (Australia)

    2013-05-10

    Highlights: •Some clinical breast cancers are associated with MCU overexpression. •MCU silencing did not alter cell death initiated with the Bcl-2 inhibitor ABT-263. •MCU silencing potentiated caspase-independent cell death initiated by ionomycin. •MCU silencing promoted ionomycin-mediated cell death without changes in bulk Ca{sup 2+}. -- Abstract: The mitochondrial calcium uniporter (MCU) transports free ionic Ca{sup 2+} into the mitochondrial matrix. We assessed MCU expression in clinical breast cancer samples using microarray analysis and the consequences of MCU silencing in a breast cancer cell line. Our results indicate that estrogen receptor negative and basal-like breast cancers are characterized by elevated levels of MCU. Silencing of MCU expression in the basal-like MDA-MB-231 breast cancer cell line produced no change in proliferation or cell viability. However, distinct consequences of MCU silencing were seen on cell death pathways. Caspase-dependent cell death initiated by the Bcl-2 inhibitor ABT-263 was not altered by MCU silencing; whereas caspase-independent cell death induced by the calcium ionophore ionomycin was potentiated by MCU silencing. Measurement of cytosolic Ca{sup 2+} levels showed that the promotion of ionomycin-induced cell death by MCU silencing occurs independently of changes in bulk cytosolic Ca{sup 2+} levels. This study demonstrates that MCU overexpression is a feature of some breast cancers and that MCU overexpression may offer a survival advantage against some cell death pathways. MCU inhibitors may be a strategy to increase the effectiveness of therapies that act through the induction of caspase-independent cell death pathways in estrogen receptor negative and basal-like breast cancers.

  15. Design of Two Feeder Three Phase Four Wire Distribution System Utilizing Multi Converter UPQC with Fuzzy Logic Controller

    Directory of Open Access Journals (Sweden)

    Chandra Babu Paduchuri

    2014-01-01

    Full Text Available This paper proposes the instantaneous p-q theory based fuzzy logic controller (FLC for multi converter unified power quality conditioner (MC-UPQC to mitigate power quality issues in two feeders three-phase four-wire distribution systems. The proposed system is extended system of the existing one feeder three-phase four-wire distribution system, which is operated with UPQC. This system is employed with three voltage source converters, which are connected commonly to two feeder distribution systems. The performance of this proposed system used to compensate voltage sag, neutral current mitigation and compensation of voltage and current harmonics under linear and nonlinear load conditions. The neutral current flowing in series transformers is zero in the implementation of the proposed system. The simulation performance analysis is carried out using MATLAB.

  16. Interactions among contaminant exposure route, kinetics and toxicity in marine deposit-feeders

    DEFF Research Database (Denmark)

    Selck, Henriette

    Many of the chemicals having the greatest likelihood of causing damage in ecological systems, such as polycyclic aromatic hydrocarbons (PAHs) and metals are of persistent and/or hydrophobic character. Deposit-feeders may be important in facilitating the removal of organic contaminants from bulk...

  17. Rat embryonic fibroblasts improve reprogramming of human keratinocytes into induced pluripotent stem cells.

    Science.gov (United States)

    Linta, Leonhard; Stockmann, Marianne; Kleinhans, Karin N; Böckers, Anja; Storch, Alexander; Zaehres, Holm; Lin, Qiong; Barbi, Gotthold; Böckers, Tobias M; Kleger, Alexander; Liebau, Stefan

    2012-04-10

    Patient-specific human induced pluripotent stem (hiPS) cells not only provide a promising tool for cellular disease models in general, but also open up the opportunity to establish cell-type-specific systems for personalized medicine. One of the crucial prerequisites for these strategies, however, is a fast and efficient reprogramming strategy from easy accessible somatic cell populations. Keratinocytes from plucked human hair had been introduced as a superior cell source for reprogramming purposes compared with the widely used skin fibroblasts. The starting cell population is, however, limited and thereby further optimization in terms of time, efficiency, and quality is inevitable. Here we show that rat embryonic fibroblasts (REFs) should replace mouse embryonic fibroblasts as feeder cells in the reprogramming process. REFs enable a significantly more efficient reprogramming procedure as shown by colony number and total amount of SSEA4-positive cells. We successfully produced keratinocyte-derived hiPS (k-hiPS) cells from various donors. The arising k-hiPS cells display the hallmarks of pluripotency such as expression of stem cell markers and differentiation into all 3 germ layers. The increased reprogramming efficiency using REFs as a feeder layer occurred independent of the proliferation rate in the parental keratinocytes and acts, at least in part, in a non-cell autonomous way by secreting factors known to facilitate pluripotency such as Tgfb1, Inhba and Grem1. Hence, we provide an easy to use and highly efficient reprogramming system that could be very useful for a broad application to generate human iPS cells. © Mary Ann Liebert, Inc.

  18. A HYBRID APPROACH FOR RURAL FEEDER DESIGN

    Directory of Open Access Journals (Sweden)

    DAMANJEET KAUR

    2012-08-01

    Full Text Available In this paper, a population based approach for conductor size selection in rural radial distribution system is presented. The proposed hybrid approach implies a particle swarm optimization (PSO approach in combination with mutant property of differential evolution (DE for conductor size selection in radial distribution system. The conductor size for each feeder segment is selected such that the total cost of capital investment and capitalized cost of energy losses is minimized while constraints of voltage at each node and current carrying capacity of conductor is within the limits. The applicability and effectiveness of the proposed method is demonstrated with the help of 32-node test system.

  19. The effect of stocking density and bin feeder space on performance ...

    African Journals Online (AJOL)

    Unknown

    The effect of stocking density and bin feeder space on performance in pigs. G.A. Lavers# and N.S. Ferguson. School of Agricultural Sciences & Agribusiness, University of Natal, P Bag X01, Scottsville 3209. #Email: lavers@agric.unp.ac.za. Introduction. Pigs housed individually have been shown to have higher feed intakes ...

  20. Transition to independent practice: a national enquiry into the educational support for newly qualified GPs.

    Science.gov (United States)

    Griffin, Ann; Abouharb, Tareq; Etherington, Clare; Bandura, Induja

    2010-09-01

    The nature of the work that NQGPs are undertaking in their transition to independent practice is changing; current training may not fully prepare them for this new peripatetic role, as indicated by rising numbers of reports of poor performance in this group. Educational support at the time of transition from general practitioner (GP) training to independent practice had previously demonstrated benefits, but many formal schemes have finished. This study aimed to map out the current provision of educational support provided by the UK deaneries for NQGPs and to explore NQGPs' perceptions of the present transition from registrar to independent practitioner. Questionnaire surveys of deanery provision and semi-structured telephone interviews of a purposeful sample of newly qualified GPs across the UK. Interviews were thematically analysed. Deanery provision of educational support to NQGPs varies across the UK. Telephone interviews highlighted the transformation as problematic; NQGPs perceived that independent practice was substantially different from being in a training post - locum work, isolation and accessing educational opportunities were concerns. NQGPs frequently expressed a desire for more formalised relationships with mentors, senior colleagues or peer groups, to support their shift. As NQGPs increasingly find themselves working as locums, lacking the opportunity for stable work-based relationships, and with an increase in medical errors being reported in this group of doctors, it is suggested that there is a need to reconsider the educational support required to facilitate the transition in the early years of independent general practice.

  1. Changes in food intake and abnormal behavior using a puzzle feeder in newly acquired sub-adult rhesus monkeys (Macaca mulatta): a short term study.

    Science.gov (United States)

    Lee, Jae-Il; Lee, Chi-Woo; Kwon, Hyouk-Sang; Kim, Young-Tae; Park, Chung-Gyu; Kim, Sang-Joon; Kang, Byeong-Cheol

    2008-10-01

    The majority of newly acquired nonhuman primates encounter serious problems adapting themselves to new environments or facilities. In particular, loss of appetite and abnormal behavior can occur in response to environmental stresses. These adaptation abnormalities can ultimately have an affect on the animal's growth and well-being. In this study, we evaluated the affects of a puzzle feeder on the food intake and abnormal behavior of newly acquired rhesus monkeys for a short period. The puzzle feeder was applied to 47- to 58-month-old animals that had never previously encountered one. We found that there was no difference in the change of food intake between the bucket condition and the puzzle feeder condition. In contrast, the time spent for consumption of food was three times longer in the puzzle feeder condition than in the bucket condition. Two monkeys initially exhibited stereotypic behavior. One showed a decreasing, and the other an increasing pattern of abnormal behavior after introduction of the puzzle feeder. In conclusion, this result suggests that over a short period, the puzzle feeder can only affect the time for food consumption since it failed to affect the food intake and did not consistently influence stereotypic behaviors in newly acquired rhesus monkeys.

  2. Independent rate and temporal coding in hippocampal pyramidal cells.

    Science.gov (United States)

    Huxter, John; Burgess, Neil; O'Keefe, John

    2003-10-23

    In the brain, hippocampal pyramidal cells use temporal as well as rate coding to signal spatial aspects of the animal's environment or behaviour. The temporal code takes the form of a phase relationship to the concurrent cycle of the hippocampal electroencephalogram theta rhythm. These two codes could each represent a different variable. However, this requires the rate and phase to vary independently, in contrast to recent suggestions that they are tightly coupled, both reflecting the amplitude of the cell's input. Here we show that the time of firing and firing rate are dissociable, and can represent two independent variables: respectively the animal's location within the place field, and its speed of movement through the field. Independent encoding of location together with actions and stimuli occurring there may help to explain the dual roles of the hippocampus in spatial and episodic memory, or may indicate a more general role of the hippocampus in relational/declarative memory.

  3. Resolving the architecture of monogenetic feeder systems from exposures of extinct volcanic fields

    Science.gov (United States)

    Muirhead, J.; Van Eaton, A. R.; Re, G.; White, J. D. L.; Ort, M. H.

    2016-12-01

    Monogenetic volcanic fields pose hazards to a number of major cities worldwide. During an eruption, the evolution of the intrusive feeder network modulates eruption behavior and location, as well as the warning signs of impending activity. However, historical examples of monogenetic eruptions are rare, particularly those monitored with the modern tools required to constrain the geometry and interconnectivity of subsurface intrusive feeders (e.g., InSAR, GPS). Geologic exposures in extinct fields around the Colorado Plateau provide clues to the geometry of shallow intrusions (<1000 m depth) that feed monogenetic volcanoes. We present field- and satellite-based observations of exposed intrusions in the Hopi Buttes volcanic field (Arizona), which reveal that many eruptions were fed by interconnected dike-sill systems. Results from the Hopi Buttes show that volcanic cone alignment studies are biased to the identification of dike intrusions, and thereby neglect the important contributions of sills to shallow feeder systems. For example, estimates of intruded volumes in fields exhumed by uplift and erosion in Utah and Arizona show that sills make up 30 - 92% of the shallow intruded volume within 1000 m of the paleosurface. By transporting magma toward and away from eruptive conduits, these sills likely played a role in modulating eruption styles (e.g., explosive vs effusive) and controlling lateral vent migrations. Sill transitions at Hopi Buttes would have produced detectable surface uplifts, and illustrate the importance of geological studies for informing interpretations of geodetic and seismological data during volcanic crises.

  4. The effects of automated scatter feeders on captive grizzly bear activity budgets.

    Science.gov (United States)

    Andrews, Nathan L P; Ha, James C

    2014-01-01

    Although captive bears are popular zoo attractions, they are known to exhibit high levels of repetitive behaviors (RBs). These behaviors have also made them particularly popular subjects for welfare research. To date, most research on ursid welfare has focused on various feeding methods that seek to increase time spent searching for, extracting, or consuming food. Prior research indicates an average of a 50% reduction in RBs when attempts are successful and, roughly, a 50% success rate across studies. This research focused on decreasing time spent in an RB while increasing the time spent active by increasing time spent searching for, extracting, and consuming food. The utility of timed, automated scatter feeders was examined for use with captive grizzly bears (Ursis arctos horribilis). Findings include a significant decrease in time spent in RB and a significant increase in time spent active while the feeders were in use. Further, the bears exhibited a wider range of behaviors and a greater use of their enclosure.

  5. Dosage and cell line dependent inhibitory effect of bFGF supplement in human pluripotent stem cell culture on inactivated human mesenchymal stem cells.

    Science.gov (United States)

    Quang, Tara; Marquez, Maribel; Blanco, Giselle; Zhao, Yuanxiang

    2014-01-01

    Many different culture systems have been developed for expanding human pluripotent stem cells (hESCs and hiPSCs). In general, 4-10 ng/ml of bFGF is supplemented in culture media in feeder-dependent systems regardless of feeder cell types, whereas in feeder-free systems, up to 100 ng/ml of bFGF is required for maintaining long-term culture on various substrates. The amount of bFGF required in native hESCs growth niche is unclear. Here we report using inactivated adipose-derived human mesenchymal stem cells as feeder cells to examine long-term parallel cultures of two hESCs lines (H1 and H9) and one hiPSCs line (DF19-9-7T) in media supplemented with 0, 0.4 or 4 ng/ml of bFGF for up to 23 passages, as well as parallel cultures of H9 and DF19 in media supplemented with 4, 20 or 100 ng/ml bFGF for up to 13 passages for comparison. Across all cell lines tested, bFGF supplement demonstrated inhibitory effect over growth expansion, single cell colonization and recovery from freezing in a dosage dependent manner. In addition, bFGF exerted differential effects on different cell lines, inducing H1 and DF19 differentiation at 4 ng/ml or higher, while permitting long-term culture of H9 at the same concentrations with no apparent dosage effect. Pluripotency was confirmed for all cell lines cultured in 0, 0.4 or 4 ng/ml bFGF excluding H1-4 ng, as well as H9 cultured in 4, 20 and 100 ng/ml bFGF. However, DF19 demonstrated similar karyotypic abnormality in both 0 and 4 ng/ml bFGF media while H1 and H9 were karyotypically normal in 0 ng/ml bFGF after long-term culture. Our results indicate that exogenous bFGF exerts dosage and cell line dependent effect on human pluripotent stem cells cultured on mesenchymal stem cells, and implies optimal use of bFGF in hESCs/hiPSCs culture should be based on specific cell line and its culture system.

  6. Mitochondrial clearance by the STK38 kinase supports oncogenic Ras-induced cell transformation

    Science.gov (United States)

    Bettoun, Audrey; Surdez, Didier; Vallerand, David; Gundogdu, Ramazan; Sharif, Ahmad A.D.; Gomez, Marta; Cascone, Ilaria; Meunier, Brigitte; White, Michael A.; Codogno, Patrice; Parrini, Maria Carla; Camonis, Jacques H.; Hergovich, Alexander

    2016-01-01

    Oncogenic Ras signalling occurs frequently in many human cancers. However, no effective targeted therapies are currently available to treat patients suffering from Ras-driven tumours. Therefore, it is imperative to identify downstream effectors of Ras signalling that potentially represent promising new therapeutic options. Particularly, considering that autophagy inhibition can impair the survival of Ras-transformed cells in tissue culture and mouse models, an understanding of factors regulating the balance between autophagy and apoptosis in Ras-transformed human cells is needed. Here, we report critical roles of the STK38 protein kinase in oncogenic Ras transformation. STK38 knockdown impaired anoikis resistance, anchorage-independent soft agar growth, and in vivo xenograft growth of Ras-transformed human cells. Mechanistically, STK38 supports Ras-driven transformation through promoting detachment-induced autophagy. Even more importantly, upon cell detachment STK38 is required to sustain the removal of damaged mitochondria by mitophagy, a selective autophagic process, to prevent excessive mitochondrial reactive oxygen species production that can negatively affect cancer cell survival. Significantly, knockdown of PINK1 or Parkin, two positive regulators of mitophagy, also impaired anoikis resistance and anchorage-independent growth of Ras-transformed human cells, while knockdown of USP30, a negative regulator of PINK1/Parkin-mediated mitophagy, restored anchorage-independent growth of STK38-depleted Ras-transformed human cells. Therefore, our findings collectively reveal novel molecular players that determine whether Ras-transformed human cells die or survive upon cell detachment, which potentially could be exploited for the development of novel strategies to target Ras-transformed cells. PMID:27283898

  7. Role of non-canonical Beclin 1-independent autophagy in cell death induced by resveratrol in human breast cancer cells.

    Science.gov (United States)

    Scarlatti, F; Maffei, R; Beau, I; Codogno, P; Ghidoni, R

    2008-08-01

    Resveratrol, a polyphenol found in grapes and other fruit and vegetables, is a powerful chemopreventive and chemotherapeutic molecule potentially of interest for the treatment of breast cancer. The human breast cancer cell line MCF-7, which is devoid of caspase-3 activity, is refractory to apoptotic cell death after incubation with resveratrol. Here we show that resveratrol arrests cell proliferation, triggers death and decreases the number of colonies of cells that are sensitive to caspase-3-dependent apoptosis (MCF-7 casp-3) and also those that are unresponsive to it (MCF-7vc). We demonstrate that resveratrol (i) acts via multiple pathways to trigger cell death, (ii) induces caspase-dependent and caspase-independent cell death in MCF-7 casp-3 cells, (iii) induces only caspase-independent cell death in MCF-7vc cells and (iv) stimulates macroautophagy. Using BECN1 and hVPS34 (human vacuolar protein sorting 34) small interfering RNAs, we demonstrate that resveratrol activates Beclin 1-independent autophagy in both cell lines, whereas cell death via this uncommon form of autophagy occurs only in MCF-7vc cells. We also show that this variant form of autophagic cell death is blocked by the expression of caspase-3, but not by its enzymatic activity. In conclusion, this study reveals that non-canonical autophagy induced by resveratrol can act as a caspase-independent cell death mechanism in breast cancer cells.

  8. harmonic load modeling: a case study of 33 kv abuja steel mill feeder

    African Journals Online (AJOL)

    HOD

    An in-depth study of the harmonic orders inherent in a power system network is required ... This paper studied the harmonic orders of the 33 kV Abuja Steel Feeder .... models for various industrial and household electrical ..... Malaysia, 2013.

  9. The social basis of separatism: explaining support for the Puerto Rican Independence Movement

    Directory of Open Access Journals (Sweden)

    Astrid Arrarás

    2007-12-01

    Full Text Available In the 1990s, particularly in the wake of the fall of the Soviet Union, comparativists rediscovered the enduring appeal of separatist movements around the globe. Many of the most significant political events of the past decade have revolved around issues of nationalism, ethnicity, irredenta, and the like. In light of these renewed preoccupations, it is surprising that U.S. scholars have largely ignored a nationalist movement closer to home. The Puerto Rican independence movement is by far the most significant separatist tendency under the U.S. flag, and is one of the few major anticolonialist movements to survive into the twenty-first century. Although supporters of Puerto Rican independence have typically drawn only three to five percent support in local elections and in plebiscites on the island’s political status, neither have they disappeared from the political scene. The remarkable durability of the independence movement demands systematic explanation. However, as strange as it may seem, to date there have been no serious scholarly studies of the social bases of the Puerto Rican independence movement.

  10. Cone-Beam Computed Tomography (CBCT) Hepatic Arteriography in Chemoembolization for Hepatocellular Carcinoma: Performance Depicting Tumors and Tumor Feeders

    Energy Technology Data Exchange (ETDEWEB)

    Lee, In Joon [National Cancer Center, Department of Radiology (Korea, Republic of); Chung, Jin Wook, E-mail: chungjw@snu.ac.kr; Yin, Yong Hu; Kim, Hyo-Cheol; Kim, Young Il; Jae, Hwan Jun; Park, Jae Hyung [Seoul National University College of Medicine, Seoul National University Hospital, Department of Radiology (Korea, Republic of)

    2015-10-15

    PurposeThis study was designed to analyze retrospectively the performance of cone-beam computed tomography (CBCT) hepatic arteriography in depicting tumors and their feeders and to investigate the related determining factors in chemoembolization for hepatocellular carcinoma (HCC).MethodsEighty-six patients with 142 tumors satisfying the imaging diagnosis criteria of HCC were included in this study. The performance of CBCT hepatic arteriography for chemoembolization per tumor and per patient was evaluated using maximum intensity projection images alone (MIP analysis) or MIP combined with multiplanar reformation images (MIP + MPR analysis) regarding the following three aspects: tumor depiction, confidence of tumor feeder detection, and trackability of tumor feeders. Tumor size, tumor enhancement, tumor location, number of feeders, diaphragmatic motion, portal vein enhancement, and hepatic artery to parenchyma enhancement ratio were regarded as potential determining factors.ResultsTumors were depicted in 125 (88.0 %) and 142 tumors (100 %) on MIP and MIP + MPR analysis, respectively. Imaging performances on MIP and MIP + MPR analysis were good enough to perform subsegmental chemoembolization without additional angiographic investigation in 88 (62.0 %) and 128 tumors (90.1 %) on per-tumor basis and in 43 (50 %) and 73 (84.9 %) on per-patient basis, respectively. Significant determining factors for performance in MIP + MPR analysis on per tumor basis were tumor size (p = 0.030), tumor enhancement (0.005), tumor location (p = 0.001), and diaphragmatic motion (p < 0.001).ConclusionsCBCT hepatic arteriography provided sufficient information for subsegmental chemoembolization by depicting tumors and their feeders in the vast majority of patients. Combined analysis of MIP and MPR images was essential to enhance the performance of CBCT hepatic arteriography.

  11. Flow visualization study of two-phase flow in a single bend outlet feeder pipe of a CANDU reactor

    International Nuclear Information System (INIS)

    Savalaxs, S.-A.; Lister, D.H.; Steward, F.R.

    2005-01-01

    In CANDU reactors, the feeder piping that is used to direct the high-temperature water coolant between the fuel channels and the steam generators is made of carbon steel. Since 1996, several CANDU stations have reported excessive corrosion of their outlet feeders. The first metre is particularity vulnerable because the piping there consists of single or double bends, which have relatively thin walls produced by the bending process. Early studies related the attack to the hydrodynamics of the coolant and verified that it was a type of flow-accelerated corrosion. In order to understand the hydrodynamics of the coolant in the outlet feeders by flow visualization, a full-scale transparent test section simulating the geometry and orientation of an outlet feeder bend with its upstream components was fabricated. The feeder consisted of a 54 mm diameter acrylic pipe with a 73 degree bend. This was connected to the upstream component with an acrylic simulation of a Grayloc flanged fitting. A test loop supplied room temperature water to the test section at flow rates up to 0.019 m3/s. Air could be injected into the water to give a mean volume fraction of up to 0.56. In this preliminary investigation, the size and velocity of air bubbles at different flow conditions and their distribution within the pipe bend were studied. Particular attention was paid to the flow pattern at the inside of the bend, where a CFD (computational fluid dynamics) code - Fluent 6.1-had failed to predict a liquid film in an earlier study. A high-speed digital video camera was used to determine the relation between bubble size and velocity. Such a relation should help to explain the discrepancy in the CFD modelling and provide the basis for accurate predictions of phase distribution in complex geometries at high flow rates. (authors)

  12. Optimalisatie performance van vleeskuikens bij mobiel voer- en drinksysteem (scanFeeder) op praktijkbedrijf

    NARCIS (Netherlands)

    Harn, van J.; Veldkamp, T.

    2007-01-01

    Het gebruik van de Ridder ScanFeeder, een mobiel voer- en drinksysteem met strooiselbeluchting voor vleeskuikens, geeft een betere strooiselkwaliteit (ruller en droger strooisel) en minder voetzoolaandoeningen. De technische resultaten bij dit systeem blijven echter achter bij die van een

  13. Manufacture and test of prototype water pipe chase barrier in ITER Magnet Feeder system

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Kun, E-mail: lukun@ipp.ac.cn [Institute of Plasma Physics, Shushan Hu Road 350, Hefei, Anhui (China); Wen, Xinjie; Liu, Chen; Song, Yuntao [Institute of Plasma Physics, Shushan Hu Road 350, Hefei, Anhui (China); Niu, Erwu [ITER China, 15B Fuxing Road, Beijing 100862 (China); Gung, Chenyu; Su, Man [ITER Organization, Route de Vinon-sur-Verdon – CS 90046, 13067 St Paul-lez-Durance Cedex (France)

    2016-11-01

    The Magnet Feeder system in the International Thermonuclear Experimental Reactor (ITER) deploys electrical currents and supercritical helium to the superconducting magnets and the magnet diagnostic signals to the operators. In the current design, the feeders located in the upper L3 level of the Tokamak gallery penetrate the Tokamak coolant water system vault, the biological shield and the cryostat. As a secondary confinement to contain the activated coolant water in the vault in the case of water pipe burst accident, a water barrier is welded between the penetration in the water pipe chase outer wall and the mid-plane of the vacuum jacket of the Feeder Coil Terminal Box (CTB). A thin-wall stainless steel diaphragm with an omega shape profile is welded around the CTB as the water barrier to endure 2 bar hydraulic pressure. In addition, the barrier is designed as a flexible compensator to withstand a maximum of 15 mm of axial displacement of the CTB in case of helium leak accident without failure. This paper presents the detail configuration, the manufacturing and assembly processes of the water barrier. Test results of the prototype water barrier under simulated accident conditions are also reported. Successful qualification of the design and manufacturing process of the water barrier lays a good foundation for the series production of this subsystem.

  14. Human embryonic stem cells and microenvironment

    Directory of Open Access Journals (Sweden)

    Banu İskender

    2014-09-01

    Full Text Available Human embryonic stem cells (hESCs possess a great potential in the field of regenerative medicine by their virtue of pluripotent potential with indefinite proliferation capabilities. They can self renew themselves and differentiate into three embryonic germ layers. Although they are conventionally grown on mitotically inactivated mouse feeder cells, there are in vitro culture systems utilizing feeder cells of human origin in order to prevent cross-species contamination. Recently established in vitro culture systems suggested that direct interaction with feeder cells is not necessary but rather attachment to a substrate is required to ensure long-term, efficient hESC culture in vitro. This substrate is usually composed of a mixture of extracellular matrix components representing in vivo natural niche. In hESC biology, the mechanism of interaction of hESCs with extracellular matrix molecules remained insufficiently explored area of research due to their transient nature of interaction with the in vivo niche. However, an in vitro culture system established using extracellular matrix molecules may provide a safer alternative to culture systems with feeder cells while paving the way to Good Manufacturing Practice-GMP production of hESCs for therapeutic purposes. Therefore, it is essential to study the interaction of extracellular matrix molecules with hESCs in order to standardize in vitro culture systems for large-scale production of hESCs in a less labor-intensive way. This would not only provide valuable information regarding the mechanisms that control pluripotency but also serve to dissect the molecular signaling pathways of directed differentiation for prospective therapeutic applications in the future. J Clin Exp Invest 2014; 5 (3: 486-495

  15. Overview and Categorization of Robots Supporting Independent Living of Elderly People: What Activities Do They Support and How Far Have They Developed.

    Science.gov (United States)

    Bedaf, Sandra; Gelderblom, Gert Jan; De Witte, Luc

    2015-01-01

    Over the past decades, many robots for the elderly have been developed, supporting different activities of elderly people. A systematic review in four scientific literature databases and a search in article references and European projects was performed in order to create an overview of robots supporting independent living of elderly people. The robots found were categorized based on their development stage, the activity domains they claim to support, and the type of support provided (i.e., physical, non-physical, and/or non-specified). In total, 107 robots for the elderly were identified. Six robots were still in a concept phase, 95 in a development phase, and six of these robots were commercially available. These robots claimed to provide support related to four activity domains: mobility, self-care, interpersonal interaction & relationships, and other activities. Of the many robots developed, only a small percentage is commercially available. Technical ambitions seem to be guiding robot development. To prolong independent living, the step towards physical support is inevitable and needs to be taken. However, it will be a long time before a robot will be capable of supporting multiple activities in a physical manner in the home of an elderly person in order to enhance their independent living.

  16. Mannosides as crucial part of bioactive supports for cultivation of human epidermal keratinocytes without feeder cells

    Czech Academy of Sciences Publication Activity Database

    Labský, Jiří; Dvořánková, B.; Smetana, Karel; Holíková, Z.; Brož, L.; Gabius, H.J.

    2003-01-01

    Roč. 24, č. 5 (2003), s. 863-872 ISSN 0142-9612 R&D Projects: GA ČR GA203/00/1310; GA MŠk LN00A065; GA MZd ND6340 Institutional research plan: CEZ:AV0Z4050913 Keywords : cell therapy * keratinocyte * mannose Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.903, year: 2003

  17. In vitro study of cell differentiation by two type mouse embryo stem cells on mono- and multilayer nanocarbon tubes

    Science.gov (United States)

    Imai, Koichi; Akasaka, Tsukasa; Watari, Fumio; Tanoue, Akito; Nakamura, Kazuaki; Suese, Kazuhiko; Takashima, Hiromasa; Nishikawa, Tetsunari; Tanaka, Akio; Takeda, Shoji

    2012-09-01

    The effects of nanomaterials on human reproduction and development remain unknown. The risks of nanomaterials for future generations should be elucidated. Thus, it is important to establish an experimental method to accurately examine embryotoxicity. We previously investigated the myocardial cell differentiation of ES-D3 cells using monolayer (SWCNTs) and multilayer (MWCNTs) nanocarbon tubes. As a result, in spite of having the same carbon composition, the effects on the cell differentiation levels differed between the tubes. We investigated their cell differentiation and cytotoxic effects on EL M3 and ES-R1-EGFP B2/EGFP cells, which require feeder cells. As a result, myocardial pulse rates differed between the presence of SWCNTs and MWCNTs even when feeder cells existed between the samples and cells. The different surface structures of SWCNTs and MWCNTs may have influenced ES cell differentiation.

  18. Effect of concentrate feeder design on performance, eating and animal behavior, welfare, ruminal health, and carcass quality in Holstein bulls fed high-concentrate diets.

    Science.gov (United States)

    Verdú, M; Bach, A; Devant, M

    2015-06-01

    A total of 240 Holstein bulls (121 ± 2.0 kg initial BW; 99 ± 1.0 d of age), from 2 consecutive fattening cycles, were randomly allocated in 1 of 6 pens and assigned to 1 of the 3 treatments consisting of different concentrate feeder designs: a control feeder with 4 feeding spaces (CF), a feeder with less concentrate capacity (CFL), and a single-space feeder with lateral protections (SF). Each pen had a straw feeder and a drinker. All animals were fed a high-concentrate diet for ad libitum intake. Concentrate consumption was recorded daily using a computerized feeder, straw consumption was recorded weekly, and BW was recorded every 14 d. Animal behavior was registered on d 1, 3, 5, 8, and 14 and every 28 d by scan sampling. Eating behavior at concentrate feeders was filmed on d 12, 125, and 206. On d 7, 120, and 204, samples of rumen contents were collected for measurement of pH and VFA and blood samples were obtained to analyze NEFA, haptoglobin, glucose, and insulin. Animals were slaughtered after 223 d, and HCW and lesions of the rumen wall and liver were recorded. The accumulative concentrate consumption per animal tended (P = 0.09) to be greater with CF than with CFL and SF. Also, CV of concentrate consumption was greater (P carcass data. Also, no differences among treatments in rumen wall evaluation and liver abscesses were observed. At 7 and 204 d of study, SF bulls had greater (P animal welfare in Holstein bulls fed high-concentrate diets. However, at the beginning, there was evidence that animals fed using SF had problems with adaptation.

  19. Some software issues in mapping of power distribution feeders

    International Nuclear Information System (INIS)

    Mufti, I.A.

    1994-01-01

    This paper is about the in-house developed software for distribution feeders mapping project. It first gives birds eye view of the project, highlight its technical complexity in management and logistics, introduced by sheer size of the project. It gives an overview of the software developed and the moves on to describe circuit tracing, circuit model, leaves isolation (for tree structured network) and backtracking in more detail among many different parts of software, description of all which is not possible because of space limitations. (author)

  20. Enamel tissue engineering using subcultured enamel organ epithelial cells in combination with dental pulp cells.

    Science.gov (United States)

    Honda, Masaki J; Shinmura, Yuka; Shinohara, Yoshinori

    2009-01-01

    We describe a strategy for the in vitro engineering of enamel tissue using a novel technique for culturing enamel organ epithelial (EOE) cells isolated from the enamel organ using 3T3-J2 cells as a feeder layer. These subcultured EOE cells retain the capacity to produce enamel structures over a period of extended culture. In brief, enamel organs from 6-month-old porcine third molars were dissociated into single cells and subcultured on 3T3-J2 feeder cell layers. These subcultured EOE cells were then seeded onto a collagen sponge in combination with primary dental pulp cells isolated at an early stage of crown formation, and these constructs were transplanted into athymic rats. After 4 weeks, complex enamel-dentin structures were detected in the implants. These results show that our culture technique maintained ameloblast lineage cells that were able to produce enamel in vivo. This novel subculture technique provides an important tool for tooth tissue engineering. Copyright 2008 S. Karger AG, Basel.

  1. Coworking Spaces: A Source of Social Support for Independent Professionals.

    Science.gov (United States)

    Gerdenitsch, Cornelia; Scheel, Tabea E; Andorfer, Julia; Korunka, Christian

    2016-01-01

    Coworking spaces are shared office environments for independent professionals. Such spaces have been increasing rapidly throughout the world, and provide, in addition to basic business infrastructure, the opportunity for social interaction. This article explores social interaction in coworking spaces and reports the results of two studies. Study 1 (N = 69 coworkers) finds that social interaction in coworking spaces can take the form of social support. Study 2 further investigates social support among coworkers (N = 154 coworkers) and contrasts these results with those of social support among colleagues in traditional work organizations (N = 609). A moderated mediation model using time pressure and self-efficacy, based on the conservation of resources theory, is tested. Social support from both sources was positively related to performance satisfaction. Self-efficacy mediated this relationship in the employee sample, while in the coworking sample, self-efficacy only mediated the relationship between social support and performance satisfaction if time pressure was high. Thus, a mobilization of social support seems necessary in coworking spaces. We conclude that coworking spaces, as modern social work environments, should align flexible work infrastructure with well-constructed opportunities for social support.

  2. Coworking Spaces: A Source of Social Support for Independent Professionals

    Directory of Open Access Journals (Sweden)

    Cornelia eGerdenitsch

    2016-04-01

    Full Text Available Coworking spaces are shared office environments for independent professionals. Such spaces have been increasing rapidly throughout the world, and provide, in addition to basic business infrastructure, the opportunity for social interaction. This article explores social interaction in coworking spaces and reports the results of two studies. Study 1 (N = 69 coworkers finds that social interaction in coworking spaces can take the form of social support. Study 2 further investigates social support among coworkers (N = 154 coworkers and contrasts these results with those of social support among colleagues in traditional work organizations (N = 609. A moderated mediation model using time pressure and self-efficacy, based on the conservation of resources theory, is tested. Social support from both sources was positively related to performance satisfaction. Self-efficacy mediated this relationship in the employee sample, while in the coworking sample, self-efficacy only mediated the relationship between social support and performance satisfaction if time pressure was high. Thus, a mobilization of social support seems necessary in coworking spaces. We conclude that coworking spaces, as modern social work environments, should align flexible work infrastructure with well-constructed opportunities for social support.

  3. Feeder use predicts both acquisition and transmission of a contagious pathogen in a North American songbird.

    Science.gov (United States)

    Adelman, James S; Moyers, Sahnzi C; Farine, Damien R; Hawley, Dana M

    2015-09-22

    Individual heterogeneity can influence the dynamics of infectious diseases in wildlife and humans alike. Thus, recent work has sought to identify behavioural characteristics that contribute disproportionately to individual variation in pathogen acquisition (super-receiving) or transmission (super-spreading). However, it remains unknown whether the same behaviours enhance both acquisition and transmission, a scenario likely to result in explosive epidemics. Here, we examined this possibility in an ecologically relevant host-pathogen system: house finches and their bacterial pathogen, Mycoplasma gallisepticum, which causes severe conjunctivitis. We examined behaviours likely to influence disease acquisition (feeder use, aggression, social network affiliations) in an observational field study, finding that the time an individual spends on bird feeders best predicted the risk of conjunctivitis. To test whether this behaviour also influences the likelihood of transmitting M. gallisepticum, we experimentally inoculated individuals based on feeding behaviour and tracked epidemics within captive flocks. As predicted, transmission was fastest when birds that spent the most time on feeders initiated the epidemic. Our results suggest that the same behaviour underlies both pathogen acquisition and transmission in this system and potentially others. Identifying individuals that exhibit such behaviours is critical for disease management. © 2015 The Author(s).

  4. Measurements of local mass transfer coefficient of Flow Accelerated Corrosion at feeder outlet pipes in CANDU using Plaster of Paris method

    International Nuclear Information System (INIS)

    Hyuk Kwon; Dong Un Seo; Goon-Cherl Park

    2005-01-01

    Full text of publication follows: Flow Accelerated Corrosion (FAC) is a corrosion mechanism that results in wall thinning in piping systems and components. FAC on carbon steels in pure water has occurred in a number of industry and power plant. A pipe wall thinning due to the FAC at nuclear power plant is just reported in confined to carbon steel pipe on the secondary system which does not equip a radioactive component. Recently, at Canadian Deuterium Uranium type reactor, it is reported that the feeder lines suffer the wall thinning on the primary system to equip 380 feeder pipes. Differently from the fast FAC rate of the secondary system on PWR, FAC rate of feeder pipes on CANDU is not more than the 0.2 mm/year. Although the FAC rate of feeder pipe is relatively slow, the narrow thickness margin of 2.6089 mm is endangered sufficiently by only the slow FAC rate. The FAC is governed by the mass transfer coefficient which is determined by the flow field. To well predict the FAC rate, mass transfer coefficient should be well measured or calculated. New measurement method of mass transfer coefficient is developed to obtain the coefficient for the complex shape like feeder pipe. The method evaluated the naphthalene sublimation method to apply it under the water flow. The naphthalene sublimation method can be used to study mass and heat transfer with confidence for a variety of applications, but with certain restrictions. At present, Plaster of paris method can modified the sublimation method to overcome the restrictions. This method is particularly useful in complex flows on geometries and for flows with large gradients in wall transport rate. The test specimen can be easily prepared by several methods, including dipping, machining, spraying, and casting. The local transfer coefficients can be determined with high accuracy and in detail by automated measurement systems that eliminate most human errors during the measurement process. To evaluate the method, the

  5. DYNAMICS OF VIBRATION FEEDERS WITH A NONLINEAR ELASTIC CHARACTERISTIC

    Directory of Open Access Journals (Sweden)

    V. I. Dyrda

    2017-04-01

    Full Text Available Purpose. Subject to the smooth and efficient operation of each production line, is the use of vehicles transporting high specification. It worked well in practice for transporting construction machines, which are used during the vibration. The use of vibration machines requires optimization of their operation modes. In the form of elastic link in them are increasingly using rubber-metallic elements, which are characterized by nonlinear damping properties. So it is necessary to search for new, more modern, methods of calculation of dynamic characteristics of the vibration machines on the properties of rubber as a cushioning material. Methodology. The dynamics of vibration machine that is as elastic rubber block units and buffer shock absorbers limiting the amplitude of the vibrations of the working body. The method of determining amplitude-frequency characteristics of the vibrating feeder is based on the principle of Voltaire, who in the calculations of the damping properties of the dampers will allow for elastic-hereditary properties of rubber. When adjusting the basic dynamic stiffness of the elastic ties and vibratory buffers, using the principle of heredity rubber properties, determine the dependence of the amplitude of the working body of the machine vibrations. This method is called integro-operator using the fractional-exponential kernels of relaxation. Findings. Using the derived formula for determining the amplitude of the resonance curve is constructed one-mass nonlinear system. It is established that the use of the proposed method of calculation will provide a sufficiently complete description of the damping parameters of rubber-metallic elements and at the same time be an effective means of calculating the amplitude-frequency characteristics of nonlinear vibration systems. Originality. The authors improved method of determining damping characteristics of rubber-metallic elements and the amplitude-frequency characteristics of nonlinear

  6. Expression of a hyperactive androgen receptor leads to androgen-independent growth of prostate cancer cells.

    Science.gov (United States)

    Hsieh, Chen-Lin; Cai, Changmeng; Giwa, Ahmed; Bivins, Aaronica; Chen, Shao-Yong; Sabry, Dina; Govardhan, Kumara; Shemshedini, Lirim

    2008-07-01

    Cellular changes that affect the androgen receptor (AR) can cause prostate cancer to transition from androgen dependent to androgen independent, which is usually lethal. One common change in prostate tumors is overexpression of the AR, which has been shown to lead to androgen-independent growth of prostate cancer cells. This led us to hypothesize that expression of a hyperactive AR would be sufficient for androgen-independent growth of prostate cancer cells. To test this hypothesis, stable lune cancer prostate (LNCaP) cell lines were generated, which express a virion phosphoprotein (VP)16-AR hybrid protein that contains full-length AR fused to the strong viral transcriptional activation domain VP16. This fusion protein elicited as much as a 20-fold stronger transcriptional activity than the natural AR. Stable expression of VP16-AR in LNCaP cells yielded androgen-independent cell proliferation, while under the same growth conditions the parental LNCaP cells exhibited only androgen-dependent growth. These results show that expression of a hyperactive AR is sufficient for androgen-independent growth of prostate cancer cells. To study the molecular basis of this enhanced growth, we measured the expression of soluble guanylyl cyclase-alpha1 (sGCalpha1), a subunit of the sGC, an androgen-regulated gene that has been shown to be involved in prostate cancer cell growth. Interestingly, the expression of sGCalpha1 is androgen independent in VP16-AR-expressing cells, in contrast to its androgen-induced expression in control LNCaP cells. RNA(I)-dependent inhibition of sGCalpha1 expression resulted in significantly reduced proliferation of VP16-AR cells, implicating an important role for sGCalpha1 in the androgen-independent growth of these cells.

  7. In Vitro T-Cell Generation From Adult, Embryonic, and Induced Pluripotent Stem Cells: Many Roads to One Destination.

    Science.gov (United States)

    Smith, Michelle J; Webber, Beau R; Mohtashami, Mahmood; Stefanski, Heather E; Zúñiga-Pflücker, Juan Carlos; Blazar, Bruce R

    2015-11-01

    T lymphocytes are critical mediators of the adaptive immune system and have the capacity to serve as therapeutic agents in the areas of transplant and cancer immunotherapy. While T cells can be isolated and expanded from patients, T cells derived in vitro from both hematopoietic stem/progenitor cells (HSPCs) and human pluripotent stem cells (hPSCs) offer great potential advantages in generating a self-renewing source of T cells that can be readily genetically modified. T-cell differentiation in vivo is a complex process requiring tightly regulated signals; providing the correct signals in vitro to induce T-cell lineage commitment followed by their development into mature, functional, single positive T cells, is similarly complex. In this review, we discuss current methods for the in vitro derivation of T cells from murine and human HSPCs and hPSCs that use feeder-cell and feeder-cell-free systems. Furthermore, we explore their potential for adoption for use in T-cell-based therapies. © 2015 AlphaMed Press.

  8. Derivation and characterization of a pig embryonic stem cell-derived exocrine pancreatic cell line

    Science.gov (United States)

    The establishment and initial characterization of a pig embryonic stem cell-derived pancreatic cell line, PICM-31, and a colony-cloned derivative cell line, PICM-31A, is described. The cell lines were propagated for several months at split ratios of 1:3 or 1:5 at each passage on STO feeder cells af...

  9. Challenges associated with the current processes for ultrasonic inspection of CANDU reactor feeder piping

    Energy Technology Data Exchange (ETDEWEB)

    Machowski, C. [Babcock & Wilcox Canada Ltd., Cambridge, Ontario (Canada)

    2012-07-01

    CANDU® PHT Feeder Piping is generally constructed from SA106 Grade B carbon steel, which is known to be susceptible to flow accelerated corrosion when exposed to certain environmental conditions. The configuration of the CANDU reactor promotes thinning of the inside surface of the pipe walls, predominantly at the outlet feeders. Inspection of this piping is currently conducted using ultrasonic techniques and is governed by the requirements established by the CANDU Owners Group (COG). There are many challenges associated with these inspections as a result of the complexity of the reactor piping configuration. Geometrical anomalies on the surface of the pipe and non-circular geometries at the tight radius bends hinder the performance of conventional ultrasonic techniques. This can cause lost signals in areas of interest, which in turn often results in rework in order to satisfy the inspection requirements and justify fitness for service of these components. There are also many inspection sites which have limited access due to physical restrictions on the reactor face; therefore in order to maximize the performance of an inspection campaign, it is paramount that the inspection personnel and the inspection technology be well integrated through training simulations prior to execution. These inspection challenges increase the complexity of the analysis process as ultrasonic signals get distorted and lost as a result of non-circular pipe geometries. In order to ensure a high level of integrity in the analysis results, a conservative process is utilized in which two analysts independently examine the data, and a third analyst reviews their results and submits the final call. A Data Management Software application (DMS) is used to input and store the three analysis results. Another important function of the DMS is to provide a communication link between the different work-groups associated with the inspection activities. The focus of this presentation discusses:

  10. Rapid and non-enzymatic in vitro retrieval of tumour cells from surgical specimens.

    Directory of Open Access Journals (Sweden)

    Brigitte Mack

    Full Text Available The study of tumourigenesis commonly involves the use of established cell lines or single cell suspensions of primary tumours. Standard methods for the generation of short-term tumour cell cultures include the disintegration of tissue based on enzymatic and mechanical stress. Here, we describe a simple and rapid method for the preparation of single cells from primary carcinomas, which is independent of enzymatic treatment and feeder cells. Tumour biopsies are processed to 1 mm(3 cubes termed explants, which are cultured 1-3 days on agarose-coated well plates in specified medium. Through incisions generated in the explants, single cells are retrieved and collected from the culture supernatant and can be used for further analysis including in vitro and in vivo studies. Collected cells retain tumour-forming capacity in xenotransplantation assays, mimic the phenotype of the primary tumour, and facilitate the generation of cell lines.

  11. Optimization of culture conditions to support long-term self-renewal of buffalo (Bubalus bubalis) embryonic stem cell-like cells.

    Science.gov (United States)

    Sharma, Ruchi; George, Aman; Kamble, Nitin Manchindra; Singh, Karn Pratap; Chauhan, Manmohan Singh; Singla, Suresh Kumar; Manik, Radhey Sham; Palta, Prabhat

    2011-12-01

    A culture system capable of sustaining self-renewal of buffalo embryonic stem (ES) cell-like cells in an undifferentiated state over a long period of time was developed. Inner cell masses were seeded on KO-DMEM+15% KO-serum replacer on buffalo fetal fibroblast feeder layer. Supplementation of culture medium with 5 ng/mL FGF-2 and 1000 IU/mL mLIF gave the highest (p<0.05) rate of primary colony formation. The ES cell-like cells' colony survival rate and increase in colony size were highest (p<0.05) following supplementation with FGF-2 and LIF compared to other groups examined. FGF-2 supplementation affected the quantitative expression of NANOG, SOX-2, ACTIVIN A, BMP 4, and TGFβ1, but not OCT4 and GREMLIN. Supplementation with SU5402, an FGFR inhibitor (≥20 μM) increased (p<0.05) the percentage of colonies that differentiated. FGFR1-3 and ERK1, K-RAS, E-RAS, and SHP-2, key signaling intermediates of FGF signaling, were detected in ES cell-like cells. Under culture conditions described, three ES cell lines were derived that, to date, have been maintained for 135, 95, and 85 passages for over 27, 19, and 17 months, respectively, whereas under other conditions examined, ES cell-like cells did not survive beyond passage 10. The ES cell-like cells were regularly monitored for expression of pluripotency markers and their potency to form embryoid bodies.

  12. Activation of CHK1 in Supporting Cells Indirectly Promotes Hair Cell Survival

    Directory of Open Access Journals (Sweden)

    Azadeh Jadali

    2017-05-01

    Full Text Available The sensory hair cells of the inner ear are exquisitely sensitive to ototoxic insults. Loss of hair cells after exposure to ototoxic agents causes hearing loss. Chemotherapeutic agents such as cisplatin causes hair cell loss. Cisplatin forms DNA mono-adducts as well as intra- and inter-strand DNA crosslinks. DNA cisplatin adducts are repaired through the DNA damage response. The decision between cell survival and cell death following DNA damage rests on factors that are involved in determining damage tolerance, cell survival and apoptosis. Cisplatin damage on hair cells has been the main focus of many ototoxic studies, yet the effect of cisplatin on supporting cells has been largely ignored. In this study, the effects of DNA damage response in cochlear supporting cells were interrogated. Supporting cells play a major role in the development, maintenance and oto-protection of hair cells. Loss of supporting cells may indirectly affect hair cell survival or maintenance. Activation of the Phosphoinositide 3-Kinase (PI3K signaling was previously shown to promote hair cell survival. To test whether activating PI3K signaling promotes supporting cell survival after cisplatin damage, cochlear explants from the neural subset (NS Cre Pten conditional knockout mice were employed. Deletion of Phosphatase and Tensin Homolog (PTEN activates PI3K signaling in multiple cell types within the cochlea. Supporting cells lacking PTEN showed increased cell survival after cisplatin damage. Supporting cells lacking PTEN also showed increased phosphorylation of Checkpoint Kinase 1 (CHK1 levels after cisplatin damage. Nearest neighbor analysis showed increased numbers of supporting cells with activated PI3K signaling in close proximity to surviving hair cells in cisplatin damaged cochleae. We propose that increased PI3K signaling promotes supporting cell survival through phosphorylation of CHK1 and increased survival of supporting cells indirectly increases hair cell

  13. Hop/STI1 modulates retinal proliferation and cell death independent of PrPC

    International Nuclear Information System (INIS)

    Arruda-Carvalho, Maithe; Njaine, Brian; Silveira, Mariana S.; Linden, Rafael; Chiarini, Luciana B.

    2007-01-01

    Hop/STI1 is a co-chaperone adaptor protein for Hsp70/Hsp90 complexes. Hop/STI1 is found extracellularly and modulates cell death and differentiation through interaction with the prion protein (PrP C ). Here, we investigated the expression of hop/STI1 and its role upon cell proliferation and cell death in the developing retina. Hop/STI1 is more expressed in developing rat retina than in the mature tissue. Hop/STI1 blocks retinal cell death in the neuroblastic layer (NBL) in a PrP C dependent manner, but failed to protect ganglion cells against axotomy-induced cell death. An antibody raised against hop/STI1 (α-STI1) blocked both ganglion cell and NBL cell death independent of PrP C . cAMP/PKA, ERK, PI3K and PKC signaling pathways were not involved in these effects. Hop/STI1 treatment reduced proliferation, while α-STI1 increased proliferation in the developing retina, both independent of PrP C . We conclude that hop/STI1 can modulate both proliferation and cell death in the developing retina independent of PrP C

  14. Antigen-specific T cell activation independently of the MHC: chimeric antigen receptor (CAR-redirected T cells.

    Directory of Open Access Journals (Sweden)

    Hinrich eAbken

    2013-11-01

    Full Text Available Adoptive T cell therapy has recently shown powerful in initiating a lasting anti-tumor response with spectacular therapeutic success in some cases. Specific T cell therapy, however, is limited since a number of cancer cells are not recognized by T cells due to various mechanisms including the limited availability of tumor-specific T cells and deficiencies in antigen processing or major histocompatibility complex (MHC expression of cancer cells. To make adoptive cell therapy applicable for the broad variety of cancer entities, patient's T cells are engineered ex vivo with pre-defined specificity by a recombinant chimeric antigen receptor (CAR which consists in the extracellular part of an antibody-derived domain for binding with a tumor-associated antigen and in the intracellular part of a TCR-derived signaling moiety for T cell activation. The specificity of CAR mediated T cell recognition is defined by the antibody domain, is independent of MHC presentation and can be extended to any target for which an antibody is available. We discuss the advantages and limitations of MHC-independent T cell targeting by an engineered CAR and review most significant progress recently made in early stage clinical trials to treat cancer.

  15. Changes in thunderstorm characteristics due to feeder cloud merging

    Science.gov (United States)

    Sinkevich, Andrei A.; Krauss, Terrence W.

    2014-06-01

    Cumulus cloud merging is a complex dynamical and microphysical process in which two convective cells merge into a single cell. Previous radar observations and numerical simulations have shown a substantial increase in the maximum area, maximum echo top and maximum reflectivity as a result of the merging process. Although the qualitative aspects of merging have been well documented, the quantitative effects on storm properties remain less defined. Therefore, a statistical assessment of changes in storm characteristics due to merging is of importance. Further investigation into the effects of cloud merging on precipitation flux (Pflux) in a statistical manner provided the motivation for this study in the Asir region of Saudi Arabia. It was confirmed that merging has a strong effect on storm development in this region. The data analysis shows that an increase in the median of the distribution of maximum reflectivity was observed just after merging and was equal to 3.9 dBZ. A detailed analysis of the individual merge cases compared the merged storm Pflux and mass to the sum of the individual Feeder and Storm portions just before merging for each case. The merged storm Pflux increased an average of 106% over the 20-min period after merging, and the mass increased on average 143%. The merged storm clearly became larger and more severe than the sum of the two parts prior to merging. One consequence of this study is that any attempts to evaluate the precipitation enhancement effects of cloud seeding must also include the issue of cloud mergers because merging can have a significant effect on the results.

  16. Human Induced Pluripotent Stem Cell-Derived Macrophages Share Ontogeny with MYB-Independent Tissue-Resident Macrophages

    Directory of Open Access Journals (Sweden)

    Julian Buchrieser

    2017-02-01

    Full Text Available Tissue-resident macrophages, such as microglia, Kupffer cells, and Langerhans cells, derive from Myb-independent yolk sac (YS progenitors generated before the emergence of hematopoietic stem cells (HSCs. Myb-independent YS-derived resident macrophages self-renew locally, independently of circulating monocytes and HSCs. In contrast, adult blood monocytes, as well as infiltrating, gut, and dermal macrophages, derive from Myb-dependent HSCs. These findings are derived from the mouse, using gene knockouts and lineage tracing, but their applicability to human development has not been formally demonstrated. Here, we use human induced pluripotent stem cells (iPSCs as a tool to model human hematopoietic development. By using a CRISPR-Cas9 knockout strategy, we show that human iPSC-derived monocytes/macrophages develop in an MYB-independent, RUNX1-, and SPI1 (PU.1-dependent fashion. This result makes human iPSC-derived macrophages developmentally related to and a good model for MYB-independent tissue-resident macrophages, such as alveolar and kidney macrophages, microglia, Kupffer cells, and Langerhans cells.

  17. Learning Algorithms for Audio and Video Processing: Independent Component Analysis and Support Vector Machine Based Approaches

    National Research Council Canada - National Science Library

    Qi, Yuan

    2000-01-01

    In this thesis, we propose two new machine learning schemes, a subband-based Independent Component Analysis scheme and a hybrid Independent Component Analysis/Support Vector Machine scheme, and apply...

  18. Differences in carotenoid accumulation among three feeder-cricket species: implications for carotenoid delivery to captive insectivores.

    Science.gov (United States)

    Ogilvy, Victoria; Fidgett, Andrea L; Preziosi, Richard F

    2012-01-01

    There are a limited number of feeder-invertebrates available to feed captive insectivores, and many are deficient in certain nutrients. Gut-loading is used to increase the diversity of nutrients present in the captive insectivore diet; however, little is known about delivery of carotenoids via gut-loading. Carotenoids may influence health and reproduction due to their roles in immune and antioxidant systems. We assessed interspecific variation in carotenoid accumulation and retention in three feeder-cricket species (Gryllus bimaculatus, Gryllodes sigillatus and Acheta domesticus) fed one of three diets (wheat-bran, fish-food based formulated diet, and fresh fruit and vegetables). Out of the three species of feeder-cricket in the fish-food-based dietary treatment group, G. bimaculatus had the greatest total carotenoid concentration. All cricket species fed the wheat-bran diet had very low carotenoid concentrations. Species on the fish-food-based diet had intermediate carotenoid concentrations, and those on the fruit and vegetable diet had the highest concentrations. Carotenoid retention was poor across all species. Overall, this study shows that, by providing captive insectivores with G. bimaculatus crickets recently fed a carotenoid-rich diet, the quantity of carotenoids in the diet can be increased. © 2011 Wiley Periodicals, Inc.

  19. Multiphoton autofluorescence lifetime imaging of induced pluripotent stem cells

    Science.gov (United States)

    Uchugonova, Aisada

    2017-06-01

    The multiphoton fluorescence lifetime imaging tomograph MPTflex with its flexible 360-deg scan head, articulated arm, and tunable femtosecond laser source was employed to study induced pluripotent stem cell (iPS) cultures. Autofluorescence (AF) lifetime imaging was performed with 250-ps temporal resolution and submicron spatial resolution using time-correlated single-photon counting. The two-photon excited AF was based on the metabolic coenzymes NAD(P)H and flavin adenine dinucleotide/flavoproteins. iPS cells generated from mouse embryonic fibroblasts (MEFs) and cocultured with growth-arrested MEFs as feeder cells have been studied. Significant differences on AF lifetime signatures were identified between iPS and feeder cells as well as between their differentiating counterparts.

  20. Calcium-independent phospholipase A₂, group VIA, is critical for RPE cell survival

    DEFF Research Database (Denmark)

    Kolko, Miriam; Vohra, Rupali; Westlund, Barbro S.

    2014-01-01

    PURPOSE: To investigate the significance of calcium-independent phospholipase A₂, group VIA (iPLA2-VIA), in RPE cell survival following responses to sodium iodate (SI) in cell cultures. METHODS: The human retinal pigment epithelium (RPE) cell line (ARPE-19) cells and primary mouse-RPE cultures were...

  1. Gremlin-1 induces BMP-independent tumor cell proliferation, migration, and invasion.

    Directory of Open Access Journals (Sweden)

    Minsoo Kim

    Full Text Available Gremlin-1, a bone morphogenetic protein (BMP antagonist, is overexpressed in various cancerous tissues but its role in carcinogenesis has not been established. Here, we report that gremlin-1 binds various cancer cell lines and this interaction is inhibited by our newly developed gremlin-1 antibody, GRE1. Gremlin-1 binding to cancer cells was unaffected by the presence of BMP-2, BMP-4, and BMP-7. In addition, the binding was independent of vascular endothelial growth factor receptor-2 (VEGFR2 expression on the cell surface. Addition of gremlin-1 to A549 cells induced a fibroblast-like morphology and decreased E-cadherin expression. In a scratch wound healing assay, A549 cells incubated with gremlin-1 or transfected with gremlin-1 showed increased migration, which was inhibited in the presence of the GRE1 antibody. Gremlin-1 transfected A549 cells also exhibited increased invasiveness as well as an increased growth rate. These effects were also inhibited by the addition of the GRE1 antibody. In conclusion, this study demonstrates that gremlin-1 directly interacts with cancer cells in a BMP- and VEGFR2-independent manner and can induce cell migration, invasion, and proliferation.

  2. Design study on an independently-tunable-cells thermionic RF gun

    International Nuclear Information System (INIS)

    Hama, H.; Tanaka, T.; Hinode, F.; Kawai, M.

    2006-01-01

    Characteristics of a thermionic RF gun have been studied by a 3-D simulation code developed using an FDTD (Finite Difference Time Domain) method as a Maxwell's equations solver. The gun is consists of two independent power feeding cavities, so that we call it independently-tunable-cells (ITC)'-RF gun. The first cell is the cathode cell and the second one is an accelerating cell. The ITC gun can be operated at various modes of different RF-power ratio and phase between two cavities. Simulation study shows a velocity-bunching like effect may be occurred in the gun, so that the short pulse beam from the thermionic RF gun is a better candidate to produce the coherent THz synchrotron radiation. Expected bunch length with a total charge of ∼20 pC (1% energy width from the top energy) is around 200 fs (fwhm). Even the beam energy extracted from the gun is varied by which the input powers are changed, almost same shape of the longitudinal phase space can be produced by tuning the phase. (author)

  3. Using automated feeders to wean calves fed large amounts of milk according to their ability to eat solid feed.

    Science.gov (United States)

    de Passillé, Anne Marie; Rushen, Jeffrey

    2016-05-01

    Dairy calves weaned off milk at an early age show signs of hunger and can lose weight. We examined whether using automated feeders to wean calves according to individual voluntary solid feed intake reduced the effects of weaning. Female Holstein calves were housed in groups of 5 to 9. All calves were fed 12 L/d milk and ad libitum grain starter and hay from automated feeders immediately after grouping, and were allocated to 3 weaning strategies: (1) early-weaned (EW; n=14): weaning began on d 40, and milk allowance gradually decreased until weaning was complete on d 48; (2) late-weaned (LW; n=14): weaning began on d 80 and was completed on d 89; (3) weaned by starter intake (WSI; n=28): weaning began when calves consumed 200g/d of starter and was completed when the calves consumed 1,400g/d. Each day, the automated feeders recorded quantities of milk, starter, and hay eaten by all calves, as well as the frequency of visits to the milk feeder; we used unrewarded visit frequency as a sign of hunger. Body weights (BW) were recorded weekly. We estimated daily digestible energy (DE) intake for each calf based on the milk, hay, and starter consumed. Average daily gains (ADG) were expressed as percent of BW. For calves in WSI, weaning began at 54.7±18.9 d (mean ± SD) of age, the duration of weaning was 21.1±10.6 d, and weaning ended at 75.8±10.7 d of age. Both LW and WSI calves had better ADG from wk 3 to 13 than EW calves. Calves in the WSI group drank less milk and ate more starter than LW calves but had similar ADG. During the period of weaning, EW calves made more unrewarded visits to the milk feeder than LW and WSI calves. Three EW calves lost weight during weaning, whereas all LW and WSI calves gained weight. Calves differ greatly in when they begin to eat solid feed and how quickly they increase the intake in response to a decrease in milk allowance. An advantage of automated feeders is that calves can be weaned at variable ages depending on their ability and

  4. Capture of cell culture-derived influenza virus by lectins: strain independent, but host cell dependent.

    Science.gov (United States)

    Opitz, Lars; Zimmermann, Anke; Lehmann, Sylvia; Genzel, Yvonne; Lübben, Holger; Reichl, Udo; Wolff, Michael W

    2008-12-01

    Strategies to control influenza outbreaks are focused mainly on prophylactic vaccination. Human influenza vaccines are trivalent blends of different virus subtypes. Therefore and due to frequent antigenic drifts, strain independent manufacturing processes are required for vaccine production. This study verifies the strain independency of a capture method based on Euonymus europaeus lectin-affinity chromatography (EEL-AC) for downstream processing of influenza viruses under various culture conditions propagated in MDCK cells. A comprehensive lectin binding screening was conducted for two influenza virus types from the season 2007/2008 (A/Wisconsin/67/2005, B/Malaysia/2506/2004) including a comparison of virus-lectin interaction by surface plasmon resonance technology. EEL-AC resulted in a reproducible high product recovery rate and a high degree of contaminant removal in the case of both MDCK cell-derived influenza virus types demonstrating clearly the general applicability of EEL-AC. In addition, host cell dependency of EEL-AC was studied with two industrial relevant cell lines: Vero and MDCK cells. However, the choice of the host cell lines is known to lead to different product glycosylation profiles. Hence, altered lectin specificities have been observed between the two cell lines, requiring process adaptations between different influenza vaccine production systems.

  5. Mammalian cochlear supporting cells can divide and trans-differentiate into hair cells.

    Science.gov (United States)

    White, Patricia M; Doetzlhofer, Angelika; Lee, Yun Shain; Groves, Andrew K; Segil, Neil

    2006-06-22

    Sensory hair cells of the mammalian organ of Corti in the inner ear do not regenerate when lost as a consequence of injury, disease, or age-related deafness. This contrasts with other vertebrates such as birds, where the death of hair cells causes surrounding supporting cells to re-enter the cell cycle and give rise to both new hair cells and supporting cells. It is not clear whether the lack of mammalian hair cell regeneration is due to an intrinsic inability of supporting cells to divide and differentiate or to an absence or blockade of regenerative signals. Here we show that post-mitotic supporting cells purified from the postnatal mouse cochlea retain the ability to divide and trans-differentiate into new hair cells in culture. Furthermore, we show that age-dependent changes in supporting cell proliferative capacity are due in part to changes in the ability to downregulate the cyclin-dependent kinase inhibitor p27(Kip1) (also known as Cdkn1b). These results indicate that postnatal mammalian supporting cells are potential targets for therapeutic manipulation.

  6. Generation of Human Induced Pluripotent Stem Cells Using RNA-Based Sendai Virus System and Pluripotency Validation of the Resulting Cell Population.

    Science.gov (United States)

    Chichagova, Valeria; Sanchez-Vera, Irene; Armstrong, Lyle; Steel, David; Lako, Majlinda

    2016-01-01

    Human induced pluripotent stem cells (hiPSCs) provide a platform for studying human disease in vitro, increase our understanding of human embryonic development, and provide clinically relevant cell types for transplantation, drug testing, and toxicology studies. Since their discovery, numerous advances have been made in order to eliminate issues such as vector integration into the host genome, low reprogramming efficiency, incomplete reprogramming and acquisition of genomic instabilities. One of the ways to achieve integration-free reprogramming is by using RNA-based Sendai virus. Here we describe a method to generate hiPSCs with Sendai virus in both feeder-free and feeder-dependent culture systems. Additionally, we illustrate methods by which to validate pluripotency of the resulting stem cell population.

  7. Effect of Co-Culturing of Mice Liver Cells and Embryonic Carcinomatous Stem Cells on the Rate of Differentiation to Hematopoietic Cells

    Directory of Open Access Journals (Sweden)

    AA Pourfatollah

    2005-10-01

    Full Text Available Introduction: Considering the importance of co-culture in differentiation of embryonic stem cells, the aim of this study was evaluation of the effect of co-culturing fetal liver stroma cells with P19 cells on the line of differentiation. Materials and Methods: For this purpose, P19 cells were cultured directly in semisolid medium. These cells proliferated and primarily differentiated to colonies know as embryoid bodies (EBs after 8-12 days. The Ebs cells were trypsinized and dissociated to single or double cells. Then these cells were co-cultured on the mouse fetal liver feeder layer in the absence of exogenous factors. After 14-18 days, the colonies were studied morphologically by benzidine and giemsa staining and also counted under invert microscope. Results: The percentages of benzidine positive (or erythroid and negative colonies were 94% and 6% respectively and also the cells of colonies were studied by Giemsa staining. Results showed that they were myeloid or lymphoid type cells. Thus, the results show that in the presence of mouse fetal liver feeder layer, the number of erythroid colonies was increased. Conclusions: Therefore, this technique may be effective for differentiation of stem cells from different sources into hematopoietic cells and can be used in future for human cell therapy.

  8. Introducing tool-based feeders to zoo-housed chimpanzees as a cognitive challenge: spontaneous acquisition of new types of tool use and effects on behaviours and use of space.

    OpenAIRE

    Yamanashi, Yumi; Matsunaga, Masayuki; Shimada, Kanae; Kado, Ryuichiro; Tanaka, Masayuki

    2016-01-01

    Cognitively challenging environments are vital to the welfare of captive animals. However, practical enrichment devices that can facilitate animals’ natural behaviours and accommodate individual variation are still limited. We created two types of feeders to facilitate tool-using behaviour in captive chimpanzees: pounding and dipping feeders. The pounding feeder was inspired by pestle-pounding behaviour observed in wild chimpanzees, and we expected that chimpanzees would pound soft foods. The...

  9. Tits recognize the potential dangers of predators and harmless birds in feeder experiments

    Czech Academy of Sciences Publication Activity Database

    Tvardíková, Kateřina; Fuchs, R.

    2012-01-01

    Roč. 30, č. 1 (2012), s. 157-165 ISSN 0289-0771 Grant - others:AV ČR(CZ) IAA601410803 Institutional research plan: CEZ:AV0Z50070508 Keywords : feeders * predator recognition * risk Subject RIV: EH - Ecology, Behaviour Impact factor: 1.000, year: 2012 http://www.springerlink.com/content/2338033t30261604/fulltext.pdf

  10. Wean-to-finish feeder space availability effects on nursery and finishing pig performance and total tract digestibility in a commercial setting when feeding dried distillers grains with solubles.

    Science.gov (United States)

    Weber, E K; Stalder, K J; Patience, J F

    2015-04-01

    The study objectives were to determine nursery phase feeder space allowance effects on pig performance when double stocked and, second, to determine feeder space allowance and dried distillers grains with solubles (DDGS) inclusion level effects on pig performance and nutrient digestibility during the growing-finishing phase. This study was performed on the same group of pigs within a commercial wean-finish system. For the nursery phase, a completely randomized design was used to compare 3 feeder space allowance treatments (2.1, 2.5, and 2.9 cm/pig). A total of 3,720 pigs were randomly allotted to same-sex pens (10 feeders/treatment) housing 62 pigs/pen. Thirty 7-hole, double-sided feeders were utilized in the study. Differing linear feeder space treatments were established by blocking off sections for the nursery and grow-finish portions of this experiment. All pigs were provided equal floor space (0.26 m2/pig). In the grow-finish phase, a total of 1,860 pigs (n = 60 pens) were utilized in a 2 × 3 factorial design with 3 feeder space allowances (4.1, 4.9, or 5.7 cm/pig) and 2 dietary DDGS treatments (30% [D30] or 60% [D60]). Fecal and diet samples were collected and analyzed to estimate apparent total tract digestibility percentage (ATTD %). In the nursery portion of the trial, there was no feeder space treatment effect on ADG, ADFI, or feed efficiency (P > 0.10) from weaning to d 56 postweaning or during any weigh period. In the grow-finish portion of the trial, feeder space allowance and DDGS inclusion level did not affect ADG, ADFI, or feed efficiency (P > 0.05) from d 57 postweaning to market. Pigs fed the D30 diet had greater HCW, percent yield, and loin depth than those on the D60 diet (P feeder space allowance did not impact pig performance during the nursery or grow-finish production phases. Inclusion of DDGS at higher levels will decrease ADFI but not ADG or efficiency when isocaloric diets are fed. The inclusion level of DDGS does impact HCW and percent

  11. Mechanical Design of the LHC Standard Half-Cell

    Science.gov (United States)

    Poncet, A.; Brunet, J. C.; Cruikshank, P.; Genet, M.; Parma, V.; Rohmig, P.; Saban, R.; Tavian, L.; Veness, R.; Vlogaert, J.; Williams, L. R.

    1997-05-01

    The LHC Conceptual Design Report issued on 20th October 1995 (CERN/AC/95-05 (LHC) - nicknamed "Yellow Book") introduced significant changes to some fundamental features of the LHC standard half-cell, composed of one quadrupole, 3 dipoles and a set of corrector magnets. A separate cryogenic distribution line was introduced, which was previously inside the main cryostat. The dipole length has been increased from 10 to 15 m and independent powering of the focusing and defocusing quadrupole magnets was chosen. Individual quench protection diodes were introduced in magnets interconnects and many auxiliary bus bars were added to feed in series the various families of correcting superconducting magnets. The various highly intricate basic systems such as: cryostats and cryogenics feeders, superconducting magnets and their electrical feeding and protection, vacuum beam screen and its cooling, support and alignment devices have been redesigned, taking into account the very tight space available. These space constraints are given by the necessity to have maximum integral bending field strength for maximum LHC energy, and the existing LHC tunnel. Finally, cryogenic and vacuum sectorisation have been introduced to reduce downtimes and facilitate commissioning.

  12. Effect of feeder space during the growing and laying periods and the rate of feed increase at the onset of lay on broiler breeder female reproductive function.

    Science.gov (United States)

    Leksrisompong, N; Romero-Sanchez, H; Oviedo-Rondón, E O; Brake, J

    2014-07-01

    A study was conducted to examine how 2 feeder space allocations during the rearing period followed by 2 feeder space allocations after photostimulation and 2 female feeding to peak programs (fast or slow) affected female broiler breeder reproductive performance and mortality. Sixteen pens of 76 breeder females each were equipped with either 4 tube feeders with a 132 cm circumference pan (7.0 cm/female) or 6 feeders (10.4 cm/female) to 21 wk of age. Thereafter, 64 females were moved to breeding pens, photostimulated, and fed sex-separate from either 3 (6.2 cm/female) or 5 (10.3 cm/female) feeders with either fast or slow feeding to peak feeding programs applied to complete a 2 × 2 × 2 factorial design. Seven males that were separately reared in a similar manner were added per pen. Individual female BW was determined at 6, 20, and 32 wk of age and BW uniformity assessed. Greater feeder space during rearing increased BW at 32 wk of age, whereas greater feeder space during lay or slow feeding to peak decreased BW at 32 wk. There were no differences in BW uniformity. Hens from the 10.4 to 10.3 cm/female combination produced a significantly greater number of eggs as compared with the 7.0 to 10.3 cm/female and 10.4 to 6.2 cm/female combinations with the 7.0 to 6.2 cm/female combination intermediate. Percentage hen-day egg production of the 10.4 to 10.3 cm/female combination hens was significantly greater than all other combinations. Livability was improved in the 10.4 to 10.3 cm/female combination relative to the 7.0 to 10.3 cm/female combination with the others intermediate. The fast feeding to peak program increased yolk weight as well as yolk:albumen ratio at 28 and 30 wk of age, but egg weight did not differ. These data indicated that increased or decreased feeder space between the growing and laying periods did not affect broiler breeder female BW, uniformity, egg weight, fertility, or hatchability. The 10.3 cm/female laying feeder space exhibited the best hen

  13. Effect of stall design on dairy calf transition to voluntary feeding on an automatic milk feeder after introduction to group housing.

    Science.gov (United States)

    Wilson, Tanya R; LeBlanc, Stephen J; DeVries, Trevor J; Haley, Derek B

    2018-06-01

    Automatic milk feeders (AMF) for young dairy calves are widely used in the dairy industry. These feeders are thought to have benefits for calf health and welfare and may reduce labor required for feeding; however, little is known about how calves adapt to feeding with AMF. The objective of this study was to observe the effects of feeding stall design on calves learning to use the AMF. The hypothesis was that solid side stalls, compared with steel bar stalls, would result in a longer latency to approach and feed from the AMF without assistance. A total of 147 Holstein calves (80 male and 67 female) were enrolled at 4 d of age, introduced to a group pen, and, at the same time, trained on an AMF. For training, calves were allowed to suck on the trainer's fingers and guided to the teat. Calves were allocated to 1 of 2 stall designs at the pen level, depending on which treatment cohort they were born into, either with steel bar stall walls (n = 46 male, 34 female calves) or with solid side stall walls (n = 34 male, 33 female calves). For 72 h after introductory training on the AMF, data from the feeders were collected and calf behavior was monitored by video. Outcomes measured included latency to first voluntary visit to the feeder and to first feeding, time spent in the feeder, amount of milk consumed over 72 h, number of retraining sessions required (retrained if linear regression models or a Poisson model for the outcome of retraining. For certain outcomes the effects of stall design interacted with difficulty of training (willingness to enter feeder and drink); for the 38% of calves that were scored as moderately difficult to train on a scale of easy, moderate, or difficult, treatment (stall design) differences were detected. These calves took 2× longer to lick or bite toward the nipple, 2× longer to first voluntarily feeding, and consumed less milk over 72 h following training when trained on the steel bar stall design. These results suggest simple features of a

  14. Development of an extruder-feeder biomass direct liquefaction process

    Energy Technology Data Exchange (ETDEWEB)

    White, D.H.; Wolf, D. (Arizona Univ., Tucson, AZ (United States). Dept. of Chemical Engineering)

    1991-10-01

    As an abundant, renewable, domestic energy resource, biomass could help the United States reduce its dependence on imported oil. Biomass is the only renewable energy technology capable of addressing the national need for liquid transportation fuels. Thus, there is an incentive to develop economic conversion processes for converting biomass, including wood, into liquid fuels. Through research sponsored by the US DOE's Biomass Thermochemical Conversion Program, the University of Arizona has developed a unique biomass direct liquefaction system. The system features a modified single-screw extruder capable of pumping solid slurries containing as high as 60 wt% wood flour in wood oil derived vacuum bottoms at pressures up to 3000 psi. The extruder-feeder has been integrated with a unique reactor by the University to form a system which offers potential for improving high pressure biomass direct liquefaction technology. The extruder-feeder acts simultaneously as both a feed preheater and a pumping device for injecting wood slurries into a high pressure reactor in the biomass liquefaction process. An experimental facility was constructed and following shakedown operations, wood crude oil was produced by mid-1985. By July 1988, a total of 57 experimental continuous biomass liquefaction runs were made using White Birch wood feedstock. Good operability was achieved at slurry feed rates up to 30 lb/hr, reactor pressures from 800 to 3000 psi and temperatures from 350{degree}C to 430{degree}C under conditions covering a range of carbon monoxide feed rates and sodium carbonate catalyst addition. Crude wood oils containing as little as 6--10 wt% residual oxygen were produced. 38 refs., 82 figs., 26 tabs.

  15. Adjusting the weaning age of calves fed by automated feeders according to individual intakes of solid feed.

    Science.gov (United States)

    de Passillé, A M; Rushen, J

    2012-09-01

    When weaned at the ages typical of commercial dairy production, dairy calves usually show reduced growth rates, lowered energy intake, and increased behavioral signs of hunger, reflecting their difficulty in switching from a milk diet to solid feed. However, large differences exist between calves in their ability to adapt to solid feed, and automated feeders allow the age of weaning to be adjusted to an individual calf's intake of solid feed. We examined the effects of weaning according to solid feed intake on age at weaning, feed intake, and behavioral signs of hunger. In a 2×2 factorial design, 60 female Holstein calves in groups of 8 and fed milk, grain starter, and hay from automated feeders began to be weaned when their voluntary intake of grain starter was either 400 g/d (high start) or 200 g/d (low start), with weaning completed when their voluntary intakes of starter were either 1,600 g/d (high end) or 800 g/d (low end). Digestible energy intakes were calculated from milk, starter and hay intakes, corrected for body weights, calves were weighed, and the frequency of visits to the milk feeders were measured each day. The main effects and interactions between treatments were tested with ANOVA. Large differences were observed between calves in the age at which weaning was complete, with weaning completed earlier for low-end calves compared with high-end calves. No treatment effects (either of start amount or end amount) on intakes of milk, starter, or hay or on weight gains occurred. However, the calves that began weaning earlier had longer durations of weaning, greater growth rates from d 20 to 87, and were heavier on d 87, and had lower milk intakes, higher starter intakes, higher hay intakes, and a greater digestible energy intake, but showed more unrewarded visits to the milk feeder. Adjusting the age at which individual calves are weaned off milk according to their ability to eat solid feed can reduce the age at which weaning occurs while reducing the

  16. In Vitro Modeling of Human Germ Cell Development Using Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Yuncheng Zhao

    2018-02-01

    Full Text Available Summary: Due to differences across species, the mechanisms of cell fate decisions determined in mice cannot be readily extrapolated to humans. In this study, we developed a feeder- and xeno-free culture protocol that efficiently induced human pluripotent stem cells (iPSCs into PLZF+/GPR125+/CD90+ spermatogonium-like cells (SLCs. These SLCs were enriched with key genes in germ cell development such as MVH, DAZL, GFRα1, NANOS3, and DMRT1. In addition, a small fraction of SLCs went through meiosis in vitro to develop into haploid cells. We further demonstrated that this chemically defined induction protocol faithfully recapitulated the features of compromised germ cell development of PSCs with NANOS3 deficiency or iPSC lines established from patients with non-obstructive azoospermia. Taken together, we established a powerful experimental platform to investigate human germ cell development and pathology related to male infertility. : In this article, Wang and colleagues established a feeder- and xeno-free system to robustly induce human pluripotent stem cells (PSCs into spermatogonia-like cells. This chemically defined induction protocol faithfully recapitulated the features of compromised germ cell development of PSCs with NANOS3 deficiency or iPSC lines established from patients with non-obstructive azoospermia. Keywords: pluripotent stem cells, spermatogonia, infertility, non-obstructive azoospermia

  17. Biomechanics of P-selectin PSGL-1 bonds: Shear threshold and integrin-independent cell adhesion

    Energy Technology Data Exchange (ETDEWEB)

    Xiao, Zhihua; Goldsmith, Harry L.; MacIntosh, Fiona A.; Shankaran, Harish; Neelamegham, Sriram

    2006-03-01

    Platelet-leukocyte adhesion may contribute to thrombosis and inflammation. We examined the heterotypic interaction between unactivated neutrophils and either thrombin receptor activating peptide (TRAP) stimulated platelets or P-selectin bearing beads (Ps-beads) in suspension. Cone-plate viscometers were used to apply controlled shear rates from 14-3000/s. Platelet-neutrophil and bead-neutrophil adhesion analysis was performed using both flow cytometry and high-speed videomicroscopy. We observed that while blocking antibodies against either P-selectin or P-selectin glycoprotein ligand-1 (PSGL-1) alone inhibited platelet-neutrophil adhesion by ~60% at 140/s, these reagents completely blocked adhesion at 3000/s. Anti-Mac-1 alone did not alter platelet-neutrophil adhesion rates at any shear rate, though in synergy with selectin antagonists it abrogated cell binding. Unstimulated neutrophils avidly bound Ps-beads and activated platelets in an integrin-independent manner, suggesting that purely selectin-dependent cell adhesion is possible. In support of this, antagonists against P-selectin or PSGL-1 dissociated previously formed platelet-neutrophil and Ps-bead neutrophil aggregates under shear in a variety of experimental systems, including in assays performed with whole blood. In studies where medium viscosity and shear rate were varied, a subtle shear threshold for P-selectin PSGL-1 binding was also noted at shear rates<100/s and at force loading rates of ~300pN/sec. Results are discussed in light of biophysical computations that characterize the collision between unequal size particles in linear shear flow. Overall, our studies reveal an integrin-independent regime for cell adhesion that may be physiologically relevant.

  18. Effect of PVRC damping with independent support motion response spectrum analysis of piping systems

    International Nuclear Information System (INIS)

    Wang, Y.K.; Bezler, P.; Shteyngart, S.

    1986-01-01

    The Technical Committee for Piping Systems of the Pressure Vessel Research Committee (PVRC) has recommended new damping values to be used in the seismic analyses of piping systems in nuclear power plants. To evaluate the effects of coupling these recommendations with the use of independent support motion analyses methods, two sets of seismic analyses have been carried out for several piping systems. One set based on the use of uniform damping as specified in Regulatory Guide 1.61, the other based on the PVRC recommendations. In each set the analyses were performed using independent support motion time history and response spectrum methods as well as the envelope spectrum method. In the independent response spectrum analyses, 14 response estimates were in fact obtained by considering different combination procedures between the support group contributions and all sequences of combinations between support groups, modes and directions. For each analysis set, the response spectrum results were compared with time history estimates of those results. Comparison tables were then prepared depicting the percentage by which the response spectrum estimates exceeded the time history estimates. By comparing the result tables between both analysis sets, the impact of PVRC damping can be observed. Preliminary results show that the degree of exceedance of the response spectrum estimates based on PVRC damping is less than that based on uniform damping for the same piping problem. Expressed differently the results obtained if ISM methods are coupled with PVRC damping are not as conservative as those obtained using uniform damping

  19. An automatic speech recognition system with speaker-independent identification support

    Science.gov (United States)

    Caranica, Alexandru; Burileanu, Corneliu

    2015-02-01

    The novelty of this work relies on the application of an open source research software toolkit (CMU Sphinx) to train, build and evaluate a speech recognition system, with speaker-independent support, for voice-controlled hardware applications. Moreover, we propose to use the trained acoustic model to successfully decode offline voice commands on embedded hardware, such as an ARMv6 low-cost SoC, Raspberry PI. This type of single-board computer, mainly used for educational and research activities, can serve as a proof-of-concept software and hardware stack for low cost voice automation systems.

  20. Human spermatogonial stem cells display limited proliferation in vitro under mouse spermatogonial stem cell culture conditions.

    Science.gov (United States)

    Medrano, Jose V; Rombaut, Charlotte; Simon, Carlos; Pellicer, Antonio; Goossens, Ellen

    2016-11-01

    To study the ability of human spermatogonial stem cells (hSSCs) to proliferate in vitro under mouse spermatogonial stem cell (mSSC) culture conditions. Experimental basic science study. Reproductive biology laboratory. Cryopreserved testicular tissue with normal spermatogenesis obtained from three donors subjected to orchiectomy due to a prostate cancer treatment. Testicular cells used to create in vitro cell cultures corresponding to the following groups: [1] unsorted human testicular cells, [2] differentially plated human testicular cells, and [3] cells enriched with major histocompatibility complex class 1 (HLA - )/epithelial cell surface antigen (EPCAM + ) in coculture with inactivated testicular feeders from the same patient. Analyses and characterization including immunocytochemistry and quantitative reverse-transcription polymerase chain reaction for somatic and germ cell markers, testosterone and inhibin B quantification, and TUNEL assay. Putative hSSCs appeared in singlets, doublets, or small groups of up to four cells in vitro only when testicular cells were cultured in StemPro-34 medium supplemented with glial cell line-derived neurotrophic factor (GDNF), leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF). Fluorescence-activated cell sorting with HLA - /EPCAM + resulted in an enrichment of 27% VASA + /UTF1 + hSSCs, compared to 13% in unsorted controls. Coculture of sorted cells with inactivated testicular feeders gave rise to an average density of 112 hSSCs/cm 2 after 2 weeks in vitro compared with unsorted cells (61 hSSCs/cm 2 ) and differentially plated cells (49 hSSCS/cm 2 ). However, putative hSSCs rarely stained positive for the proliferation marker Ki67, and their presence was reduced to the point of almost disappearing after 4 weeks in vitro. We found that hSSCs show limited proliferation in vitro under mSSC culture conditions. Coculture of HLA - /EPCAM + sorted cells with testicular

  1. Conceptualizing Parental Autonomy Support: Adolescent Perceptions of Promotion of Independence versus Promotion of Volitional Functioning

    Science.gov (United States)

    Soenens, Bart; Vansteenkiste, Maarten; Lens, Willy; Luyckx, Koen; Goossens, Luc; Beyers, Wim; Ryan, Richard M.

    2007-01-01

    In current research on parenting, 2 ways of conceptualizing perceived parental autonomy support can be distinguished. Parental autonomy support can be defined in terms of promotion of independence (PI) or in terms of promotion of volitional functioning (PVF). This study aimed to establish the empirical distinctiveness of both conceptualizations…

  2. Nitrogen fixed by cyanobacteria is utilized by deposit-feeders.

    Science.gov (United States)

    Karlson, Agnes M L; Gorokhova, Elena; Elmgren, Ragnar

    2014-01-01

    Benthic communities below the photic zone depend for food on allochthonous organic matter derived from seasonal phytoplankton blooms. In the Baltic Sea, the spring diatom bloom is considered the most important input of organic matter, whereas the contribution of the summer bloom dominated by diazotrophic cyanobacteria is less understood. The possible increase in cyanobacteria blooms as a consequence of eutrophication and climate change calls for evaluation of cyanobacteria effects on benthic community functioning and productivity. Here, we examine utilization of cyanobacterial nitrogen by deposit-feeding benthic macrofauna following a cyanobacteria bloom at three stations during two consecutive years and link these changes to isotopic niche and variations in body condition (assayed as C:N ratio) of the animals. Since nitrogen-fixing cyanobacteria have δ(15)N close to -2‰, we expected the δ(15)N in the deposit-feeders to decrease after the bloom if their assimilation of cyanobacteria-derived nitrogen was substantial. We also expected the settled cyanobacteria with their associated microheterotrophic community and relatively high nitrogen content to increase the isotopic niche area, trophic diversity and dietary divergence between individuals (estimated as the nearest neighbour distance) in the benthic fauna after the bloom. The three surface-feeding species (Monoporeia affinis, Macoma balthica and Marenzelleria arctia) showed significantly lower δ(15)N values after the bloom, while the sub-surface feeder Pontoporeia femorata did not. The effect of the bloom on isotopic niche varied greatly between stations; populations which increased niche area after the bloom had better body condition than populations with reduced niche, regardless of species. Thus, cyanobacterial nitrogen is efficiently integrated into the benthic food webs in the Baltic, with likely consequences for their functioning, secondary production, transfer efficiency, trophic interactions, and

  3. Nitrogen fixed by cyanobacteria is utilized by deposit-feeders.

    Directory of Open Access Journals (Sweden)

    Agnes M L Karlson

    Full Text Available Benthic communities below the photic zone depend for food on allochthonous organic matter derived from seasonal phytoplankton blooms. In the Baltic Sea, the spring diatom bloom is considered the most important input of organic matter, whereas the contribution of the summer bloom dominated by diazotrophic cyanobacteria is less understood. The possible increase in cyanobacteria blooms as a consequence of eutrophication and climate change calls for evaluation of cyanobacteria effects on benthic community functioning and productivity. Here, we examine utilization of cyanobacterial nitrogen by deposit-feeding benthic macrofauna following a cyanobacteria bloom at three stations during two consecutive years and link these changes to isotopic niche and variations in body condition (assayed as C:N ratio of the animals. Since nitrogen-fixing cyanobacteria have δ(15N close to -2‰, we expected the δ(15N in the deposit-feeders to decrease after the bloom if their assimilation of cyanobacteria-derived nitrogen was substantial. We also expected the settled cyanobacteria with their associated microheterotrophic community and relatively high nitrogen content to increase the isotopic niche area, trophic diversity and dietary divergence between individuals (estimated as the nearest neighbour distance in the benthic fauna after the bloom. The three surface-feeding species (Monoporeia affinis, Macoma balthica and Marenzelleria arctia showed significantly lower δ(15N values after the bloom, while the sub-surface feeder Pontoporeia femorata did not. The effect of the bloom on isotopic niche varied greatly between stations; populations which increased niche area after the bloom had better body condition than populations with reduced niche, regardless of species. Thus, cyanobacterial nitrogen is efficiently integrated into the benthic food webs in the Baltic, with likely consequences for their functioning, secondary production, transfer efficiency, trophic

  4. Optimal distribution feeder reconfiguration for increasing the penetration of plug-in electric vehicles and minimizing network costs

    International Nuclear Information System (INIS)

    Kavousi-Fard, Abdollah; Abbasi, Alireza; Rostami, Mohammad-Amin; Khosravi, Abbas

    2015-01-01

    Appearance of PEVs (Plug-in Electric Vehicles) in future transportation sector brings forward opportunities and challenges from grid perspective. Increased utilization of PEVs will result in problems such as greater total loss, unbalanced load factor, feeder congestion and voltage drop. PEVs are mobile energy storages dispersed all over the network with benefits to both owners and utilities in case of V2G (Vehicle-to-Grid) possibility. The intelligent bidirectional power flow between grid and large number of vehicles adds complexity to the system and requires operative tools to schedule V2G energy and subdue PEV impacts. In this paper, DFR (Distribution Feeder Reconfiguration) is utilized to optimally coordinate PEV operation in a stochastic framework. Uncertainty in PEVs characteristics can be due to several sources from location and time of grid connection to driving pattern and battery SoC (State-of-Charge). The proposed stochastic problem is solved with a self-adaptive evolutionary swarm algorithm based on SSO (Social Spider Optimization) algorithm. Numerical studies verify the efficacy of the proposed DFR to improve the system performance and optimal dispatch of V2G. - Highlights: • Consideration effect of PEVS on the distribution feeder reconfiguration. • Increasing the penetration of PEVS. • Introducing a new artificial optimization algorithm. • Modeling the uncertainty in network. • Investigating the degradation cost of batteries in V2G technology.

  5. Cool Down Experiences with the SST-1 Helium Cryogenics System before and after Current Feeders System Modification

    Science.gov (United States)

    Patel, R.; Panchal, P.; Panchal, R.; Tank, J.; Mahesuriya, G.; Sonara, D.; Srikanth, G. L. N.; Garg, A.; Bairagi, N.; Christian, D.; Patel, K.; Shah, P.; Nimavat, H.; Sharma, R.; Patel, J. C.; Gupta, N. C.; Prasad, U.; Sharma, A. N.; Tanna, V. L.; Pradhan, S.

    The SST-1 machine comprises a superconducting magnet system (SCMS), which includes TF and PF magnets. In order to charge the SCMS, we need superconducting current feeders consisting of SC feeders and vapor cooled current leads (VCCLs). We have installed all 10 (+/-) pairs of VCCLs for the TF and PF systems. While conducting initial engineering validation of the SST-1 machine, our prime objective was to produce circular plasma using only the TF system. During the SST-1 campaign I to VI, we have to stop the PF magnets cooling in order to get the cryo- stable conditions for current charging of the TF magnets system. In that case, the cooling of the PF current leads is not essential. It has been also observed that after aborting the PF system cooling, there was a limited experimental window of TF operation. Therefore, in the recent SST-1 campaign-VII, we removed the PF current leads (9 pairs) and kept only single (+/-) pair of the 10,000 A rated VCCLs to realize the charging of the TF system for the extended window of operation. We have observed a better cryogenic stability in the TF magnets after modifications in the CFS. In this paper, we report the comparison of the cool down performance for the SST-1 machine operation before and after modifications of the current feeders system.

  6. Jaspine B induces nonapoptotic cell death in gastric cancer cells independently of its inhibition of ceramide synthase.

    Science.gov (United States)

    Cingolani, Francesca; Simbari, Fabio; Abad, Jose Luis; Casasampere, Mireia; Fabrias, Gemma; Futerman, Anthony H; Casas, Josefina

    2017-08-01

    Sphingolipids (SLs) have been extensively investigated in biomedical research due to their role as bioactive molecules in cells. Here, we describe the effect of a SL analog, jaspine B (JB), a cyclic anhydrophytosphingosine found in marine sponges, on the gastric cancer cell line, HGC-27. JB induced alterations in the sphingolipidome, mainly the accumulation of dihydrosphingosine, sphingosine, and their phosphorylated forms due to inhibition of ceramide synthases. Moreover, JB provoked atypical cell death in HGC-27 cells, characterized by the formation of cytoplasmic vacuoles in a time and dose-dependent manner. Vacuoles appeared to originate from macropinocytosis and triggered cytoplasmic disruption. The pan-caspase inhibitor, z-VAD, did not alter either cytotoxicity or vacuole formation, suggesting that JB activates a caspase-independent cell death mechanism. The autophagy inhibitor, wortmannin, did not decrease JB-stimulated LC3-II accumulation. In addition, cell vacuolation induced by JB was characterized by single-membrane vacuoles, which are different from double-membrane autophagosomes. These findings suggest that JB-induced cell vacuolation is not related to autophagy and it is also independent of its action on SL metabolism. Copyright © 2017 by the American Society for Biochemistry and Molecular Biology, Inc.

  7. Apoptosis-Dependent and Apoptosis-Independent Functions Bim in Prostate Cancer Cells

    National Research Council Canada - National Science Library

    Liu, Junwei

    2004-01-01

    ... (death, survival, proliferation/division, etc). Our hypothesis is that, under normal, unstimulated conditions, with its apoptotic function blocked, the upregulated Bim in PCa cells play an apoptosis-independent function...

  8. A Frequency-Based Assignment Model under Day-to-Day Information Evolution of Oversaturated Conditions on a Feeder Bus Service

    Directory of Open Access Journals (Sweden)

    Silin Zhang

    2017-02-01

    Full Text Available Day-to-day information is increasingly being implemented in transit networks worldwide. Feeder bus service (FBS plays a vital role in a public transit network by providing feeder access to hubs and rails. As a feeder service, a space-time path for frequent passengers is decided by its dynamic strategy procedure, in which a day-to-day information self-learning mechanism is identified and analyzed from our survey data. We formulate a frequency-based assignment model considering day-to-day evolution under oversaturated conditions, which takes into account the residual capacity of bus and the comfort feelings of sitting or standing. The core of our proposed model is to allocate the passengers on each segment belonging to their own paths according to multi-utilities transformed from the time values and parametric demands, such as frequency, bus capacity, seat comfort, and stop layout. The assignment method, albeit general, allows us to formulate an equivalent optimization problem in terms of interaction between the FBS’ operation and frequent passengers’ rational behaviors. Finally, a real application case is generated to test the ability of the modeling framework capturing the theoretical consequents, serving the passengers’ dynamic externalities.

  9. Surgical management of dural arteriovenous fistulas with transosseous arterial feeders involving the jugular bulb.

    Science.gov (United States)

    Tirakotai, W; Benes, L; Kappus, C; Sure, U; Farhoud, A; Bien, S; Bertalanffy, H

    2007-01-01

    Dural arteriovenous fistulas located in the vicinity of the jugular foramen are complex vascular malformations and belong to the most challenging skull base lesions to treat. The authors comprehensively analyze multiple features in a series of dural arteriovenous fistulas with transosseous arterial feeders involving the jugular bulb. Four patients who underwent surgery via the transcondylar approach to treat dural arteriovenous fistulas around the jugular foramen were retrospectively reviewed. Previously, endovascular treatment was attempted in all patients. The success of the surgical treatment was examined with postoperative angiography. Complete obliteration of the dural arteriovenous fistulas (DAVFs) was achieved in three patients, and significant flow reduction in one individual. All patients had a good postoperative outcome, and only one experienced mild hypoglossal nerve palsy. Despite extensive bone drilling, an occipitocervical fusion was necessary in only one patient with bilateral lesions. The use of an individually tailored transcondylar approach to treat dural arteriovenous fistulas at the region of the jugular foramen is most effective. This approach allows for complete obliteration of the connecting arterial feeders, and removal of bony structures containing pathological vessels.

  10. Caspase-1 from Human Myeloid-Derived Suppressor Cells Can Promote T Cell-Independent Tumor Proliferation.

    Science.gov (United States)

    Zeng, Qi; Fu, Juan; Korrer, Michael; Gorbounov, Mikhail; Murray, Peter J; Pardoll, Drew; Masica, David L; Kim, Young J

    2018-05-01

    Immunosuppressive myeloid-derived suppressive cells (MDSCs) are characterized by their phenotypic and functional heterogeneity. To better define their T cell-independent functions within the tumor, sorted monocytic CD14 + CD11b + HLA-DR low/- MDSCs (mMDSC) from squamous cell carcinoma patients showed upregulated caspase-1 activity, which was associated with increased IL1β and IL18 expression. In vitro studies demonstrated that mMDSCs promoted caspase-1-dependent proliferation of multiple squamous carcinoma cell lines in both human and murine systems. In vivo , growth rates of B16, MOC1, and Panc02 were significantly blunted in chimeric mice adoptively transferred with caspase-1 null bone marrow cells under T cell-depleted conditions. Adoptive transfer of wild-type Gr-1 + CD11b + MDSCs from tumor-bearing mice reversed this antitumor response, whereas caspase-1 inhibiting thalidomide-treated MDSCs phenocopied the antitumor response found in caspase-1 null mice. We further hypothesized that MDSC caspase-1 activity could promote tumor-intrinsic MyD88-dependent carcinogenesis. In mice with wild-type caspase-1, MyD88-silenced tumors displayed reduced growth rate, but in chimeric mice with caspase-1 null bone marrow cells, MyD88-silenced tumors did not display differential tumor growth rate. When we queried the TCGA database, we found that caspase-1 expression is correlated with overall survival in squamous cell carcinoma patients. Taken together, our findings demonstrated that caspase-1 in MDSCs is a direct T cell-independent mediator of tumor proliferation. Cancer Immunol Res; 6(5); 566-77. ©2018 AACR . ©2018 American Association for Cancer Research.

  11. Flow visualization study of two phase flow in a single bend outlet feeder pipe and horizontal annulus of outlet end-fitting of a CANDU reactor

    International Nuclear Information System (INIS)

    Supa-Amornkul, S.; Lister, D.H.; Steward, F.R.

    2005-01-01

    'Full text:' In CANDU-6 reactors, the pressurized high-temperature coolant flows through 380 fuel channels passing horizontally through the core. Each end of a fuel channel has a stainless steel annular end-fitting connected to a carbon steel feeder pipe. The outlet coolant, which is at 310 o C with up to 0.30 steam voidage, turns through 90 o as it passes from flow in the annular end-fitting to pipe flow in the feeder via a Grayloc connector. Since 1996, several CANDU stations have reported excessive corrosion of their outlet feeder pipes; especially between the first metre, which consisted of single or double bends. Early studies related the attack to the hydrodynamics of the coolant and verified that it was a type of flow accelerated corrosion. In order to understand the hydrodynamics of the coolant in the outlet feeders by flow-visualization, a full-scale transparent test section simulating the geometry and orientation of an outlet feeder bend with its upstream annular end-fitting were fabricated. The feeder consisted of a 54 mm inside diameter acrylic pipe with a 73 o bend, connecting to an acrylic simulation of a Grayloc flanged fitting and annular end-fitting. The annular end-fitting consisted of an inner pipe, 110 mm outer diameter, and an outer pipe, 150 mm inner diameter, both 190.7 cm long in length. The tests were performed with water and air at atmospheric pressure and room temperature. The maximum water volumetric flow rate was 19 L/s and the volume fraction of air varied from 0.05 to 0.56. The phase distributions within the feeder pipe and along the length of the annulus were investigated with a digital video recorder. Size, concentration and velocity of the air bubbles at particular locations were studied with a high-speed digital still camera and a high-speed digital video camera. Phase distributions and variations in bubble size with velocity were determined. Particular attention was paid to the flow pattern at the inside of the bend, where a CFD

  12. Flow visualization study of two phase flow in a single bend outlet feeder pipe and horizontal annulus of outlet end-fitting of a CANDU reactor

    Energy Technology Data Exchange (ETDEWEB)

    Supa-Amornkul, S.; Lister, D.H.; Steward, F.R. [Univ. of New Brunswick, Fredericton, New Brunswick (Canada)]. E-mail: h796e@unb.ca; dlister@unb.ca; fsteward@unb.ca

    2005-07-01

    'Full text:' In CANDU-6 reactors, the pressurized high-temperature coolant flows through 380 fuel channels passing horizontally through the core. Each end of a fuel channel has a stainless steel annular end-fitting connected to a carbon steel feeder pipe. The outlet coolant, which is at 310{sup o}C with up to 0.30 steam voidage, turns through 90{sup o} as it passes from flow in the annular end-fitting to pipe flow in the feeder via a Grayloc connector. Since 1996, several CANDU stations have reported excessive corrosion of their outlet feeder pipes; especially between the first metre, which consisted of single or double bends. Early studies related the attack to the hydrodynamics of the coolant and verified that it was a type of flow accelerated corrosion. In order to understand the hydrodynamics of the coolant in the outlet feeders by flow-visualization, a full-scale transparent test section simulating the geometry and orientation of an outlet feeder bend with its upstream annular end-fitting were fabricated. The feeder consisted of a 54 mm inside diameter acrylic pipe with a 73{sup o} bend, connecting to an acrylic simulation of a Grayloc flanged fitting and annular end-fitting. The annular end-fitting consisted of an inner pipe, 110 mm outer diameter, and an outer pipe, 150 mm inner diameter, both 190.7 cm long in length. The tests were performed with water and air at atmospheric pressure and room temperature. The maximum water volumetric flow rate was 19 L/s and the volume fraction of air varied from 0.05 to 0.56. The phase distributions within the feeder pipe and along the length of the annulus were investigated with a digital video recorder. Size, concentration and velocity of the air bubbles at particular locations were studied with a high-speed digital still camera and a high-speed digital video camera. Phase distributions and variations in bubble size with velocity were determined. Particular attention was paid to the flow pattern at the inside

  13. A genetic screen for anchorage-independent proliferation in mammalian cells identifies a membrane-bound neuregulin.

    Directory of Open Access Journals (Sweden)

    Davide Danovi

    2010-07-01

    Full Text Available Anchorage-independent proliferation is a hallmark of oncogenic transformation and is thought to be conducive to proliferation of cancer cells away from their site of origin. We have previously reported that primary Schwann cells expressing the SV40 Large T antigen (LT are not fully transformed in that they maintain a strict requirement for attachment, requiring a further genetic change, such as oncogenic Ras, to gain anchorage-independence. Using the LT-expressing cells, we performed a genetic screen for anchorage-independent proliferation and identified Sensory and Motor Neuron Derived Factor (SMDF, a transmembrane class III isoform of Neuregulin 1. In contrast to oncogenic Ras, SMDF induced enhanced proliferation in normal primary Schwann cells but did not trigger cellular senescence. In cooperation with LT, SMDF drove anchorage-independent proliferation, loss of contact inhibition and tumourigenicity. This transforming ability was shared with membrane-bound class III but not secreted class I isoforms of Neuregulin, indicating a distinct mechanism of action. Importantly, we show that despite being membrane-bound signalling molecules, class III neuregulins transform via a cell intrinsic mechanism, as a result of constitutive, elevated levels of ErbB signalling at high cell density and in anchorage-free conditions. This novel transforming mechanism may provide new targets for cancer therapy.

  14. DeltaFosB induces osteosclerosis and decreases adipogenesis by two independent cell-autonomous mechanisms

    DEFF Research Database (Denmark)

    Kveiborg, Marie; Sabatakos, George; Chiusaroli, Riccardo

    2004-01-01

    establishes that the skeletal phenotype is cell autonomous to the osteoblast lineage and independent of adipocyte formation. It also strongly suggests that the decreased fat phenotype of NSE-DeltaFosB mice is independent of the changes in the osteoblast lineage. In vitro, overexpression of Delta......Osteoblasts and adipocytes may develop from common bone marrow mesenchymal precursors. Transgenic mice overexpressing DeltaFosB, an AP-1 transcription factor, under the control of the neuron-specific enolase (NSE) promoter show both markedly increased bone formation and decreased adipogenesis...... of DeltaFosB on adipocyte differentiation appears to occur at early stages of stem cell commitment, affecting C/EBPbeta functions. It is concluded that the changes in osteoblast and adipocyte differentiation in DeltaFosB transgenic mice result from independent cell-autonomous mechanisms....

  15. Changes in reward contingency modulate the trial-to-trial variability of hippocampal place cells.

    Science.gov (United States)

    Wikenheiser, Andrew M; Redish, A David

    2011-08-01

    Pyramidal cells in the rodent hippocampus often exhibit clear spatial tuning. Theories of hippocampal function suggest that these "place cells" implement multiple, independent neural representations of position (maps), based on different reference frames or environmental features. Consistent with the "multiple maps" theory, previous studies have shown that manipulating spatial factors related to task performance modulates the within-session variability (overdispersion) of cells in the hippocampus. However, the influence of changes in reward contingency on overdispersion has not been examined. To test this, we first trained rats to collect food from three feeders positioned around a circular track (task(1)). When subjects were proficient, the reward contingency was altered such that every other feeder delivered food (task(2)). We recorded ensembles of hippocampal neurons as rats performed both tasks. Place cell overdispersion was high during task(1) but decreased significantly during task(2), and this increased reliability could not be accounted for by changes in running speed or familiarity with the task. Intuitively, decreased variability might be expected to improve neural representations of position. To test this, we used Bayesian decoding of hippocampal spike trains to estimate subjects' location. Neither the amount of probability decoded to subjects' position (local probability) nor the difference between estimated position and true location (decoding accuracy) differed between tasks. However, we found that hippocampal ensembles were significantly more self-consistent during task(2) performance. These results suggest that changes in task demands can affect the firing statistics of hippocampal neurons, leading to changes in the properties of decoded neural representations.

  16. A Double-Resistive Active Power Filter System to Attenuate Harmonic Voltages of a Radial Power Distribution Feeder

    DEFF Research Database (Denmark)

    Sun, Xiaofeng; Han, Ruijing; Shen, Hong

    2016-01-01

    voltages. This paper proposes a double-resistive active power filter (D-RAPF) system consisting of a terminal-RAPF and an attenuation-RAPF for each individual harmonic. The terminal-RAPF operating as the characteristic impedance of the feeder is installed at the end bus to suppress harmonic propagation......Harmonic propagation between power-factor correction capacitors and system inductors seriously deteriorates power quality in a radial power distribution feeder. Installation of a resistive active power filter (RAPF) at the end bus only suppresses harmonic propagation, not attenuates harmonic...... at all harmonic frequencies. The attenuation-RAPF, whose control gain can be set according to the requirements of the harmonic voltage distortion limit, is installed at a specific position for each individual harmonic to attenuate the corresponding harmonic voltages. The D-RAPF system not only suppresses...

  17. Why drivers use cell phones and support legislation to restrict this practice.

    Science.gov (United States)

    Sanbonmatsu, David M; Strayer, David L; Behrends, Arwen A; Ward, Nathan; Watson, Jason M

    2016-07-01

    The use of cell phones while driving is ubiquitous, particularly in countries where the practice is legal. However, surveys indicate that most drivers favor legislation to limit the use of mobile devices during the operation of a vehicle. A study was conducted to understand this inconsistency between what drivers do and what they advocate for others. Participants completed a survey about their driving attitudes, abilities, and behaviors. Following previous research, drivers reported using cell phones for benefits such as getting work done. The hypocrisy of using cell phones while advocating restrictions appears to stem from differences in the perceived safety risks of self vs. others' use of cell phones. Many if not most drivers believe they can drive safely while using mobile devices. However, they lack confidence in others' ability to drive safely while distracted and believe that others' use of cell phones is dangerous. The threat to public safety of others' usage of mobile devices was one of the strongest independent predictors of support for legislation to restrict cell phone use. Published by Elsevier Ltd.

  18. Multimodal and ubiquitous computing systems: supporting independent-living older users.

    Science.gov (United States)

    Perry, Mark; Dowdall, Alan; Lines, Lorna; Hone, Kate

    2004-09-01

    We document the rationale and design of a multimodal interface to a pervasive/ubiquitous computing system that supports independent living by older people in their own homes. The Millennium Home system involves fitting a resident's home with sensors--these sensors can be used to trigger sequences of interaction with the resident to warn them about dangerous events, or to check if they need external help. We draw lessons from the design process and conclude the paper with implications for the design of multimodal interfaces to ubiquitous systems developed for the elderly and in healthcare, as well as for more general ubiquitous computing applications.

  19. BAX/BAK–Independent Mitoptosis during Cell Death Induced by Proteasome Inhibition?

    OpenAIRE

    Lomonosova, Elena; Ryerse, Jan; Chinnadurai, G.

    2009-01-01

    Proteasome inhibitors induce rapid death of cancer cells. We show that in epithelial cancer cells, such death is associated with dramatic and simultaneous up-regulation of several BH3-only proteins, including BIK, BIM, MCL-1S, NOXA, and PUMA, as well as p53. Elevated levels of these proteins seem to be the result of direct inhibition of their proteasomal degradation, induction of transcription, and active translation. Subsequent cell death is independent of BAX, and probably BAK, and proceeds...

  20. Utilisation of symmetrical components in a communication-based protection for loop MV feeders with variable short-circuit power

    DEFF Research Database (Denmark)

    Ciontea, Catalin-Iosif; Bak, Claus Leth; Blaabjerg, Frede

    2018-01-01

    -circuit power is presented. It relies on utilisation of symmetrical components of the short-circuit currents and on communication between the protection relays. The proposed method addresses the Single Phase to Ground (SPG) faults occurring in directly grounded distribution networks, with focus on closed......Variability of the available short-circuit power also implies variation of the fault level, which can potentially cause several protection problems in the electric networks. In this paper, a novel protection method that is insensitive to the fault level changes caused by variable short......-loop Medium Voltage (MV) feeders. Case studies are presented, which demonstrate that the proposed protection scheme is capable of effectively detecting the SPG faults in closed-loop feeders with variable short-circuit power....

  1. Highly flexible SRAM cells based on novel tri-independent-gate FinFET

    Science.gov (United States)

    Liu, Chengsheng; Zheng, Fanglin; Sun, Yabin; Li, Xiaojin; Shi, Yanling

    2017-10-01

    In this paper, a novel tri-independent-gate (TIG) FinFET is proposed for highly flexible SRAM cells design. To mitigate the read-write conflict, two kinds of SRAM cells based on TIG FinFETs are designed, and high tradeoff are obtained between read stability and speed. Both cells can offer multi read operations for frequency requirement with single voltage supply. In the first TIG FinFET SRAM cell, the strength of single-fin access transistor (TIG FinFET) can be flexibly adjusted by selecting five different modes to meet the needs of dynamic frequency design. Compared to the previous double-independent-gate (DIG) FinFET SRAM cell, 12.16% shorter read delay can be achieved with only 1.62% read stability decrement. As for the second TIG FinFET SRAM cell, pass-gate feedback technology is applied and double-fin TIG FinFETs are used as access transistors to solve the severe write-ability degradation. Three modes exist to flexibly adjust read speed and stability, and 68.2% larger write margin and 51.7% shorter write delay are achieved at only the expense of 26.2% increase in leakage power, with the same layout area as conventional FinFET SRAM cell.

  2. Role of CSL-dependent and independent Notch signaling pathways in cell apoptosis.

    Science.gov (United States)

    Zeng, Chong; Xing, Rui; Liu, Jing; Xing, Feiyue

    2016-01-01

    Apoptosis is a normally biological phenomenon in various organisms, involving complexly molecular mechanisms with a series of signaling processes. Notch signaling is found evolutionarily conserved in many species, playing a critical role in embryonic development, normal tissue homeostasis, angiogenesis and immunoregulation. The focus of this review is on currently novel advances about roles of CSL-dependent and independent Notch signaling pathways in cell apoptosis. The CSL can bind Notch intracellular domain (NIC) to act as a switch in mediating transcriptional activation or inactivation of the Notch signaling pathway downstream genes in the nucleus. It shows that CSL-dependent signaling regulates the cell apoptosis through Hes-1-PTEN-AKT-mTOR signaling, but rather the CSL-independent signaling mediates the cell apoptosis possibly via NIC-mTORC2-AKT-mTOR signaling, providing a new insight into apoptotic mechanisms.

  3. T cell regulation of the thymus-independent antibody response to trinitrophenylated-Brucella abortus (TNP-BA)

    Energy Technology Data Exchange (ETDEWEB)

    Tanay, A.; Strober, S.

    1985-06-01

    The authors have previously observed a reduction of the T cell-dependent primary antibody response to dinitrophenylated keyhole limpet hemocyanin, and an enhancement of the T cell-independent response to trinitrophenylated Brucella abortus (TNP-BA) in BALB/c mice after treatment with total lymphoid irradiation (TLI). To elucidate the relative contribution of T and B cells to the enhanced T cell-independent antibody responses after TLI, a syngeneic primary adoptive transfer system was utilized whereby irradiated hosts were reconstituted with unfractionated spleen cells or a combination of purified T and B cells from TLI-treated and untreated control mice. Antibody responses of purified splenic B cells from TLI-treated BALB/c mice (TLI/B) to TNP-BA were enhanced 10-fold as compared with those of unfractionated (UF) spleen cells or B cells from normal (NL) BALB/c mice (NL/UF and NL/B, respectively). Splenic T cells from normal animals (NL/T) suppressed the anti-TNP-BA response of TLI/B by more than 100-fold. NL/T neither suppressed nor enhanced the response of NL/B. On the other hand, T cells from TLI-treated mice (TLI/T) enhanced by 100-fold the anti-TNP-BA response of NL/B, but neither suppressed nor enhanced the response of TLI/B. Thus, T cells can regulate the T cell-independent antibody response to TNP-BA. However, experimental manipulation of the T and B cell populations is needed to demonstrate the regulatory functions.

  4. T cell regulation of the thymus-independent antibody response to trinitrophenylated-Brucella abortus (TNP-BA)

    International Nuclear Information System (INIS)

    Tanay, A.; Strober, S.

    1985-01-01

    The authors have previously observed a reduction of the T cell-dependent primary antibody response to dinitrophenylated keyhole limpet hemocyanin, and an enhancement of the T cell-independent response to trinitrophenylated Brucella abortus (TNP-BA) in BALB/c mice after treatment with total lymphoid irradiation (TLI). To elucidate the relative contribution of T and B cells to the enhanced T cell-independent antibody responses after TLI, a syngeneic primary adoptive transfer system was utilized whereby irradiated hosts were reconstituted with unfractionated spleen cells or a combination of purified T and B cells from TLI-treated and untreated control mice. Antibody responses of purified splenic B cells from TLI-treated BALB/c mice (TLI/B) to TNP-BA were enhanced 10-fold as compared with those of unfractionated (UF) spleen cells or B cells from normal (NL) BALB/c mice (NL/UF and NL/B, respectively). Splenic T cells from normal animals (NL/T) suppressed the anti-TNP-BA response of TLI/B by more than 100-fold. NL/T neither suppressed nor enhanced the response of NL/B. On the other hand, T cells from TLI-treated mice (TLI/T) enhanced by 100-fold the anti-TNP-BA response of NL/B, but neither suppressed nor enhanced the response of TLI/B. Thus, T cells can regulate the T cell-independent antibody response to TNP-BA. However, experimental manipulation of the T and B cell populations is needed to demonstrate the regulatory functions

  5. Applicability of ANSYS ELBOW290 element for flexibility calculation of tight radius bends on feeder pipes in CANDU reactors

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, X., E-mail: Xuan.Zhang@candu.com [Candu Energy Inc, Mississauga, ON (Canada)

    2015-07-01

    A curved pipe element, ELBOW290, became available in ANSYS 12. This element was developed based on a simplified shell theory, and maintains the ability to capture cross-sectional deformations of elbows. Numerical testing on the applicability of this element for the flexibility calculation of the tight radius bends in CANDU reactors is carried out to determine the usability of this element in completing stress analyses for feeder pipes. Comparisons are made between the ELBOW290 and the shell element for various feeder bend types found in domestic and overseas CANDU reactors. The comparisons show that the ELBOW290 element is suitable for calculating the flexibility of the tight radius bends. (author)

  6. Effects of an increasing filter feeder stock on larval abundance in the Oosterschelde estuary (SW Netherlands)

    NARCIS (Netherlands)

    Troost, Karin; Gelderman, Edzard; Kamermans, Pauline; Smaal, Aad C.; Wolff, Wim J.

    Predation by adult bivalves oil bivalve larvae has been suggested to reduce larval abundance in areas with high bivalve filter-feeder biomass. Although the occurrence of larviphagy is well-studied in the laboratory, its effects in the field have scarcely been studied. We studied larviphagy at

  7. Effects of an increasing filter feeder stock on larval abundance in the Oosterschelde estuary (SW Netherlands)

    NARCIS (Netherlands)

    Troost, K.; Gelderman, E.A.C.; Kamermans, P.; Smaal, A.C.; Wolff, W.

    2009-01-01

    Predation by adult bivalves on bivalve larvae has been suggested to reduce larval abundance in areas with high bivalve filter-feeder biomass. Although the occurrence of larviphagy is well-studied in the laboratory, its effects in the field have scarcely been studied. We studied larviphagy at

  8. Engineered artificial antigen presenting cells facilitate direct and efficient expansion of tumor infiltrating lymphocytes

    Directory of Open Access Journals (Sweden)

    Coukos George

    2011-08-01

    Full Text Available Abstract Background Development of a standardized platform for the rapid expansion of tumor-infiltrating lymphocytes (TILs with anti-tumor function from patients with limited TIL numbers or tumor tissues challenges their clinical application. Methods To facilitate adoptive immunotherapy, we applied genetically-engineered K562 cell-based artificial antigen presenting cells (aAPCs for the direct and rapid expansion of TILs isolated from primary cancer specimens. Results TILs outgrown in IL-2 undergo rapid, CD28-independent expansion in response to aAPC stimulation that requires provision of exogenous IL-2 cytokine support. aAPCs induce numerical expansion of TILs that is statistically similar to an established rapid expansion method at a 100-fold lower feeder cell to TIL ratio, and greater than those achievable using anti-CD3/CD28 activation beads or extended IL-2 culture. aAPC-expanded TILs undergo numerical expansion of tumor antigen-specific cells, remain amenable to secondary aAPC-based expansion, and have low CD4/CD8 ratios and FOXP3+ CD4+ cell frequencies. TILs can also be expanded directly from fresh enzyme-digested tumor specimens when pulsed with aAPCs. These "young" TILs are tumor-reactive, positively skewed in CD8+ lymphocyte composition, CD28 and CD27 expression, and contain fewer FOXP3+ T cells compared to parallel IL-2 cultures. Conclusion Genetically-enhanced aAPCs represent a standardized, "off-the-shelf" platform for the direct ex vivo expansion of TILs of suitable number, phenotype and function for use in adoptive immunotherapy.

  9. Effect of Antimicrobial Dosage Regimen on Salmonella and Escherichia coli Isolates from Feeder Swine▿

    Science.gov (United States)

    Wagner, Bruce A.; Straw, Barbara E.; Fedorka-Cray, Paula J.; Dargatz, David A.

    2008-01-01

    A body of evidence exists that suggests that antimicrobial use in food animals leads to resistance in both pathogenic and commensal bacteria. This study focused on the impact of three different antimicrobial regimes (low-level continuous, pulse, and no antimicrobial) for two antimicrobials (chlortetracycline and tylosin) on the presence of Salmonella spp. and on the prevalence of antimicrobial resistance of both Salmonella spp. and nonspecific Escherichia coli in fecal samples from feeder swine. The prevalence of fecal samples positive for Salmonella spp. significantly decreased between the samples taken at feeder placement compared to samples taken when the animals were close to market weight. Differences in resistance of Salmonella spp. did not appear to be influenced by dosing treatment including the control. Analysis of antimicrobial resistance examining both susceptibility and resistance, as well as MIC outcomes, demonstrated that only resistance to cephalothin increased in E. coli under the pulse chlortetracycline treatment. These results suggest that the dosing regimes examined in this study did not lead to an increase in either the prevalence of Salmonella spp. or the prevalence of antimicrobial resistance in isolates of Salmonella spp. or E. coli. PMID:18223115

  10. Enhanced Particle Swarm Optimization-Based Feeder Reconfiguration Considering Uncertain Large Photovoltaic Powers and Demands

    Directory of Open Access Journals (Sweden)

    Ying-Yi Hong

    2014-01-01

    Full Text Available The Kyoto protocol recommended that industrialized countries limit their green gas emissions in 2012 to 5.2% below 1990 levels. Photovoltaic (PV arrays provide clear and sustainable renewable energy to electric power systems. Solar PV arrays can be installed in distribution systems of rural and urban areas, as opposed to wind-turbine generators, which cause noise in surrounding environments. However, a large PV array (several MW may incur several operation problems, for example, low power quality and reverse power. This work presents a novel method to reconfigure the distribution feeders in order to prevent the injection of reverse power into a substation connected to the transmission level. Moreover, a two-stage algorithm is developed, in which the uncertain bus loads and PV powers are clustered by fuzzy-c-means to gain representative scenarios; optimal reconfiguration is then achieved by a novel mean-variance-based particle swarm optimization. The system loss is minimized while the operational constraints, including reverse power and voltage variation, are satisfied due to the optimal feeder reconfiguration. Simulation results obtained from a 70-bus distribution system with 4 large PV arrays validate the proposed method.

  11. Striking the Right Balance: Police Experience, Perceptions and Use of Independent Support Persons During Interviews Involving People with Intellectual Disability.

    Science.gov (United States)

    Henshaw, Marie; Spivak, Benjamin; Thomas, Stuart D M

    2018-03-01

    Several jurisdictions mandate the presence of an independent support person during police interviews with vulnerable people. The current study investigated police officers' experiences and perceptions of these volunteers during interviews with people with intellectual disability(ies) (ID). The sample comprised 229 police officers who attended a mandatory firearms training course in Melbourne, Australia, in 2010. Participants commonly reported utilizing independent support persons and displayed a fair understanding of their role. Overall, volunteers were engaged more frequently than family/friends; police considered the volunteers to be more impartial during interviews, whereas family/friends provided a greater level of emotional support to interviewees. Independent support persons need to demonstrate two quite different types of support to people with intellectual disability(ies) during police interviews; these require quite different skill sets and suggest the need for more tailored training and support for these volunteers. Implications for future research and policy are discussed. © 2016 John Wiley & Sons Ltd.

  12. Conceptual design of independently tunable cells RF gun with external injecting structure

    International Nuclear Information System (INIS)

    Liang Junjun; Feng Guangyao; Pei Yuanji; Pang Jian

    2012-01-01

    To obtain the micro-pulse bunch with the order of hundred femtoseconds length and high repetition rate, the pa- per proposes the independently tunable cells (ITC) RF gun, which has a double-cell structure with the cells being power fed independently. By choosing appropriate feeding power and phase of the two cells, this ITC-RF gun can achieve bunches of excellent characteristics. Additionally, the application of a-magnet and laser system can be avoided, which leads to more compact layout. An external injecting ITC-RF gun (DC-ITC-RF gun) structure is designed accordingly. The external injecting structure can increase beam current, decrease energy spread, and cancel the back-bombardment effect almost completely. By means of 1-D and 3- D beam dynamics calculation with different structure parameters, a group of RF parameters are obtained for better beam characteristics. Then the paper designs a pre-injector so that particles can be accelerated to 10 MeV. By choosing appropriate feeding power and incident particle phase for the pre-injector, the bunch length can be further compressed. (authors)

  13. Stability-index based method for optimal Var planning in distribution feeders

    International Nuclear Information System (INIS)

    Hamouda, Abdellatif; Zehar, Khaled

    2011-01-01

    Research highlights: → Optimal Var planning is modelled using heuristic methods. → Capacitor sizes and location are determined by a two stage method. → Capacitor locations are determined using nodes stability-indices. → Their sizes are calculated subject to a new constraint on the branches reactive currents. → The solution is fast and leads to better results without over compensation. -- Abstract: The problem of the reactive energy optimal planning can be solved in a fast and efficient way using heuristic techniques. The latter reduce the number of the control variables to be determined and lead to a near global optimal solution. The capacitor appropriate locations are firstly determined by decisive indices then, their optimal sizes are calculated. In this paper a stability-index based method is presented. The nodes stability-indices are calculated for identifying the most sensitive nodes to be candidate for receiving near optimal standard capacitors that, reduce the feeder power losses, improve the voltage profile and maximise the economic saving (objective function). In this multi-objective optimisation problem, the commonly used voltage constraint is substituted by a new constraint on the branch reactive currents. This new constraint, allows overcoming the over compensation phenomenon by setting positive branch reactive currents. The solution is further improved by regulating the source node voltage. The proposed approach has been tested on several feeder examples and its effectiveness has been demonstrated through comparative studies. The obtained results have shown that the proposed approach leads to a promising and feasible solution.

  14. Stability-index based method for optimal Var planning in distribution feeders

    Energy Technology Data Exchange (ETDEWEB)

    Hamouda, Abdellatif, E-mail: a_hamouda1@yahoo.f [QUERE Laboratory, Optics and Mechanics Institut, University Ferhat Abbas, Setif 19000 (Algeria); Zehar, Khaled [QUERE Laboratory, Department of Electrical and Electronics Engineering, University of Bahrain, Isa Town (Bahrain)

    2011-05-15

    Research highlights: {yields} Optimal Var planning is modelled using heuristic methods. {yields} Capacitor sizes and location are determined by a two stage method. {yields} Capacitor locations are determined using nodes stability-indices. {yields} Their sizes are calculated subject to a new constraint on the branches reactive currents. {yields} The solution is fast and leads to better results without over compensation. -- Abstract: The problem of the reactive energy optimal planning can be solved in a fast and efficient way using heuristic techniques. The latter reduce the number of the control variables to be determined and lead to a near global optimal solution. The capacitor appropriate locations are firstly determined by decisive indices then, their optimal sizes are calculated. In this paper a stability-index based method is presented. The nodes stability-indices are calculated for identifying the most sensitive nodes to be candidate for receiving near optimal standard capacitors that, reduce the feeder power losses, improve the voltage profile and maximise the economic saving (objective function). In this multi-objective optimisation problem, the commonly used voltage constraint is substituted by a new constraint on the branch reactive currents. This new constraint, allows overcoming the over compensation phenomenon by setting positive branch reactive currents. The solution is further improved by regulating the source node voltage. The proposed approach has been tested on several feeder examples and its effectiveness has been demonstrated through comparative studies. The obtained results have shown that the proposed approach leads to a promising and feasible solution.

  15. Cell death is induced by ciglitazone, a peroxisome proliferator-activated receptor γ (PPARγ) agonist, independently of PPARγ in human glioma cells

    International Nuclear Information System (INIS)

    Lee, Myoung Woo; Kim, Dae Seong; Kim, Hye Ryung; Kim, Hye Jin; Yang, Jin Mo; Ryu, Somi; Noh, Yoo Hun; Lee, Soo Hyun; Son, Meong Hi; Jung, Hye Lim; Yoo, Keon Hee; Koo, Hong Hoe; Sung, Ki Woong

    2012-01-01

    Highlights: ► Greater than 30 μM ciglitazone induces cell death in glioma cells. ► Cell death by ciglitazone is independent of PPARγ in glioma cells. ► CGZ induces cell death by the loss of MMP via decreased Akt. -- Abstract: Peroxisome proliferator-activated receptor γ (PPARγ) regulates multiple signaling pathways, and its agonists induce apoptosis in various cancer cells. However, their role in cell death is unclear. In this study, the relationship between ciglitazone (CGZ) and PPARγ in CGZ-induced cell death was examined. At concentrations of greater than 30 μM, CGZ, a synthetic PPARγ agonist, activated caspase-3 and induced apoptosis in T98G cells. Treatment of T98G cells with less than 30 μM CGZ effectively induced cell death after pretreatment with 30 μM of the PPARγ antagonist GW9662, although GW9662 alone did not induce cell death. This cell death was also observed when cells were co-treated with CGZ and GW9662, but was not observed when cells were treated with CGZ prior to GW9662. In cells in which PPARγ was down-regulated cells by siRNA, lower concentrations of CGZ (<30 μM) were sufficient to induce cell death, although higher concentrations of CGZ (⩾30 μM) were required to induce cell death in control T98G cells, indicating that CGZ effectively induces cell death in T98G cells independently of PPARγ. Treatment with GW9662 followed by CGZ resulted in a down-regulation of Akt activity and the loss of mitochondrial membrane potential (MMP), which was accompanied by a decrease in Bcl-2 expression and an increase in Bid cleavage. These data suggest that CGZ is capable of inducing apoptotic cell death independently of PPARγ in glioma cells, by down-regulating Akt activity and inducing MMP collapse.

  16. Intrinsic Cell Stress is Independent of Organization in Engineered Cell Sheets.

    Science.gov (United States)

    van Loosdregt, Inge A E W; Dekker, Sylvia; Alford, Patrick W; Oomens, Cees W J; Loerakker, Sandra; Bouten, Carlijn V C

    2018-06-01

    Understanding cell contractility is of fundamental importance for cardiovascular tissue engineering, due to its major impact on the tissue's mechanical properties as well as the development of permanent dimensional changes, e.g., by contraction or dilatation of the tissue. Previous attempts to quantify contractile cellular stresses mostly used strongly aligned monolayers of cells, which might not represent the actual organization in engineered cardiovascular tissues such as heart valves. In the present study, therefore, we investigated whether differences in organization affect the magnitude of intrinsic stress generated by individual myofibroblasts, a frequently used cell source for in vitro engineered heart valves. Four different monolayer organizations were created via micro-contact printing of fibronectin lines on thin PDMS films, ranging from strongly anisotropic to isotropic. Thin film curvature, cell density, and actin stress fiber distribution were quantified, and subsequently, intrinsic stress and contractility of the monolayers were determined by incorporating these data into sample-specific finite element models. Our data indicate that the intrinsic stress exerted by the monolayers in each group correlates with cell density. Additionally, after normalizing for cell density and accounting for differences in alignment, no consistent differences in intrinsic contractility were found between the different monolayer organizations, suggesting that the intrinsic stress exerted by individual myofibroblasts is independent of the organization. Consequently, this study emphasizes the importance of choosing proper architectural properties for scaffolds in cardiovascular tissue engineering, as these directly affect the stresses in the tissue, which play a crucial role in both the functionality and remodeling of (engineered) cardiovascular tissues.

  17. Cathode-supported hybrid direct carbon fuel cells

    DEFF Research Database (Denmark)

    Gil, Vanesa; Gurauskis, Jonas; Deleebeeck, Lisa

    2017-01-01

    The direct conversion of coal to heat and electricity by a hybrid direct carbon fuel cell (HDCFC) is a highly efficient and cleaner technology than the conventional combustion power plants. HDCFC is defined as a combination of solid oxide fuel cell and molten carbonate fuel cell. This work...... investigates cathode-supported cells as an alternative configuration for HDCFC, with better catalytic activity and performance. This study aims to define the best processing route to manufacture highly efficient cathode-supported cells based on La0.75Sr0.25MnO3/yttria-stabilized zirconia infiltrated backbones...

  18. Apoptosis-inducing factor plays a critical role in caspase-independent, pyknotic cell death in hydrogen peroxide-exposed cells.

    Science.gov (United States)

    Son, Young-Ok; Jang, Yong-Suk; Heo, Jung-Sun; Chung, Wan-Tae; Choi, Ki-Choon; Lee, Jeong-Chae

    2009-06-01

    It has been proposed that continuously generated hydrogen peroxide (H(2)O(2)) inhibits typical apoptosis and instead initiates an alternate, apoptosis-inducing factor (AIF)-dependent process. Aside from the role of AIF, however, the detailed morphological characterization of H(2)O(2)-induced cell death is not complete. This study examined the cellular mechanism(s) by which the continuous presence of H(2)O(2) induces cell death. We also further analyzed the precise role of AIF by inhibiting its expression with siRNA. Exposure of cells to H(2)O(2) generated by glucose oxidase caused mitochondrion-mediated, caspase-independent cell death. In addition, H(2)O(2) exposure resulted in cell shrinkage and chromatin condensation without nuclear fragmentation, indicating that H(2)O(2) stimulates a pyknotic cell death. Further analysis of AIF-transfected cells clearly demonstrated that nuclear translocation of AIF is the most important event required for nuclear condensation, phosphatidyl serine translocation, and ultimately cell death in H(2)O(2)-exposed cells. Furthermore, ATP was rapidly and severely depleted in cells exposed to H(2)O(2) generated by glucose oxidase but not by H(2)O(2) added as a bolus. Suppression of the H(2)O(2)-mediated ATP depletion by 3-aminobenzamide led to a significant increase of nuclear fragmentation in glucose oxidase-exposed cells. Collectively, these findings suggest that an acute energy reduction by H(2)O(2) causes caspase-independent and AIF-dependent cell death.

  19. Coordinated control of distributed energy resources to support load frequency control

    International Nuclear Information System (INIS)

    Ravikumar Pandi, V.; Al-Hinai, A.; Feliachi, Ali

    2015-01-01

    Highlights: • We aims to maintain feeder power flow by the coordination of DER units. • The error in feeder flow with respect to scheduled value is used by the controller. • The particle swarm optimization is employed to minimize the error in feeder flow. • Implemented on a transmission system along with 37 bus distribution feeder. • The results of proposed feeder control is analyzed with no feeder control scheme. - Abstract: The control of generating resources to follow the unscheduled load changes is considered to be an essential process in the power system in order to maintain the frequency of power supply. This load frequency control (LFC) problem has been given more importance in the recent smart grid environment because of the impact from high penetration of distributed energy resources (DER) installed at the distribution level. The renewable sources are highly intermittent in nature, so it is required to coordinate and control the DER units to maintain the feeder power flow at substation bus bar which is seen by transmission system operator during the LFC process. This paper aims to identify the impact of distributed generation and its control method to reduce the deviation of feeder power flow from the scheduled value in real time operation. The error in feeder power flow with respect to scheduled value is utilized by the PI controller to estimate the change in power reference of all DER units. The power output of DER units are maintained to reference values by the individual PI controllers. The particle swarm optimization algorithm is employed to minimize the error in feeder power flow by optimally tuning the gain values of all PI controllers. The proposed method is examined on a small transmission system along with the feeder of IEEE 37 bus distribution system with balanced loading condition. The complete system along with DER units is implemented in the MATLAB based stability package named Power Analysis Toolbox (PAT) for performing time domain

  20. Apoptosis-Dependent and Apoptosis-Independent Functions of Bim in Prostate Cancer Cells

    National Research Council Canada - National Science Library

    Tang, Dean; Liu, Junwei

    2005-01-01

    ... (death, survival, proliferation/division, etc.). Our hypothesis is that, under normal, unstimulated conditions, with its apoptotic function blocked, the upregulated Bim in PCa cells plays an apoptosis-independent function...

  1. 47 CFR 25.147 - Licensing provision for NGSO MSS feeder downlinks in the band 6700-6875 MHz.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Licensing provision for NGSO MSS feeder downlinks in the band 6700-6875 MHz. 25.147 Section 25.147 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Applications and Licenses Space...

  2. Cancer Exosomes Perform Cell-Independent MicroRNA Biogenesis and Promote Tumorigenesis

    Science.gov (United States)

    Melo, Sonia A.; Sugimoto, Hikaru; O’Connell, Joyce T.; Kato, Noritoshi; Villanueva, Alberto; Vidal, August; Qiu, Le; Vitkin, Edward; Perelman, Lev T.; Melo, Carlos A.; Lucci, Anthony; Ivan, Cristina; Calin, George A.; Kalluri, Raghu

    2014-01-01

    SUMMARY Exosomes are secreted by all cell types and contain proteins and nucleic acids. Here, we report that breast cancer associated exosomes contain microRNAs (miRNAs) associated with the RISC Loading Complex (RLC) and display cell-independent capacity to process precursor microRNAs (pre-miRNAs) into mature miRNAs. Pre-miRNAs, along with Dicer, AGO2, and TRBP, are present in exosomes of cancer cells. CD43 mediates the accumulation of Dicer specifically in cancer exosomes. Cancer exosomes mediate an efficient and rapid silencing of mRNAs to reprogram the target cell transcriptome. Exosomes derived from cells and sera of patients with breast cancer instigate non-tumorigenic epithelial cells to form tumors in a Dicer-dependent manner. These findings offer opportunities for the development of exosomes based biomarkers and therapies. PMID:25446899

  3. Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions.

    Science.gov (United States)

    Saha, Krishanu; Mei, Ying; Reisterer, Colin M; Pyzocha, Neena Kenton; Yang, Jing; Muffat, Julien; Davies, Martyn C; Alexander, Morgan R; Langer, Robert; Anderson, Daniel G; Jaenisch, Rudolf

    2011-11-15

    The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.

  4. Selective interactions between epithelial tumour cells and bone marrow mesenchymal stem cells

    OpenAIRE

    Hombauer, H; Minguell, J J

    2000-01-01

    This work is a comparative study on the features displayed by an epithelial metastatic breast cancer cell line (MCF-7) when set in co-culture with human bone marrow mesenchymal stem cells (MSC) or a feeder layer of 3T3 fibroblasts. MSC, a subset of non-haematopoietic cells in the marrow stroma, display a potential for self-renewal, proliferation and differentiation into precursors for bone, cartilage, connective and muscular tissue. Adhesion of MCF-7 cells to monolayers of MSC or 3T3 was high...

  5. Improvement of the auto wire feeder machine in a de-soldering process

    Directory of Open Access Journals (Sweden)

    Niramon Nonkhukhetkhong

    2016-10-01

    Full Text Available This paper presents the methodology of the de-soldering process for rework of disk drive Head Stack Assembly (HSA units. The auto wire feeder is a machine that generates Tin (Sn on the product. This machine was determined to be one of the major sources of excess Sn on the HSA. The defect rate due to excess Sn is more than 30%, which leads to increased processing time and cost to perform additional cleaning steps. From process analysis, the major causes of excess Sn are as follows: 1 The machine cannot cut the wire all the way into the flux core area; 2 The sizes and types of soldering irons are not appropriate for the unit parts; and, 3 There are variations introduced into the de-soldering process by the workforce. This paper proposes a methodology to address all three of these causes. First, the auto wire feeder machine in the de-solder process will be adjusted in order to cut wires into flux core. Second, the types of equipment and material used in de-soldering will be optimized. Finally, a new standard method for operators, which can be controlled more easily, will be developed in order to reduce defects due to workforce related variation. After these process controls and machine adjustments were implemented, the overall Sn related problems were significantly improved. Sn contamination was reduced by 41% and cycle time was reduced by an average of 15 seconds.

  6. Cell-type independent MYC target genes reveal a primordial signature involved in biomass accumulation.

    Directory of Open Access Journals (Sweden)

    Hongkai Ji

    Full Text Available The functions of key oncogenic transcription factors independent of context have not been fully delineated despite our richer understanding of the genetic alterations in human cancers. The MYC oncogene, which produces the Myc transcription factor, is frequently altered in human cancer and is a major regulatory hub for many cancers. In this regard, we sought to unravel the primordial signature of Myc function by using high-throughput genomic approaches to identify the cell-type independent core Myc target gene signature. Using a model of human B lymphoma cells bearing inducible MYC, we identified a stringent set of direct Myc target genes via chromatin immunoprecipitation (ChIP, global nuclear run-on assay, and changes in mRNA levels. We also identified direct Myc targets in human embryonic stem cells (ESCs. We further document that a Myc core signature (MCS set of target genes is shared in mouse and human ESCs as well as in four other human cancer cell types. Remarkably, the expression of the MCS correlates with MYC expression in a cell-type independent manner across 8,129 microarray samples, which include 312 cell and tissue types. Furthermore, the expression of the MCS is elevated in vivo in Eμ-Myc transgenic murine lymphoma cells as compared with premalignant or normal B lymphocytes. Expression of the MCS in human B cell lymphomas, acute leukemia, lung cancers or Ewing sarcomas has the highest correlation with MYC expression. Annotation of this gene signature reveals Myc's primordial function in RNA processing, ribosome biogenesis and biomass accumulation as its key roles in cancer and stem cells.

  7. Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture

    Science.gov (United States)

    Pijuan-Galitó, Sara; Tamm, Christoffer; Schuster, Jens; Sobol, Maria; Forsberg, Lars; Merry, Catherine L. R.; Annerén, Cecilia

    2016-01-01

    Reliable, scalable and time-efficient culture methods are required to fully realize the clinical and industrial applications of human pluripotent stem (hPS) cells. Here we present a completely defined, xeno-free medium that supports long-term propagation of hPS cells on uncoated tissue culture plastic. The medium consists of the Essential 8 (E8) formulation supplemented with inter-α-inhibitor (IαI), a human serum-derived protein, recently demonstrated to activate key pluripotency pathways in mouse PS cells. IαI efficiently induces attachment and long-term growth of both embryonic and induced hPS cell lines when added as a soluble protein to the medium at seeding. IαI supplementation efficiently supports adaptation of feeder-dependent hPS cells to xeno-free conditions, clonal growth as well as single-cell survival in the absence of Rho-associated kinase inhibitor (ROCKi). This time-efficient and simplified culture method paves the way for large-scale, high-throughput hPS cell culture, and will be valuable for both basic research and commercial applications. PMID:27405751

  8. p53-Independent thermosensitization by mitomycin C in human non-small cell lung carcinoma cells

    International Nuclear Information System (INIS)

    Jin, Z.-H.; Matsumoto, H.; Hayashi, S.; Shioura, H.; Kitai, R.; Kano, E.; Hatashita, M.

    2003-01-01

    The combined treatment with hyperthermia and chemotherapeutic drugs such as cisplatin (CDDP), doxorubicin (DOX) and mitomycin C (MMC) has been widely adopted as a strategy of interdisciplinary cancer therapy to obtain greater therapeutic benefits. However, the involved mechanisms of the interactive cytotoxic effects of hyperthermia and MMC remain unclear. To elucidate the relationship between p53 functions and the interactive effects of the combined treatment with mild-hyperthermia and MMC, we examined the potentiation of cytotoxic effects, the induction of apoptosis, the changes in cell cycles and the accumulation of Hsp72 after the combined treatment with hyperthermia at 42 degree C and MMC using human non-small cell lung carcinoma H1299 transfectants with either null, wild-type (wt) or mutant (m) p53 gene. H1299/null, H1299/wtp53 and H1299/mp53 cells showed similar sensitivities to either hyperthermia at 42 degree C alone or MMC alone. The combined treatment resulted in a synergistically enhanced cytotoxicity in H1299 transfectants in a p53-independent manner. The mechanisms involved an enhancement of heat-induced apoptosis and a modulation of the cell cycle distribution by the combined treatment. The accumulation of Hsp72 was not suppressed by the combined treatment, as is not the case of the combined treatment with hyperthermia and either CDDP (1) or bleomycin (2). Our findings demonstrate a p53-independent mechanism for a synergistically cytotoxic enhancement by the combined treatment with mild-hyperthermia and MMC

  9. Layered double hydroxide nanoparticles promote self-renewal of mouse embryonic stem cells through the PI3K signaling pathway

    Science.gov (United States)

    Wu, Youjun; Zhu, Rongrong; Zhou, Yang; Zhang, Jun; Wang, Wenrui; Sun, Xiaoyu; Wu, Xianzheng; Cheng, Liming; Zhang, Jing; Wang, Shilong

    2015-06-01

    Embryonic stem cells (ESCs) hold great potential for regenerative medicine due to their two unique characteristics: self-renewal and pluripotency. Several groups of nanoparticles have shown promising applications in directing the stem cell fate. Herein, we investigated the cellular effects of layered double hydroxide nanoparticles (LDH NPs) on mouse ESCs (mESCs) and the associated molecular mechanisms. Mg-Al-LDH NPs with an average diameter of ~100 nm were prepared by hydrothermal methods. To determine the influences of LDH NPs on mESCs, cellular cytotoxicity, self-renewal, differentiation potential, and the possible signaling pathways were explored. Evaluation of cell viability, lactate dehydrogenase release, ROS generation and apoptosis demonstrated the low cytotoxicity of LDH NPs. The alkaline phosphatase activity and the expression of pluripotency genes in mESCs were examined, which indicated that exposure to LDH NPs could support self-renewal and inhibit spontaneous differentiation of mESCs under feeder-free culture conditions. The self-renewal promotion was further proved to be independent of the leukemia inhibitory factor (LIF). Furthermore, cells treated with LDH NPs maintained the potential to differentiate into all three germ layers both in vitro and in vivo through formation of embryoid bodies and teratomas. In addition, we observed that LDH NPs initiated the activation of the PI3K/Akt pathway, while treatment with the PI3K inhibitor LY294002 could block the effects of LDH NPs on mESCs. The results confirmed that the promotion of self-renewal by LDH NPs was associated with activation of the PI3K/Akt signaling pathway. Altogether, our studies identified a new role of LDH NPs in maintaining self-renewal of mouse ES cells which could potentially be applied in stem cell research.Embryonic stem cells (ESCs) hold great potential for regenerative medicine due to their two unique characteristics: self-renewal and pluripotency. Several groups of nanoparticles

  10. Short communication: Milk meal pattern of dairy calves is affected by computer-controlled milk feeder set-up

    DEFF Research Database (Denmark)

    Jensen, Margit Bak

    2009-01-01

    for a minimum of 2 and 4 portions, respectively, whereas low-fed calves ingested their milk in 2.4 and 4.4 meals for a minimum of 2 and 4 portions, respectively. Calves on a high milk allowance had fewer milk meals over time, whereas calves on a low milk allowance had the same number of milk meals throughout...... milk portions, whereas the other half could ingest the milk in 4 or more daily portions. Data were collected during 3 successive 14-d periods, the first period starting the day after introduction to the feeder at minimum 12 d of age. High-fed calves ingested their milk in 4.0 and 4.9 meals....... Thus, the development from small and frequent milk meals to fewer and larger meals reported by studies of natural suckling was also found among high-fed calves on a computer-controlled milk feeder. Irrespectively of minimum number of milk portions, the low-fed calves had more unrewarded visits...

  11. A look at one of the world`s largest apron feeder drives - Alberta Oil Sands Project

    Energy Technology Data Exchange (ETDEWEB)

    Persson, O. [Hagglunds Drives Canada Inc., Vancouver, BC (Canada)

    1999-10-01

    Various types of equipment to transport tar sands to processing plants are discussed, with special attention to the advantages of hydraulic direct drives over conventional electro-mechanical drives. A hydraulic direct drive such as the Hagglund Drive has exceptional starting torque capacity due to the high torque capability of the hydraulic motor. As such, it can be particularly useful in applications where shock loads occur with some frequency, or where many starts and stops are needed. Application of the Hagglund drive to power one of the world`s largest apron feeders in the Alberta Oil Sands is described as an illustration of the exceptional reliability, productivity and performance of this equipment. It has about one five-hundredth of the inertia of an equivalent high speed drive with gear reducer, a feature which is particularly significant in the case of feeders which are known to suffer much downtime due to chain related problems. These types of drives have also been used to great advantage in the process industries like pulp and paper, chemical, rubber and plastics, recycling and steel. 1 tab., 1 fig.

  12. Handling system for nuclear fuel cans to a fuel pellet feeder

    International Nuclear Information System (INIS)

    Vere, B.; Mathevon, P.

    1985-01-01

    The handling system comprises a first array of conveyors which takes a batch of casings from a delivery rack, alters the spacing between the casings, and delivers them to a vibrating table feeder, a second array of conveyors which readjusts the spacing between casing to its initial value and transfers the casings to a removal rack, and automatic and synchronized control means for ensuring the displacements of casings always in the same direction. The increase of spacing between casings can be used, before feeding, to allow them to be weighed one after the other, and after feeding, for cleaning the end part of fuel cans [fr

  13. Practice on Protection and Improvement of Withstand Voltage Weighing Belt Coal Feeder%耐压称重式皮带给煤机保护改进实践

    Institute of Scientific and Technical Information of China (English)

    王良

    2016-01-01

    This paper introduces the traits of withstand voltage weighing belt coal feeder. According to the modification and practice of parameters of the frequency converter, when there is a malfunction, the automatic recovery of the frequency converter of coal feeder can be achieved. The reliability of the coal feeder ’s operation will be improved, and the effects of the accident power supply perturbation on the coal feeder can be eliminated. Eventually, the safety and stability of the boilers can be guaranteed.%文章介绍了耐压称重式皮带给煤机的特性,通过对变频器参数修改实践,实现给煤机变频器故障保护自动恢复,提高给煤机运行的可靠性,消除电源意外扰动对给煤机的影响,保障锅炉安全持续稳定运行。

  14. Radiation induced bystander signals are independent of DNA damage and DNA repair capacity of the irradiated cells

    Energy Technology Data Exchange (ETDEWEB)

    Kashino, Genro [Gray Cancer Institute, P.O. Box 100, Mount Vernon Hospital, Northwood, Middlesex HA6 2JR (United Kingdom); Particle Radiation Oncology Research Center, Research Reactor Institute, Kyoto University, 2-1010 Asashiro-nishi, Kumatori-cho, Sennan-gun, Osaka 590-0494 (Japan); Suzuki, Keiji [Division of Radiation Biology, Department of Radiology and Radiation Biology, Course of Life Sciences and Radiation Research, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521 (Japan); Matsuda, Naoki [Division of Radiation Biology and Protection, Center for Frontier Life Sciences, Nagasaki University, Nagasaki 852-8102 (Japan); Kodama, Seiji [Radiation Biology Laboratory, Radiation Research Center, Frontier Science Innovation Center, Organization for University-Industry-Government Cooperation, Osaka Prefecture University, 1-2 Gakuen-cho, Sakai, Osaka 599-8570 (Japan); Ono, Koji [Particle Radiation Oncology Research Center, Research Reactor Institute, Kyoto University, 2-1010 Asashiro-nishi, Kumatori-cho, Sennan-gun, Osaka 590-0494 (Japan); Watanabe, Masami [Laboratory of Radiation Biology, Division of Radiation Life Science, Department of Radiation Life Science and Radiation Medical Science, Kyoto University Research Reactor Institute, 2-1010 Asashiro-nishi, Kumatori-cho, Sennan-gun, Osaka 590-0494 (Japan); Prise, Kevin M [Gray Cancer Institute, P.O. Box 100, Mount Vernon Hospital, Northwood, Middlesex HA6 2JR (United Kingdom) and Centre for Cancer Research and Cell Biology, Queen' s University Belfast, Lisburn Road, Belfast BT9 7AB (United Kingdom)]. E-mail: prise@gci.ac.uk

    2007-06-01

    Evidence is accumulating that irradiated cells produce signals, which interact with non-exposed cells in the same population. Here, we analysed the mechanism for bystander signal arising in wild-type CHO cells and repair deficient varients, focussing on the relationship between DNA repair capacity and bystander signal arising in irradiated cells. In order to investigate the bystander effect, we carried out medium transfer experiments after X-irradiation where micronuclei were scored in non-targeted DSB repair deficient xrs5 cells. When conditioned medium from irradiated cells was transferred to unirradiated xrs5 cells, the level of induction was independent of whether the medium came from irradiated wild-type, ssb or dsb repair deficient cells. This result suggests that the activation of a bystander signal is independent of the DNA repair capacity of the irradiated cells. Also, pre-treatment of the irradiated cells with 0.5% DMSO, which suppresses micronuclei induction in CHO but not in xrs5 cells, suppressed bystander effects completely in both conditioned media, suggesting that DMSO is effective for suppression of bystander signal arising independently of DNA damage in irradiated cells. Overall the work presented here adds to the understanding that it is the repair phenotype of the cells receiving bystander signals, which determines overall response rather than that of the cell producing the bystander signal.

  15. A new hybrid evolutionary algorithm based on new fuzzy adaptive PSO and NM algorithms for Distribution Feeder Reconfiguration

    International Nuclear Information System (INIS)

    Niknam, Taher; Azadfarsani, Ehsan; Jabbari, Masoud

    2012-01-01

    Highlights: ► Network reconfiguration is a very important way to save the electrical energy. ► This paper proposes a new algorithm to solve the DFR. ► The algorithm combines NFAPSO with NM. ► The proposed algorithm is tested on two distribution test feeders. - Abstract: Network reconfiguration for loss reduction in distribution system is a very important way to save the electrical energy. This paper proposes a new hybrid evolutionary algorithm to solve the Distribution Feeder Reconfiguration problem (DFR). The algorithm is based on combination of a New Fuzzy Adaptive Particle Swarm Optimization (NFAPSO) and Nelder–Mead simplex search method (NM) called NFAPSO–NM. In the proposed algorithm, a new fuzzy adaptive particle swarm optimization includes two parts. The first part is Fuzzy Adaptive Binary Particle Swarm Optimization (FABPSO) that determines the status of tie switches (open or close) and second part is Fuzzy Adaptive Discrete Particle Swarm Optimization (FADPSO) that determines the sectionalizing switch number. In other side, due to the results of binary PSO(BPSO) and discrete PSO(DPSO) algorithms highly depends on the values of their parameters such as the inertia weight and learning factors, a fuzzy system is employed to adaptively adjust the parameters during the search process. Moreover, the Nelder–Mead simplex search method is combined with the NFAPSO algorithm to improve its performance. Finally, the proposed algorithm is tested on two distribution test feeders. The results of simulation show that the proposed method is very powerful and guarantees to obtain the global optimization.

  16. Genetic Validation of Cell Proliferation via Ras-Independent Activation of the Raf/Mek/Erk Pathway.

    Science.gov (United States)

    Lechuga, Carmen G; Simón-Carrasco, Lucía; Jacob, Harrys K C; Drosten, Matthias

    2017-01-01

    Signaling transmitted by the Ras family of small GTPases (H-, N-, and K-Ras) is essential for proliferation of mouse embryonic fibroblasts (MEFs). However, constitutive activation of the downstream Raf/Mek/Erk pathway can bypass the requirement for Ras proteins and allow cells to proliferate in the absence of the three Ras isoforms. Here we describe a protocol for a colony formation assay that permits evaluating the role of candidate proteins that are positive or negative regulators of cell proliferation mediated via Ras-independent Raf/Mek/Erk pathway activation. K-Ras lox (H-Ras -/- , N-Ras -/- , K-Ras lox/lox , RERT ert/ert ) MEFs are infected with retro- or lentiviral vectors expressing wild-type or constitutively activated candidate cDNAs, shRNAs, or sgRNAs in combination with Cas9 to ascertain the possibility of candidate proteins to function either as an activator or inhibitor of Ras-independent Raf/Mek/Erk activation. These cells are then seeded in the absence or presence of 4-Hydroxytamoxifen (4-OHT), which activates the resident CreERT2 alleles resulting in elimination of the conditional K-Ras alleles and ultimately generating Rasless cells. Colony formation in the presence of 4-OHT indicates cell proliferation via Ras-independent Raf/Mek/Erk activation.

  17. Effects of Home Energy Management Systems on Distribution Utilities and Feeders Under Various Market Structure; NREL (National Renewable Energy Laboratory)

    Energy Technology Data Exchange (ETDEWEB)

    Ruth, M.; Pratt, A.; Lunacek, M.; Mittal, S.; Wu, H.; Jones, W.

    2015-06-15

    The combination of distributed energy resources (DER) and retail tariff structures to provide benefits to both utility consumers and the utilities is not well understood. To improve understanding, an Integrated Energy System Model (IESM) is being developed to simulate the physical and economic aspects of DER technologies, the buildings where they reside, and feeders servicing them. The IESM was used to simulate 20 houses with home energy management systems on a single feeder under a time-of-use (TOU) tariff to estimate economic and physical impacts on both the households and the distribution utilities. Home energy management systems (HEMS) reduce consumers’ electric bills by precooling houses in the hours before peak electricity pricing. Utilization of HEMS reduce peak loads during high price hours but shifts it to hours with off-peak and shoulder prices, resulting in a higher peak load. used to simulate 20 houses with home energy management systems on a single feeder under a time-of-use (TOU) tariff to estimate economic and physical impacts on both the households and the distribution utilities. Home energy management systems (HEMS) reduce consumers’ electric bills by precooling houses in the hours before peak electricity pricing. Utilization of HEMS reduce peak loads during high price hours but shifts it to hours with off-peak and shoulder prices, resulting in a higher peak load.

  18. A review of human cell radiosensitivity in vitro

    International Nuclear Information System (INIS)

    Deschavanne, Patrick J.; Fertil, Bernard

    1996-01-01

    The survival curves of 694 human cell lines irradiated in exponentially growing phase in vitro were collected from the literature. Among them, 271 were derived from tumors, 423 were nontransformed fibroblasts and other normal cell strains from healthy people or people with some genetic disorders. Seventy-six different cell types are identified, and a specific radiosensitivity could be associated with each, using D-bar and surviving fraction at 2 Gy. Technical factors such as culture medium, feeder cells, and scoring method were found to affect intrinsic radiosensitivity. In particular, the cell type is not a discriminating factor when cells are studied in agar. Results obtained with cells irradiated in agar must be used cautiously, depending on how the cells were prepared for the experiments. The use of feeder cells narrows the range of radiosensitivity of human cells. For cells irradiated as monolayer, it was possible to build a scale of radiosensitivity according to cell type, ranging, in terms of D-bar from 0.6 Gy for the most sensitive cell lines to more than 4 Gy for the most resistant. Considering that, in most cases, we could estimate the variation of radiosensitivity within each cell type, our classification among cell types can be used by researchers to place their results in the context of the literature

  19. The Linehaul-Feeder Vehicle Routing Problem with Virtual Depots and Time Windows

    Directory of Open Access Journals (Sweden)

    Huey-Kuo Chen

    2011-01-01

    Full Text Available This paper addresses the linehaul-feeder vehicle routing problem with virtual depots and time windows (LFVRPTW. Small and large vehicles deliver services to customers within time constraints; small vehicles en route may reload commodities from either the physical depot or from the larger vehicle at a virtual depot before continuing onward. A two-stage solution heuristic involving Tabu search is proposed to solve this problem. The test results show that the LFVRPTW performs better than the vehicle routing problem with time windows in terms of both objective value and the number of small vehicles dispatched.

  20. Is perceived emotional support beneficial? Well-being and health in independent and interdependent cultures.

    Science.gov (United States)

    Uchida, Yukiko; Kitayama, Shinobu; Mesquita, Batja; Reyes, Jose Alberto S; Morling, Beth

    2008-06-01

    Previous studies show there is little or no association between perceived emotional support and well-being in European American culture. The authors hypothesized that this paradoxical absence of any benefit of perceived support is unique to cultural contexts that privilege independence rather than interdependence of the self. Study 1 tested college students and found, as predicted, that among Euro-Americans a positive effect of perceived emotional support on subjective well-being (positive affect) was weak and, moreover, it disappeared entirely once self-esteem was statistically controlled. In contrast, among Asians in Asia (Japanese and Filipinos) perceived emotional support positively predicted subjective well-being even after self-esteem was controlled. Study 2 extended Study 1 by testing both Japanese and American adults in midlife with respect to multiple indicators of well-being and physical health. Overall, the evidence underscores the central significance of culture as a moderator of the effectiveness of perceived emotional support.

  1. Microenvironmental independence associated with tumor progression.

    Science.gov (United States)

    Anderson, Alexander R A; Hassanein, Mohamed; Branch, Kevin M; Lu, Jenny; Lobdell, Nichole A; Maier, Julie; Basanta, David; Weidow, Brandy; Narasanna, Archana; Arteaga, Carlos L; Reynolds, Albert B; Quaranta, Vito; Estrada, Lourdes; Weaver, Alissa M

    2009-11-15

    Tumor-microenvironment interactions are increasingly recognized to influence tumor progression. To understand the competitive dynamics of tumor cells in diverse microenvironments, we experimentally parameterized a hybrid discrete-continuum mathematical model with phenotypic trait data from a set of related mammary cell lines with normal, transformed, or tumorigenic properties. Surprisingly, in a resource-rich microenvironment, with few limitations on proliferation or migration, transformed (but not tumorigenic) cells were most successful and outcompeted other cell types in heterogeneous tumor simulations. Conversely, constrained microenvironments with limitations on space and/or growth factors gave a selective advantage to phenotypes derived from tumorigenic cell lines. Analysis of the relative performance of each phenotype in constrained versus unconstrained microenvironments revealed that, although all cell types grew more slowly in resource-constrained microenvironments, the most aggressive cells were least affected by microenvironmental constraints. A game theory model testing the relationship between microenvironment resource availability and competitive cellular dynamics supports the concept that microenvironmental independence is an advantageous cellular trait in resource-limited microenvironments.

  2. Independent prognostic value of eosinophil and mast cell infiltration in colorectal cancer tissue

    DEFF Research Database (Denmark)

    Nielsen, Hans Jørgen; Hansen, Ulla; Christensen, Ib Jarle

    1999-01-01

    -assisted microscope, which allowed semi-automated quantification of cells within a fixed area. Total white cells and individual counts of eosinophils, neutrophils, mast cells, lymphocytes, and plasma cells were evaluated in every tumour specimen. Stratification into four groups with similar numbers of events was used...... age ( p=0.0003), and tumour location in the rectum predicted poor survival, while high counts of eosinophils ( p=0.006) and mast cells ( p=0.02) predicted good survival. Tumour-associated eosinophilia and mastocytosis appear to be independent prognostic variables in colorectal cancer. Future studies...... should investigate the potential biological role of tumour tissue eosinophils and mast cells in the modulation of tumour growth....

  3. CCR9 interactions support ovarian cancer cell survival and resistance to cisplatin-induced apoptosis in a PI3K-dependent and FAK-independent fashion

    Directory of Open Access Journals (Sweden)

    Johnson Erica L

    2010-06-01

    Full Text Available Abstract Background Cisplatin is more often used to treat ovarian cancer (OvCa, which provides modest survival advantage primarily due to chemo-resistance and up regulated anti-apoptotic machineries in OvCa cells. Therefore, targeting the mechanisms responsible for cisplatin resistance in OvCa cell may improve therapeutic outcomes. We have shown that ovarian cancer cells express CC chemokine receptor-9 (CCR9. Others have also shown that CCL25, the only natural ligand for CCR9, up regulates anti-apoptotic proteins in immature T lymphocytes. Hence, it is plausible that CCR9-mediated cell signals might be involved in OvCa cell survival and inhibition of cisplatin-induced apoptosis. In this study, we investigated the potential role and molecular mechanisms of CCR9-mediated inhibition of cisplatin-induced apoptosis in OvCa cells. Methods Cell proliferation, vibrant apoptosis, and TUNEL assays were performed with or without cisplatin treatment in presence or absence of CCL25 to determine the role of the CCR9-CCL25 axis in cisplatin resistance. In situ Fast Activated cell-based ELISA (FACE assays were performed to determine anti-apoptotic signaling molecules responsible for CCL25-CCR9 mediated survival. Results Our results show interactions between CCR9 and CCL25 increased anti-apoptotic signaling cascades in OvCa cells, which rescued cells from cisplatin-induced cell death. Specifically, CCL25-CCR9 interactions mediated Akt, activation as well as GSK-3β and FKHR phosphorylation in a PI3K-dependent and FAK-independent fashion. Conclusions Our results suggest the CCR9-CCL25 axis plays an important role in reducing cisplatin-induced apoptosis of OvCa cells.

  4. Rapid generation of mitochondrial superoxide induces mitochondrion-dependent but caspase-independent cell death in hippocampal neuronal cells that morphologically resembles necroptosis

    International Nuclear Information System (INIS)

    Fukui, Masayuki; Choi, Hye Joung; Zhu, Bao Ting

    2012-01-01

    Studies in recent years have revealed that excess mitochondrial superoxide production is an important etiological factor in neurodegenerative diseases, resulting from oxidative modifications of cellular lipids, proteins, and nucleic acids. Hence, it is important to understand the mechanism by which mitochondrial oxidative stress causes neuronal death. In this study, the immortalized mouse hippocampal neuronal cells (HT22) in culture were used as a model and they were exposed to menadione (also known as vitamin K 3 ) to increase intracellular superoxide production. We found that menadione causes preferential accumulation of superoxide in the mitochondria of these cells, along with the rapid development of mitochondrial dysfunction and cellular ATP depletion. Neuronal death induced by menadione is independent of the activation of the MAPK signaling pathways and caspases. The lack of caspase activation is due to the rapid depletion of cellular ATP. It was observed that two ATP-independent mitochondrial nucleases, namely, AIF and Endo G, are released following menadione exposure. Silencing of their expression using specific siRNAs results in transient suppression (for ∼ 12 h) of mitochondrial superoxide-induced neuronal death. While suppression of the mitochondrial superoxide dismutase expression markedly sensitizes neuronal cells to mitochondrial superoxide-induced cytotoxicity, its over-expression confers strong protection. Collectively, these findings showed that many of the observed features associated with mitochondrial superoxide-induced cell death, including caspase independency, rapid depletion of ATP level, mitochondrial release of AIF and Endo G, and mitochondrial swelling, are distinctly different from those of apoptosis; instead they resemble some of the known features of necroptosis. -- Highlights: ► Menadione causes mitochondrial superoxide accumulation and injury. ► Menadione-induced cell death is caspase-independent, due to rapid depletion of ATP

  5. Rapid generation of mitochondrial superoxide induces mitochondrion-dependent but caspase-independent cell death in hippocampal neuronal cells that morphologically resembles necroptosis

    Energy Technology Data Exchange (ETDEWEB)

    Fukui, Masayuki; Choi, Hye Joung; Zhu, Bao Ting, E-mail: BTZhu@kumc.edu

    2012-07-15

    Studies in recent years have revealed that excess mitochondrial superoxide production is an important etiological factor in neurodegenerative diseases, resulting from oxidative modifications of cellular lipids, proteins, and nucleic acids. Hence, it is important to understand the mechanism by which mitochondrial oxidative stress causes neuronal death. In this study, the immortalized mouse hippocampal neuronal cells (HT22) in culture were used as a model and they were exposed to menadione (also known as vitamin K{sub 3}) to increase intracellular superoxide production. We found that menadione causes preferential accumulation of superoxide in the mitochondria of these cells, along with the rapid development of mitochondrial dysfunction and cellular ATP depletion. Neuronal death induced by menadione is independent of the activation of the MAPK signaling pathways and caspases. The lack of caspase activation is due to the rapid depletion of cellular ATP. It was observed that two ATP-independent mitochondrial nucleases, namely, AIF and Endo G, are released following menadione exposure. Silencing of their expression using specific siRNAs results in transient suppression (for ∼ 12 h) of mitochondrial superoxide-induced neuronal death. While suppression of the mitochondrial superoxide dismutase expression markedly sensitizes neuronal cells to mitochondrial superoxide-induced cytotoxicity, its over-expression confers strong protection. Collectively, these findings showed that many of the observed features associated with mitochondrial superoxide-induced cell death, including caspase independency, rapid depletion of ATP level, mitochondrial release of AIF and Endo G, and mitochondrial swelling, are distinctly different from those of apoptosis; instead they resemble some of the known features of necroptosis. -- Highlights: ► Menadione causes mitochondrial superoxide accumulation and injury. ► Menadione-induced cell death is caspase-independent, due to rapid depletion of

  6. Elimination of remaining undifferentiated induced pluripotent stem cells in the process of human cardiac cell sheet fabrication using a methionine-free culture condition.

    Science.gov (United States)

    Matsuura, Katsuhisa; Kodama, Fumiko; Sugiyama, Kasumi; Shimizu, Tatsuya; Hagiwara, Nobuhisa; Okano, Teruo

    2015-03-01

    Cardiac tissue engineering is a promising method for regenerative medicine. Although we have developed human cardiac cell sheets by integration of cell sheet-based tissue engineering and scalable bioreactor culture, the risk of contamination by induced pluripotent stem (iPS) cells in cardiac cell sheets remains unresolved. In the present study, we established a novel culture method to fabricate human cardiac cell sheets with a decreased risk of iPS cell contamination while maintaining viabilities of iPS cell-derived cells, including cardiomyocytes and fibroblasts, using a methionine-free culture condition. When cultured in the methionine-free condition, human iPS cells did not survive without feeder cells and could not proliferate or form colonies on feeder cells or in coculture with cells for cardiac cell sheet fabrication. When iPS cell-derived cells after the cardiac differentiation were transiently cultured in the methionine-free condition, gene expression of OCT3/4 and NANOG was downregulated significantly compared with that in the standard culture condition. Furthermore, in fabricated cardiac cell sheets, spontaneous and synchronous beating was observed in the whole area while maintaining or upregulating the expression of various cardiac and extracellular matrix genes. These findings suggest that human iPS cells are methionine dependent and a methionine-free culture condition for cardiac cell sheet fabrication might reduce the risk of iPS cell contamination.

  7. Antibody-independent control of gamma-herpesvirus latency via B cell induction of anti-viral T cell responses.

    Directory of Open Access Journals (Sweden)

    Kelly B McClellan

    2006-06-01

    Full Text Available B cells can use antibody-dependent mechanisms to control latent viral infections. It is unknown whether this represents the sole function of B cells during chronic viral infection. We report here that hen egg lysozyme (HEL-specific B cells can contribute to the control of murine gamma-herpesvirus 68 (gammaHV68 latency without producing anti-viral antibody. HEL-specific B cells normalized defects in T cell numbers and proliferation observed in B cell-/- mice during the early phase of gammaHV68 latency. HEL-specific B cells also reversed defects in CD8 and CD4 T cell cytokine production observed in B cell-/- mice, generating CD8 and CD4 T cells necessary for control of latency. Furthermore, HEL-specific B cells were able to present virally encoded antigen to CD8 T cells. Therefore, B cells have antibody independent functions, including antigen presentation, that are important for control of gamma-herpesvirus latency. Exploitation of this property of B cells may allow enhanced vaccine responses to chronic virus infection.

  8. The Adder Phenomenon Emerges from Independent Control of Pre- and Post-Start Phases of the Budding Yeast Cell Cycle.

    Science.gov (United States)

    Chandler-Brown, Devon; Schmoller, Kurt M; Winetraub, Yonatan; Skotheim, Jan M

    2017-09-25

    Although it has long been clear that cells actively regulate their size, the molecular mechanisms underlying this regulation have remained poorly understood. In budding yeast, cell size primarily modulates the duration of the cell-division cycle by controlling the G1/S transition known as Start. We have recently shown that the rate of progression through Start increases with cell size, because cell growth dilutes the cell-cycle inhibitor Whi5 in G1. Recent phenomenological studies in yeast and bacteria have shown that these cells add an approximately constant volume during each complete cell cycle, independent of their size at birth. These results seem to be in conflict, as the phenomenological studies suggest that cells measure the amount they grow, rather than their size, and that size control acts over the whole cell cycle, rather than specifically in G1. Here, we propose an integrated model that unifies the adder phenomenology with the molecular mechanism of G1/S cell-size control. We use single-cell microscopy to parameterize a full cell-cycle model based on independent control of pre- and post-Start cell-cycle periods. We find that our model predicts the size-independent amount of cell growth during the full cell cycle. This suggests that the adder phenomenon is an emergent property of the independent regulation of pre- and post-Start cell-cycle periods rather than the consequence of an underlying molecular mechanism measuring a fixed amount of growth. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Knock-in of large reporter genes in human cells via CRISPR/Cas9-induced homology-dependent and independent DNA repair.

    Science.gov (United States)

    He, Xiangjun; Tan, Chunlai; Wang, Feng; Wang, Yaofeng; Zhou, Rui; Cui, Dexuan; You, Wenxing; Zhao, Hui; Ren, Jianwei; Feng, Bo

    2016-05-19

    CRISPR/Cas9-induced site-specific DNA double-strand breaks (DSBs) can be repaired by homology-directed repair (HDR) or non-homologous end joining (NHEJ) pathways. Extensive efforts have been made to knock-in exogenous DNA to a selected genomic locus in human cells; which, however, has focused on HDR-based strategies and was proven inefficient. Here, we report that NHEJ pathway mediates efficient rejoining of genome and plasmids following CRISPR/Cas9-induced DNA DSBs, and promotes high-efficiency DNA integration in various human cell types. With this homology-independent knock-in strategy, integration of a 4.6 kb promoterless ires-eGFP fragment into the GAPDH locus yielded up to 20% GFP+ cells in somatic LO2 cells, and 1.70% GFP+ cells in human embryonic stem cells (ESCs). Quantitative comparison further demonstrated that the NHEJ-based knock-in is more efficient than HDR-mediated gene targeting in all human cell types examined. These data support that CRISPR/Cas9-induced NHEJ provides a valuable new path for efficient genome editing in human ESCs and somatic cells. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  10. Recombinant human albumin supports single cell cloning of CHO cells in chemically defined media.

    Science.gov (United States)

    Zhu, Jiang; Wooh, Jong Wei; Hou, Jeff Jia Cheng; Hughes, Benjamin S; Gray, Peter P; Munro, Trent P

    2012-01-01

    Biologic drugs, such as monoclonal antibodies, are commonly made using mammalian cells in culture. The cell lines used for manufacturing should ideally be clonal, meaning derived from a single cell, which represents a technically challenging process. Fetal bovine serum is often used to support low cell density cultures, however, from a regulatory perspective, it is preferable to avoid animal-derived components to increase process consistency and reduce the risk of contamination from adventitious agents. Chinese hamster ovary (CHO) cells are the most widely used cell line in industry and a large number of serum-free, protein-free, and fully chemically defined growth media are commercially available, although these media alone do not readily support efficient single cell cloning. In this work, we have developed a simple, fully defined, single-cell cloning media, specifically for CHO cells, using commercially available reagents. Our results show that a 1:1 mixture of CD-CHO™ and DMEM/F12 supplemented with 1.5 g/L of recombinant albumin (Albucult®) supports single cell cloning. This formulation can support recovery of single cells in 43% of cultures compared to 62% in the presence of serum. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

  11. Extracellular Matrix-Dependent Generation of Integration- and Xeno-Free iPS Cells Using a Modified mRNA Transfection Method

    Directory of Open Access Journals (Sweden)

    Kang-In Lee

    2016-01-01

    Full Text Available Human induced pluripotent stem cells (iPS cells hold great promise in the field of regenerative medicine, especially immune-compatible cell therapy. The most important safety-related issues that must be resolved before the clinical use of iPS cells include the generation of “footprint-free” and “xeno-free” iPS cells. In this study, we sought to examine whether an extracellular matrix- (ECM- based xeno-free culture system that we recently established could be used together with a microRNA-enhanced mRNA reprogramming method for the generation of clinically safe iPS cells. The notable features of this method are the use of a xeno-free/feeder-free culture system for the generation and expansion of iPS cells rather than the conventional labor-intensive culture systems using human feeder cells or human feeder-conditioned medium and the enhancement of mRNA-mediated reprogramming via the delivery of microRNAs. Strikingly, we observed the early appearance of iPS cell colonies (~11 days, substantial reprogramming efficiency (~0.2–0.3%, and a high percentage of ESC-like colonies among the total colonies (~87.5%, indicating enhanced kinetics and reprogramming efficiency. Therefore, the combined method established in this study provides a valuable platform for the generation and expansion of clinically safe (i.e., integration- and xeno-free iPS cells, facilitating immune-matched cell therapy in the near future.

  12. CD40 signaling synergizes with TLR-2 in the BCR independent activation of resting B cells.

    Directory of Open Access Journals (Sweden)

    Shweta Jain

    Full Text Available Conventionally, signaling through BCR initiates sequence of events necessary for activation and differentiation of B cells. We report an alternative approach, independent of BCR, for stimulating resting B (RB cells, by involving TLR-2 and CD40--molecules crucial for innate and adaptive immunity. CD40 triggering of TLR-2 stimulated RB cells significantly augments their activation, proliferation and differentiation. It also substantially ameliorates the calcium flux, antigen uptake capacity and ability of B cells to activate T cells. The survival of RB cells was improved and it increases the number of cells expressing activation induced deaminase (AID, signifying class switch recombination (CSR. Further, we also observed increased activation rate and decreased threshold period required for optimum stimulation of RB cells. These results corroborate well with microarray gene expression data. This study provides novel insights into coordination between the molecules of innate and adaptive immunity in activating B cells, in a BCR independent manner. This strategy can be exploited to design vaccines to bolster B cell activation and antigen presenting efficiency, leading to faster and better immune response.

  13. CD40 signaling synergizes with TLR-2 in the BCR independent activation of resting B cells.

    Science.gov (United States)

    Jain, Shweta; Chodisetti, Sathi Babu; Agrewala, Javed N

    2011-01-01

    Conventionally, signaling through BCR initiates sequence of events necessary for activation and differentiation of B cells. We report an alternative approach, independent of BCR, for stimulating resting B (RB) cells, by involving TLR-2 and CD40--molecules crucial for innate and adaptive immunity. CD40 triggering of TLR-2 stimulated RB cells significantly augments their activation, proliferation and differentiation. It also substantially ameliorates the calcium flux, antigen uptake capacity and ability of B cells to activate T cells. The survival of RB cells was improved and it increases the number of cells expressing activation induced deaminase (AID), signifying class switch recombination (CSR). Further, we also observed increased activation rate and decreased threshold period required for optimum stimulation of RB cells. These results corroborate well with microarray gene expression data. This study provides novel insights into coordination between the molecules of innate and adaptive immunity in activating B cells, in a BCR independent manner. This strategy can be exploited to design vaccines to bolster B cell activation and antigen presenting efficiency, leading to faster and better immune response.

  14. A look at one of the world's largest apron feeder drives - Alberta Oil Sands Project

    Energy Technology Data Exchange (ETDEWEB)

    Persson, O. (Hagglunds Drives Canada Inc., Vancouver, BC (Canada))

    1999-01-01

    Various types of equipment to transport tar sands to processing plants are discussed, with special attention to the advantages of hydraulic direct drives over conventional electro-mechanical drives. A hydraulic direct drive such as the Hagglund Drive has exceptional starting torque capacity due to the high torque capability of the hydraulic motor. As such, it can be particularly useful in applications where shock loads occur with some frequency, or where many starts and stops are needed. Application of the Hagglund drive to power one of the world's largest apron feeders in the Alberta Oil Sands is described as an illustration of the exceptional reliability, productivity and performance of this equipment. It has about one five-hundredth of the inertia of an equivalent high speed drive with gear reducer, a feature which is particularly significant in the case of feeders which are known to suffer much downtime due to chain related problems. These types of drives have also been used to great advantage in the process industries like pulp and paper, chemical, rubber and plastics, recycling and steel. 1 tab., 1 fig.

  15. The PPARγ ligand ciglitazone regulates androgen receptor activation differently in androgen-dependent versus androgen-independent human prostate cancer cells

    International Nuclear Information System (INIS)

    Moss, Patrice E.; Lyles, Besstina E.; Stewart, LaMonica V.

    2010-01-01

    The androgen receptor (AR) regulates growth and progression of androgen-dependent as well as androgen-independent prostate cancer cells. Peroxisome proliferator-activated receptor gamma (PPARγ) agonists have been reported to reduce AR activation in androgen-dependent LNCaP prostate cancer cells. To determine whether PPARγ ligands are equally effective at inhibiting AR activity in androgen-independent prostate cancer, we examined the effect of the PPARγ ligands ciglitazone and rosiglitazone on C4-2 cells, an androgen- independent derivative of the LNCaP cell line. Luciferase-based reporter assays and Western blot analysis demonstrated that PPARγ ligand reduced dihydrotestosterone (DHT)-induced increases in AR activity in LNCaP cells. However, in C4-2 cells, these compounds increased DHT-induced AR driven luciferase activity. In addition, ciglitazone did not significantly alter DHT-mediated increases in prostate specific antigen (PSA) protein or mRNA levels within C4-2 cells. siRNA-based experiments demonstrated that the ciglitazone-induced regulation of AR activity observed in C4-2 cells was dependent on the presence of PPARγ. Furthermore, overexpression of the AR corepressor cyclin D1 inhibited the ability of ciglitazone to induce AR luciferase activity in C4-2 cells. Thus, our data suggest that both PPARγ and cyclin D1 levels influence the ability of ciglitazone to differentially regulate AR signaling in androgen-independent C4-2 prostate cancer cells.

  16. High Temperature Electrolysis using Electrode-Supported Cells

    International Nuclear Information System (INIS)

    O'Brien, J.E.; Stoots, C.M.

    2010-01-01

    An experimental study is under way to assess the performance of electrode-supported solid-oxide cells operating in the steam electrolysis mode for hydrogen production. The cells currently under study were developed primarily for the fuel cell mode of operation. Results presented in this paper were obtained from single cells, with an active area of 16 cm2 per cell. The electrolysis cells are electrode-supported, with yttria-stabilized zirconia (YSZ) electrolytes (∼10 (micro)m thick), nickel-YSZ steam/hydrogen electrodes (∼1400 (micro)m thick), and manganite (LSM) air-side electrodes (∼90 (micro)m thick). The purpose of the present study was to document and compare the performance and degradation rates of these cells in the fuel cell mode and in the electrolysis mode under various operating conditions. Initial performance was documented through a series of DC potential sweeps and AC impedance spectroscopy measurements. Degradation was determined through long-duration testing, first in the fuel cell mode, then in the electrolysis mode over more than 500 hours of operation. Results indicate accelerated degradation rates in the electrolysis mode compared to the fuel cell mode, possibly due to electrode delamination. The paper also includes details of the single-cell test apparatus developed specifically for these experiments.

  17. Ibrutinib synergizes with poly(ADP-ribose) glycohydrolase inhibitors to induce cell death in AML cells via a BTK-independent mechanism.

    Science.gov (United States)

    Rotin, Lianne E; Gronda, Marcela; MacLean, Neil; Hurren, Rose; Wang, XiaoMing; Lin, Feng-Hsu; Wrana, Jeff; Datti, Alessandro; Barber, Dwayne L; Minden, Mark D; Slassi, Malik; Schimmer, Aaron D

    2016-01-19

    Targeting Bruton's tyrosine kinase (BTK) with the small molecule BTK inhibitor ibrutinib has significantly improved patient outcomes in several B-cell malignancies, with minimal toxicity. Given the reported expression and constitutive activation of BTK in acute myeloid leukemia (AML) cells, there has been recent interest in investigating the anti-AML activity of ibrutinib. We noted that ibrutinib had limited single-agent toxicity in a panel of AML cell lines and primary AML samples, and therefore sought to identify ibrutinib-sensitizing drugs. Using a high-throughput combination chemical screen, we identified that the poly(ADP-ribose) glycohydrolase (PARG) inhibitor ethacridine lactate synergized with ibrutinib in TEX and OCI-AML2 leukemia cell lines. The combination of ibrutinib and ethacridine induced a synergistic increase in reactive oxygen species that was functionally important to explain the observed cell death. Interestingly, synergistic cytotoxicity of ibrutinib and ethacridine was independent of the inhibitory effect of ibrutinib against BTK, as knockdown of BTK did not sensitize TEX and OCI-AML2 cells to ethacridine treatment. Thus, our findings indicate that ibrutinib may have a BTK-independent role in AML and that PARG inhibitors may have utility as part of a combination therapy for this disease.

  18. Restrictions in cell cycle progression of adult vestibular supporting cells in response to ectopic cyclin D1 expression.

    Directory of Open Access Journals (Sweden)

    Heidi Loponen

    Full Text Available Sensory hair cells and supporting cells of the mammalian inner ear are quiescent cells, which do not regenerate. In contrast, non-mammalian supporting cells have the ability to re-enter the cell cycle and produce replacement hair cells. Earlier studies have demonstrated cyclin D1 expression in the developing mouse supporting cells and its downregulation along maturation. In explant cultures of the mouse utricle, we have here focused on the cell cycle control mechanisms and proliferative potential of adult supporting cells. These cells were forced into the cell cycle through adenoviral-mediated cyclin D1 overexpression. Ectopic cyclin D1 triggered robust cell cycle re-entry of supporting cells, accompanied by changes in p27(Kip1 and p21(Cip1 expressions. Main part of cell cycle reactivated supporting cells were DNA damaged and arrested at the G2/M boundary. Only small numbers of mitotic supporting cells and rare cells with signs of two successive replications were found. Ectopic cyclin D1-triggered cell cycle reactivation did not lead to hyperplasia of the sensory epithelium. In addition, a part of ectopic cyclin D1 was sequestered in the cytoplasm, reflecting its ineffective nuclear import. Combined, our data reveal intrinsic barriers that limit proliferative capacity of utricular supporting cells.

  19. Restrictions in cell cycle progression of adult vestibular supporting cells in response to ectopic cyclin D1 expression.

    Science.gov (United States)

    Loponen, Heidi; Ylikoski, Jukka; Albrecht, Jeffrey H; Pirvola, Ulla

    2011-01-01

    Sensory hair cells and supporting cells of the mammalian inner ear are quiescent cells, which do not regenerate. In contrast, non-mammalian supporting cells have the ability to re-enter the cell cycle and produce replacement hair cells. Earlier studies have demonstrated cyclin D1 expression in the developing mouse supporting cells and its downregulation along maturation. In explant cultures of the mouse utricle, we have here focused on the cell cycle control mechanisms and proliferative potential of adult supporting cells. These cells were forced into the cell cycle through adenoviral-mediated cyclin D1 overexpression. Ectopic cyclin D1 triggered robust cell cycle re-entry of supporting cells, accompanied by changes in p27(Kip1) and p21(Cip1) expressions. Main part of cell cycle reactivated supporting cells were DNA damaged and arrested at the G2/M boundary. Only small numbers of mitotic supporting cells and rare cells with signs of two successive replications were found. Ectopic cyclin D1-triggered cell cycle reactivation did not lead to hyperplasia of the sensory epithelium. In addition, a part of ectopic cyclin D1 was sequestered in the cytoplasm, reflecting its ineffective nuclear import. Combined, our data reveal intrinsic barriers that limit proliferative capacity of utricular supporting cells.

  20. Assessment of Fuel Analysis Methodology and Fission Product Release for 37-Element Fuel by Using the Latest IST Codes during Stagnation Feeder Break in CANDU

    International Nuclear Information System (INIS)

    Park, Joo Hwan; Jung, Jong Yeob

    2009-09-01

    Feeder break accident is regarded as one of the design basis accident in CANDU reactor which results in a fuel failure. For a particular range of inlet feeder break sizes, the flow in the channel is reduced sufficiently that the fuel and fuel channel integrity can be significantly affected to have damage in the affected channel, while the remainder of the core remains adequately cooled. The flow in the downstream channel can be more or less stagnated due to a balance between pressure at the break on the upstream side and the reverse driving pressure between the break and the downstream end. In the extreme, this can lead to rapid fuel heatup and fuel damage and failure of the fuel channel similar to that associated with a severe channel flow blockage. Such an inlet feeder break scenario is called a stagnation break. In this report, the fuel analysis methodology and the assessment results of fission product inventory and release during the stagnation feeder break are described for conservatively assumed limiting channel. The accident was assumed to be occurred in the refurbished Wolsong unit 1 and the latest safety codes were used in the analysis. Fission product inventories during the steady state were calculated by using ELESTRES-IST 1.2 code. The whole analysis process was carried out by a script file which was programmed by Perl language. The perl script file was programmed to make all ELESTRES input files for each bundle and each ring based on the given power-burnup history and thermal-hydraulic conditions of the limiting channel and to perform the fuel analysis automatically. The fission product release during the transient period of stagnation feeder break was evaluated by applying Gehl model. The amounts of each isotope's release are conservatively evaluated for additional 2 seconds after channel failure. The calculated fission product releases are provided to the following dose assessment as a source term

  1. Retinoic Acid Signaling in B Cells Is Required for the Generation of an Effective T-Independent Immune Response.

    Science.gov (United States)

    Marks, Ellen; Ortiz, Carla; Pantazi, Eirini; Bailey, Charlotte S; Lord, Graham M; Waldschmidt, Thomas J; Noelle, Randolph J; Elgueta, Raul

    2016-01-01

    Retinoic acid (RA) plays an important role in the balance of inflammation and tolerance in T cells. Furthermore, it has been demonstrated that RA facilitates IgA isotype switching in B cells in vivo . However, it is unclear whether RA has a direct effect on T-independent B cell responses in vivo . To address this question, we generated a mouse model where RA signaling is specifically silenced in the B cell lineage. This was achieved through the overexpression of a dominant negative receptor α for RA (dnRARα) in the B cell lineage. In this model, we found a dramatic reduction in marginal zone (MZ) B cells and accumulation of transitional 2 B cells in the spleen. We also observed a reduction in B1 B cells in the peritoneum with a defect in the T-independent B cell response against 2,4,6-trinitrophenyl. This was not a result of inhibited development of B cells in the bone marrow, but likely the result of both defective expression of S1P 1 in MZ B cells and a defect in the development of MZ and B1 B cells. This suggests that RARα expression in B cells is important for B cell frequency in the MZ and peritoneum, which is crucial for the generation of T-independent humoral responses.

  2. Retinoic Acid Signaling in B Cells Is Required for the Generation of an Effective T-Independent Immune Response

    Science.gov (United States)

    Marks, Ellen; Ortiz, Carla; Pantazi, Eirini; Bailey, Charlotte S.; Lord, Graham M.; Waldschmidt, Thomas J.; Noelle, Randolph J.; Elgueta, Raul

    2016-01-01

    Retinoic acid (RA) plays an important role in the balance of inflammation and tolerance in T cells. Furthermore, it has been demonstrated that RA facilitates IgA isotype switching in B cells in vivo. However, it is unclear whether RA has a direct effect on T-independent B cell responses in vivo. To address this question, we generated a mouse model where RA signaling is specifically silenced in the B cell lineage. This was achieved through the overexpression of a dominant negative receptor α for RA (dnRARα) in the B cell lineage. In this model, we found a dramatic reduction in marginal zone (MZ) B cells and accumulation of transitional 2 B cells in the spleen. We also observed a reduction in B1 B cells in the peritoneum with a defect in the T-independent B cell response against 2,4,6-trinitrophenyl. This was not a result of inhibited development of B cells in the bone marrow, but likely the result of both defective expression of S1P1 in MZ B cells and a defect in the development of MZ and B1 B cells. This suggests that RARα expression in B cells is important for B cell frequency in the MZ and peritoneum, which is crucial for the generation of T-independent humoral responses. PMID:28066447

  3. The effect of a hay grid feeder on feed consumption and measurement of the gastric pH using an intragastric electrode device in horses: a preliminary report.

    Science.gov (United States)

    Aristizabal, F; Nieto, J; Yamout, S; Snyder, J

    2014-07-01

    Obesity and gastric ulceration are highly prevalent in horses. Management modifications for preventing squamous gastric ulceration include frequent feeding and free access to pasture; however, these practices may predispose horses to obesity. To compare the percentage of hay consumed, intragastric pH and horse activity between feeding from the ground and a hay grid feeder. Crossover experimental study. A pH electrode was inserted into the stomach to record the intragastric pH for 48 h. Horses received 1% of their body weight in grass hay twice a day. Horses were assigned to be fed from the ground or a commercial hay grid feeder for 24 h and then switched to the opposite protocol for an additional 24 h. Horses were continuously video-recorded and the percentage of time spent eating or drinking, walking or standing, and lying down were calculated. Two point data were compared by paired t test and pH over time was compared by repeated measures ANOVA. Horses consumed significantly greater amounts of grass hay when fed on the ground compared with a hay grid feeder (n = 9; PpH values (n = 6; P = 0.97), mean intragastric pH over time (n = 6; P = 0.45) the length of time the pH was below 4.0 (n = 6; P = 0.54), and the percentage of time horses spent eating or drinking (n = 9; P = 0.52), walking or standing (n = 9; P = 0.3), or lying down (n = 9; P = 0.4). Within each group horses spent more time eating during the day compared with the night (n = 9; hay grid feeder P = 0.003; ground feeding P = 0.007). The hay grid feeder studied may be used to reduce the amount of hay ingested by horses without reducing the time horses spend eating. © 2013 EVJ Ltd.

  4. EPO-independent functional EPO receptor in breast cancer enhances estrogen receptor activity and promotes cell proliferation

    International Nuclear Information System (INIS)

    Reinbothe, Susann; Larsson, Anna-Maria; Vaapil, Marica; Wigerup, Caroline; Sun, Jianmin; Jögi, Annika; Neumann, Drorit; Rönnstrand, Lars; Påhlman, Sven

    2014-01-01

    Highlights: • New anti-human EPOR antibody confirms full-length EPOR expression in breast cancer cells. • Proliferation of breast cancer cells is not affected by rhEPO treatment in vitro. • EPOR knockdown impairs proliferation of ERa positive breast cancer cells. • EPOR knockdown reduces AKT phosphorylation and ERa activity. - Abstract: The main function of Erythropoietin (EPO) and its receptor (EPOR) is the stimulation of erythropoiesis. Recombinant human EPO (rhEPO) is therefore used to treat anemia in cancer patients. However, clinical trials have indicated that rhEPO treatment might promote tumor progression and has a negative effect on patient survival. In addition, EPOR expression has been detected in several cancer forms. Using a newly produced anti-EPOR antibody that reliably detects the full-length isoform of the EPOR we show that breast cancer tissue and cells express the EPOR protein. rhEPO stimulation of cultured EPOR expressing breast cancer cells did not result in increased proliferation, overt activation of EPOR (receptor phosphorylation) or a consistent activation of canonical EPOR signaling pathway mediators such as JAK2, STAT3, STAT5, or AKT. However, EPOR knockdown experiments suggested functional EPO receptors in estrogen receptor positive (ERα + ) breast cancer cells, as reduced EPOR expression resulted in decreased proliferation. This effect on proliferation was not seen in ERα negative cells. EPOR knockdown decreased ERα activity further supports a mechanism by which EPOR affects proliferation via ERα-mediated mechanisms. We show that EPOR protein is expressed in breast cancer cells, where it appears to promote proliferation by an EPO-independent mechanism in ERα expressing breast cancer cells

  5. EPO-independent functional EPO receptor in breast cancer enhances estrogen receptor activity and promotes cell proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Reinbothe, Susann; Larsson, Anna-Maria; Vaapil, Marica; Wigerup, Caroline [Department of Laboratory Medicine, Translational Cancer Research, Medicon Village, Lund University, SE-223 81 Lund (Sweden); CREATE Health, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv (Israel); Sun, Jianmin [Department of Laboratory Medicine, Translational Cancer Research, Medicon Village, Lund University, SE-223 81 Lund (Sweden); Jögi, Annika [Department of Laboratory Medicine, Translational Cancer Research, Medicon Village, Lund University, SE-223 81 Lund (Sweden); CREATE Health, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv (Israel); Neumann, Drorit [Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv (Israel); Rönnstrand, Lars [Department of Laboratory Medicine, Translational Cancer Research, Medicon Village, Lund University, SE-223 81 Lund (Sweden); Påhlman, Sven, E-mail: sven.pahlman@med.lu.se [Department of Laboratory Medicine, Translational Cancer Research, Medicon Village, Lund University, SE-223 81 Lund (Sweden); CREATE Health, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv (Israel)

    2014-02-28

    Highlights: • New anti-human EPOR antibody confirms full-length EPOR expression in breast cancer cells. • Proliferation of breast cancer cells is not affected by rhEPO treatment in vitro. • EPOR knockdown impairs proliferation of ERa positive breast cancer cells. • EPOR knockdown reduces AKT phosphorylation and ERa activity. - Abstract: The main function of Erythropoietin (EPO) and its receptor (EPOR) is the stimulation of erythropoiesis. Recombinant human EPO (rhEPO) is therefore used to treat anemia in cancer patients. However, clinical trials have indicated that rhEPO treatment might promote tumor progression and has a negative effect on patient survival. In addition, EPOR expression has been detected in several cancer forms. Using a newly produced anti-EPOR antibody that reliably detects the full-length isoform of the EPOR we show that breast cancer tissue and cells express the EPOR protein. rhEPO stimulation of cultured EPOR expressing breast cancer cells did not result in increased proliferation, overt activation of EPOR (receptor phosphorylation) or a consistent activation of canonical EPOR signaling pathway mediators such as JAK2, STAT3, STAT5, or AKT. However, EPOR knockdown experiments suggested functional EPO receptors in estrogen receptor positive (ERα{sup +}) breast cancer cells, as reduced EPOR expression resulted in decreased proliferation. This effect on proliferation was not seen in ERα negative cells. EPOR knockdown decreased ERα activity further supports a mechanism by which EPOR affects proliferation via ERα-mediated mechanisms. We show that EPOR protein is expressed in breast cancer cells, where it appears to promote proliferation by an EPO-independent mechanism in ERα expressing breast cancer cells.

  6. Replication of cultured lung epithelial cells

    International Nuclear Information System (INIS)

    Guzowski, D.; Bienkowski, R.

    1986-01-01

    The authors have investigated the conditions necessary to support replication of lung type 2 epithelial cells in culture. Cells were isolated from mature fetal rabbit lungs (29d gestation) and cultured on feeder layers of mitotically inactivated 3T3 fibroblasts. The epithelial nature of the cells was demonstrated by indirect immunofluorescent staining for keratin and by polyacid dichrome stain. Ultrastructural examination during the first week showed that the cells contained myofilaments, microvilli and lamellar bodies (markers for type 2 cells). The following changes were observed after the first week: increase in cell size; loss of lamellar bodies and appearance of multivesicular bodies; increase in rough endoplasmic reticulum and golgi; increase in tonafilaments and well-defined junctions. General cell morphology was good for up to 10 wk. Cells cultured on plastic surface degenerated after 1 wk. Cell replication was assayed by autoradiography of cultures exposed to ( 3 H)-thymidine and by direct cell counts. The cells did not replicate during the first week; however, between 2-10 wk the cells incorporated the label and went through approximately 6 population doublings. They have demonstrated that lung alveolar epithelial cells can replicate in culture if they are maintained on an appropriate substrate. The coincidence of ability to replicate and loss of markers for differentiation may reflect the dichotomy between growth and differentiation commonly observed in developing systems

  7. The use of multiple imputation for the accurate measurements of individual feed intake by electronic feeders.

    Science.gov (United States)

    Jiao, S; Tiezzi, F; Huang, Y; Gray, K A; Maltecca, C

    2016-02-01

    Obtaining accurate individual feed intake records is the key first step in achieving genetic progress toward more efficient nutrient utilization in pigs. Feed intake records collected by electronic feeding systems contain errors (erroneous and abnormal values exceeding certain cutoff criteria), which are due to feeder malfunction or animal-feeder interaction. In this study, we examined the use of a novel data-editing strategy involving multiple imputation to minimize the impact of errors and missing values on the quality of feed intake data collected by an electronic feeding system. Accuracy of feed intake data adjustment obtained from the conventional linear mixed model (LMM) approach was compared with 2 alternative implementations of multiple imputation by chained equation, denoted as MI (multiple imputation) and MICE (multiple imputation by chained equation). The 3 methods were compared under 3 scenarios, where 5, 10, and 20% feed intake error rates were simulated. Each of the scenarios was replicated 5 times. Accuracy of the alternative error adjustment was measured as the correlation between the true daily feed intake (DFI; daily feed intake in the testing period) or true ADFI (the mean DFI across testing period) and the adjusted DFI or adjusted ADFI. In the editing process, error cutoff criteria are used to define if a feed intake visit contains errors. To investigate the possibility that the error cutoff criteria may affect any of the 3 methods, the simulation was repeated with 2 alternative error cutoff values. Multiple imputation methods outperformed the LMM approach in all scenarios with mean accuracies of 96.7, 93.5, and 90.2% obtained with MI and 96.8, 94.4, and 90.1% obtained with MICE compared with 91.0, 82.6, and 68.7% using LMM for DFI. Similar results were obtained for ADFI. Furthermore, multiple imputation methods consistently performed better than LMM regardless of the cutoff criteria applied to define errors. In conclusion, multiple imputation

  8. Taxifolin synergizes Andrographolide-induced cell death by attenuation of autophagy and augmentation of caspase dependent and independent cell death in HeLa cells.

    Directory of Open Access Journals (Sweden)

    Mazen Alzaharna

    Full Text Available Andrographolide (Andro has emerged recently as a potential and effective anticancer agent with induction of apoptosis in some cancer cell lines while induction of G2/M arrest with weak apoptosis in others. Few studies have proved that Andro is also effective in combination therapy. The flavonoid Taxifolin (Taxi has showed anti-oxidant and antiproliferative effects against different cancer cells. Therefore, the present study investigated the cytotoxic effects of Andro alone or in combination with Taxi on HeLa cells. The combination of Andro with Taxi was synergistic at all tested concentrations and combination ratios. Andro alone induced caspase-dependent apoptosis which was enhanced by the combination with Taxi and attenuated partly by using Z-Vad-Fmk. Andro induced a protective reactive oxygen species (ROS-dependent autophagy which was attenuated by Taxi. The activation of p53 was involved in Andro-induced autophagy where the use of Taxi or pifithrin-α (PFT-α decreased it while the activation of JNK was involved in the cell death of HeLa cells but not in the induction of autophagy. The mitochondrial outer-membrane permeabilization (MOMP plays an important role in Andro-induced cell death in HeLa cells. Andro alone increased the MOMP which was further increased in the case of combination. This led to the increase in AIF and cytochrome c release from mitochondria which consequently increased caspase-dependent and independent cell death. In conclusion, Andro induced a protective autophagy in HeLa cells which was reduced by Taxi and the cell death was increased by increasing the MOMP and subsequently the caspase-dependent and independent cell death.

  9. Taxifolin synergizes Andrographolide-induced cell death by attenuation of autophagy and augmentation of caspase dependent and independent cell death in HeLa cells

    Science.gov (United States)

    Alzaharna, Mazen; Alqouqa, Iyad; Cheung, Hon-Yeung

    2017-01-01

    Andrographolide (Andro) has emerged recently as a potential and effective anticancer agent with induction of apoptosis in some cancer cell lines while induction of G2/M arrest with weak apoptosis in others. Few studies have proved that Andro is also effective in combination therapy. The flavonoid Taxifolin (Taxi) has showed anti-oxidant and antiproliferative effects against different cancer cells. Therefore, the present study investigated the cytotoxic effects of Andro alone or in combination with Taxi on HeLa cells. The combination of Andro with Taxi was synergistic at all tested concentrations and combination ratios. Andro alone induced caspase-dependent apoptosis which was enhanced by the combination with Taxi and attenuated partly by using Z-Vad-Fmk. Andro induced a protective reactive oxygen species (ROS)-dependent autophagy which was attenuated by Taxi. The activation of p53 was involved in Andro-induced autophagy where the use of Taxi or pifithrin-α (PFT-α) decreased it while the activation of JNK was involved in the cell death of HeLa cells but not in the induction of autophagy. The mitochondrial outer-membrane permeabilization (MOMP) plays an important role in Andro-induced cell death in HeLa cells. Andro alone increased the MOMP which was further increased in the case of combination. This led to the increase in AIF and cytochrome c release from mitochondria which consequently increased caspase-dependent and independent cell death. In conclusion, Andro induced a protective autophagy in HeLa cells which was reduced by Taxi and the cell death was increased by increasing the MOMP and subsequently the caspase-dependent and independent cell death. PMID:28182713

  10. Role of the NRP-1-mediated VEGFR2-independent pathway on radiation sensitivity of non-small cell lung cancer cells.

    Science.gov (United States)

    Hu, Chenxi; Zhu, Panrong; Xia, Youyou; Hui, Kaiyuan; Wang, Mei; Jiang, Xiaodong

    2018-07-01

    To determine if inhibiting neuropilin-1 (NRP-1) affects the radiosensitivity of NSCLC cells through a vascular endothelial growth factor receptor 2 (VEGFR2)-independent pathway, and to assess the underlying mechanisms. The expression of VEGFR2, NRP-1, related signaling molecules, abelson murine leukemia viral oncogene homolog 1 (ABL-1), and RAD51 were determined by RT-PCR and Western blotting, respectively. Radiosensitivity was assessed using the colony-forming assay, and the cell apoptosis were analyzed by flow cytometry. We selected two cell lines with high expression levels of VEGFR2, including Calu-1 cells that have high NRP-1 expression, and H358 cells that have low NRP-1 expression. Upon inhibition of p-VEGFR2 by apatinib in Calu-1 cells, the expression of NRP-1 protein and other related proteins in the pathway was still high. Upon NRP-1 siRNA treatment, the expression of both NRP-1 and RAD51 decreased (p cells treated with NRP-1 siRNA exhibited significantly higher apoptosis and radiation sensitivity in radiation therapy compared to Calu-1 cells treated with apatinib alone (p cells with NRP-1 overexpression was similar to the control group regardless of VEGFR2 inhibition. We demonstrated that when VEGFR2 was inhibited, NRP-1 appeared to regulate RAD51 expression through the VEGFR2-independent ABL-1 pathway, consequently regulating radiation sensitivity. In addition, the combined inhibition of VEGFR2 and NRP-1 appears to sensitize cancer cells to radiation.

  11. Cell death is induced by ciglitazone, a peroxisome proliferator-activated receptor γ (PPARγ) agonist, independently of PPARγ in human glioma cells.

    Science.gov (United States)

    Lee, Myoung Woo; Kim, Dae Seong; Kim, Hye Ryung; Kim, Hye Jin; Yang, Jin Mo; Ryu, Somi; Noh, Yoo Hun; Lee, Soo Hyun; Son, Meong Hi; Jung, Hye Lim; Yoo, Keon Hee; Koo, Hong Hoe; Sung, Ki Woong

    2012-01-06

    Peroxisome proliferator-activated receptor γ (PPARγ) regulates multiple signaling pathways, and its agonists induce apoptosis in various cancer cells. However, their role in cell death is unclear. In this study, the relationship between ciglitazone (CGZ) and PPARγ in CGZ-induced cell death was examined. At concentrations of greater than 30 μM, CGZ, a synthetic PPARγ agonist, activated caspase-3 and induced apoptosis in T98G cells. Treatment of T98G cells with less than 30 μM CGZ effectively induced cell death after pretreatment with 30 μM of the PPARγ antagonist GW9662, although GW9662 alone did not induce cell death. This cell death was also observed when cells were co-treated with CGZ and GW9662, but was not observed when cells were treated with CGZ prior to GW9662. In cells in which PPARγ was down-regulated cells by siRNA, lower concentrations of CGZ (death, although higher concentrations of CGZ (≥30 μM) were required to induce cell death in control T98G cells, indicating that CGZ effectively induces cell death in T98G cells independently of PPARγ. Treatment with GW9662 followed by CGZ resulted in a down-regulation of Akt activity and the loss of mitochondrial membrane potential (MMP), which was accompanied by a decrease in Bcl-2 expression and an increase in Bid cleavage. These data suggest that CGZ is capable of inducing apoptotic cell death independently of PPARγ in glioma cells, by down-regulating Akt activity and inducing MMP collapse. Copyright © 2011 Elsevier Inc. All rights reserved.

  12. Autologous bone marrow Th cells can support multiple myeloma cell proliferation in vitro and in xenografted mice.

    Science.gov (United States)

    Wang, D; Fløisand, Y; Myklebust, C V; Bürgler, S; Parente-Ribes, A; Hofgaard, P O; Bogen, B; Taskén, K; Tjønnfjord, G E; Schjesvold, F; Dalgaard, J; Tveita, A; Munthe, L A

    2017-10-01

    Multiple myeloma (MM) is a plasma cell malignancy where MM cell growth is supported by the bone marrow (BM) microenvironment with poorly defined cellular and molecular mechanisms. MM cells express CD40, a receptor known to activate autocrine secretion of cytokines and elicit proliferation. Activated T helper (Th) cells express CD40 ligand (CD40L) and BM Th cells are significantly increased in MM patients. We hypothesized that activated BM Th cells could support MM cell growth. We here found that activated autologous BM Th cells supported MM cell growth in a contact- and CD40L-dependent manner in vitro. MM cells had retained the ability to activate Th cells that reciprocated and stimulated MM cell proliferation. Autologous BM Th cells supported MM cell growth in xenografted mice and were found in close contact with MM cells. MM cells secreted chemokines that attracted Th cells, secretion was augmented by CD40-stimulation. Within 14 days of culture of whole BM aspirates in autologous serum, MM cells and Th cells mutually stimulated each other, and MM cells required Th cells for further expansion in vitro and in mice. The results suggest that Th cells may support the expansion of MM cells in patients.

  13. ERK-dependent and -independent pathways trigger human neural progenitor cell migration

    International Nuclear Information System (INIS)

    Moors, Michaela; Cline, Jason E.; Abel, Josef; Fritsche, Ellen

    2007-01-01

    Besides differentiation and apoptosis, cell migration is a basic process in brain development in which neural cells migrate several centimeters within the developing brain before reaching their proper positions and forming the right connections. For identifying signaling events that control neural migration and are therefore potential targets of chemicals to disturb normal brain development, we developed a human neurosphere-based migration assay based on normal human neural progenitor (NHNP) cells, in which the distance is measured that cells wander over time. Applying this assay, we investigated the role of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) in the regulation of NHNP cell migration. Exposure to model substances like ethanol or phorbol 12-myristate 13-acetate (PMA) revealed a correlation between ERK1/2 activation and cell migration. The participation of phospho-(P-) ERK1/2 was confirmed by exposure of the cells to the MEK inhibitor PD98059, which directly prohibits ERK1/2 phosphorylation and inhibited cell migration. We identified protein kinase C (PKC) and epidermal growth factor receptor (EGFR) as upstream signaling kinases governing ERK1/2 activation, thereby controlling NHNP cell migration. Additionally, treatments with src kinase inhibitors led to a diminished cell migration without affecting ERK1/2 phosphorylation. Based on these results, we postulate that migration of NHNP cells is controlled via ERK1/2-dependent and -independent pathways

  14. The Regulation of Lactogenic Differentiation in Mammary Epithelial Cells by Ras-Dependent and -Independent Signal Transduction

    Science.gov (United States)

    2006-01-06

    Lynch, 2002). In normal cells, programmed cell 19 death controls excessive proliferation. Most cancer cells, however, have devised methods for...also phosphorylates and inactivates caspase 9, a cell death -promoting protease. Phosphorylation of another Akt target, FKHR1, prevents its nuclear...Cancer Res 50:4204–4208. Gao J, Horseman N. 1999. Prolactin-independent modulation of the beta-casein response element by Erk2 Map kinase. Cell

  15. Enhanced adhesion of osteoblastic cells on polystyrene films by independent control of surface topography and wettability

    International Nuclear Information System (INIS)

    Yang, Seung Yun; Kim, Eung-Sam; Jeon, Gumhye; Choi, Kwan Yong; Kim, Jin Kon

    2013-01-01

    We independently controlled surface topography and wettability of polystyrene (PS) films by CF 4 and oxygen plasma treatments, respectively, to evaluate the adhesion and proliferation of human fetal osteoblastic (hFOB) cells on the films. Among the CF 4 plasma-treated PS films with the average surface roughness ranging from 0.9 to 70 nm, the highest adhesion of hFOB cells was observed on a PS film with roughness of ∼ 11 nm. When this film was additionally treated by oxygen plasma to provide a hydrophilic surface with a contact angle less than 10°, the proliferation of bone-forming cell was further enhanced. Thus, the plasma-based independent modification of PS film into an optimum nanotexture for human osteoblast cells could be appplied to materials used in bone tissue engineering. Highlights: ► New approach based on plasma treatment to independently control the surface topography and wettability ► The adhesion of human fetal osteoblast (hFOB) was enhanced on a surface with an average roughness of ∼ 11 nm. ► The adhesion and proliferation of hFOB was maximized when nanotextured surface became highly hydrophilic

  16. Enhanced adhesion of osteoblastic cells on polystyrene films by independent control of surface topography and wettability

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Seung Yun [National Creative Research Center for Block Copolymer Self-Assembly, Departments of Environmental Science and Engineering and Chemical Engineering, Pohang University of Science and Technology, Pohang, 790-784 (Korea, Republic of); Kim, Eung-Sam [School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, 790-784 (Korea, Republic of); Jeon, Gumhye [National Creative Research Center for Block Copolymer Self-Assembly, Departments of Environmental Science and Engineering and Chemical Engineering, Pohang University of Science and Technology, Pohang, 790-784 (Korea, Republic of); Choi, Kwan Yong, E-mail: kchoi@postech.ac.kr [School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, 790-784 (Korea, Republic of); Department of Life Science, Division of Molecular and Life Science, Pohang University of Science and Technology, Pohang, 790-784 (Korea, Republic of); Kim, Jin Kon, E-mail: jkkim@postech.ac.kr [National Creative Research Center for Block Copolymer Self-Assembly, Departments of Environmental Science and Engineering and Chemical Engineering, Pohang University of Science and Technology, Pohang, 790-784 (Korea, Republic of)

    2013-04-01

    We independently controlled surface topography and wettability of polystyrene (PS) films by CF{sub 4} and oxygen plasma treatments, respectively, to evaluate the adhesion and proliferation of human fetal osteoblastic (hFOB) cells on the films. Among the CF{sub 4} plasma-treated PS films with the average surface roughness ranging from 0.9 to 70 nm, the highest adhesion of hFOB cells was observed on a PS film with roughness of ∼ 11 nm. When this film was additionally treated by oxygen plasma to provide a hydrophilic surface with a contact angle less than 10°, the proliferation of bone-forming cell was further enhanced. Thus, the plasma-based independent modification of PS film into an optimum nanotexture for human osteoblast cells could be appplied to materials used in bone tissue engineering. Highlights: ► New approach based on plasma treatment to independently control the surface topography and wettability ► The adhesion of human fetal osteoblast (hFOB) was enhanced on a surface with an average roughness of ∼ 11 nm. ► The adhesion and proliferation of hFOB was maximized when nanotextured surface became highly hydrophilic.

  17. Impacts of the IBM Cell Processor to Support Climate Models

    Science.gov (United States)

    Zhou, Shujia; Duffy, Daniel; Clune, Tom; Suarez, Max; Williams, Samuel; Halem, Milt

    2008-01-01

    NASA is interested in the performance and cost benefits for adapting its applications to the IBM Cell processor. However, its 256KB local memory per SPE and the new communication mechanism, make it very challenging to port an application. We selected the solar radiation component of the NASA GEOS-5 climate model, which: (1) is representative of column physics (approximately 50% computational time), (2) has a high computational load relative to transferring data from and to main memory, (3) performs independent calculations across multiple columns. We converted the baseline code (single-precision, Fortran) to C and ported it with manually SIMDizing 4 independent columns and found that a Cell with 8 SPEs can process 2274 columns per second. Compared with the baseline results, the Cell is approximately 5.2X, approximately 8.2X, approximately 15.1X faster than a core on Intel Woodcrest, Dempsey, and Itanium2, respectively. We believe this dramatic performance improvement makes a hybrid cluster with Cell and traditional nodes competitive.

  18. HAMLET triggers apoptosis but tumor cell death is independent of caspases, Bcl-2 and p53.

    Science.gov (United States)

    Hallgren, O; Gustafsson, L; Irjala, H; Selivanova, G; Orrenius, S; Svanborg, C

    2006-02-01

    HAMLET (Human alpha-lactalbumin Made Lethal to Tumor cells) triggers selective tumor cell death in vitro and limits tumor progression in vivo. Dying cells show features of apoptosis but it is not clear if the apoptotic response explains tumor cell death. This study examined the contribution of apoptosis to cell death in response to HAMLET. Apoptotic changes like caspase activation, phosphatidyl serine externalization, chromatin condensation were detected in HAMLET-treated tumor cells, but caspase inhibition or Bcl-2 over-expression did not prolong cell survival and the caspase response was Bcl-2 independent. HAMLET translocates to the nuclei and binds directly to chromatin, but the death response was unrelated to the p53 status of the tumor cells. p53 deletions or gain of function mutations did not influence the HAMLET sensitivity of tumor cells. Chromatin condensation was partly caspase dependent, but apoptosis-like marginalization of chromatin was also observed. The results show that tumor cell death in response to HAMLET is independent of caspases, p53 and Bcl-2 even though HAMLET activates an apoptotic response. The use of other cell death pathways allows HAMLET to successfully circumvent fundamental anti-apoptotic strategies that are present in many tumor cells.

  19. Computer support for physiological cell modelling using an ontology on cell physiology.

    Science.gov (United States)

    Takao, Shimayoshi; Kazuhiro, Komurasaki; Akira, Amano; Takeshi, Iwashita; Masanori, Kanazawa; Tetsuya, Matsuda

    2006-01-01

    The development of electrophysiological whole cell models to support the understanding of biological mechanisms is increasing rapidly. Due to the complexity of biological systems, comprehensive cell models, which are composed of many imported sub-models of functional elements, can get quite complicated as well, making computer modification difficult. Here, we propose a computer support to enhance structural changes of cell models, employing the markup languages CellML and our original PMSML (physiological model structure markup language), in addition to a new ontology for cell physiological modelling. In particular, a method to make references from CellML files to the ontology and a method to assist manipulation of model structures using markup languages together with the ontology are reported. Using these methods three software utilities, including a graphical model editor, are implemented. Experimental results proved that these methods are effective for the modification of electrophysiological models.

  20. Development of video probe system for inspection of feeder pipe support in calandria reactor

    International Nuclear Information System (INIS)

    Cho, Jai Wan; Lee, Nam Ho; Choi, Young Soo

    2000-07-01

    There are 760 feederpipes, which they are connected to inlet/outlet of the 380 pressure tube channels on the front of the calandria, in CANDU-type Reactor of Wolsung Nuclear Power Plant. As an ISI(In-Service Inspection) and PSI (Post- Service Inspection) requirements, maintenance activities of measuring the thickness of curvilinear part of feederpipe and inspecting the feederpipe support area within calandria are needed to ensure continued reliable operation of nuclear power plant. And untrasonic probe is used to measure the thickness of curvilinear part of feederpipe, however workers are exposed to radioactivity irradiation during the measurement period. But, it is impossible to inspect feederpipe support area thoroughlv because of narrow and confined accessibility, that is, an inspection space between the pressure tube channels is less than 100mm and pipes in feederpipe support area are congested. And also, workers involved in inspecting feederpipe support area are under the jeopardy of high-level radiation exposure. Concerns about sliding home, which make the move of feederpipe connected to pressure tube channel smooth as pressure tube expands and contracts in its axial direction, stuck to feederpipe support and some of the structural components have made necessary the development of video inspection probe system with narrow and confined accessibility to observe and inspect feederpipe support area more close. Using video inspection probe system, it is possible to inspect and repair abnormality of feederpipe support connected to pressure tube channels of the calandria more accurate and quantative than naked eye. Therefore, that will do much for ensuring safety of CANDU-type nuclear power plant

  1. Pluripotency transcription factors and Tet1/2 maintain Brd4-independent stem cell identity.

    Science.gov (United States)

    Finley, Lydia W S; Vardhana, Santosha A; Carey, Bryce W; Alonso-Curbelo, Direna; Koche, Richard; Chen, Yanyang; Wen, Duancheng; King, Bryan; Radler, Megan R; Rafii, Shahin; Lowe, Scott W; Allis, C David; Thompson, Craig B

    2018-05-01

    A robust network of transcription factors and an open chromatin landscape are hallmarks of the naive pluripotent state. Recently, the acetyllysine reader Brd4 has been implicated in stem cell maintenance, but the relative contribution of Brd4 to pluripotency remains unclear. Here, we show that Brd4 is dispensable for self-renewal and pluripotency of embryonic stem cells (ESCs). When maintained in their ground state, ESCs retain transcription factor binding and chromatin accessibility independent of Brd4 function or expression. In metastable ESCs, Brd4 independence can be achieved by increased expression of pluripotency transcription factors, including STAT3, Nanog or Klf4, so long as the DNA methylcytosine oxidases Tet1 and Tet2 are present. These data reveal that Brd4 is not essential for ESC self-renewal. Rather, the levels of pluripotency transcription factor abundance and Tet1/2 function determine the extent to which bromodomain recognition of protein acetylation contributes to the maintenance of gene expression and cell identity.

  2. Care and support for older adults in The Netherlands living independently.

    Science.gov (United States)

    Verver, Didi; Merten, Hanneke; Robben, Paul; Wagner, Cordula

    2018-05-01

    The growth in the numbers of older adults needing long-term care has resulted in rising costs which have forced the Dutch government to change its long-term care system. Now, the local authorities have greater responsibility for supporting older adults and in prolonging independent living with increased support provided by the social network. However, it is unclear whether these older adults have such a network to rely upon. The objective of this study was to gain insight into the providers of formal and informal care to older adults, and to assess possible differences between older adults who are frail and those who are not. In addition, we investigated their care and support needs. We used data from a quantitative survey using a cross-sectional design in different regions of the Netherlands from July until September 2014 (n = 181). Frailty was measured using the Tilburg Frailty indicator. To analyse the data chi-square tests, crosstabs and odds ratios were used for dichotomous data and the Mann-Whitney U-Test for nominal data. The number of formal care providers involved was significantly higher (median = 2) for those deemed frail than for those not deemed frail (median = 1), U = 2,130, p older adults deemed to be frail did not have their needs sufficiently addressed by their care network. For a substantial part of this group of older adults, the informal network seems to be unable to support them sufficiently. Additional attention for their needs and wishes is required to implement the policy reforms successfully. © 2018 John Wiley & Sons Ltd.

  3. Field evaluation of cutter and feeder mechanism of chickpea harvester for lentil harvesting

    Directory of Open Access Journals (Sweden)

    S Kamgar

    2016-09-01

    Full Text Available Introduction The main producers of lentil are Canada, India, Nepal and China, respectively and Iran is the ninth producer in the world. The hand pulling is the usual method of lentil harvesting. Use of conventional combine because of short leg varieties, wide combine head in dry land and grain losses by cutter bar vibrations is impossible. So a mechanism should be designed to harvest the lentil plants with minimum damage. This mechanism should be evaluated under different tests of crop and machines such as forward speed (FS, grain moisture content (GMC, different varieties and other parameters. Some researchers studied the effects of GMC (Andrews and et al., 1993; Huitink, 2005; Adisa, 2009; Abdi and Jalali, 2013 and FS on grain losses (Geng et al., 1984; Swapan et al., 2001; Mostafavand and Kamgar, 2014; Hunt, 1995. Field tests were conducted at three levels of FS 1.5, 3 and 4.5 km.h-1; three levels of cutting height (CH 4, 8 and 13 cm and two levels of GMC, 8 and 14% on two varieties of lentils including Flip and Shiraz with three replications. Materials and Methods The feeder and cutter mechanism for chickpea harvesting that was the base design of device which is notched wheel and counter shear, was used. The other components of device were dividers, slat and chain feeders, belt and pulleys, chassis, elevator conveyor and storage. Two split plot design based on a randomized complete design was used to determine the effects of above treatments on lentil losses. Results and Discussion The ANOVA results indicated that the all studied factors; FS of feeder and cutter mechanism, CH and GMC had significant effect on losses of Shiraz variety (P0.05. The ranges of losses of Flip variety at 8% GMC were 8.6 to 10% for FS of 1.5 km.h-1, 9.1 to 10.4% for FS of 3 km.h-1and 10.4 to 11.4% for FS of 4.5 km h-1. These ranges at 14% GMC were 7.9 to 8.9% for FS of 1.5 km.h-1, 8.4 to 9.2% for FS of 3 km.h-1and 8.5 to 10% for FS of 4.5 km h-1. The ranges of

  4. Strength of Anode‐Supported Solid Oxide Fuel Cells

    DEFF Research Database (Denmark)

    Faes, A.; Frandsen, Henrik Lund; Kaiser, Andreas

    2011-01-01

    Nickel oxide and yttria doped zirconia composite strength is crucial for anode‐supported solid oxide fuel cells, especially during transient operation, but also for the initial stacking process, where cell curvature after sintering can cause problems. This work first compares tensile and ball....... Even though the electrolyte is to the tensile side, it is found that the anode support fails due to the thermo‐mechanical residual stresses....

  5. Manufacture and testing of the CTB&SBB thermal shield for the ITER magnet feeder system

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Kun; Liu, Chen; Song, Yuntao; Feng, Hansheng; Ding, Kaizhong, E-mail: kzding@ipp.ac.cn; Wang, Tanbin; Ji, Hui

    2015-10-15

    The system of International Thermonuclear Experimental Reactor (ITER) feeders is responsible for the power, helium cooling, and instrumentation of the magnets of the coil terminal box and S-bend box (CTB&SBB) thermal shield outside the cryostat. An 80-K rectangular Al thermal shield is hung inside the CTB&SBB to reduce the thermal radiation heat loads of 4.5-K helium. The American Society of Interventional Pain Physicians (ASIPP) will supply all the 31 sets of feeders for ITER. A manufactured prototype of CTB&SBB thermal shield is first quality-tested before the commencement of the series production. First, a detailed configuration of the rectangular Al thermal shield is presented in this article. The paper also presents more information on the manufacturing process of the thermal shield, especially the welding process, the procedure for ensuring good weld quality, and the use of a specially designed tool to ensure <5-mm deformation on such a 7.3-m-long thermal shield during welding. In addition, the cold test and results, including the cooling process with 13-bar and 17.5-g/s 80-K He gas, and the temperature distribution on different panels of the thermal shield are presented. The whole process of manufacture and testing lays a good foundation for the series production of the thermal shield.

  6. Modeling and Control of Low Voltage Flexible Units for Enhanced Operation of Distribution Feeders

    DEFF Research Database (Denmark)

    Raboni, Pietro; Hu, Weihao; Chaudhary, Sanjay

    2013-01-01

    In some networks Distributed Generators (DGs) are phasing out conventional power plants in terms of power production but still large efforts are required for providing ancillary services. In this paper the usage of fast response units like a Micro Turbines (MT) and a stationary Electric Vehicle...... Battery (EVB) is proposed for providing primary regulation in grid connected mode and for hierarchically manage an islanded LV distribution feeder. The unit models are described and a novel EVB model directly based on manufacturer’s data is proposed and evaluated comparing its performances with Sim...

  7. Signal transduction of p53-independent apoptotic pathway induced by hexavalent chromium in U937 cells

    International Nuclear Information System (INIS)

    Hayashi, Yoko; Kondo, Takashi; Zhao Qingli; Ogawa Ryohei; Cui Zhengguo; Feril, Loreto B.; Teranishi, Hidetoyo; Kasuya, Minoru

    2004-01-01

    It has been reported that the hexavalent chromium compound (Cr(VI)) can induce both p53-dependent and p53-independent apoptosis. While a considerable amount of information is available on the p53-dependent pathway, only little is known about the p53-independent pathway. To elucidate the p53-independent mechanism, the roles of the Ca 2+ -calpain- and mitochondria-caspase-dependent pathways in apoptosis induced by Cr(VI) were investigated. When human lymphoma U937 cells, p53 mutated cells, were treated with 20 μM Cr(VI) for 24 h, nuclear morphological changes and DNA fragmentation were observed. Production of hydroxyl radicals revealed by electron paramagnetic resonance (EPR)-spin trapping, and increase of intracellular calcium ion concentration monitored by digital imaging were also observed in Cr(VI)-treated cells. An intracellular Ca 2+ chelator, BAPTA-AM, and calpain inhibitors suppressed the Cr(VI)-induced DNA fragmentation. The number of cells showing low mitochondrial membrane potential (MMP), high level of superoxide anion radicals (O 2 - ), and high activity of caspase-3, which are indicators of mitochondria-caspase-dependent pathway, increased significantly in Cr(VI)-treated cells. An antioxidant, N-acetyl-L-cysteine (NAC), decreased DNA fragmentation and inhibited the changes in MMP, O 2 - formation, and activation of caspase-3 induced by Cr(VI). No increase of the expressions of Fas and phosphorylated JNK was observed after Cr(VI) treatment. Cell cycle analysis revealed that the fraction of G2/M phase tended to increase after 24 h of treatment, suggesting that Cr(VI)-induced apoptosis is related to the G2 block. These results indicate that Ca 2+ -calpain- and mitochondria-caspase-dependent pathways play significant roles in the Cr(VI)-induced apoptosis via the G2 block, which are independent of JNK and Fas activation. The inhibition of apoptosis and all its signal transductions by NAC suggests that intracellular reactive oxygen species (ROS) are

  8. Enhanced adhesion of osteoblastic cells on polystyrene films by independent control of surface topography and wettability.

    Science.gov (United States)

    Yang, Seung Yun; Kim, Eung-Sam; Jeon, Gumhye; Choi, Kwan Yong; Kim, Jin Kon

    2013-04-01

    We independently controlled surface topography and wettability of polystyrene (PS) films by CF4 and oxygen plasma treatments, respectively, to evaluate the adhesion and proliferation of human fetal osteoblastic (hFOB) cells on the films. Among the CF4 plasma-treated PS films with the average surface roughness ranging from 0.9 to 70 nm, the highest adhesion of hFOB cells was observed on a PS film with roughness of ~11 nm. When this film was additionally treated by oxygen plasma to provide a hydrophilic surface with a contact angle less than 10°, the proliferation of bone-forming cell was further enhanced. Thus, the plasma-based independent modification of PS film into an optimum nanotexture for human osteoblast cells could be appplied to materials used in bone tissue engineering. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. A model for strategic selection of feeder management systems: A case study

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Hsing Hung [Faculty of Management and Administration, Macau University of Science and Technology, Avenue Wei Long, Taipa, Macau (China); Lee, Amy H.I. [Department of Industrial Management, Chung Hua University, No. 707, Sec. 2, Wu Fu Rd., Hsinchu 300 (China); Kang, He-Yau [Department of Industrial Engineering and Management, National Chin-Yi University of Technology, 35, Lane 215, Sec. 1, Chung San Rd., Taiping, Taichung 411 (China)

    2010-06-15

    The move to integrating distribution management system (DMS) and feeder management system (FMS) in China is becoming the main trend in recent years, in addition to upgrading and rebuilding existing energy management system (EMS) and DMS. However, with increasing complexity in the social environments along with rapidly changing technologies, how to select a suitable contractor and a FMS project is becoming an important issue for electric power companies. This paper first briefly introduces FMS and then lists its critical success criteria. A model that applies a multi-criteria decision-making (MCDM) method, an analytic network process (ANP) associated with benefits, opportunities, costs and risks (BOCR), is constructed to help power companies to select the most suitable FMS project. (author)

  10. CbtA toxin of Escherichia coli inhibits cell division and cell elongation via direct and independent interactions with FtsZ and MreB.

    Science.gov (United States)

    Heller, Danielle M; Tavag, Mrinalini; Hochschild, Ann

    2017-09-01

    The toxin components of toxin-antitoxin modules, found in bacterial plasmids, phages, and chromosomes, typically target a single macromolecule to interfere with an essential cellular process. An apparent exception is the chromosomally encoded toxin component of the E. coli CbtA/CbeA toxin-antitoxin module, which can inhibit both cell division and cell elongation. A small protein of only 124 amino acids, CbtA, was previously proposed to interact with both FtsZ, a tubulin homolog that is essential for cell division, and MreB, an actin homolog that is essential for cell elongation. However, whether or not the toxic effects of CbtA are due to direct interactions with these predicted targets is not known. Here, we genetically separate the effects of CbtA on cell elongation and cell division, showing that CbtA interacts directly and independently with FtsZ and MreB. Using complementary genetic approaches, we identify the functionally relevant target surfaces on FtsZ and MreB, revealing that in both cases, CbtA binds to surfaces involved in essential cytoskeletal filament architecture. We show further that each interaction contributes independently to CbtA-mediated toxicity and that disruption of both interactions is required to alleviate the observed toxicity. Although several other protein modulators are known to target FtsZ, the CbtA-interacting surface we identify represents a novel inhibitory target. Our findings establish CbtA as a dual function toxin that inhibits both cell division and cell elongation via direct and independent interactions with FtsZ and MreB.

  11. Numerical evaluation of micro-structural parameters of porous supports in metal-supported solid oxide fuel cells

    DEFF Research Database (Denmark)

    Reiss, Georg; Frandsen, Henrik Lund; Brandstätter, Wilhelm

    2014-01-01

    Metallic supported Solid Oxide Fuel Cells (SOFCs) are considered as a durable and cost effective alternative to the state-of-the-art ceramic supported cell designs. In order to understand the mass and charge transport in the metal-support of this new type of cell a novel technique involving X......-ray tomography and micro-structural modelling is presented in this work. The simulation technique comprises a novel treatment of the boundary conditions, which leads to more accurate effective transport parameters compared to those, which can be achieved with the conventional homogenisation procedures....... Furthermore, the porosity distribution in the metal-support was determined, which provided information about the inhomogeneous nature of the material. In addition to that, transport parameters for two identified, different dense layers of the metal-support are evaluated separately. The results...

  12. Pyruvate carboxylase is required for glutamine-independent growth of tumor cells

    Science.gov (United States)

    Cheng, Tzuling; Sudderth, Jessica; Yang, Chendong; Mullen, Andrew R.; Jin, Eunsook S.; Matés, José M.; DeBerardinis, Ralph J.

    2011-01-01

    Tumor cells require a constant supply of macromolecular precursors, and interrupting this supply has been proposed as a therapeutic strategy in cancer. Precursors for lipids, nucleic acids, and proteins are generated in the tricarboxylic acid (TCA) cycle and removed from the mitochondria to participate in biosynthetic reactions. Refilling the pool of precursor molecules (anaplerosis) is therefore crucial to maintain cell growth. Many tumor cells use glutamine to feed anaplerosis. Here we studied how “glutamine-addicted” cells react to interruptions of glutamine metabolism. Silencing of glutaminase (GLS), which catalyzes the first step in glutamine-dependent anaplerosis, suppressed but did not eliminate the growth of glioblastoma cells in culture and in vivo. Profiling metabolic fluxes in GLS-suppressed cells revealed induction of a compensatory anaplerotic mechanism catalyzed by pyruvate carboxylase (PC), allowing the cells to use glucose-derived pyruvate rather than glutamine for anaplerosis. Although PC was dispensable when glutamine was available, forcing cells to adapt to low-glutamine conditions rendered them absolutely dependent on PC for growth. Furthermore, in other cell lines, measuring PC activity in nutrient-replete conditions predicted dependence on specific anaplerotic enzymes. Cells with high PC activity were resistant to GLS silencing and did not require glutamine for survival or growth, but displayed suppressed growth when PC was silenced. Thus, PC-mediated, glucose-dependent anaplerosis allows cells to achieve glutamine independence. Induction of PC during chronic suppression of glutamine metabolism may prove to be a mechanism of resistance to therapies targeting glutaminolysis. PMID:21555572

  13. PAX2 regulates ADAM10 expression and mediates anchorage-independent cell growth of melanoma cells.

    Directory of Open Access Journals (Sweden)

    Sophia Boyoung Lee

    Full Text Available PAX transcription factors play an important role during development and carcinogenesis. In this study, we investigated PAX2 protein levels in melanocytes and melanoma cells by Western Blot and immunofluorescence analysis and characterized the role of PAX2 in the pathogenesis of melanoma. In vitro we found weak PAX2 protein expression in keratinocytes and melanocytes. Compared to melanocytes increased PAX2 protein levels were detectable in melanoma cell lines. Interestingly, in tissue sections of melanoma patients nuclear PAX2 expression strongly correlated with nuclear atypia and the degree of prominent nucleoli, indicating an association of PAX2 with a more atypical cellular phenotype. In addition, with chromatin immunoprecipitation assay, PAX2 overexpression and PAX2 siRNA we present compelling evidence that PAX2 can regulate ADAM10 expression, a metalloproteinase known to play important roles in melanoma metastasis. In human tissue samples we found co-expression of PAX2 and ADAM10 in melanocytes of benign nevi and in melanoma cells of patients with malignant melanoma. Importantly, the downregulation of PAX2 by specific siRNA inhibited the anchorage independent cell growth and decreased the migratory and invasive capacity of melanoma cells. Furthermore, the downregulation of PAX2 abrogated the chemoresistance of melanoma cells against cisplatin, indicating that PAX2 expression mediates cell survival and plays important roles during melanoma progression.

  14. Derivation of transgene-free human induced pluripotent stem cells from human peripheral T cells in defined culture conditions.

    Directory of Open Access Journals (Sweden)

    Yoshikazu Kishino

    Full Text Available Recently, induced pluripotent stem cells (iPSCs were established as promising cell sources for revolutionary regenerative therapies. The initial culture system used for iPSC generation needed fetal calf serum in the culture medium and mouse embryonic fibroblast as a feeder layer, both of which could possibly transfer unknown exogenous antigens and pathogens into the iPSC population. Therefore, the development of culture systems designed to minimize such potential risks has become increasingly vital for future applications of iPSCs for clinical use. On another front, although donor cell types for generating iPSCs are wide-ranging, T cells have attracted attention as unique cell sources for iPSCs generation because T cell-derived iPSCs (TiPSCs have a unique monoclonal T cell receptor genomic rearrangement that enables their differentiation into antigen-specific T cells, which can be applied to novel immunotherapies. In the present study, we generated transgene-free human TiPSCs using a combination of activated human T cells and Sendai virus under defined culture conditions. These TiPSCs expressed pluripotent markers by quantitative PCR and immunostaining, had a normal karyotype, and were capable of differentiating into cells from all three germ layers. This method of TiPSCs generation is more suitable for the therapeutic application of iPSC technology because it lowers the risks associated with the presence of undefined, animal-derived feeder cells and serum. Therefore this work will lead to establishment of safer iPSCs and extended clinical application.

  15. Demonstration of the presence of independent pre-osteoblastic and pre-adipocytic cell populations in bone marrow-derived mesenchymal stem cells

    DEFF Research Database (Denmark)

    Post, S; Abdallah, B M; Bentzon, J F

    2008-01-01

    differentiation into one particular lineage. However, this inverse relationship between bone and fat is not consistent and under certain in vivo conditions, bone and fat can change independently suggesting separate precursor cell populations. In order to test for this hypothesis, we extensively characterized two...... of mature adipocytes visualized by Oil Red O staining. On the other hand, mMSC2 and not mMSC1 differentiated to osteoblast lineage as demonstrated by up-regulation of osteoblastic makers (CBFA1/RUNX2, Osterix, alkaline phosphatase, bone sialoprotein and osteopontin) and formation of alizarin red stained...... that are committed to either osteoblast or adipocyte lineage. These cell populations may undergo independent changes during aging and in bone diseases and thus represent important targets for therapy....

  16. Manufacture and test of seismic bellows for ITER magnet feeder

    International Nuclear Information System (INIS)

    Liu, Chen; Lu, Kun; Sheng, Liang; Song, Yuntao; Su, Jinjin; Su, Man; Gung, Chenyu

    2016-01-01

    Highlights: • The design of the double bellows was iterated with the results of analysis based on the Expansion Joint Manufacturers Association (EJMA) standard. • The seismic bellows was tested with cyclic pressurization of the interlayer space to 2 bars absolute pressure for 5 cycles. • 200 cycles of tensile fatigue test with 90 mm of stretching from the nominal design length. • A full tensile test with 315 mm of stretching from its nominal length was conducted. • The prototype bellows was qualified for its leak tightness (less than 1 × 10 −9 Pam 3 /s of helium) at all time during the qualification test. - Abstract: This paper presents the key manufacturing and testing processes of the prototype ITER feeder seismic bellows. The design of the double bellows was iterated with the results of analysis based on the Expansion Joint Manufacturers Association (EJMA) standard. Each inner and outer bellows was supported in dedicated molds and formed by a hydraulic pressure machine rated at 800 tons. The double bellows were constructed by welding individual collars to the end flanges. The seismic bellows was tested with cyclic pressurization of the interlayer space to 2 bars absolute pressure for 5 cycles. This was followed by 200 cycles of tensile fatigue test with 90 mm of stretching from the nominal design length. After the mechanical fatigue test, a full tensile test with 315 mm of stretching from its nominal length was conducted. Helium leak tests, with the sensitivity of the helium leak detector set to 1 × 10 −9 Pa m 3 /s of helium, were performed at different stages of pressure and mechanical tests. The prototype bellows was qualified for its leak tightness at all time during the qualification test.

  17. Manufacture and test of seismic bellows for ITER magnet feeder

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Chen, E-mail: liuchen@ipp.ac.cn [Institute of Plasma Physics Chinese Academy of Sciences, ShuShanhu Road No. 350, Hefei (China); Lu, Kun [Institute of Plasma Physics Chinese Academy of Sciences, ShuShanhu Road No. 350, Hefei (China); Sheng, Liang [AEROSUN-TOLA Expansion Joint Co., Ltd., Jiangjun Road No. 199, Nanjing (China); Song, Yuntao; Su, Jinjin [Institute of Plasma Physics Chinese Academy of Sciences, ShuShanhu Road No. 350, Hefei (China); Su, Man; Gung, Chenyu [ITER Organization, Route de Vinon-sur-Verdon, CS 90 046, 13067 St. Paul-lez-Durance (France)

    2016-11-01

    Highlights: • The design of the double bellows was iterated with the results of analysis based on the Expansion Joint Manufacturers Association (EJMA) standard. • The seismic bellows was tested with cyclic pressurization of the interlayer space to 2 bars absolute pressure for 5 cycles. • 200 cycles of tensile fatigue test with 90 mm of stretching from the nominal design length. • A full tensile test with 315 mm of stretching from its nominal length was conducted. • The prototype bellows was qualified for its leak tightness (less than 1 × 10{sup −9} Pam{sup 3}/s of helium) at all time during the qualification test. - Abstract: This paper presents the key manufacturing and testing processes of the prototype ITER feeder seismic bellows. The design of the double bellows was iterated with the results of analysis based on the Expansion Joint Manufacturers Association (EJMA) standard. Each inner and outer bellows was supported in dedicated molds and formed by a hydraulic pressure machine rated at 800 tons. The double bellows were constructed by welding individual collars to the end flanges. The seismic bellows was tested with cyclic pressurization of the interlayer space to 2 bars absolute pressure for 5 cycles. This was followed by 200 cycles of tensile fatigue test with 90 mm of stretching from the nominal design length. After the mechanical fatigue test, a full tensile test with 315 mm of stretching from its nominal length was conducted. Helium leak tests, with the sensitivity of the helium leak detector set to 1 × 10{sup −9} Pa m{sup 3}/s of helium, were performed at different stages of pressure and mechanical tests. The prototype bellows was qualified for its leak tightness at all time during the qualification test.

  18. Preliminary analysis of accident in SST-1 current feeder system

    International Nuclear Information System (INIS)

    Roy, Swati; Kanabar, Deven; Garg, Atul; Singh, Amit; Tanna, Vipul; Prasad, Upendra; Srinivasan, R.

    2017-01-01

    Steady-state Tokamak-1 (SST-1) has 16 superconducting Toroidal field (TF) and 9 superconducting poloidal field (PF) coils rated for 10kA DC. All the TF are connected in series and are operated in DC condition whereas PF coils are individually operated in pulse mode during SST-1 campaigns. SST-1 current feeder system (CFS) houses 9 pairs of PF current leads and 1 pair of TF current leads. During past SST-1 campaign, there were arcing incidents within SST-1 CFS chamber which caused significant damage to PF superconducting current leads as well as its Helium cooling lines of the current leads. This paper brings out the preliminary analysis of the mentioned arcing incident, possible reasons and its investigation thereby laying out the sequence of events. From this analysis and observations, various measures to avoid such arcing incidents have also been proposed. (author)

  19. The PPAR{gamma} ligand ciglitazone regulates androgen receptor activation differently in androgen-dependent versus androgen-independent human prostate cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Moss, Patrice E.; Lyles, Besstina E.; Stewart, LaMonica V., E-mail: lstewart@mmc.edu

    2010-12-10

    The androgen receptor (AR) regulates growth and progression of androgen-dependent as well as androgen-independent prostate cancer cells. Peroxisome proliferator-activated receptor gamma (PPAR{gamma}) agonists have been reported to reduce AR activation in androgen-dependent LNCaP prostate cancer cells. To determine whether PPAR{gamma} ligands are equally effective at inhibiting AR activity in androgen-independent prostate cancer, we examined the effect of the PPAR{gamma} ligands ciglitazone and rosiglitazone on C4-2 cells, an androgen- independent derivative of the LNCaP cell line. Luciferase-based reporter assays and Western blot analysis demonstrated that PPAR{gamma} ligand reduced dihydrotestosterone (DHT)-induced increases in AR activity in LNCaP cells. However, in C4-2 cells, these compounds increased DHT-induced AR driven luciferase activity. In addition, ciglitazone did not significantly alter DHT-mediated increases in prostate specific antigen (PSA) protein or mRNA levels within C4-2 cells. siRNA-based experiments demonstrated that the ciglitazone-induced regulation of AR activity observed in C4-2 cells was dependent on the presence of PPAR{gamma}. Furthermore, overexpression of the AR corepressor cyclin D1 inhibited the ability of ciglitazone to induce AR luciferase activity in C4-2 cells. Thus, our data suggest that both PPAR{gamma} and cyclin D1 levels influence the ability of ciglitazone to differentially regulate AR signaling in androgen-independent C4-2 prostate cancer cells.

  20. Generation of an iPS cell line via a non-integrative method using urine-derived cells from a patient with USH2A-associated retinitis pigmentosa

    Directory of Open Access Journals (Sweden)

    Yonglong Guo

    2018-05-01

    Full Text Available We have established an induced pluripotent stem (iPS cell line using urine-derived cells from a 27-year-old male patient with retinitis pigmentosa associated with point mutations in the USH2A gene. Feeder-free culture conditions and the integration-free CytoTune™-iPS 2.0 Sendai Reprogramming Kit were used.

  1. In vitro differentiation and maturation of mouse embryonic stem cells into hepatocytes

    International Nuclear Information System (INIS)

    Ishii, Takamichi; Yasuchika, Kentaro; Fujii, Hideaki; Hoppo, Toshitaka; Baba, Shinji; Naito, Masato; Machimoto, Takafumi; Kamo, Naoko; Suemori, Hirofumi; Nakatsuji, Norio; Ikai, Iwao

    2005-01-01

    It is difficult to induce the maturation of embryonic stem (ES) cells into hepatocytes in vitro. We previously reported that Thy1-positive mesenchymal cells derived from the mouse fetal liver promote the maturation of hepatic progenitor cells. Here, we isolated alpha-fetoprotein (AFP)-producing cells from mouse ES cells for subsequent differentiation into hepatocytes in vitro by coculture with Thy1-positive cells. ES cells expressing green fluorescent protein (GFP) under the control of an AFP promoter were cultured under serum- and feeder layer-free culture conditions. The proportion of GFP-positive cells plateaued at 41.6 ± 12.2% (means ± SD) by day 7. GFP-positive cells, isolated by flow cytometry, were cultured in the presence or absence of Thy1-positive cells as a feeder layer. Isolated GFP-positive cells were stained for AFP, Foxa2, and albumin. The expression of mRNAs encoding tyrosine amino transferase, tryptophan 2,3-dioxygenase, and glucose-6-phosphatase were only detected following coculture with Thy1-positive cells. Following coculture with Thy1-positive cells, the isolated cells produced and stored glycogen. Ammonia clearance activity was also enhanced following coculture. Electron microscopic analysis indicated that the cocultured cells exhibited the morphologic features of mature hepatocytes. In conclusion, coculture with Thy1-positive cells in vitro induced the maturation of AFP-producing cells isolated from ES cell cultures into hepatocytes

  2. Pseudomonas aeruginosa forms Biofilms in Acute InfectionIndependent of Cell-to-Cell Signaling

    Energy Technology Data Exchange (ETDEWEB)

    Schaber, J. Andy; Triffo, W.J.; Suh, Sang J.; Oliver, Jeffrey W.; Hastert, Mary C.; Griswold, John A.; Auer, Manfred; Hamood, Abdul N.; Rumbaugh, Kendra P.

    2006-09-20

    Biofilms are bacterial communities residing within a polysaccharide matrix that are associated with persistence and antibiotic resistance in chronic infections. We show that the opportunistic pathogen Pseudomonas aeruginosa forms biofilms within 8 hours of infection in thermally-injured mice, demonstrating that biofilms contribute to bacterial colonization in acute infections. P. aeruginosa biofilms were visualized within burned tissue surrounding blood vessels and adipose cells. Although quorum sensing (QS), a bacterial signaling mechanism, coordinates differentiation of biofilms in vitro, wild type and QS-deficient P. aeruginosa formed similar biofilms in vivo. Our findings demonstrate that P. aeruginosa forms biofilms on specific host tissues independent of QS.

  3. Research on communication system of underground safety management based on leaky feeder cable

    Institute of Scientific and Technical Information of China (English)

    CHEN Jian-hong; ZHANG Tao; CHENG Yun-cai; ZHANG Han

    2007-01-01

    According to the current working status of underground safety management and production scheduling, the importance and existed problem of underground mine radio communication were summarized, and the basic principle and classification of leaky feeder cable were introduced and the characteristics of cable were analyzed specifically in depth, and the application model of radio communication system for underground mine safety management was put forward. Meanwhile, the research explanation of the system component, function and evaluation was provided. The discussion result indicates that communication system of underground mine safety management which is integrated two-way relay amplifier and other equipment has many communication functions, and underground mine mobile communication can be achieved well.

  4. Development of buffalo (Bubalus bubalis embryonic stem cell lines from somatic cell nuclear transferred blastocysts

    Directory of Open Access Journals (Sweden)

    Syed Mohmad Shah

    2015-11-01

    Full Text Available We developed buffalo embryonic stem cell lines from somatic cell nuclear transfer derived blastocysts, produced by hand-guided cloning technique. The inner cell mass of the blastocyst was cut mechanically using a Microblade and cultured onto feeder cells in buffalo embryonic stem (ES cell culture medium at 38 °C in a 5% CO2 incubator. The stem cell colonies were characterized for alkaline phosphatase activity, karyotype, pluripotency and self-renewal markers like OCT4, NANOG, SOX2, c-Myc, FOXD3, SSEA-1, SSEA-4, TRA-1-60, TRA-1-81 and CD90. The cell lines also possessed the capability to differentiate across all the three germ layers under spontaneous differentiation conditions.

  5. Fast neutrons-induced apoptosis is Fas-independent in lymphoblastoid cells

    International Nuclear Information System (INIS)

    Fischer, Barbara; Benzina, Sami; Jeannequin, Pierre; Dufour, Patrick; Bergerat, Jean-Pierre; Denis, Jean-Marc; Gueulette, John; Bischoff, Pierre L.

    2005-01-01

    We have previously shown that ionizing radiation-induced apoptosis in human lymphoblastoid cells differs according to their p53 status, and that caspase 8-mediated cleavage of BID is involved in the p53-dependent pathway. In the present study, we investigated the role of Fas signaling in caspase 8 activation induced by fast neutrons irradiation in these cells. Fas and FasL expression was assessed by flow cytometry and by immunoblot. We also measured Fas aggregation after irradiation by fluorescence microscopy. We found a decrease of Fas expression after irradiation, but no change in Fas ligand expression. We also showed that, in contrast to the stimulation of Fas by an agonistic antibody, Fas aggregation did not occur after irradiation. Altogether, our data strongly suggest that fast neutrons induced-apoptosis is Fas-independent, even in p53-dependent apoptosis

  6. Brief Report: An Independent Replication and Extension of Psychometric Evidence Supporting the Theory of Mind Inventory

    Science.gov (United States)

    Greenslade, Kathryn J.; Coggins, Truman E.

    2016-01-01

    This study presents an independent replication and extension of psychometric evidence supporting the "Theory of Mind Inventory" ("ToMI"). Parents of 20 children with ASD (4; 1-6; 7 years; months) and 20 with typical development (3; 1-6; 5), rated their child's theory of mind abilities in everyday situations. Other parent report…

  7. Cell line name recognition in support of the identification of synthetic lethality in cancer from text

    Science.gov (United States)

    Kaewphan, Suwisa; Van Landeghem, Sofie; Ohta, Tomoko; Van de Peer, Yves; Ginter, Filip; Pyysalo, Sampo

    2016-01-01

    Motivation: The recognition and normalization of cell line names in text is an important task in biomedical text mining research, facilitating for instance the identification of synthetically lethal genes from the literature. While several tools have previously been developed to address cell line recognition, it is unclear whether available systems can perform sufficiently well in realistic and broad-coverage applications such as extracting synthetically lethal genes from the cancer literature. In this study, we revisit the cell line name recognition task, evaluating both available systems and newly introduced methods on various resources to obtain a reliable tagger not tied to any specific subdomain. In support of this task, we introduce two text collections manually annotated for cell line names: the broad-coverage corpus Gellus and CLL, a focused target domain corpus. Results: We find that the best performance is achieved using NERsuite, a machine learning system based on Conditional Random Fields, trained on the Gellus corpus and supported with a dictionary of cell line names. The system achieves an F-score of 88.46% on the test set of Gellus and 85.98% on the independently annotated CLL corpus. It was further applied at large scale to 24 302 102 unannotated articles, resulting in the identification of 5 181 342 cell line mentions, normalized to 11 755 unique cell line database identifiers. Availability and implementation: The manually annotated datasets, the cell line dictionary, derived corpora, NERsuite models and the results of the large-scale run on unannotated texts are available under open licenses at http://turkunlp.github.io/Cell-line-recognition/. Contact: sukaew@utu.fi PMID:26428294

  8. Flavopiridol induces apoptosis in glioma cell lines independent of retinoblastoma and p53 tumor suppressor pathway alterations by a caspase-independent pathway.

    Science.gov (United States)

    Alonso, Michelle; Tamasdan, Cristina; Miller, Douglas C; Newcomb, Elizabeth W

    2003-02-01

    Flavopiridol is a synthetic flavone, which inhibits growth in vitro and in vivo of several solid malignancies such as renal, prostate, and colon cancers. It is a potent cyclin-dependent kinase inhibitor presently in clinical trials. In this study, we examined the effect of flavopiridol on a panel of glioma cell lines having different genetic profiles: five of six have codeletion of p16(INK4a) and p14(ARF); three of six have p53 mutations; and one of six shows overexpression of mouse double minute-2 (MDM2) protein. Independent of retinoblastoma and p53 tumor suppressor pathway alterations, flavopiridol induced apoptosis in all cell lines but through a caspase-independent mechanism. No cleavage products for caspase 3 or its substrate poly(ADP-ribose) polymerase or caspase 8 were detected. The pan-caspase inhibitor Z-VAD-fmk did not inhibit flavopiridol-induced apoptosis. Mitochondrial damage measured by cytochrome c release and transmission electron microscopy was not observed in drug-treated glioma cells. In contrast, flavopiridol treatment induced translocation of apoptosis-inducing factor from the mitochondria to the nucleus. The proteins cyclin D(1) and MDM2 involved in the regulation of retinoblastoma and p53 activity, respectively, were down-regulated early after flavopiridol treatment. Given that MDM2 protein can confer oncogenic properties under certain circumstances, loss of MDM2 expression in tumor cells could promote increased chemosensitivity. After drug treatment, a low Bcl-2/Bax ratio was observed, a condition that may favor apoptosis. Taken together, the data indicate that flavopiridol has activity against glioma cell lines in vitro and should be considered for clinical development in the treatment of glioblastoma multiforme.

  9. Pumpkin seed extract: Cell growth inhibition of hyperplastic and cancer cells, independent of steroid hormone receptors.

    Science.gov (United States)

    Medjakovic, Svjetlana; Hobiger, Stefanie; Ardjomand-Woelkart, Karin; Bucar, Franz; Jungbauer, Alois

    2016-04-01

    Pumpkin seeds have been known in folk medicine as remedy for kidney, bladder and prostate disorders since centuries. Nevertheless, pumpkin research provides insufficient data to back up traditional beliefs of ethnomedical practice. The bioactivity of a hydro-ethanolic extract of pumpkin seeds from the Styrian pumpkin, Cucurbita pepo L. subsp. pepo var. styriaca, was investigated. As pumpkin seed extracts are standardized to cucurbitin, this compound was also tested. Transactivational activity was evaluated for human androgen receptor, estrogen receptor and progesterone receptor with in vitro yeast assays. Cell viability tests with prostate cancer cells, breast cancer cells, colorectal adenocarcinoma cells and a hyperplastic cell line from benign prostate hyperplasia tissue were performed. As model for non-hyperplastic cells, effects on cell viability were tested with a human dermal fibroblast cell line (HDF-5). No transactivational activity was found for human androgen receptor, estrogen receptor and progesterone receptor, for both, extract and cucurbitin. A cell growth inhibition of ~40-50% was observed for all cell lines, with the exception of HDF-5, which showed with ~20% much lower cell growth inhibition. Given the receptor status of some cell lines, a steroid-hormone receptor independent growth inhibiting effect can be assumed. The cell growth inhibition for fast growing cells together with the cell growth inhibition of prostate-, breast- and colon cancer cells corroborates the ethnomedical use of pumpkin seeds for a treatment of benign prostate hyperplasia. Moreover, due to the lack of androgenic activity, pumpkin seed applications can be regarded as safe for the prostate. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  10. Kalimantan energy resource management to support energy independence and industry growth

    International Nuclear Information System (INIS)

    Rizki Firmansyah Setya Budi; Wiku Lulus Widodo; Djati Hoesen Salimy

    2014-01-01

    There are a large number of energy resource in Kalimantan such as coal, oil, CBM, gas and nuclear. While the electricity consumption still low. That condition caused by the bad energy planning. The aim of the study are to know the number and the ability of energy resource to supply the energy demand that support the growth of Kalimantan industry. The methodology are collecting and processing data through calculation using MESSAGE Program. The result is energy resource in Kalimantan can support Kalimantan energy independence and industry growth in Kalimantan. The coal resource is 34,814 million ton consumption 835 million ton, gas resource is 31,814 BSCF consumption 3,281 BSCF, Oil resource is 920 MMSTB consumption 4406 MMSTB, CBM resource is 210 TCF consumption 2.1 TCF, U 3 O 8 resource is 12,409 ton consumption zero. Whereas for hydro and biomass, the resource are 256 and 138 MWyr, the maximum consumption 185 and 126 MWyr every year. Oil consumption will exceed the resource so need import from other island or replaced by others energy that have large resource such as gas, CBM, or coal. Potency to make cleaner environment can be done by used nuclear energy. (author)

  11. Saikosaponin d induces cell death through caspase-3-dependent, caspase-3-independent and mitochondrial pathways in mammalian hepatic stellate cells

    International Nuclear Information System (INIS)

    Chen, Ming-Feng; Huang, S. Joseph; Huang, Chao-Cheng; Liu, Pei-Shan; Lin, Kun-I; Liu, Ching-Wen; Hsieh, Wen-Chuan; Shiu, Li-Yen; Chen, Chang-Han

    2016-01-01

    Saikosaponin d (SSd) is one of the main active triterpene saponins in Bupleurum falcatum. It has a steroid-like structure, and is reported to have pharmacological activities, including liver protection in rat, cell cycle arrest and apoptosis induction in several cancer cell lines. However, the biological functions and molecular mechanisms of mammalian cells under SSd treatment are still unclear. The cytotoxicity and apoptosis of hepatic stellate cells (HSCs) upon SSd treatment were discovered by MTT assay, colony formation assay and flow cytometry. The collage I/III, caspase activity and apoptotic related genes were examined by quantitative PCR, Western blotting, immunofluorescence and ELISA. The mitochondrial functions were monitored by flow cytometry, MitoTracker staining, ATP production and XF24 bioenergetic assay. This study found that SSd triggers cell death via an apoptosis path. An example of this path might be typical apoptotic morphology, increased sub-G1 phase cell population, inhibition of cell proliferation and activation of caspase-3 and caspase-9. However, the apoptotic effects induced by SSd are partially blocked by the caspase-3 inhibitor, Z-DEVD-FMK, suggesting that SSd may trigger both HSC-T6 and LX-2 cell apoptosis through caspase-3-dependent and independent pathways. We also found that SSd can trigger BAX and BAK translocation from the cytosol to the mitochondria, resulting in mitochondrial function inhibition, membrane potential disruption. Finally, SSd also increases the release of apoptotic factors. The overall analytical data indicate that SSd-elicited cell death may occur through caspase-3-dependent, caspase-3-independent and mitochondrial pathways in mammalian HSCs, and thus can delay the formation of liver fibrosis by reducing the level of HSCs

  12. Bacterial CpG-DNA activates dendritic cells in vivo: T helper cell-independent cytotoxic T cell responses to soluble proteins.

    Science.gov (United States)

    Sparwasser, T; Vabulas, R M; Villmow, B; Lipford, G B; Wagner, H

    2000-12-01

    Receptors for conserved molecular patterns associated with microbial pathogens induce synthesis of co-stimulatory molecules and cytokines in immature dendritic cells (DC), as do antigen-reactive CD4 T helper cells via CD40 signaling. Once activated, antigen-presenting DC may activate CD8 T cell responses in a T helper cell-independent fashion. Using immunostimulatory CpG-oligonucleotides (ODN) mimicking bacterial CpG-DNA, we tested whether CpG-DNA bypasses the need for T helper cells in CTL responses towards proteins by directly activating antigen-presenting DC to transit into professional APC. We describe that immature DC in situ constitutively process soluble proteins and generate CD8 T cell determinants yet CD8 T cell responses remain abortive. Induction of primary antigen-specific CD8 cytotoxic T lymphocyte (CTL)-mediated responses becomes initiated in wild-type as well as T helper cell-deficient mice, provided soluble protein and CpG-ODN are draining into the same lymph node. Specifically we show that CpG-ODN trigger antigen-presenting immature DC within the draining lymph node to acutely up-regulate co-stimulatory molecules and produce IL-12. These results provide new insights for generating in vivo efficient CTL responses to soluble proteins which may influence vaccination strategies.

  13. Quercetin and epigallocatechin gallate inhibit glucose uptake and metabolism by breast cancer cells by an estrogen receptor-independent mechanism

    Energy Technology Data Exchange (ETDEWEB)

    Moreira, Liliana, E-mail: lilianam87@gmail.com [Department of Biochemistry (U38-FCT), Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto (Portugal); Araújo, Isabel, E-mail: isa.araujo013@gmail.com [Department of Biochemistry (U38-FCT), Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto (Portugal); Costa, Tito, E-mail: tito.fmup16@gmail.com [Department of Biochemistry (U38-FCT), Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto (Portugal); Correia-Branco, Ana, E-mail: ana.clmc.branco@gmail.com [Department of Biochemistry (U38-FCT), Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto (Portugal); Faria, Ana, E-mail: anafaria@med.up.pt [Department of Biochemistry (U38-FCT), Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto (Portugal); Chemistry Investigation Centre (CIQ), Faculty of Sciences of University of Porto, Rua Campo Alegre, 4169-007 Porto (Portugal); Faculty of Nutrition and Food Sciences of University of Porto, Rua Dr. Roberto Frias, 4200-465 Porto (Portugal); Martel, Fátima, E-mail: fmartel@med.up.pt [Department of Biochemistry (U38-FCT), Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto (Portugal); Keating, Elisa, E-mail: keating@med.up.pt [Department of Biochemistry (U38-FCT), Faculty of Medicine of University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto (Portugal)

    2013-07-15

    In this study we characterized {sup 3}H-2-deoxy-D-glucose ({sup 3}H -DG) uptake by the estrogen receptor (ER)-positive MCF7 and the ER-negative MDA-MB-231 human breast cancer cell lines and investigated the effect of quercetin (QUE) and epigallocatechin gallate (EGCG) upon {sup 3}H-DG uptake, glucose metabolism and cell viability and proliferation. In both MCF7 and MDA-MB-231 cells {sup 3}H-DG uptake was (a) time-dependent, (b) saturable with similar capacity (V{sub max}) and affinity (K{sub m}), (c) potently inhibited by cytochalasin B, an inhibitor of the facilitative glucose transporters (GLUT), (d) sodium-independent and (e) slightly insulin-stimulated. This suggests that {sup 3}H-DG uptake by both cell types is mediated by members of the GLUT family, including the insulin-responsive GLUT4 or GLUT12, while being independent of the sodium-dependent glucose transporter (SGLT1). QUE and EGCG markedly and concentration-dependently inhibited {sup 3}H-DG uptake by MCF7 and by MDA-MB-231 cells, and both compounds blocked lactate production by MCF7 cells. Additionally, a 4 h-treatment with QUE or EGCG decreased MCF7 cell viability and proliferation, an effect that was more potent when glucose was available in the extracellular medium. Our results implicate QUE and EGCG as metabolic antagonists in breast cancer cells, independently of estrogen signalling, and suggest that these flavonoids could serve as therapeutic agents/adjuvants even for ER-negative breast tumors. -- Highlights: • Glucose uptake by MCF7 and MDA-MB-231 cells is mainly mediated by GLUT1. • QUE and EGCG inhibit cellular glucose uptake thus abolishing the Warburg effect. • This process induces cytotoxicity and proliferation arrest in MCF7 cells. • The flavonoids’ effects are independent of estrogen receptor signalling.

  14. Quercetin and epigallocatechin gallate inhibit glucose uptake and metabolism by breast cancer cells by an estrogen receptor-independent mechanism

    International Nuclear Information System (INIS)

    Moreira, Liliana; Araújo, Isabel; Costa, Tito; Correia-Branco, Ana; Faria, Ana; Martel, Fátima; Keating, Elisa

    2013-01-01

    In this study we characterized 3 H-2-deoxy-D-glucose ( 3 H -DG) uptake by the estrogen receptor (ER)-positive MCF7 and the ER-negative MDA-MB-231 human breast cancer cell lines and investigated the effect of quercetin (QUE) and epigallocatechin gallate (EGCG) upon 3 H-DG uptake, glucose metabolism and cell viability and proliferation. In both MCF7 and MDA-MB-231 cells 3 H-DG uptake was (a) time-dependent, (b) saturable with similar capacity (V max ) and affinity (K m ), (c) potently inhibited by cytochalasin B, an inhibitor of the facilitative glucose transporters (GLUT), (d) sodium-independent and (e) slightly insulin-stimulated. This suggests that 3 H-DG uptake by both cell types is mediated by members of the GLUT family, including the insulin-responsive GLUT4 or GLUT12, while being independent of the sodium-dependent glucose transporter (SGLT1). QUE and EGCG markedly and concentration-dependently inhibited 3 H-DG uptake by MCF7 and by MDA-MB-231 cells, and both compounds blocked lactate production by MCF7 cells. Additionally, a 4 h-treatment with QUE or EGCG decreased MCF7 cell viability and proliferation, an effect that was more potent when glucose was available in the extracellular medium. Our results implicate QUE and EGCG as metabolic antagonists in breast cancer cells, independently of estrogen signalling, and suggest that these flavonoids could serve as therapeutic agents/adjuvants even for ER-negative breast tumors. -- Highlights: • Glucose uptake by MCF7 and MDA-MB-231 cells is mainly mediated by GLUT1. • QUE and EGCG inhibit cellular glucose uptake thus abolishing the Warburg effect. • This process induces cytotoxicity and proliferation arrest in MCF7 cells. • The flavonoids’ effects are independent of estrogen receptor signalling

  15. Trophodynamics of inorganic pollutants in a wide-range feeder: The relevance of dietary inputs and biomagnification in the Yellow-legged gull (Larus michahellis)

    International Nuclear Information System (INIS)

    Ramos, Raül; Ramírez, Francisco; Jover, Lluís

    2013-01-01

    The suitability of sentinel species to monitor environmental pollution is often hampered by an insufficient knowledge on pollutant trophodynamics. We simultaneously evaluated the influence of individuals' trophic position (as revealed by δ 15 N values) and dietary exploitation of particular systems (using δ 13 C and δ 34 S as proxies) on inorganic pollutant concentrations measured on fledglings' feathers of a wide-range feeder, the Yellow-legged Gull (Larus michahellis), sampled at four locations throughout the Western Mediterranean. Concentrations of total Hg and Se in fledgling feathers (2.43 ± 1.30 and 1.16 ± 0.43 μg/g, respectively) were under the threshold points for deleterious effects on seabirds. On the contrary, alarming Pb concentrations were found in one colony (mean: 1.57 ± 2.46 μg/g, range: 0.16–12.13). With the exception of Pb, pollutant concentrations were positively influenced by consumption of marine resources (as suggested by the positive relationship with δ 34 S values), whereas trophic position played a minor role in determining pollutant body burdens. - Graphical abstract: Individual feeding behaviours in wide-range feeders may represent unavoidable knowledge for an appropriate understanding of contaminant acquisition, adding complexity to the study of the dynamic of contaminants throughout food chains. In this case study, pollutant body burden of Yellow-legged gull (Larus michahellis) chicks was highly influenced by dietary exposure, whereas the influence of trophic position was relatively low. Display Omitted Highlights: ► Diet origin (δ 34 S) and trophic position (δ 15 N) were tested relative to Hg, Se, and Pb levels. ► Hg and Se concentrations in gull feathers were highly influenced by δ 34 S signatures. ► Exploited habitat influenced pollutant levels to a greater extent than trophic position. ► Biomagnification processes contribute less to contamination exposure in wide-range feeders. - Pollutant body burden of a wide

  16. Autoradiographic studies on the kinetics of fetal supporting cells and wall cells in rats 19 days after conception

    International Nuclear Information System (INIS)

    Lugani-Mehta, S.

    1980-01-01

    The duration of the S-phase of supporting cells and wall cells of rat fetuses aged 19 days was determined by the ''labelled mitosis'' method. The supporting cells are predecessors of the sertoli cells while the wall cells are predecessors of the boundary tissue and, possibly, of part of the peritubular Leydig cell system. The S-phase of the supporting cells was found to last 10.1 h while the S-phase of the wall cells lasted 9.2 h. The data were not in agreement with the data of other authors. (orig./MG) [de

  17. Microplastic in a macro filter feeder: Humpback whale Megaptera novaeangliae.

    Science.gov (United States)

    Besseling, E; Foekema, E M; Van Franeker, J A; Leopold, M F; Kühn, S; Bravo Rebolledo, E L; Heße, E; Mielke, L; IJzer, J; Kamminga, P; Koelmans, A A

    2015-06-15

    Marine filter feeders are exposed to microplastic because of their selection of small particles as food source. Baleen whales feed by filtering small particles from large water volumes. Macroplastic was found in baleen whales before. This study is the first to show the presence of microplastic in intestines of a baleen whale (Megaptera novaeangliae). Contents of its gastrointestinal tract were sieved, dissolved in 10% potassium hydroxide and washed. From the remaining dried material, potential synthetic polymer particles were selected based on density and appearance, and analysed by Fourier transform infrared (FTIR) spectroscopy. Several polymer types (polyethylene, polypropylene, polyvinylchloride, polyethylene terephthalate, nylon) were found, in varying particle shapes: sheets, fragments and threads with a size of 1mm to 17cm. This diversity in polymer types and particle shapes, can be interpreted as a representation of the varying characteristics of marine plastic and the unselective way of ingestion by M. novaeangliae. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Cycloheximide Can Induce Bax/Bak Dependent Myeloid Cell Death Independently of Multiple BH3-Only Proteins.

    Directory of Open Access Journals (Sweden)

    Katharine J Goodall

    Full Text Available Apoptosis mediated by Bax or Bak is usually thought to be triggered by BH3-only members of the Bcl-2 protein family. BH3-only proteins can directly bind to and activate Bax or Bak, or indirectly activate them by binding to anti-apoptotic Bcl-2 family members, thereby relieving their inhibition of Bax and Bak. Here we describe a third way of activation of Bax/Bak dependent apoptosis that does not require triggering by multiple BH3-only proteins. In factor dependent myeloid (FDM cell lines, cycloheximide induced apoptosis by a Bax/Bak dependent mechanism, because Bax-/-Bak-/- lines were profoundly resistant, whereas FDM lines lacking one or more genes for BH3-only proteins remained highly sensitive. Addition of cycloheximide led to the rapid loss of Mcl-1 but did not affect the expression of other Bcl-2 family proteins. In support of these findings, similar results were observed by treating FDM cells with the CDK inhibitor, roscovitine. Roscovitine reduced Mcl-1 abundance and caused Bax/Bak dependent cell death, yet FDM lines lacking one or more genes for BH3-only proteins remained highly sensitive. Therefore Bax/Bak dependent apoptosis can be regulated by the abundance of anti-apoptotic Bcl-2 family members such as Mcl-1, independently of several known BH3-only proteins.

  19. Increased expression of heparin binding EGF (HB-EGF), amphiregulin, TGF alpha and epiregulin in androgen-independent prostate cancer cell lines.

    DEFF Research Database (Denmark)

    Tørring, Niels; Sørensen, Boe Sandahl; Nexø, Ebba

    2000-01-01

    BACKGROUND: The proliferation of androgen-independent prostate cancer cell lines has previously been shown to be influenced by an autocrine loop of the epidermal growth factor (EGF) system. This observation has alerted us to study the expression of ligands and receptors from the EGF......-system in prostate cell lines. METHODS: The expression of the EGF system was determined by quantitative RT-PCR and ELISA in the normal prostate epithelial cell line (PNT1A), in the androgen sensitive-(LNCaP), and the androgen-independent (DU145 and PC3) prostate cancer cell lines. RESULTS: The expression of m...... which exhibit low expression of HER1. Similar results were obtained by ELISA. CONCLUSIONS: The data indicates a selective up-regulation of a subclass of ligands of the EGF-system in androgen-independent prostate cancer cell lines. We suggest this could be a mechanism to escape androgen dependence...

  20. Fluorescent labelling of intestinal epithelial cells reveals independent long-lived intestinal stem cells in a crypt

    Energy Technology Data Exchange (ETDEWEB)

    Horita, Nobukatsu [Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University (Japan); Tsuchiya, Kiichiro, E-mail: kii.gast@tmd.ac.jp [Department of Advanced Therapeutics for Gastrointestinal Diseases, Graduate School, Tokyo Medical and Dental University (Japan); Hayashi, Ryohei [Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University (Japan); Department of Gastroenterology and Metabolism, Hiroshima University (Japan); Fukushima, Keita; Hibiya, Shuji; Fukuda, Masayoshi; Kano, Yoshihito; Mizutani, Tomohiro; Nemoto, Yasuhiro; Yui, Shiro [Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University (Japan); Okamoto, Ryuichi; Nakamura, Tetsuya [Department of Advanced Therapeutics for Gastrointestinal Diseases, Graduate School, Tokyo Medical and Dental University (Japan); Watanabe, Mamoru [Department of Gastroenterology and Hepatology, Graduate School, Tokyo Medical and Dental University (Japan)

    2014-11-28

    Highlights: • Lentivirus mixed with Matrigel enables direct infection of intestinal organoids. • Our original approach allows the marking of a single stem cell in a crypt. • Time-lapse imaging shows the dynamics of a single stem cell. • Our lentivirus transgene system demonstrates plural long-lived stem cells in a crypt. - Abstract: Background and aims: The dynamics of intestinal stem cells are crucial for regulation of intestinal function and maintenance. Although crypt stem cells have been identified in the intestine by genetic marking methods, identification of plural crypt stem cells has not yet been achieved as they are visualised in the same colour. Methods: Intestinal organoids were transferred into Matrigel® mixed with lentivirus encoding mCherry. The dynamics of mCherry-positive cells was analysed using time-lapse imaging, and the localisation of mCherry-positive cells was analysed using 3D immunofluorescence. Results: We established an original method for the introduction of a transgene into an organoid generated from mouse small intestine that resulted in continuous fluorescence of the mCherry protein in a portion of organoid cells. Three-dimensional analysis using confocal microscopy showed a single mCherry-positive cell in an organoid crypt that had been cultured for >1 year, which suggested the presence of long-lived mCherry-positive and -negative stem cells in the same crypt. Moreover, a single mCherry-positive stem cell in a crypt gave rise to both crypt base columnar cells and transit amplifying cells. Each mCherry-positive and -negative cell contributed to the generation of organoids. Conclusions: The use of our original lentiviral transgene system to mark individual organoid crypt stem cells showed that long-lived plural crypt stem cells might independently serve as intestinal epithelial cells, resulting in the formation of a completely functional villus.

  1. Fluorescent labelling of intestinal epithelial cells reveals independent long-lived intestinal stem cells in a crypt

    International Nuclear Information System (INIS)

    Horita, Nobukatsu; Tsuchiya, Kiichiro; Hayashi, Ryohei; Fukushima, Keita; Hibiya, Shuji; Fukuda, Masayoshi; Kano, Yoshihito; Mizutani, Tomohiro; Nemoto, Yasuhiro; Yui, Shiro; Okamoto, Ryuichi; Nakamura, Tetsuya; Watanabe, Mamoru

    2014-01-01

    Highlights: • Lentivirus mixed with Matrigel enables direct infection of intestinal organoids. • Our original approach allows the marking of a single stem cell in a crypt. • Time-lapse imaging shows the dynamics of a single stem cell. • Our lentivirus transgene system demonstrates plural long-lived stem cells in a crypt. - Abstract: Background and aims: The dynamics of intestinal stem cells are crucial for regulation of intestinal function and maintenance. Although crypt stem cells have been identified in the intestine by genetic marking methods, identification of plural crypt stem cells has not yet been achieved as they are visualised in the same colour. Methods: Intestinal organoids were transferred into Matrigel® mixed with lentivirus encoding mCherry. The dynamics of mCherry-positive cells was analysed using time-lapse imaging, and the localisation of mCherry-positive cells was analysed using 3D immunofluorescence. Results: We established an original method for the introduction of a transgene into an organoid generated from mouse small intestine that resulted in continuous fluorescence of the mCherry protein in a portion of organoid cells. Three-dimensional analysis using confocal microscopy showed a single mCherry-positive cell in an organoid crypt that had been cultured for >1 year, which suggested the presence of long-lived mCherry-positive and -negative stem cells in the same crypt. Moreover, a single mCherry-positive stem cell in a crypt gave rise to both crypt base columnar cells and transit amplifying cells. Each mCherry-positive and -negative cell contributed to the generation of organoids. Conclusions: The use of our original lentiviral transgene system to mark individual organoid crypt stem cells showed that long-lived plural crypt stem cells might independently serve as intestinal epithelial cells, resulting in the formation of a completely functional villus

  2. An efficient hybrid evolutionary algorithm based on PSO and HBMO algorithms for multi-objective Distribution Feeder Reconfiguration

    Energy Technology Data Exchange (ETDEWEB)

    Niknam, Taher [Electronic and Electrical Engineering Department, Shiraz University of Technology, Shiraz (Iran)

    2009-08-15

    This paper introduces a robust searching hybrid evolutionary algorithm to solve the multi-objective Distribution Feeder Reconfiguration (DFR). The main objective of the DFR is to minimize the real power loss, deviation of the nodes' voltage, the number of switching operations, and balance the loads on the feeders. Because of the fact that the objectives are different and no commensurable, it is difficult to solve the problem by conventional approaches that may optimize a single objective. This paper presents a new approach based on norm3 for the DFR problem. In the proposed method, the objective functions are considered as a vector and the aim is to maximize the distance (norm2) between the objective function vector and the worst objective function vector while the constraints are met. Since the proposed DFR is a multi objective and non-differentiable optimization problem, a new hybrid evolutionary algorithm (EA) based on the combination of the Honey Bee Mating Optimization (HBMO) and the Discrete Particle Swarm Optimization (DPSO), called DPSO-HBMO, is implied to solve it. The results of the proposed reconfiguration method are compared with the solutions obtained by other approaches, the original DPSO and HBMO over different distribution test systems. (author)

  3. Water quality in Pearl Harbor and feeder streams during 1971 - 2001 collected primarily by oceanography students from Leeward Community College (NODC Accession 0000590)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Water quality data were collected in Pearl Harbor and surrounding feeder streams from 30 December 1971 to 24 August 2001. Data were collected by Leeward Community...

  4. Simvastatin induces caspase-independent apoptosis in LPS-activated RAW264.7 macrophage cells

    International Nuclear Information System (INIS)

    Kim, Yong Chan; Song, Seok Bean; Lee, Mi Hee; Kang, Kwang Il; Lee, Hayyoung; Paik, Sang-Gi; Kim, Kyoon Eon; Kim, Young Sang

    2006-01-01

    Macrophages participate in several inflammatory pathologies such as sepsis and arthritis. We examined the effect of simvastatin on the LPS-induced proinflammatory macrophage RAW264.7 cells. Co-treatment of LPS and a non-toxic dose of simvastatin induced cell death in RAW264.7 cells. The cell death was accompanied by disruption of mitochondrial membrane potential (MMP), genomic DNA fragmentation, and caspase-3 activation. Surprisingly, despite caspase-dependent apoptotic cascade being completely blocked by Z-VAD-fmk, a pan-caspase inhibitor, the cell death was only partially repressed. In the presence of Z-VAD-fmk, DNA fragmentation was blocked, but DNA condensation, disruption of MMP, and nuclear translocation of apoptosis inducing factor were obvious. The cell death by simvastatin and LPS was effectively decreased by both the FPP and GGPP treatments as well as mevalonate. Our findings indicate that simvastatin triggers the cell death of LPS-treated RAW264.7 cells through both caspase-dependent and -independent apoptotic pathways, suggesting a novel mechanism of statins for the severe inflammatory disease therapy

  5. Cloning of a novel cell type from human fetal liver expressing cytoplasmic CD3 delta and epsilon but not membrane CD3

    NARCIS (Netherlands)

    Hori, T.; de Waal Malefyt, R.; Duncan, B. W.; Harrison, M. R.; Roncarolo, M. G.; Spits, H.

    1991-01-01

    Seventeen-week human fetal liver cells cultured with a feeder cell mixture of irradiated PBL, irradiated JY cells (an EBV-transformed B cell line) and PHA contained a subpopulation of CD3- cells in addition to a major population of T cells with the mature phenotype. After 12 days in culture, CD3-

  6. Oxygen enhancement ratios in synchronous HeLa cells exposed to low-LET radiation

    International Nuclear Information System (INIS)

    Sapozink, M.D.

    1977-01-01

    HeLa cells were synchronized by the mitotic selection method and rendered hypoxic by coincubation with an excess of heavily irradiated, but metabolically active, feeder cells. An oxygen enhancement ratio (OER) of about 3 was obtained in interphase HeLa cells irradiated with x or gamma rays. A significantly lower OER was obtained with cells in, or close to, mitosis. The significance of this decrease in the oxygen effect in mitotic cells is discussed

  7. Enterococcus faecalis Infection Causes Inflammation, Intracellular Oxphos-Independent ROS Production, and DNA Damage in Human Gastric Cancer Cells

    DEFF Research Database (Denmark)

    Strickertsson, Jesper A. B; Desler, Claus; Martin-Bertelsen, Tomas

    2013-01-01

    therefore wanted to study the impact of E. faecalis infection on inflammatory response, reactive oxygen species (ROS) formation, mitochondrial respiration, and mitochondrial genetic stability in gastric mucosal cells. Methods To separate the changes induced by bacteria from those of the inflammatory cells...... intracellular ROS production through a pathway independent of oxidative phosphorylation (oxphos). Furthermore, E. faecalis infection induced mitochondrial DNA instability. Following infection, genes coding for inflammatory response proteins were transcriptionally up-regulated while DNA damage repair and cell...... cycle control genes were down-regulated. Cell growth slowed down when infected with viable E. faecalis and responded in a dose dependent manner to E. faecalis lysate. Conclusions Infection by E. faecalis induced an oxphos-independent intracellular ROS response and damaged the mitochondrial genome...

  8. Exposure time independent summary statistics for assessment of drug dependent cell line growth inhibition.

    Science.gov (United States)

    Falgreen, Steffen; Laursen, Maria Bach; Bødker, Julie Støve; Kjeldsen, Malene Krag; Schmitz, Alexander; Nyegaard, Mette; Johnsen, Hans Erik; Dybkær, Karen; Bøgsted, Martin

    2014-06-05

    In vitro generated dose-response curves of human cancer cell lines are widely used to develop new therapeutics. The curves are summarised by simplified statistics that ignore the conventionally used dose-response curves' dependency on drug exposure time and growth kinetics. This may lead to suboptimal exploitation of data and biased conclusions on the potential of the drug in question. Therefore we set out to improve the dose-response assessments by eliminating the impact of time dependency. First, a mathematical model for drug induced cell growth inhibition was formulated and used to derive novel dose-response curves and improved summary statistics that are independent of time under the proposed model. Next, a statistical analysis workflow for estimating the improved statistics was suggested consisting of 1) nonlinear regression models for estimation of cell counts and doubling times, 2) isotonic regression for modelling the suggested dose-response curves, and 3) resampling based method for assessing variation of the novel summary statistics. We document that conventionally used summary statistics for dose-response experiments depend on time so that fast growing cell lines compared to slowly growing ones are considered overly sensitive. The adequacy of the mathematical model is tested for doxorubicin and found to fit real data to an acceptable degree. Dose-response data from the NCI60 drug screen were used to illustrate the time dependency and demonstrate an adjustment correcting for it. The applicability of the workflow was illustrated by simulation and application on a doxorubicin growth inhibition screen. The simulations show that under the proposed mathematical model the suggested statistical workflow results in unbiased estimates of the time independent summary statistics. Variance estimates of the novel summary statistics are used to conclude that the doxorubicin screen covers a significant diverse range of responses ensuring it is useful for biological

  9. Epigenetic repression of regulator of G-protein signaling 2 promotes androgen-independent prostate cancer cell growth.

    Science.gov (United States)

    Wolff, Dennis W; Xie, Yan; Deng, Caishu; Gatalica, Zoran; Yang, Mingjie; Wang, Bo; Wang, Jincheng; Lin, Ming-Fong; Abel, Peter W; Tu, Yaping

    2012-04-01

    G-protein-coupled receptor (GPCR)-stimulated androgen-independent activation of androgen receptor (AR) contributes to acquisition of a hormone-refractory phenotype by prostate cancer. We previously reported that regulator of G-protein signaling (RGS) 2, an inhibitor of GPCRs, inhibits androgen-independent AR activation (Cao et al., Oncogene 2006;25:3719-34). Here, we show reduced RGS2 protein expression in human prostate cancer specimens compared to adjacent normal or hyperplastic tissue. Methylation-specific PCR analysis and bisulfite sequencing indicated that methylation of the CpG island in the RGS2 gene promoter correlated with RGS2 downregulation in prostate cancer. In vitro methylation of this promoter suppressed reporter gene expression in transient transfection studies, whereas reversal of this promoter methylation with 5-aza-2'-deoxycytidine (5-Aza-dC) induced RGS2 reexpression in androgen-independent prostate cancer cells and inhibited their growth under androgen-deficient conditions. Interestingly, the inhibitory effect of 5-Aza-dC was significantly reduced by an RGS2-targeted short hairpin RNA, indicating that reexpressed RGS2 contributed to this growth inhibition. Restoration of RGS2 levels by ectopic expression in androgen-independent prostate cancer cells suppressed growth of xenografts in castrated mice. Thus, RGS2 promoter hypermethylation represses its expression and unmasks a latent pathway for AR transactivation in prostate cancer cells. Targeting this reversible process may provide a new strategy for suppressing prostate cancer progression by reestablishing its androgen sensitivity. Copyright © 2011 UICC.

  10. Rapid generation of mitochondrial superoxide induces mitochondrion-dependent but caspase-independent cell death in hippocampal neuronal cells that morphologically resembles necroptosis☆

    Science.gov (United States)

    Fukui, Masayuki; Choi, Hye Joung; Zhu, Bao Ting

    2013-01-01

    Studies in recent years have revealed that excess mitochondrial superoxide production is an important etiological factor in neurodegenerative diseases, resulting from oxidative modifications of cellular lipids, proteins, and nucleic acids. Hence, it is important to understand the mechanism by which mitochondrial oxidative stress causes neuronal death. In this study, the immortalized mouse hippocampal neuronal cells (HT22) in culture were used as a model and they were exposed to menadione (also known as vitamin K3) to increase intracellular superoxide production. We found that menadione causes preferential accumulation of superoxide in the mitochondria of these cells, along with the rapid development of mitochondrial dysfunction and cellular ATP depletion. Neuronal death induced by menadione is independent of the activation of the MAPK signaling pathways and caspases. The lack of caspase activation is due to the rapid depletion of cellular ATP. It was observed that two ATP-independent mitochondrial nucleases, namely, AIF and Endo G, are released following menadione exposure. Silencing of their expression using specific siRNAs results in transient suppression (for ~12 h) of mitochondrial superoxide-induced neuronal death. While suppression of the mitochondrial superoxide dismutase expression markedly sensitizes neuronal cells to mitochondrial superoxide-induced cytotoxicity, its over-expression confers strong protection. Collectively, these findings showed that many of the observed features associated with mitochondrial superoxide-induced cell death, including caspase independency, rapid depletion of ATP level, mitochondrial release of AIF and Endo G, and mitochondrial swelling, are distinctly different from those of apoptosis; instead they resemble some of the known features of necroptosis. PMID:22575170

  11. Automatic feeder for small fish held in tanks

    Science.gov (United States)

    Joeris, Leonard S.

    1965-01-01

    The Northville (Michigan) Biological Station has been a center for study of the developmental morphology of coregonid fishes. This work requires the production of numerous individual series of lake herring, lake whitefish, and several species of chubs from parent fish of positively known identity. The offspring of individual pairs or groups of fish must be held in individual tanks from the time they hatch until they reach maturity. One of the important problems in this project has been the poor growth of most fish. Though some have grown well, their growth has been less than that of the same species in nature, and a few fish from each hatch have grown very slowly. Irregularity of feeding may contribute to the slow growth of laboratory fish. The hatchery caretaker feeds them several times during his 8-hour workday, but they must go without food during the remaining 16 hours. The high metabolic rate of small fish, however, appears to make them strongly inclined toward almost continual feeding. Belief that greater, more regular food consumption would result from a mechanical feeder providing a continuous supply of food over a longer period of the day led to development of the equipment described in this paper.

  12. Establishment of cell lines from adult T-cell leukemia cells dependent on negatively charged polymers.

    Science.gov (United States)

    Kagami, Yoshitoyo; Uchiyama, Susumu; Kato, Harumi; Okada, Yasutaka; Seto, Masao; Kinoshita, Tomohiro

    2017-07-05

    Growing adult T-cell leukemia/lymphoma (ATLL) cells in vitro is difficult. Here, we examined the effects of static electricity in the culture medium on the proliferation of ATLL cells. Six out of 10 ATLL cells did not proliferate in vitro and thus had to be cultured in a medium containing negatively charged polymers. In the presence of poly-γ-glutamic acid (PGA) or chondroitin sulfate (CDR), cell lines (HKOX3-PGA, HKOX3-CDR) were established from the same single ATLL case using interleukin (IL)-2, IL-4, and feeder cells expressing OX40L (OX40L + HK). Dextran sulfate inhibited growth in both HKOX3 cell lines. Both PGA and OX40L + HK were indispensable for HKOX3-PGA growth, but HKOX3-CDR could proliferate in the presence of CDR or OX40L + HK alone. Thus, the specific action of each negatively charged polymer promoted the growth of specific ATLL cells in vitro.

  13. Service tailoring: a method and tool for user-centric creation of integrated IT-based homecare services to support independent living of elderly

    NARCIS (Netherlands)

    Zarifi Eslami, Mohammed

    2013-01-01

    This thesis addresses the problem of supporting independent living of elderly people through IT-based homecare services. Independent living is seen as one way to deal with the consequences of an aging population (especially in industrialized countries), which include rising healthcare expenditures

  14. Prolonged exposure of resveratrol induces reactive superoxide species-independent apoptosis in murine prostate cells.

    Science.gov (United States)

    Kumar, Sanjay; Stokes, James; Singh, Udai P; Scissum-Gunn, Karyn; Singh, Rajesh; Manne, Upender; Mishra, Manoj K

    2017-10-01

    Nitric oxide, a signaling molecule, inhibits mitochondrial respiration by binding with cytochrome c oxidase, resulting in elevated production of reactive superoxide species (reactive oxygen and nitrogen) in the mitochondria and increased susceptibility to cell death. Generation of mitochondrial superoxide species can be suppressed by natural compounds such as resveratrol, a dietary polyphenol found in the skin of red fruits. In various cancer cells, resveratrol shows anti-oxidant and cancer preventive properties. Since, the effect of resveratrol on reactive superoxide species-independent apoptosis in prostate cancer cells is not well illustrated; therefore, we investigated this phenomenon in TRAMP murine prostate cancer cells. To accomplish this, TRAMP cells were incubated with resveratrol, resveratrol + DETA-NONOate, DETA-NONOate (nitric oxide donor), resveratrol + L-NMMA, or L-NMMA (nitric oxide inhibitor) for 48 h, and reactive superoxide species in the mitochondria and culture supernatant were measured. In addition, the mitochondrial membrane potential, cell viability, expression of apoptotic markers (Bax and Bcl2), γ-H2A.x, p53, and caspase-3 was determined. We found that resveratrol suppressed reactive superoxide species such as reactive oxygen species in the mitochondria and nitric oxide in culture supernatant when compared to the DETA-NONOate treatment and disrupted the mitochondrial membrane potential. Resveratrol also reduced cell viability, altered the expression of apoptotic markers (Bax and Bcl2), and increased expression of γ-H2A.x (indicative marker of DNA fragmentation) and p53 (a critical DNA damage response protein). However, there was no appreciable modulation of the caspase-3. Therefore, our data suggest that resveratrol induces superoxide species-independent apoptosis and may act as a therapeutic agent against prostate cancer.

  15. 47 CFR 25.223 - Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS. 25.223 Section 25.223 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Technical Standards § 25...

  16. Pre-differentiation of mesenchymal stromal cells in combination with a microstructured nerve guide supports peripheral nerve regeneration in the rat sciatic nerve model.

    Science.gov (United States)

    Boecker, Arne Hendrik; van Neerven, Sabien Geraldine Antonia; Scheffel, Juliane; Tank, Julian; Altinova, Haktan; Seidensticker, Katrin; Deumens, Ronald; Tolba, Rene; Weis, Joachim; Brook, Gary Anthony; Pallua, Norbert; Bozkurt, Ahmet

    2016-02-01

    Many bioartificial nerve guides have been investigated pre-clinically for their nerve regeneration-supporting function, often in comparison to autologous nerve transplantation, which is still regarded as the current clinical gold standard. Enrichment of these scaffolds with cells intended to support axonal regeneration has been explored as a strategy to boost axonal regeneration across these nerve guides Ansselin et al. (1998). In the present study, 20 mm rat sciatic nerve defects were implanted with a cell-seeded microstructured collagen nerve guide (Perimaix) or an autologous nerve graft. Under the influence of seeded, pre-differentiated mesenchymal stromal cells, axons regenerated well into the Perimaix nerve guide. Myelination-related parameters, like myelin sheath thickness, benefitted from an additional seeding with pre-differentiated mesenchymal stromal cells. Furthermore, both the number of retrogradely labelled sensory neurons and the axon density within the implant were elevated in the cell-seeded scaffold group with pre-differentiated mesenchymal stromal cells. However, a pre-differentiation had no influence on functional recovery. An additional cell seeding of the Perimaix nerve guide with mesenchymal stromal cells led to an extent of functional recovery, independent of the differentiation status, similar to autologous nerve transplantation. These findings encourage further investigations on pre-differentiated mesenchymal stromal cells as a cellular support for peripheral nerve regeneration. © 2015 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

  17. Feeding current characteristics of three morphologically different bivalve suspension feeders, Crassostrea gigas, Mytilus edulis and Cerastoderma edule, in relation to food competition

    NARCIS (Netherlands)

    Troost, K.; Stamhuis, E.J.; van Duren, L.A; Wolff, W.J.

    Introduced Pacific oysters (Crassostrea gigas) have shown rapid expansion in the Oosterschelde estuary, while stocks of native bivalves declined slightly or remained stable. This indicates that they might have an advantage over native bivalve filter feeders. Hence, at the scale of individual

  18. Feeding current characteristics of three morphologically different bivalve suspension feeders, Crassostrea gigas Mytilus edulis and Cerastoderma edule , in relation to food competition

    NARCIS (Netherlands)

    Troost, K.; Stamhuis, E.J.; Duren, L.A.; Wolff, W.

    2009-01-01

    Introduced Pacific oysters (Crassostrea gigas) have shown rapid expansion in the Oosterschelde estuary, while stocks of native bivalves declined slightly or remained stable. This indicates that they might have an advantage over native bivalve filter feeders. Hence, at the scale of individual

  19. On the preparation of fire-wood splitting machines equipped with feeder devices

    International Nuclear Information System (INIS)

    Jaeaeskelaeinen, M.

    1995-01-01

    As part of the Bionergy research programme, the TTS Institute's Department of Forestry conducted a study on firewood splitting machines manufactured by Bilke Oy and Peurala Oy. The said machines are the first of their kind in Finland to be equipped with a feeder device. The Bilke unit is fed by a conveyor belt arrangement while the Peurala unit is equipped with a feeder roll. In addition, both machine types are equipped with a conveyor belt discharging device (e.g. for discharging split wood directly into a trailer). The Bilke unit can also be operated on extraction racks in the woods as the stems due to be split are fed into it from the side and the conveyor discharges the split wood rearwards into the trailer. The trailer can be hooked onto the splitting unit's draw-hook. At the landing, the Bilke unit was used to split a total of 14.2 m 3 and the Peurala unit 8.0 m 3 of approx. 3-metrelong stems averaging 9 cm in diameter. In addition to this, the Bilke unit was used to split ca. 17 m 3 of 3.5 metre-long stems with on average diameter of ca. 6 cm; the stems were in bunches alongside the extraction racks. The person operating the machines for the purposes of this study was a 27-year- old forest worker. The mean productivity achieved at the landing when using the Bilke unit was 7.7 m 3 while that of the Peurala unit was 5.8 m 3 per effective hour. When operated in the woods, the productivity achieved with the Bilke unit (excluding felling-cross cutting-delimbing-bunching and forest haulage work) was 2.8 m 3 /h. When applying a productivity of 1.7 m 3 for felling-cross cutting- delimbing-bunching work and 5.5. m 3 for forest haulage, the entire harvesting chain's productivity was 0.9 m 3 per effective hour

  20. Autocrine growth induced by the insulin-related factor in the insulin-independent teratoma cell line 1246-3A

    International Nuclear Information System (INIS)

    Yamada, Yukio; Serrero, G.

    1988-01-01

    An insulin-independent teratoma-derived cell line, called 1246-3A, has been isolated from the adipogenic cell line 1246, which stringently requires insulin for proliferation. The 1246-3A cell line, which can proliferate in the absence of exogenous insulin, produces in its conditioned medium a growth factor similar to pancreatic insulin by its biological and immunological properties. This factor, called insulin-related factor (IRF), was purified and iodinated to study its binding to cell surface receptors. 125 I-labeled IRF binding to intact 1246-3A cells is lower than to 1246 cells. Cell surface binding can be restored by culturing the 1246-3A cells in the presence of an anti-porcine insulin monoclonal antibody of by acid prewash of the cells prior to performing the binding. Scatchard analysis of binding indicates that IRF secreted by the 1246-3A cells partially occupies high-affinity binding sites on the producer cells. Moreover, insulin monoclonal antibody inhibits the proliferation of the IRF-producing 1246-3A cells, suggesting that these cells are dependent on the secreted IRF for growth in culture. The authors conclude that the insulin-related factor secreted by the insulin-independent 1246-3A cells stimulates their proliferation in an autocrine fashion

  1. In Vitro Differentiation of Human Mesenchymal Stem Cells into Functional Cardiomyocyte-like Cells.

    Science.gov (United States)

    Szaraz, Peter; Gratch, Yarden S; Iqbal, Farwah; Librach, Clifford L

    2017-08-09

    Myocardial infarction and the subsequent ischemic cascade result in the extensive loss of cardiomyocytes, leading to congestive heart failure, the leading cause of mortality worldwide. Mesenchymal stem cells (MSCs) are a promising option for cell-based therapies to replace current, invasive techniques. MSCs can differentiate into mesenchymal lineages, including cardiac cell types, but complete differentiation into functional cells has not yet been achieved. Previous methods of differentiation were based on pharmacological agents or growth factors. However, more physiologically relevant strategies can also enable MSCs to undergo cardiomyogenic transformation. Here, we present a differentiation method using MSC aggregates on cardiomyocyte feeder layers to produce cardiomyocyte-like contracting cells. Human umbilical cord perivascular cells (HUCPVCs) have been shown to have a greater differentiation potential than commonly investigated MSC types, such as bone marrow MSCs (BMSCs). As an ontogenetically younger source, we investigated the cardiomyogenic potential of first-trimester (FTM) HUCPVCs compared to older sources. FTM HUCPVCs are a novel, rich source of MSCs that retain their in utero immunoprivileged properties when cultured in vitro. Using this differentiation protocol, FTM and term HUCPVCs achieved significantly increased cardiomyogenic differentiation compared to BMSCs, as indicated by the increased expression of cardiomyocyte markers (i.e., myocyte enhancer factor 2C, cardiac troponin T, heavy chain cardiac myosin, signal regulatory protein α, and connexin 43). They also maintained significantly lower immunogenicity, as demonstrated by their lower HLA-A expression and higher HLA-G expression. Applying aggregate-based differentiation, FTM HUCPVCs showed increased aggregate formation potential and generated contracting cells clusters within 1 week of co-culture on cardiac feeder layers, becoming the first MSC type to do so. Our results demonstrate that this

  2. Complex Mobile Independent Power Station for Urban Areas

    Science.gov (United States)

    Tunik, A. A.; Tolstoy, M. Y.

    2017-11-01

    A new type of a complex mobile independent power station developed in the Department of Engineering Communications and Life-Support Systems of Irkutsk National Research Technical University, is presented in this article. This station contains only solar panel, wind turbine, accumulator, diesel generator and microbial fuel cell for to produce electric energy, heat pump and solar collector to generate heat energy and also wastewater treatment plant and new complex control system. The complex mobile independent power station is intended for full power supply of a different kind of consumers located even in remote areas thus reducing their dependence from centralized energy supply systems, decrease the fossil fuel consumption, improve the environment of urban areas and solve the problems of the purification of industrial and municipal wastewater.

  3. Unexpected T cell regulatory activity of anti-histone H1 autoantibody: Its mode of action in regulatory T cell-dependent and -independent manners

    Energy Technology Data Exchange (ETDEWEB)

    Takaoka, Yuki [Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima (Japan); Kawamoto, Seiji, E-mail: skawa@hiroshima-u.ac.jp [Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima (Japan); Katayama, Akiko [Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima (Japan); Nakano, Toshiaki [Liver Transplantation Program, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan (China); Yamanaka, Yasushi; Takahashi, Miki [Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima (Japan); Shimada, Yayoi; Chiang, Kuei-Chen [Kazusa Institute for Drug Discovery, Josai International University, Kisarazu (Japan); Ohmori, Naoya [Kazusa Institute for Drug Discovery, Josai International University, Kisarazu (Japan); Faculty of Nursing, Josai International University, Togane (Japan); Aki, Tsunehiro [Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima (Japan); Goto, Takeshi; Sato, Shuji [Kazusa Institute for Drug Discovery, Josai International University, Kisarazu (Japan); Faculty of Nursing, Josai International University, Togane (Japan); Goto, Shigeru [Liver Transplantation Program, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan (China); Iwao Hospital, Yufuin (Japan); Chen, Chao-Long [Liver Transplantation Program, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan (China); Ono, Kazuhisa [Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima (Japan)

    2013-02-08

    Highlights: ► Anti-histone H1 autoantibody (anti-H1) acts on T cells to inhibit their activation. ► Anti-H1 suppresses T cell activation in Treg cell-dependent and -independent manners. ► Suboptimal dose of anti-H1 enhances suppressor function of Treg cells. ► High dose of anti-H1 directly inhibits T cell receptor signaling. -- Abstract: Induction of anti-nuclear antibodies against DNA or histones is a hallmark of autoimmune disorders, but their actual contribution to disease predisposition remains to be clarified. We have previously reported that autoantibodies against histone H1 work as a critical graft survival factor in a rat model of tolerogeneic liver transplantation. Here we show that an immunosuppressive anti-histone H1 monoclonal antibody (anti-H1 mAb) acts directly on T cells to inhibit their activation in response to T cell receptor (TCR) ligation. Intriguingly, the T cell activation inhibitory activity of anti-H1 mAb under suboptimal dosages required regulatory T (Treg) cells, while high dose stimulation with anti-H1 mAb triggered a Treg cell-independent, direct negative regulation of T cell activation upon TCR cross-linking. In the Treg cell-dependent mode of immunosuppressive action, anti-H1 mAb did not induce the expansion of CD4{sup +}Foxp3{sup +} Treg cells, but rather potentiated their regulatory capacity. These results reveal a previously unappreciated T cell regulatory role of anti-H1 autoantibody, whose overproduction is generally thought to be pathogenic in the autoimmune settings.

  4. Unexpected T cell regulatory activity of anti-histone H1 autoantibody: Its mode of action in regulatory T cell-dependent and -independent manners

    International Nuclear Information System (INIS)

    Takaoka, Yuki; Kawamoto, Seiji; Katayama, Akiko; Nakano, Toshiaki; Yamanaka, Yasushi; Takahashi, Miki; Shimada, Yayoi; Chiang, Kuei-Chen; Ohmori, Naoya; Aki, Tsunehiro; Goto, Takeshi; Sato, Shuji; Goto, Shigeru; Chen, Chao-Long; Ono, Kazuhisa

    2013-01-01

    Highlights: ► Anti-histone H1 autoantibody (anti-H1) acts on T cells to inhibit their activation. ► Anti-H1 suppresses T cell activation in Treg cell-dependent and -independent manners. ► Suboptimal dose of anti-H1 enhances suppressor function of Treg cells. ► High dose of anti-H1 directly inhibits T cell receptor signaling. -- Abstract: Induction of anti-nuclear antibodies against DNA or histones is a hallmark of autoimmune disorders, but their actual contribution to disease predisposition remains to be clarified. We have previously reported that autoantibodies against histone H1 work as a critical graft survival factor in a rat model of tolerogeneic liver transplantation. Here we show that an immunosuppressive anti-histone H1 monoclonal antibody (anti-H1 mAb) acts directly on T cells to inhibit their activation in response to T cell receptor (TCR) ligation. Intriguingly, the T cell activation inhibitory activity of anti-H1 mAb under suboptimal dosages required regulatory T (Treg) cells, while high dose stimulation with anti-H1 mAb triggered a Treg cell-independent, direct negative regulation of T cell activation upon TCR cross-linking. In the Treg cell-dependent mode of immunosuppressive action, anti-H1 mAb did not induce the expansion of CD4 + Foxp3 + Treg cells, but rather potentiated their regulatory capacity. These results reveal a previously unappreciated T cell regulatory role of anti-H1 autoantibody, whose overproduction is generally thought to be pathogenic in the autoimmune settings

  5. T-cell-independent immune responses do not require CXC ligand 13-mediated B1 cell migration.

    Science.gov (United States)

    Colombo, Matthew J; Sun, Guizhi; Alugupalli, Kishore R

    2010-09-01

    The dynamic movement of B cells increases the probability of encountering specific antigen and facilitates cell-cell interactions required for mounting a rapid antibody response. B1a and B1b cells are enriched in the coelomic cavity, contribute to T-cell-independent (TI) antibody responses, and increase in number upon antigen exposure. B1 cell movement is largely governed by Cxc ligand 13 (Cxcl13), and mice deficient in this chemokine have a severe reduction in peritoneal B1 cells. In this study, we examined the role of Cxcl13-dependent B cell migration using Borrelia hermsii infection or intraperitoneal immunization with pneumococcal polysaccharide or 4-hydroxy-3-nitrophenyl-acetyl (NP)-Ficoll, all of which induce robust antibody responses from B1b cells. Surprisingly, we found that antibody responses to B. hermsii or to FhbA, an antigenic target of B1b cells, and the resolution of bacteremia were indistinguishable between wild-type and Cxcl13-/- mice. Importantly, we did not observe an expansion of peritoneal B1b cell numbers in Cxcl13-/- mice. Nonetheless, mice that had resolved infection were resistant to reinfection, indicating that the peritoneal B1b cell reservoir is not required for controlling B. hermsii. Furthermore, despite a reduced peritoneal B1b compartment, immunization with pneumococcal polysaccharide vaccine yielded comparable antigen-specific antibody responses in wild-type and Cxcl13-/- mice and conferred protection against Streptococcus pneumoniae. Likewise, immunization with NP-Ficoll elicited similar antibody responses in wild-type and Cxcl13-/- mice. These data demonstrate that homing of B1 cells into the coelomic cavity is not a requirement for generating protective TI antibody responses, even when antigen is initially localized to this anatomical compartment.

  6. Cordycepin-induced apoptosis and autophagy in breast cancer cells are independent of the estrogen receptor

    International Nuclear Information System (INIS)

    Choi, Sunga; Lim, Mi-Hee; Kim, Ki Mo; Jeon, Byeong Hwa; Song, Won O.; Kim, Tae Woong

    2011-01-01

    Cordycepin (3-deoxyadenosine), found in Cordyceps spp., has been known to have many therapeutic effects including immunomodulatory, anti-inflammatory, antimicrobial, and anti-aging effects. Moreover, anti-tumor and anti-metastatic effects of cordycepin have been reported, but the mechanism causing cancer cell death is poorly characterized. The present study was designed to investigate whether the mechanisms of cordycepin-induced cell death were associated with estrogen receptor in breast cancer cells. Exposure of both MDA-MB-231 and MCF-7 human breast cancer cells to cordycepin resulted in dose-responsive inhibition of cell growth and reduction in cell viability. The cordycepin-induced cell death in MDA-MB-231 cells was associated with several specific features of the mitochondria-mediated apoptotic pathway, which was confirmed by DNA fragmentation, TUNEL, and biochemical assays. Cordycepin also caused a dose-dependent increase in mitochondrial translocation of Bax, triggering cytosolic release of cytochrome c and activation of caspases-9 and -3. Interestingly, MCF-7 cells showed autophagy-associated cell death, as observed by the detection of an autophagosome-specific protein and large membranous vacuole ultrastructure morphology in the cytoplasm. Cordycepin-induced autophagic cell death has applications in treating MCF-7 cells with apoptotic defects, irrespective of the ER response. Although autophagy has a survival function in tumorigenesis of some cancer cells, autophagy may be important for cordycepin-induced MCF-7 cell death. In conclusion, the results of our study demonstrate that cordycepin effectively kills MDA-MB-231 and MCF-7 human breast cancer cell lines in culture. Hence, further studies should be conducted to determine whether cordycepin will be a clinically useful, ER-independent, chemotherapeutic agent for human breast cancer. -- Highlights: ► We studied the mechanism which cordycepin-induced cell death association with estrogen receptor (ER) in

  7. Putative porcine embryonic stem cell lines derived from aggregated four-celled cloned embryos produced by oocyte bisection cloning.

    Science.gov (United States)

    Siriboon, Chawalit; Lin, Yu-Hsuan; Kere, Michel; Chen, Chun-Da; Chen, Lih-Ren; Chen, Chien-Hong; Tu, Ching-Fu; Lo, Neng-Wen; Ju, Jyh-Cherng

    2015-01-01

    We attempted to isolate ES cell lines using inner cell masses from high-quality cloned porcine blastocysts. After being seeded onto feeders, embryos had better (P cloned embryos (62.8, 42.6 and 12.8% vs. 76.2, 55.2 and 26.2%, respectively) compared to the non-aggregated group (41.6, 23.4 and 3.9%). Effects of feeder types (STO vs. MEF) and serum sources (FBS vs. KSR) on extraction of cloned embryo-derived porcine ES cells were examined. More (17.1%) ntES cell lines over Passage 3 were generated in the MEF/KSR group. However, ntES cells cultured in KSR-supplemented medium had a low proliferation rate with defective morphology, and eventually underwent differentiation or apoptosis subsequently. Approximately 26.1, 22.7 and 35.7% of primary colonies were formed after plating embryos in DMEM, DMEM/F12 and α-MEM media, respectively. Survival rates of ntES cells cultured in α-MEM, DMEM and DMEM/F12 were 16.7, 4.3 and 6.8%, respectively (P > 0.05). We further examined the beneficial effect of TSA treatment of 3× aggregated cloned embryos on establishment of ntES cell lines. Primary colony numbers and survival rates of ntES cells beyond passage 3 were higher (P cells, remaining undifferentiated over 25 passages, had alkaline phosphatase activity and expressed ES specific markers Oct4, Nanog, Sox2, and Rex01. Moreover, these ntES cells successfully differentiated into embryoid bodies (EBs) that expressed specific genes of all three germ layers after being cultured in LIF-free medium. In conclusion, we have successfully derived putative porcine ntES cells with high efficiency from quality cloned embryos produced by embryo aggregation, and optimized the ES cell culture system suitable for establishing and maintaining ntES cell lines in undifferentiated state.

  8. A New Optimization Framework To Solve The Optimal Feeder Reconfiguration And Capacitor Placement Problems

    Directory of Open Access Journals (Sweden)

    Mohammad-Reza Askari

    2015-07-01

    Full Text Available Abstract This paper introduces a new stochastic optimization framework based bat algorithm BA to solve the optimal distribution feeder reconfiguration DFR as well as the shunt capacitor placement and sizing in the distribution systems. The objective functions to be investigated are minimization of the active power losses and minimization of the total network costs an. In order to consider the uncertainties of the active and reactive loads in the problem point estimate method PEM with 2m scheme is employed as the stochastic tool. The feasibility and good performance of the proposed method are examined on the IEEE 69-bus test system.

  9. Generating a non-integrating human induced pluripotent stem cell bank from urine-derived cells.

    Directory of Open Access Journals (Sweden)

    Yanting Xue

    Full Text Available Induced pluripotent stem cell (iPS cell holds great potential for applications in regenerative medicine, drug discovery, and disease modeling. We describe here a practical method to generate human iPS cells from urine-derived cells (UCs under feeder-free, virus-free, serum-free condition and without oncogene c-MYC. We showed that this approach could be applied in a large population with different genetic backgrounds. UCs are easily accessible and exhibit high reprogramming efficiency, offering advantages over other cell types used for the purpose of iPS generation. Using the approach described in this study, we have generated 93 iPS cell lines from 20 donors with diverse genetic backgrounds. The non-viral iPS cell bank with these cell lines provides a valuable resource for iPS cells research, facilitating future applications of human iPS cells.

  10. Definition and dynamic control of a continuous chromatography process independent of cell culture titer and impurities.

    Science.gov (United States)

    Chmielowski, Rebecca A; Mathiasson, Linda; Blom, Hans; Go, Daniel; Ehring, Hanno; Khan, Heera; Li, Hong; Cutler, Collette; Lacki, Karol; Tugcu, Nihal; Roush, David

    2017-12-01

    Advances in cell culture technology have enabled the production of antibody titers upwards of 30g/L. These highly productive cell culture systems can potentially lead to productivity bottlenecks in downstream purification due to lower column loadings, especially in the primary capture chromatography step. Alternative chromatography solutions to help remedy this bottleneck include the utilization of continuous processing systems such as periodic counter-current chromatography (PCC). Recent studies have provided methods to optimize and improve the design of PCC for cell culture titers up to about 3g/L. This paper defines a continuous loading strategy for PCC that is independent of cell culture background and encompasses cell culture titers up to about 31g/L. Initial experimentation showed a challenge with determining a difference in change in UV280nm signal (ie. ΔUV) between cell culture feed and monoclonal antibody (mAb) concentration. Further investigation revealed UV280nm absorbance of the cell culture feedstock without antibody was outside of the linear range of detection for a given cell pathlength. Additional experimentation showed the difference in ΔUV for various cell culture feeds can be either theoretically predicted by Beer's Law given a known absorbance of the media background and impurities or experimentally determined using various UV280nm cell pathlengths. Based on these results, a 0.35mm pathlength at UV280nm was chosen for dynamic control to overcome the background signal. The pore diffusion model showed good agreement with the experimental frontal analysis data, which resulted in definition of a ΔUV setpoint range between 20 and 70% for 3C-PCC experiments. Product quality of the elution pools was acceptable between various cell culture feeds and titers up to about 41g/L. Results indicated the following ΔUV setpoints to achieve robust dynamic control and maintain 3C-PCC yield: ∼20-45% for titers greater than 10g/L depending on UV absorbance of

  11. Hawaiian Electric Advanced Inverter Grid Support Function Laboratory Validation and Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Nelson, Austin [National Renewable Energy Lab. (NREL), Golden, CO (United States); Nagarajan, Adarsh [National Renewable Energy Lab. (NREL), Golden, CO (United States); Prabakar, Kumar [National Renewable Energy Lab. (NREL), Golden, CO (United States); Gevorgian, Vahan [National Renewable Energy Lab. (NREL), Golden, CO (United States); Lundstrom, Blake [National Renewable Energy Lab. (NREL), Golden, CO (United States); Nepal, Shaili [National Renewable Energy Lab. (NREL), Golden, CO (United States); Hoke, Anderson [National Renewable Energy Lab. (NREL), Golden, CO (United States); Asano, Marc [Hawaiian Electric Company, Honolulu, HI (United States); Ueda, Reid [Hawaiian Electric Company, Honolulu, HI (United States); Shindo, Jon [Hawaiian Electric Company, Honolulu, HI (United States); Kubojiri, Kandice [Hawaiian Electric Company, Honolulu, HI (United States); Ceria, Riley [Hawaiian Electric Company, Honolulu, HI (United States); Ifuku, Earle [Hawaiian Electric Company, Honolulu, HI (United States)

    2016-12-01

    The objective for this test plan was to better understand how to utilize the performance capabilities of advanced inverter functions to allow the interconnection of distributed energy resource (DER) systems to support the new Customer Self-Supply, Customer Grid-Supply, and other future DER programs. The purpose of this project was: 1) to characterize how the tested grid supportive inverters performed the functions of interest, 2) to evaluate the grid supportive inverters in an environment that emulates the dynamics of O'ahu's electrical distribution system, and 3) to gain insight into the benefits of the grid support functions on selected O'ahu island distribution feeders. These goals were achieved through laboratory testing of photovoltaic inverters, including power hardware-in-the-loop testing.

  12. Eddy current array probe for detection of surface breaking cracks in the extrados of feeder bends

    International Nuclear Information System (INIS)

    Obrutsky, L.S.; Cassidy, R.A.; Chaplin, K.; Martin, P.; Bureau, J.F.

    2006-01-01

    A new eddy current array probe has been implemented as a straightforward and promising technique for detection of outer diameter (OD) surface-breaking cracks on the extrados of feeder bends. The design is based on previous work performed at AECL, which had demonstrated that eddy current probes with laterally displaced transmit-receive coils can overcome some of the limitations of inspecting ferritic steel components for surface-breaking cracks. The Feeder Integrity Joint Program-CANDU Owners Group Inc. (FIJP-COG) Non-Destructive Evaluation (NDE) Team members commissioned AECL to work in collaboration with the probe manufacturer ZETEC, to develop a field usable eddy current array probe. The objective was to acquire a technique with the following capabilities: fast scanning non-contact inspection technique for surface breaking discontinuities; full inspection of the bend extrados OD surface in a single scan; ability to inspect first and second bends with similar settings and capabilities; permanent record for future reference; axial and circumferential crack detection in a single scan; capability to detect OD surface-breaking cracks, which can provide additional information to that provided by ultrasonic testing (UT) for flaw characterization, and detection threshold: Surface breaking cracks equivalent to a 0.5 mm deep, 10 mm long EDM notch located on the OD of the bend extrados. This paper discusses the basis for probe design, summarizes the experimental work to evaluate probe capabilities and analyzes the results from the field trial. (author)

  13. Eddy current array probe for detection of surface breaking cracks in the extrados of feeder bends

    Energy Technology Data Exchange (ETDEWEB)

    Obrutsky, L.S.; Cassidy, R.A.; Chaplin, K. [Atomic Energy of Canada Limited, Chalk River, Ontario (Canada)]. E-mail: obrutskyl@aecl.ca; Martin, P. [NB Power, Point Lepreau NGS, Point Lepreau, New Brunswick (Canada)]. E-mail: PMartin@nbpower.com; Bureau, J.F. [Zetec, Quebec, Quebec (Canada)]. E-mail: jean-francois.bureau@zetec.com

    2006-07-01

    A new eddy current array probe has been implemented as a straightforward and promising technique for detection of outer diameter (OD) surface-breaking cracks on the extrados of feeder bends. The design is based on previous work performed at AECL, which had demonstrated that eddy current probes with laterally displaced transmit-receive coils can overcome some of the limitations of inspecting ferritic steel components for surface-breaking cracks. The Feeder Integrity Joint Program-CANDU Owners Group Inc. (FIJP-COG) Non-Destructive Evaluation (NDE) Team members commissioned AECL to work in collaboration with the probe manufacturer ZETEC, to develop a field usable eddy current array probe. The objective was to acquire a technique with the following capabilities: fast scanning non-contact inspection technique for surface breaking discontinuities; full inspection of the bend extrados OD surface in a single scan; ability to inspect first and second bends with similar settings and capabilities; permanent record for future reference; axial and circumferential crack detection in a single scan; capability to detect OD surface-breaking cracks, which can provide additional information to that provided by ultrasonic testing (UT) for flaw characterization, and detection threshold: Surface breaking cracks equivalent to a 0.5 mm deep, 10 mm long EDM notch located on the OD of the bend extrados. This paper discusses the basis for probe design, summarizes the experimental work to evaluate probe capabilities and analyzes the results from the field trial. (author)

  14. SIRT1 overexpression is an independent prognosticator for patients with esophageal squamous cell carcinoma.

    Science.gov (United States)

    Ma, Ming-Chun; Chiu, Tai-Jan; Lu, Hung-I; Huang, Wan-Ting; Lo, Chien-Ming; Tien, Wan-Yu; Lan, Ya-Chun; Chen, Yen-Yang; Chen, Chang-Han; Li, Shau-Hsuan

    2018-04-10

    Sirtuin 1 (SIRT1) regulates DNA repair and metabolism by deacetylating target proteins. SIRT1 may be oncogenic because its overexpression has been detected in many cancers. The aim of the present study was to clarify the prognostic role of SIRT1 in patients with esophageal squamous cell carcinoma (ESCC) and evaluate the effect of SIRT1 inhibitor in vitro. The expression of SIRT1 was evaluated immunohistochemically in 155 surgically resected ESCC and the staining results were evaluated semiquantitatively by the Immunoreactive Scoring System. The clinical features and treatment outcome were analyzed. The effect of SIRT1 inhibitor, SIRT 1 inhibitor IV, (S)-35, was investigated in vitro on ESCC cell lines. The expression of SIRT1 on ESCC did not correlate with age, gender, tumor location, stage, T classification, N classification, surgical margin or histology. Univariate analysis showed that SIRT1 overexpression was associated with inferior overall survival (P = 0.004) and disease-free survival (P = 0.004). In multivariate comparison, SIRT1 overexpression remained independently associated with worse overall survival (P = 0.009, hazard ratio = 1.776) and disease-free survival (P = 0.017, hazard ratio = 1.642). In cell lines, SIRT1 inhibitor inhibited ESCC growth. Our study suggests that SIRT1 overexpression is an independent prognosticator for patients with ESCC and the SIRT1 inhibitor suppressed cell proliferation of ESCC cell lines. Our findings suggest that inhibition of SIRT1 signaling may be a promising novel target for ESCC.

  15. A Review of Power Distribution Test Feeders in the United States and the Need for Synthetic Representative Networks

    Directory of Open Access Journals (Sweden)

    Fernando E. Postigo Marcos

    2017-11-01

    Full Text Available Under the increasing penetration of distributed energy resources and new smart network technologies, distribution utilities face new challenges and opportunities to ensure reliable operations, manage service quality, and reduce operational and investment costs. Simultaneously, the research community is developing algorithms for advanced controls and distribution automation that can help to address some of these challenges. However, there is a shortage of realistic test systems that are publically available for development, testing, and evaluation of such new algorithms. Concerns around revealing critical infrastructure details and customer privacy have severely limited the number of actual networks published and that are available for testing. In recent decades, several distribution test feeders and US-featured representative networks have been published, but the scale, complexity, and control data vary widely. This paper presents a first-of-a-kind structured literature review of published distribution test networks with a special emphasis on classifying their main characteristics and identifying the types of studies for which they have been used. This both aids researchers in choosing suitable test networks for their needs and highlights the opportunities and directions for further test system development. In particular, we highlight the need for building large-scale synthetic networks to overcome the identified drawbacks of current distribution test feeders.

  16. Rapid and sustained CD4(+) T-cell-independent immunity from adenovirus-encoded vaccine antigens

    DEFF Research Database (Denmark)

    Holst, Peter J; Bartholdy, Christina; Buus, Anette Stryhn

    2007-01-01

    -linked lymphocytic choriomeningitis virus (LCMV)-derived epitopes was long-lived and protective. Notably, in contrast to full-length protein, the response elicited with the beta(2)-microglobulin-linked LCMV-derived epitope was CD4(+) T-cell independent. Furthermore, virus-specific CD8(+) T cells primed...... in the absence of CD4(+) T-cell help were sustained in the long term and able to expand and control a secondary challenge with LCMV. Our results demonstrate that modifications to the antigen used in adenovirus vaccines may be used to improve the induced T-cell response. Such a strategy for CD4(+) T-cell...... to that elicited with an adenovirus-encoded minimal epitope covalently linked to beta(2)-microglobulin. We demonstrate that the beta(2)-microglobulin-linked epitope induced an accelerated and augmented CD8(+) T-cell response. Furthermore, the immunity conferred by vaccination with beta(2)-microglobulin...

  17. NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells

    Directory of Open Access Journals (Sweden)

    Jennifer Ann Foltz

    2016-11-01

    Full Text Available Canines spontaneously develop many cancers similar to humans - including osteosarcoma, leukemia, and lymphoma - offering the opportunity to study immune therapies in a genetically heterogeneous and immunocompetent environment. However, a lack of antibodies recognizing canine NK cell markers has resulted in suboptimal characterization and unknown purity of NK cell products, hindering the development of canine models of NK cell adoptive immunotherapy. To this end, we generated a novel antibody to canine NCR1 (NKp46, the putative species-wide marker of NK cells, enabling purification of NK cells for further characterization. We demonstrate that CD3-/NKp46+ cells in healthy and osteosarcoma-bearing canines have phenotypic similarity to human CD3-/NKp46+ NK cells, expressing mRNA for CD16 and the natural cytotoxicity receptors NKp30, NKp44, and NKp80. Functionally, we demonstrate with the calcein release assay that canine CD3-/NKp46+ cells kill canine tumor cell lines without prior sensitization and secrete IFN-γ, TNF-α, IL-8, IL-10, and GM-CSF as measured by Luminex. Like human NK cells, CD3-/NKp46+ cells expand rapidly on feeder cells expressing 4-1BBL and membrane-bound IL-21 (median= 20,283-fold in 21 days. Further, we identify a minor Null population (CD3-/CD21-/CD14-/NKp46- with reduced cytotoxicity against osteosarcoma cells, but similar cytokine secretion as CD3-/NKp46+ cells. Null cells in canines and humans have reduced expression of NKG2D, NKp44, and CD16 compared to NKp46+ NK cells, and can be induced to express NKp46 with further expansion on feeder cells. In conclusion, we have identified and characterized canine NK cells, including an NKp46- subset of canine and human NK cells, using a novel anti-canine NKp46 antibody, and report robust ex vivo expansion of canine NK cells sufficient for adoptive immunotherapy.

  18. Granzyme A Cleaves a Mitochondrial Complex I Protein to Initiate Caspase-Independent Cell Death

    Science.gov (United States)

    Martinvalet, Denis; Dykxhoorn, Derek M.; Ferrini, Roger; Lieberman, Judy

    2010-01-01

    SUMMARY The killer lymphocyte protease granzyme A (GzmA) triggers caspase-independent target cell death with morphological features of apoptosis. We previously showed that GzmA acts directly on mitochondria to generate reactive oxygen species (ROS) and disrupt the transmembrane potential (ΔΨm) but does not permeabilize the mitochondrial outer membrane. Mitochondrial damage is critical to GzmA-induced cell death since cells treated with superoxide scavengers are resistant to GzmA. Here we find that GzmA accesses the mitochondrial matrix to cleave the complex I protein NDUFS3, an iron-sulfur subunit of the NADH:ubiquinone oxidoreductase complex I, after Lys56 to interfere with NADH oxidation and generate superoxide anions. Target cells expressing a cleavage site mutant of NDUFS3 are resistant to GzmA-mediated cell death but remain sensitive to GzmB. PMID:18485875

  19. CD34-positive cells as stem cell support after high dose therapy

    International Nuclear Information System (INIS)

    Kvalheim, G.; Pharo, A.; Holte, H.

    1996-01-01

    Six patients, five with breast cancer and one with non-Hodgkin's lymphoma, were mobilized by chemotherapy and G-CSF. CD34-positive cells were isolated by means of immunomagnetic beads and Isolex 300 Cell Separator. Mean purity of isolated CD34-positive cells was 97% and mean yield was 54%. Three patients were treated with high dose therapy followed by reinfusion of CD34-positive cells as stem cell support. Recovery of neutrophils occurred at day 8, 11 and 13 and of platelets at day 9, 14 and 32. It is concluded that immunomagnetic isolated CD34-positive cells give high purity and yield. Although use of CD34-positive cells reduces the content of contaminating tumours cells in the graft, breast cancer cells were still detectable in two out of five CD34-positive cell products. 20 refs., 2 figs., 1 tab

  20. From steep feeders to tabular plutons - Emplacement controls of syntectonic granitoid plutons in the Damara Belt, Namibia

    Science.gov (United States)

    Hall, Duncan; Kisters, Alexander

    2016-01-01

    Granitoid plutons in the deeply eroded south Central Zone of the Damara Belt in Namibia commonly show tabular geometries and pronounced stratigraphic controls on their emplacement. Subhorizontal, sheet-like pluton geometries record emplacement during regional subhorizontal shortening, but the intrusion of spatially and temporally closely-related granitoid plutons at different structural levels and in distinct structural settings suggests independent controls on their levels of emplacement. We describe and evaluate the controls on the loci of the dyke-to-sill transition that initiated the emplacement of three syntectonic (560-530 Ma) plutons in the basement-cover stratigraphy of the Erongo region. Intrusive relationships highlight the significance of (1) rigidity anisotropies associated with competent sedimentary packages or pre-existing subhorizontal granite sheets and (2) rheological anisotropies associated with the presence of thick ductile marble horizons. These mechanical anisotropies may lead to the initial deflection of steep feeder conduits as well as subsequent pluton assembly by the repeated underaccretion of later magma batches. The upward displacement of regional isotherms due to the heat advection associated with granite emplacement is likely to have a profound effect on the mechanical stratification of the upper crust and, consequently, on the level at which granitoid pluton emplacement is initiated. In this way, pluton emplacement at progressively shallower crustal depths may have resulted in the unusually high apparent geothermal gradients recorded in the upper crustal levels of the Damara Belt during its later evolution.

  1. Ecdysone-dependent and ecdysone-independent programmed cell death in the developing optic lobe of Drosophila.

    Science.gov (United States)

    Hara, Yusuke; Hirai, Keiichiro; Togane, Yu; Akagawa, Hiromi; Iwabuchi, Kikuo; Tsujimura, Hidenobu

    2013-02-01

    The adult optic lobe of Drosophila develops from the primordium during metamorphosis from mid-3rd larval stage to adult. Many cells die during development of the optic lobe with a peak of the number of dying cells at 24 h after puparium formation (h APF). Dying cells were observed in spatio-temporal specific clusters. Here, we analyzed the function of a component of the insect steroid hormone receptor, EcR, in this cell death. We examined expression patterns of two EcR isoforms, EcR-A and EcR-B1, in the optic lobe. Expression of each isoform altered during development in isoform-specific manner. EcR-B1 was not expressed in optic lobe neurons from 0 to 6h APF, but was expressed between 9 and 48 h APF and then disappeared by 60 h APF. In each cortex, its expression was stronger in older glia-ensheathed neurons than in younger ones. EcR-B1 was also expressed in some types of glia. EcR-A was expressed in optic lobe neurons and many types of glia from 0 to 60 h APF in a different pattern from EcR-B1. Then, we genetically analyzed EcR function in the optic lobe cell death. At 0 h APF, the optic lobe cell death was independent of any EcR isoforms. In contrast, EcR-B1 was required for most optic lobe cell death after 24 h APF. It was suggested that cell death cell-autonomously required EcR-B1 expressed after puparium formation. βFTZ-F1 was also involved in cell death in many dying-cell clusters, but not in some of them at 24 h APF. Altogether, the optic lobe cell death occurred in ecdysone-independent manner at prepupal stage and ecdysone-dependent manner after 24 h APF. The acquisition of ecdysone-dependence was not directly correlated with the initiation or increase of EcR-B1 expression. Copyright © 2012 Elsevier Inc. All rights reserved.

  2. The Obligations on Government and Society in our Constitutional State to Respect and Support Independent Constitutional Structures

    Directory of Open Access Journals (Sweden)

    LWH Ackermann

    2000-05-01

    these institutions is vitally important for creating and sustaining an ethos of constitutionalism among the inhabitants of the Republic.The Constitution makes explicit provision for the protection of the judiciary and the other independent state institutions and thereby indirectly for the development of habits of constitutionalism. The constitutional protection and support given to the independent state institutions are very similar to that given to the courts. One important distinction is to be noted. In the case of the courts, the Constitution provides that they "are subject only to the Constitution and the law" and no provision is made for them to be accountable to any other organ of state or any other institution or person, for that matter. By contrast, the independent institutions envisaged in section 181 of the Constitution are expressly made accountable to the National Assembly and are obliged to report on their activities and the performance of their functions to the Assembly at least once a year. While the formal independence of state institutions may at all times be scrupulously recognised by the legislature and the executive, their substantive independence can easily be undermined by fiscal starvation and their ability to function properly impeded by bureaucratic administrative obstruction or obfuscation or even, quite innocently, by a lack of appreciation of what the Constitution demands from public administration in support of these institutions. Adequate financial and administrative resources are required to achieve aspects of judicial independence. All South Africans must still be vigilant to ensure, from the outset, that all state organs develop habits and practices of constitutionalism and that they do not, whether by omission, error, or otherwise endanger the independence of our independent state institutions by neglecting their constitutional obligations.For the Constitutional Court to fulfil its role as the ultimate guardian of the Constitution, it

  3. Expression of death receptor 4 induces caspase-independent cell death in MMS-treated yeast.

    Science.gov (United States)

    Kang, Mi-Sun; Lee, Sung-Keun; Park, Chang-Shin; Kang, Ju-Hee; Bae, Sung-Ho; Yu, Sung-Lim

    2008-11-14

    DR4, a tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor, is a key element in the extrinsic pathway of TRAIL/TRAIL receptor-related apoptosis that exerts a preferential toxic effect against tumor cells. However, TRAIL and DR4 are expressed in various normal cells, and recent studies indicate that DR4 has a number of non-apoptotic functions. In this study, we evaluated the effects of human DR4 expression in yeast to determine the function of DR4 in normal cells. The expression of DR4 in yeast caused G1 arrest, which resulted in transient growth inhibition. Moreover, treatment of DR4-expressing yeast with a DNA damaging agent, MMS, elicited drastic, and sustained cell growth inhibition accompanied with massive apoptotic cell death. Further analysis revealed that cell death in the presence of DNA damage and DR4 expression was not dependent on the yeast caspase, YCA1. Taken together, these results indicate that DR4 triggers caspase-independent programmed cell death during the response of normal cells to DNA damage.

  4. Cordycepin-induced apoptosis and autophagy in breast cancer cells are independent of the estrogen receptor

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Sunga [Department of Physiology, School of Medicine, Chungnam National University, Daejeon, 301747 (Korea, Republic of); Lim, Mi-Hee [Department of Biochemistry, Kangwon National University, Gangwon-do, 200701 (Korea, Republic of); Kim, Ki Mo [Diabetic Complications Research Center, Division of Traditional Korean Medicine (TKM) Integrated Research, Korea Institute of Oriental Medicine (KIOM), 305811, Daejeon (Korea, Republic of); Jeon, Byeong Hwa [Department of Physiology, School of Medicine, Chungnam National University, Daejeon, 301747 (Korea, Republic of); Song, Won O. [Department of Food Science and Human Nutrition, Michigan State University, East Lansing, MI 48824 (United States); Kim, Tae Woong, E-mail: tawkim@kangwon.ac.kr [Department of Biochemistry, Kangwon National University, Gangwon-do, 200701 (Korea, Republic of)

    2011-12-15

    Cordycepin (3-deoxyadenosine), found in Cordyceps spp., has been known to have many therapeutic effects including immunomodulatory, anti-inflammatory, antimicrobial, and anti-aging effects. Moreover, anti-tumor and anti-metastatic effects of cordycepin have been reported, but the mechanism causing cancer cell death is poorly characterized. The present study was designed to investigate whether the mechanisms of cordycepin-induced cell death were associated with estrogen receptor in breast cancer cells. Exposure of both MDA-MB-231 and MCF-7 human breast cancer cells to cordycepin resulted in dose-responsive inhibition of cell growth and reduction in cell viability. The cordycepin-induced cell death in MDA-MB-231 cells was associated with several specific features of the mitochondria-mediated apoptotic pathway, which was confirmed by DNA fragmentation, TUNEL, and biochemical assays. Cordycepin also caused a dose-dependent increase in mitochondrial translocation of Bax, triggering cytosolic release of cytochrome c and activation of caspases-9 and -3. Interestingly, MCF-7 cells showed autophagy-associated cell death, as observed by the detection of an autophagosome-specific protein and large membranous vacuole ultrastructure morphology in the cytoplasm. Cordycepin-induced autophagic cell death has applications in treating MCF-7 cells with apoptotic defects, irrespective of the ER response. Although autophagy has a survival function in tumorigenesis of some cancer cells, autophagy may be important for cordycepin-induced MCF-7 cell death. In conclusion, the results of our study demonstrate that cordycepin effectively kills MDA-MB-231 and MCF-7 human breast cancer cell lines in culture. Hence, further studies should be conducted to determine whether cordycepin will be a clinically useful, ER-independent, chemotherapeutic agent for human breast cancer. -- Highlights: Black-Right-Pointing-Pointer We studied the mechanism which cordycepin-induced cell death association with

  5. Intermittent Hypoxia Disrupts Glucose Homeostasis in Liver Cells in an Insulin-Dependent and Independent Manner

    Directory of Open Access Journals (Sweden)

    Chen Juan Gu

    2018-05-01

    Full Text Available Background/Aims: Obstructive sleep apnea is associated with diabetes and insulin resistance, but the underlying mechanisms remain unclear. The purpose of the current study was to determine the molecular effects of intermittent hypoxia (IH on hepatic insulin signaling and glucose homeostasis, and whether c-Jun NH2-terminal-kinase (JNK contributed to metabolic responses to IH in liver cells. Methods: The human HepG2 cells and rat FAO cells were exposed to 10, 30, 120, 240 or 360 cycles of IH (1% O2 for 60 s followed by 21% O2 for 60s, 7.5 cycles per hour or normoxia as a control. In a subgroup, we exposed cells to 360 cycles of IH with the JNK inhibitor SP600125. After IH exposure, cell glycogen content and glucose output were measured using colorimetric assay kits. Canonical insulin signaling and gluconeogenic genes were measured by western blot and quantitative polymerase chain reaction. Results: IH decreased insulin-stimulated protein kinase B (AKT/glycogen synthase kinase-3β (GSK-3β phosphorylation in a time-dependent manner, while inhibiting forkhead box protein O1 (FOXO1 expression and phosphoenolpyruvate carboxykinase (PEPCK transcription independent of insulin signaling. JNK inhibitor SP600125 partially restored AKT/ GSK-3β phosphorylation and glycogen synthesis, but did not affect other IH-induced glucose metabolic changes. Conclusion: IH in vitro impaired insulin signal transduction in liver cells as assessed by inhibited AKT/GSK-3β phosphorylation via JNK activation. IH inhibited FOXO1 and gluconeogenesis in an insulin-independent manner.

  6. Effect of feeders in 3D modeling of low impedance multilayer CPW transmission line

    Science.gov (United States)

    Zaini, R. I.; Kyabaggu, P. B. K.; Sinulingga, E. P.

    2018-02-01

    Improved characteristics with low dissipation loss MMICs are highly desirable for wireless communications. However, the current industrial MMIC design is mainly based on microstrip concept which suffered from parasitic and unwanted phenomenon especially at higher frequency (>20 GHz). On the other hand, for future wireless technology, higher frequency operation is required and on-wafer microwave characterizations as well as precise modeling of 3D Multilayer CPW components are vital. This project concerns with understanding of the microwave characteristics behavior of Multilayer CPW components in MMIC applications. Feeder effect as unwanted parts in the characteristics has been investigated to determine its relation with the half wavelength resonance of the Multilayer CPW Low Impedance Transmission Line.

  7. UVC-induced apoptosis in Dubca cells is independent of JNK activation and p53Ser-15 phosphorylation

    International Nuclear Information System (INIS)

    Chathoth, Shahanas; Thayyullathil, Faisal; Hago, Abdulkader; Shahin, Allen; Patel, Mahendra; Galadari, Sehamuddin

    2009-01-01

    Ultraviolet C (UVC) irradiation in mammalian cell lines activates a complex signaling network that leads to apoptosis. By using Dubca cells as a model system, we report the presence of a UVC-induced apoptotic pathway that is independent of c-Jun N-terminal kinases (JNKs) activation and p53 phosphorylation at Ser 15 . Irradiation of Dubca cells with UVC results in a rapid JNK activation and phosphorylation of its downstream target c-Jun, as well as, phosphorylation of activating transcription factor 2 (ATF2). Pre-treatment with JNK inhibitor, SP600125, inhibited UVC-induced c-Jun phosphorylation without preventing UVC-induced apoptosis. Similarly, inhibition of UVC-induced p53 phosphorylation did not prevent Dubca cell apoptosis, suggesting that p53 Ser-15 phosphorylation is not associated with UVC-induced apoptosis signaling. The pan-caspase inhibitor z-VAD-fmk inhibited UVC-induced PARP cleavage, DNA fragmentation, and ultimately apoptosis of Dubca cells. Altogether, our study clearly indicates that UVC-induced apoptosis is independent of JNK and p53 activation in Dubca cells, rather, it is mediated through a caspase dependent pathway. Our findings are not in line with the ascribed critical role for JNKs activation, and downstream phosphorylation of targets such as c-Jun and ATF2 in UVC-induced apoptosis.

  8. Effects of catalyst-support materials on the performance of fuel cells

    CSIR Research Space (South Africa)

    Ejikeme, PM

    2016-07-01

    Full Text Available The operating life of a fuel cell is expected to be thousands of hours. One of the critical components of the fuel cell that will allow for such long-life cycle is the catalyst-support material. The support material is expected, amongst others...

  9. Independent Activation of Hepatitis B Virus Biosynthesis by Retinoids, Peroxisome Proliferators, and Bile Acids

    Science.gov (United States)

    Reese, Vanessa C.; Oropeza, Claudia E.

    2013-01-01

    In the human hepatoma cell line HepG2, retinoic acid, clofibric acid, and bile acid treatment can only modestly increase hepatitis B virus (HBV) biosynthesis. Utilizing the human embryonic kidney cell line 293T, it was possible to demonstrate that the retinoid X receptor α (RXRα) plus its ligand can support viral biosynthesis independently of additional nuclear receptors. In addition, RXRα/peroxisome proliferator-activated receptor α (PPARα) and RXRα/farnesoid X receptor α (FXRα) heterodimeric nuclear receptors can also mediate ligand-dependent HBV transcription and replication when activated by clofibric acid and bile acid, respectively, independently of a requirement for the ligand-dependent activation of RXRα. These observations indicate that there are at least three possible modes of ligand-mediated activation of HBV transcription and replication existing within hepatocytes, suggesting that multiple independent mechanisms control viral production in the livers of infected individuals. PMID:23135717

  10. (−)-Xanthatin Selectively Induces GADD45γ and Stimulates Caspase-Independent Cell Death in Human Breast Cancer MDA-MB-231 Cells

    Science.gov (United States)

    Takeda, Shuso; Matsuo, Kazumasa; Yaji, Kentaro; Okajima-Miyazaki, Shunsuke; Harada, Mari; Miyoshi, Hiroko; Okamoto, Yoshiko; Amamoto, Toshiaki; Shindo, Mitsuru; Omiecinski, Curtis J.; Aramaki, Hironori

    2014-01-01

    exo-Methylene lactone group-containing compounds, such as (−)-xanthatin, are present in a large variety of biologically active natural products, including extracts of Xanthium strumarium (Cocklebur). These substances are reported to possess diverse functional activities, exhibiting anti-inflammatory, antimalarial, and anticancer potential. In this study, we synthesized six structurally related xanthanolides containing exo-methylene lactone moieties, including (−)-xanthatin and (+)-8-epi-xanthatin, and examined the effects of these chemically defined substances on the highly aggressive and farnesyltransferase inhibitor (FTI)-resistant MDA-MB-231 cancer cell line. The results obtained demonstrate that (−)-xanthatin was a highly effective inhibitor of MDA-MB-231 cell growth, inducing caspase-independent cell death, and that these effects were independent of FTase inhibition. Further, our results show that among the GADD45 isoforms, GADD45γ was selectively induced by (−)-xanthatin and that GADD45γ-primed JNK and p38 signaling pathways are, at least in part, involved in mediating the growth inhibition and potential anticancer activities of this agent. Given that GADD45γ is becoming increasingly recognized for its tumor suppressor function, the results presented here suggest the novel possibility that (−)-xanthatin may have therapeutic value as a selective inducer of GADD45γ in human cancer cells, in particular in FTI-resistant aggressive breast cancers. PMID:21568272

  11. Support structure concept for integration of ITER diagnostics in the port cell

    Energy Technology Data Exchange (ETDEWEB)

    Udintsev, V.S., E-mail: victor.udintsev@iter.org [ITER Organization, Route de Vinon sur Verdon, 13115 St. Paul-Lez-Durance (France); Portalès, M.; Giacomin, T.; Darcourt, O.; Direz, M.-F.; Martins, J.P.; Penot, C.; Arumugam, A.P.; Drevon, J.-M.; Friconneau, J.P.; Levesy, B.; Maquet, P.; Patel, K.M.; Pitcher, C.S. [ITER Organization, Route de Vinon sur Verdon, 13115 St. Paul-Lez-Durance (France); Popova, E. [Russian Federation Domestic Agency, Moscow (Russian Federation); Proust, M. [ITER Organization, Route de Vinon sur Verdon, 13115 St. Paul-Lez-Durance (France); Ronden, D.M.S. [DIFFER, Nieuwegein (Netherlands); Walker, C.I.; Walsh, M.J.; Watts, C. [ITER Organization, Route de Vinon sur Verdon, 13115 St. Paul-Lez-Durance (France)

    2013-10-15

    Highlights: ► An interspace support structure to support the diagnostic systems from the back of the upper and equatorial port plugs to the biological shield plug. ► Port cell support structures are foreseen to handle the equipment in the port cell. ► Both ISS and PCSS will be supported by means of RH rail system. ► The structures will be positioned with a certain tolerance. ► The proposed concepts are found to fulfil the needs for support of the diagnostics in ITER. -- Abstract: Development of the diagnostics for ITER tokamak, which is presently under construction by several international partners at Cadarache in France, is a major challenge because of severe environment, strict engineering requirements, and the need for high reliability in the measurements. The diagnostic systems in the upper, equatorial and lower port cells on ITER are designed to be integrated within the interspace and port cell support structures. These structures are interfacing with remote handling rail system for the cask operations, thus facilitating the removal and installation of the diagnostics in the port and hence minimizing time for working close to the tokamak. In this paper, the challenges associated with the integration of the diagnostics in the port interspace and port cell, as well as their solutions will be addressed and presented. The interspace and the port cell support structures, as well as their interfaces with the biological shield, will be discussed.

  12. Weaning age of calves fed a high milk allowance by automated feeders: effects on feed, water, and energy intake, behavioral signs of hunger, and weight gains.

    Science.gov (United States)

    de Passillé, A M; Borderas, T F; Rushen, J

    2011-03-01

    Dairy calves are increasingly fed large volumes of milk, which reduces feeding motivation and improves weight gain. However, calves often show signs of hunger and lose weight when weaned off milk due to low starter intake. We examined whether delaying the age at weaning would reduce responses to weaning. Calves were raised in groups of 9 and fed milk, starter, hay, and water with automated feeders. In each group, 3 calves were randomly assigned to 1 of 3 treatments: (1) low-milk, early-weaned: fed 6 L/d of milk and weaned at 47 d of age; (2) high-milk early-weaned: fed 12 L/d of milk and weaned at 47 d; (3) high-milk later-weaned: fed 12 L/d of milk and weaned at 89 d of age. Milk, starter, and hay intakes were recorded daily and digestible energy (DE) intake estimated. Feeder visits were recorded. Before weaning, the high-milk calves drank more milk, ate less starter and hay, but had higher DE intakes, gained more weight, and made fewer visits to the milk feeder than the low-milk, early-weaned calves. During and immediately after weaning, the high-fed, early-weaned calves ate less starter and hay, had lower DE intakes, and gained less weight than the low-milk, early-weaned calves and lost their body weight advantage 7 d after weaning. During and immediately after weaning, the high-milk, later-weaned calves ate more starter and hay and had higher DE intakes, higher weight gains, and made fewer visits to the milk feeder than the high-milk, early-weaned calves. They were still heavier than the low-milk, early-weaned calves 18 d after weaning. Delaying the age at which calves are weaned off milk reduces the drop in energy intake and behavioral signs of hunger that result from weaning. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  13. Retinoic Acid Signaling Mediates Hair Cell Regeneration by Repressing p27kip and sox2 in Supporting Cells.

    Science.gov (United States)

    Rubbini, Davide; Robert-Moreno, Àlex; Hoijman, Esteban; Alsina, Berta

    2015-11-25

    During development, otic sensory progenitors give rise to hair cells and supporting cells. In mammalian adults, differentiated and quiescent sensory cells are unable to generate new hair cells when these are lost due to various insults, leading to irreversible hearing loss. Retinoic acid (RA) has strong regenerative capacity in several organs, but its role in hair cell regeneration is unknown. Here, we use genetic and pharmacological inhibition to show that the RA pathway is required for hair cell regeneration in zebrafish. When regeneration is induced by laser ablation in the inner ear or by neomycin treatment in the lateral line, we observe rapid activation of several components of the RA pathway, with dynamics that position RA signaling upstream of other signaling pathways. We demonstrate that blockade of the RA pathway impairs cell proliferation of supporting cells in the inner ear and lateral line. Moreover, in neuromast, RA pathway regulates the transcription of p27(kip) and sox2 in supporting cells but not fgf3. Finally, genetic cell-lineage tracing using Kaede photoconversion demonstrates that de novo hair cells derive from FGF-active supporting cells. Our findings reveal that RA has a pivotal role in zebrafish hair cell regeneration by inducing supporting cell proliferation, and shed light on the underlying transcriptional mechanisms involved. This signaling pathway might be a promising approach for hearing recovery. Hair cells are the specialized mechanosensory cells of the inner ear that capture auditory and balance sensory input. Hair cells die after acoustic trauma, ototoxic drugs or aging diseases, leading to progressive hearing loss. Mammals, in contrast to zebrafish, lack the ability to regenerate hair cells. Here, we find that retinoic acid (RA) pathway is required for hair cell regeneration in vivo in the zebrafish inner ear and lateral line. RA pathway is activated very early upon hair cell loss, promotes cell proliferation of progenitor cells

  14. A decision-support framework for promoting independent living and ageing well.

    Science.gov (United States)

    Billis, Antonis S; Papageorgiou, Elpiniki I; Frantzidis, Christos A; Tsatali, Marianna S; Tsolaki, Anthoula C; Bamidis, Panagiotis D

    2015-01-01

    Artificial intelligence and decision support systems offer a plethora of health monitoring capabilities in ambient assisted living environment. Continuous assessment of health indicators for elderly people living on their own is of utmost importance, so as to prolong their independence and quality of life. Slow varying, long-term deteriorating health trends are not easily identifiable in seniors. Thus, early sign detection of a specific condition, as well as, any likely transition from a healthy state to a pathological one are key problems that the herein proposed framework aims at resolving. Statistical process control concepts offer a personalized approach toward identification of trends that are away from the atypical behavior or state of the seniors, while fuzzy cognitive maps knowledge representation and inference schema have proved to be efficient in terms of disease classification. Geriatric depression is used as a case study throughout the paper, so to prove the validity of the framework, which is planned to be pilot tested with a series of lone-living seniors in their own homes.

  15. Spheroid Culture of Head and Neck Cancer Cells Reveals an Important Role of EGFR Signalling in Anchorage Independent Survival.

    Science.gov (United States)

    Braunholz, Diana; Saki, Mohammad; Niehr, Franziska; Öztürk, Merve; Borràs Puértolas, Berta; Konschak, Robert; Budach, Volker; Tinhofer, Ingeborg

    2016-01-01

    In solid tumours millions of cells are shed into the blood circulation each day. Only a subset of these circulating tumour cells (CTCs) survive, many of them presumable because of their potential to form multi-cellular clusters also named spheroids. Tumour cells within these spheroids are protected from anoikis, which allows them to metastasize to distant organs or re-seed at the primary site. We used spheroid cultures of head and neck squamous cell carcinoma (HNSCC) cell lines as a model for such CTC clusters for determining the role of the epidermal growth factor receptor (EGFR) in cluster formation ability and cell survival after detachment from the extra-cellular matrix. The HNSCC cell lines FaDu, SCC-9 and UT-SCC-9 (UT-SCC-9P) as well as its cetuximab (CTX)-resistant sub-clone (UT-SCC-9R) were forced to grow in an anchorage-independent manner by coating culture dishes with the anti-adhesive polymer poly-2-hydroxyethylmethacrylate (poly-HEMA). The extent of apoptosis, clonogenic survival and EGFR signalling under such culture conditions was evaluated. The potential of spheroid formation in suspension culture was found to be positively correlated with the proliferation rate of HNSCC cell lines as well as their basal EGFR expression levels. CTX and gefitinib blocked, whereas the addition of EGFR ligands promoted anchorage-independent cell survival and spheroid formation. Increased spheroid formation and growth were associated with persistent activation of EGFR and its downstream signalling component (MAPK/ERK). Importantly, HNSCC cells derived from spheroid cultures retained their clonogenic potential in the absence of cell-matrix contact. Addition of CTX under these conditions strongly inhibited colony formation in CTX-sensitive cell lines but not their resistant subclones. Altogether, EGFR activation was identified as crucial factor for anchorage-independent survival of HNSCC cells. Targeting EGFR in CTC cluster formation might represent an attractive anti

  16. HOXB13 promotes androgen independent growth of LNCaP prostate cancer cells by the activation of E2F signaling

    Directory of Open Access Journals (Sweden)

    Choi Chan

    2010-05-01

    Full Text Available Abstract Background Androgen signaling plays a critical role in the development of prostate cancer and its progression. However, androgen-independent prostate cancer cells emerge after hormone ablation therapy, resulting in significant clinical problems. We have previously demonstrated that the HOXB13 homeodomain protein functions as a prostate cancer cell growth suppressor by inhibiting androgen-mediated signals. However, the role of the HOXB13 in androgen-independent growth of prostate cancer cells remains unexplained. Results In this report, we first demonstrated that HOXB13 was highly overexpressed in hormone-refractory tumors compared to tumors without prostate-specific antigen after initial treatment. Functionally, in an androgen-free environment minimal induction of HOXB13 in LNCaP prostate cancer cells, to the level of the normal prostate, markedly promoted cell proliferation while suppression inhibited cell proliferation. The HOXB13-mediated cell growth promotion in the absence of androgen, appears to be mainly accomplished through the activation of RB-E2F signaling by inhibiting the expression of the p21waf tumor suppressor. Indeed, forced expression of HOXB13 dramatically decreased expression of p21waf; this inhibition largely affected HOXB13-mediated promotion of E2F signaling. Conclusions Taken together, the results of this study demonstrated the presence of a novel pathway that helps understand androgen-independent survival of prostate cancer cells. These findings suggest that upregulation of HOXB13 is associated with an additive growth advantage of prostate cancer cells in the absence of or low androgen concentrations, by the regulation of p21-mediated E2F signaling.

  17. 47 CFR 25.258 - Sharing between NGSO MSS Feeder links Stations and GSO FSS services in the 29.25-29.5 GHz Bands.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Sharing between NGSO MSS Feeder links Stations and GSO FSS services in the 29.25-29.5 GHz Bands. 25.258 Section 25.258 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Technical Standards § 25...

  18. The anterior lens capsule used as support material in RPE cell-transplantation

    DEFF Research Database (Denmark)

    Nicolini, J; Kiilgaard, Jens Folke; Wiencke, A K

    2000-01-01

    To investigate the use of an ocular basement membrane as support material for transplanted porcine RPE cells.......To investigate the use of an ocular basement membrane as support material for transplanted porcine RPE cells....

  19. Temporal Regulation of Natural Killer T Cell Interferon Gamma Responses by β-Catenin-Dependent and -Independent Wnt Signaling

    Directory of Open Access Journals (Sweden)

    Jessica C. Kling

    2018-03-01

    Full Text Available Natural killer T (NKT cells are prominent innate-like lymphocytes in the liver with critical roles in immune responses during infection, cancer, and autoimmunity. Interferon gamma (IFN-γ and IL-4 are key cytokines rapidly produced by NKT cells upon recognition of glycolipid antigens presented by antigen-presenting cells (APCs. It has previously been reported that the transcriptional coactivator β-catenin regulates NKT cell differentiation and functionally biases NKT cell responses toward IL-4, at the expense of IFN-γ production. β-Catenin is not only a central effector of Wnt signaling but also contributes to other signaling networks. It is currently unknown whether Wnt ligands regulate NKT cell functions. We thus investigated how Wnt ligands and β-catenin activity shape liver NKT cell functions in vivo in response to the glycolipid antigen, α-galactosylceramide (α-GalCer using a mouse model. Pharmacologic targeting of β-catenin activity with ICG001, as well as myeloid-specific genetic ablation of Wntless (Wls, to specifically target Wnt protein release by APCs, enhanced early IFN-γ responses. By contrast, within several hours of α-GalCer challenge, myeloid-specific Wls deficiency, as well as pharmacologic targeting of Wnt release using the small molecule inhibitor IWP-2 impaired α-GalCer-induced IFN-γ responses, independent of β-catenin activity. These data suggest that myeloid cell-derived Wnt ligands drive early Wnt/β-catenin signaling that curbs IFN-γ responses, but that, subsequently, Wnt ligands sustain IFN-γ expression independent of β-catenin activity. Our analyses in ICG001-treated mice confirmed a role for β-catenin activity in driving early IL-4 responses by liver NKT cells. However, neither pharmacologic nor genetic perturbation of Wnt production affected the IL-4 response, suggesting that IL-4 production by NKT cells in response to α-GalCer is not driven by released Wnt ligands. Collectively, these data reveal

  20. Temporal Regulation of Natural Killer T Cell Interferon Gamma Responses by β-Catenin-Dependent and -Independent Wnt Signaling.

    Science.gov (United States)

    Kling, Jessica C; Jordan, Margaret A; Pitt, Lauren A; Meiners, Jana; Thanh-Tran, Thao; Tran, Le Son; Nguyen, Tam T K; Mittal, Deepak; Villani, Rehan; Steptoe, Raymond J; Khosrotehrani, Kiarash; Berzins, Stuart P; Baxter, Alan G; Godfrey, Dale I; Blumenthal, Antje

    2018-01-01

    Natural killer T (NKT) cells are prominent innate-like lymphocytes in the liver with critical roles in immune responses during infection, cancer, and autoimmunity. Interferon gamma (IFN-γ) and IL-4 are key cytokines rapidly produced by NKT cells upon recognition of glycolipid antigens presented by antigen-presenting cells (APCs). It has previously been reported that the transcriptional coactivator β-catenin regulates NKT cell differentiation and functionally biases NKT cell responses toward IL-4, at the expense of IFN-γ production. β-Catenin is not only a central effector of Wnt signaling but also contributes to other signaling networks. It is currently unknown whether Wnt ligands regulate NKT cell functions. We thus investigated how Wnt ligands and β-catenin activity shape liver NKT cell functions in vivo in response to the glycolipid antigen, α-galactosylceramide (α-GalCer) using a mouse model. Pharmacologic targeting of β-catenin activity with ICG001, as well as myeloid-specific genetic ablation of Wntless (Wls) , to specifically target Wnt protein release by APCs, enhanced early IFN-γ responses. By contrast, within several hours of α-GalCer challenge, myeloid-specific Wls deficiency, as well as pharmacologic targeting of Wnt release using the small molecule inhibitor IWP-2 impaired α-GalCer-induced IFN-γ responses, independent of β-catenin activity. These data suggest that myeloid cell-derived Wnt ligands drive early Wnt/β-catenin signaling that curbs IFN-γ responses, but that, subsequently, Wnt ligands sustain IFN-γ expression independent of β-catenin activity. Our analyses in ICG001-treated mice confirmed a role for β-catenin activity in driving early IL-4 responses by liver NKT cells. However, neither pharmacologic nor genetic perturbation of Wnt production affected the IL-4 response, suggesting that IL-4 production by NKT cells in response to α-GalCer is not driven by released Wnt ligands. Collectively, these data reveal complex temporal

  1. Ontogeny of thymic independent antibody responses in vitro in normal mice and mice with an x-linked B cell defect

    Energy Technology Data Exchange (ETDEWEB)

    Mosier, D.E.; Mond, J.J.; Goldings, E.A.

    1977-12-01

    The primary in vitro antibody response of neonatal spleen cells to three thymic independent antigens has been examined. The time of onset of responsiveness to TNP-Brucella abortus and TNP-lipopolysaccharide was significantly earlier than the onset of responsiveness to TNP-Ficoll. This ontologic sequence was not affected by T cell depletion or antigen presentation on adult macrophages. In neonatal mice bearing the X-linked CBA/N defect, the response to TNP-Brucella abortus and TNP-lipopolysaccharide was much delayed and no response to TNP-Ficoll developed. We conclude that different thymic independent antigens address different subpopulations of B cells, one of which appears earlier in ontogeny than the other.

  2. Ontogeny of thymic independent antibody responses in vitro in normal mice and mice with an x-linked B cell defect

    International Nuclear Information System (INIS)

    Mosier, D.E.; Mond, J.J.; Goldings, E.A.

    1977-01-01

    The primary in vitro antibody response of neonatal spleen cells to three thymic independent antigens has been examined. The time of onset of responsiveness to TNP-Brucella abortus and TNP-lipopolysaccharide was significantly earlier than the onset of responsiveness to TNP-Ficoll. This ontologic sequence was not affected by T cell depletion or antigen presentation on adult macrophages. In neonatal mice bearing the X-linked CBA/N defect, the response to TNP-Brucella abortus and TNP-lipopolysaccharide was much delayed and no response to TNP-Ficoll developed. We conclude that different thymic independent antigens address different subpopulations of B cells, one of which appears earlier in ontogeny than the other

  3. Implementation of the resonant vibratory feeders control algorithm on Simatic S7-1200 from MATLAB Simulink enviroment

    Directory of Open Access Journals (Sweden)

    Mitrović Radomir B.

    2016-01-01

    Full Text Available Simulink is an important tool for modeling and simulation of process and control algorithms. It's expansion, PLC Coder, enables direct conversion of model subsystem into SCL, structured text code, which is then used by PLC IDE to create function blocks. This shortens developing time of algorithms for PLC controller. Also, this reduces possibility for a coding error. This paper describes Simulink PLC Coder and workflow for developing PID control algorithm for Siemens Simatic S7-1200 PLC. Control object used here is resonant vibratory feeder having electromagnetic drive.

  4. Weight loss independent changes in adipose tissue macrophage and T cell populations after sleeve gastrectomy in mice

    Directory of Open Access Journals (Sweden)

    Henriette Frikke-Schmidt

    2017-04-01

    Full Text Available Objective: In addition to adipocytes, adipose tissue contains large numbers of immune cells. A wide range of evidence links the activity of these cells to regulation of adipocyte and systemic metabolic function. Bariatric surgery improves several aspects of metabolic derangements and at least some of these effects occur in a weight-loss independent manner. We sought to investigate the impact of vertical sleeve gastrectomy (VSG on adipose immune cell frequencies. Methods: We analyzed the frequencies of immune cells within distinct adipose tissue depots in obese mice that had VSG or sham surgery with a portion of the latter group pair-fed such that their body mass was matched to the VSG animals. Results: We demonstrate that VSG induced a shift in the epididymal adipose tissue leukocyte profile including increased frequencies of CD11c− macrophages, increased frequencies of T cells (CD4+, CD8+, and CD4−/CD8− T cells all increased, but a significantly decreased frequency of adipose tissue dendritic cells (ATDC that, despite the continued high fat feeding of the VSG group, dropped below control diet levels. Conclusions: These results indicate that VSG induces substantial changes in the immune populations residing in the adipose depots independent of weight loss. Author Video: Author Video Watch what authors say about their articles Keywords: Sleeve gastrectomy, Adipose tissue, Macrophages, T cells, Dendritic cells, Abbreviations: ATDC, adipose tissue dendritic cell, ATM, adipose tissue macrophage, eWAT, epididymal white adipose tissue, FFA, free fatty acids, HFS, high fat sham, iWAT, inguinal white adipose tissue, SVC, stromal vascular cells, VSG, vertical sleeve gastrectomy

  5. Derivation of Stromal (Skeletal, Mesenchymal) Stem-like cells from Human Embryonic Stem Cells

    DEFF Research Database (Denmark)

    Mahmood, Amer; Harkness, Linda; Abdallah, Basem

    2012-01-01

    EBs using BMP2 (bone morphogenic protein 2) combined with standard osteoblast induction medium led to weak osteoblastic induction. Conversely, subcutaneous in vivo implantation of day 20 hEBs in immune deficient mice, mixed with hydroxyapatite/tricalcium phosphate (HA/TCP) as an osteoconductive scaffold......Derivation of bone forming cells (osteoblasts) from human embryonic stem cells (hESC) is a pre-requisite for their use in clinical applications. However, there is no standard protocol for differentiating hESC into osteoblastic cells. The aim of this study was to identify the emergence of a human...... stromal (mesenchymal, skeletal) stem cell (hMSC)-like population, known to be osteoblastic cell precursors and to test their osteoblastic differentiation capacity in ex vivo cultures and in vivo. We cultured hESC in a feeder-free environment using serum replacement and as suspension aggregates (embryoid...

  6. NF-κB dependent and independent mechanisms of quartz-induced proinflammatory activation of lung epithelial cells

    Directory of Open Access Journals (Sweden)

    Schins Roel PF

    2010-05-01

    Full Text Available Abstract In the initiation and progression of pulmonary inflammation, macrophages have classically been considered as a crucial cell type. However, evidence for the role of epithelial type II cells in pulmonary inflammation has been accumulating. In the current study, a combined in vivo and in vitro approach has been employed to investigate the mechanisms of quartz-induced proinflammatory activation of lung epithelial cells. In vivo, enhanced expression of the inflammation- and oxidative stress-related genes HO-1 and iNOS was found on the mRNA level in rat lungs after instillation with DQ12 respirable quartz. Activation of the classical NF-κB pathway in macrophages and type II pneumocytes was indicated by enhanced immunostaining of phospho-IκBα in these specific lung cell types. In vitro, the direct, particle-mediated effect on proinflammatory signalling in a rat lung epithelial (RLE cell line was compared to the indirect, macrophage product-mediated effect. Treatment with quartz particles induced HO-1 and COX-2 mRNA expression in RLE cells in an NF-κB independent manner. Supernatant from quartz-treated macrophages rapidly activated the NF-κB signalling pathway in RLE cells and markedly induced iNOS mRNA expression up to 2000-fold compared to non-treated control cells. Neutralisation of TNFα and IL-1β in macrophage supernatant did not reduce its ability to elicit NF-κB activation of RLE cells. In addition the effect was not modified by depletion or supplementation of intracellular glutathione. The results from the current work suggest that although both oxidative stress and NF-κB are likely involved in the inflammatory effects of toxic respirable particles, these phenomena can operate independently on the cellular level. This might have consequences for in vitro particle hazard testing, since by focusing on NF-κB signalling one might neglect alternative inflammatory pathways.

  7. Dipeptidyl peptidase-IV inhibits glioma cell growth independent of its enzymatic activity

    Czech Academy of Sciences Publication Activity Database

    Busek, P.; Stremeňová, J.; Sromová, L.; Hilser, M.; Balaziová, E.; Kosek, D.; Trylcová, J.; Strnad, Hynek; Křepela, E.; Šedo, A.

    2012-01-01

    Roč. 44, č. 5 (2012), s. 738-747 ISSN 1357-2725 Institutional support: RVO:68378050 Keywords : protease * tumour suppression * primary cell cultures * astrocytoma Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.152, year: 2012

  8. Does proximity to urban centres affect the dietary regime of marine benthic filter feeders?

    Science.gov (United States)

    Puccinelli, Eleonora; Noyon, Margaux; McQuaid, Christopher D.

    2016-02-01

    Threats to marine ecosystems include habitat destruction and degradation of water quality, resulting from land- and ocean-based human activities. Anthropogenic input causing modification of water quality, can affect primary productivity and thus food availability and quality for higher trophic levels. This is especially important for sedentary benthic intertidal communities, which rely on local food availability. We investigated the effect of urbanization on the dietary regime of four species of intertidal filter feeders (three barnacles and one mussel) at sites close to high-density cities and at sites far from heavily urbanized areas using fatty acid and stable isotope techniques. δ15N was significantly higher at urbanized sites compared to their corresponding control sites for all species with few exceptions, while no effect on δ13C was recorded. Barnacle fatty acid profiles were not affected by cities, while mussels from sites close to cities had fatty acid signatures with a higher proportion of polyunsaturated fatty acids (PUFA). We suggest that the enrichment in δ15N at urbanised sites reflects the influence of anthropogenically derived nitrogen directly linked to wastewater input from domestic and industrial sewage. Linked to this, the high proportion of PUFA in mussels at urbanized sites may reflect the influence of increased nitrogen concentrations on primary production and enhanced growth of large phytoplankton cells. The results indicate that anthropogenic effects can strongly influence the diets of benthic organisms, but these effects differ among taxa. Changes in the diet of such habitat forming species can affect their fitness and survival with potential effects on the populations associated with them.

  9. Plasma position control device for thermonuclear device

    Energy Technology Data Exchange (ETDEWEB)

    Onozuka, Masanori [Mitsubishi Heavy Industries Ltd., Tokyo (Japan); Fujita, Jun-ya; Ioki, Kimihiro

    1995-10-03

    The present invention concerns plasma position control coils having a feeder line structure not requiring high strength for the support portion. Namely, the coils are formed by twisting feeder lines extended from plasma position control coils in a vacuum vessel. The twisted feeder lines are supported using an appropriate structural member. Electromagnetic load is generated to the feeder lines being extended from the position control coils and traversing toroidal fields at a current introduction lines and at current delivery lines respectively. However, since the feeder lines have substantially spiral shape consisting of two twisted lines, the electromagnetic load and the moment caused by the generated load which are inversed to each other are off set. Accordingly, only extremely small force is exerted on the fittings which support the feeder lines. Therefore, small strength may suffice for the fittings and the gaps of mounting the fittings may be made longer. (I.S.).

  10. Plasma position control device for thermonuclear device

    International Nuclear Information System (INIS)

    Onozuka, Masanori; Fujita, Jun-ya; Ioki, Kimihiro.

    1995-01-01

    The present invention concerns plasma position control coils having a feeder line structure not requiring high strength for the support portion. Namely, the coils are formed by twisting feeder lines extended from plasma position control coils in a vacuum vessel. The twisted feeder lines are supported using an appropriate structural member. Electromagnetic load is generated to the feeder lines being extended from the position control coils and traversing toroidal fields at a current introduction lines and at current delivery lines respectively. However, since the feeder lines have substantially spiral shape consisting of two twisted lines, the electromagnetic load and the moment caused by the generated load which are inversed to each other are off set. Accordingly, only extremely small force is exerted on the fittings which support the feeder lines. Therefore, small strength may suffice for the fittings and the gaps of mounting the fittings may be made longer. (I.S.)

  11. Transplanting Retinal Cells using Bucky Paper for Support

    Science.gov (United States)

    Loftus, David J.; Cinke, Martin; Meyyappan, Meyya; Fishman, Harvey; Leng, Ted; Huie, Philip; Bilbao, Kalayaan

    2004-01-01

    A novel treatment for retinal degenerative disorders involving transplantation of cells into the eye is currently under development at NASA Ames Research Center and Stanford University School of Medicine. The technique uses bucky paper as a support material for retinal pigment epithelial (RPE) cells, iris pigment epithelial (IPE) cells, and/or stem cells. This technology is envisioned as a treatment for age-related macular degeneration, which is the leading cause of blindness in persons over age 65 in Western nations. Additionally, patients with other retinal degenerative disorders, such as retinitis pigmentosa, may be treated by this strategy. Bucky paper is a mesh of carbon nanotubes (CNTs), as shown in Figure 1, that can be made from any of the commercial sources of CNTs. Bucky paper is biocompatible and capable of supporting the growth of biological cells. Because bucky paper is highly porous, nutrients, oxygen, carbon dioxide, and waste can readily diffuse through it. The thickness, density, and porosity of bucky paper can be tailored in manufacturing. For transplantation of cells into the retina, bucky paper serves simultaneously as a substrate for cell growth and as a barrier for new blood vessel formation, which can be a problem in the exudative type of macular degeneration. Bucky paper is easily handled during surgical implantation into the eye. Through appropriate choice of manufacturing processes, bucky paper can be made relatively rigid yet able to conform to the retina when the bucky paper is implanted. Bucky paper offers a distinct advantage over other materials that have been investigated for retinal cell transplantation - lens capsule and Descemet's membrane - which are difficult to handle during surgery because they are flimsy and do not stay flat.

  12. Identifying off-target effects of etomoxir reveals that carnitine palmitoyltransferase I is essential for cancer cell proliferation independent of β-oxidation.

    Directory of Open Access Journals (Sweden)

    Cong-Hui Yao

    2018-03-01

    Full Text Available It has been suggested that some cancer cells rely upon fatty acid oxidation (FAO for energy. Here we show that when FAO was reduced approximately 90% by pharmacological inhibition of carnitine palmitoyltransferase I (CPT1 with low concentrations of etomoxir, the proliferation rate of various cancer cells was unaffected. Efforts to pharmacologically inhibit FAO more than 90% revealed that high concentrations of etomoxir (200 μM have an off-target effect of inhibiting complex I of the electron transport chain. Surprisingly, however, when FAO was reduced further by genetic knockdown of CPT1, the proliferation rate of these same cells decreased nearly 2-fold and could not be restored by acetate or octanoic acid supplementation. Moreover, CPT1 knockdowns had altered mitochondrial morphology and impaired mitochondrial coupling, whereas cells in which CPT1 had been approximately 90% inhibited by etomoxir did not. Lipidomic profiling of mitochondria isolated from CPT1 knockdowns showed depleted concentrations of complex structural and signaling lipids. Additionally, expression of a catalytically dead CPT1 in CPT1 knockdowns did not restore mitochondrial coupling. Taken together, these results suggest that transport of at least some long-chain fatty acids into the mitochondria by CPT1 may be required for anabolic processes that support healthy mitochondrial function and cancer cell proliferation independent of FAO.

  13. NASA Alternative Orion Small Cell Battery Design Support

    Science.gov (United States)

    Haynes, Chuck

    2016-01-01

    The NASA Orion Crew Module Reference Design was produced to address large scale thermal runaway (TR) hazard with specific safety controls for the Orion Spacecraft. The design presented provides the description of a full scale battery design reference for implementation as a drop in replacement to meet all spacecraft energy requirements with compatible 120 Vdc electrical and mechanical interface using small cell technology (18650) packaging. The 32V SuperBrick incorporates unique support features and an electrical bus bar arrangement that allows cells negative can insertion into heat sink that is compressively coupled to the battery enclosure to promote good thermal management. The housing design also provides an internal flame suppression "filter tray" and positive venting path internal to the enclosure to allow hot effluent ejecta to escape in the event of single cell TR. Virtual cells (14P Banks) that are supported to provide cell spacing with interstitial materials to prevent side can failures that can produce cell to cell TR propagation. These features were successfully test in four separate TR run with the full scale DTA1 test article in February 2016. Successfully Completed Test Objectives - Four separate TR test runs with Full-Scale DTA1 housing with Two SuperBricks, Two SuperBrick Emulators All Tests resulted in "clean" gas with less than 6 C rise at Battery vent All Tests resulted in less than 2 C temperature rise on cold-plate outlet All Tests resulted in less than 6 psi pressure rise in the battery housing Test Run 1 -One neighbor cell TR, highest remaining neighbor 139 C. Ejecta shorted to bus caused prolonged additional heating, One shorted cell did experience TR after 12 minutes, remaining cells had adequate thermal margin Test Run 2 - No cell to cell propagation, highest neighbor cell 112 C; Test Run 3 - No cell to cell propagation, highest neighbor cell 96 C; Test Run 4 - No cell to cell propagation, highest neighbor cell 101 C; Primary TR testing

  14. FGF7 supports hematopoietic stem and progenitor cells and niche-dependent myeloblastoma cells via autocrine action on bone marrow stromal cells in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Ishino, Ruri; Minami, Kaori; Tanaka, Satowa [Laboratory of Hematology, Division of Medical Biophysics, Kobe University Graduate School of Health Sciences, 7-10-2 Tomogaoka, Suma-ku, Kobe 654-0142 (Japan); Nagai, Mami [Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 159-8555 (Japan); Matsui, Keiji; Hasegawa, Natsumi [Laboratory of Hematology, Division of Medical Biophysics, Kobe University Graduate School of Health Sciences, 7-10-2 Tomogaoka, Suma-ku, Kobe 654-0142 (Japan); Roeder, Robert G. [Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10065 (United States); Asano, Shigetaka [Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 159-8555 (Japan); Ito, Mitsuhiro, E-mail: itomi@med.kobe-u.ac.jp [Laboratory of Hematology, Division of Medical Biophysics, Kobe University Graduate School of Health Sciences, 7-10-2 Tomogaoka, Suma-ku, Kobe 654-0142 (Japan); Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10065 (United States); Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 159-8555 (Japan); Department of Family and Community Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 654-0142 (Japan)

    2013-10-11

    Highlights: •FGF7 is downregulated in MED1-deficient mesenchymal cells. •FGF7 produced by mesenchymal stromal cells is a novel hematopoietic niche molecule. •FGF7 supports hematopoietic progenitor cells and niche-dependent leukemia cells. •FGF7 activates FGFR2IIIb of bone marrow stromal cells in an autocrine manner. •FGF7 indirectly acts on hematopoietic cells lacking FGFR2IIIb via stromal cells. -- Abstract: FGF1 and FGF2 support hematopoietic stem and progenitor cells (HSPCs) under stress conditions. In this study, we show that fibroblast growth factor (FGF7) may be a novel niche factor for HSPC support and leukemic growth. FGF7 expression was attenuated in mouse embryonic fibroblasts (MEFs) deficient for the MED1 subunit of the Mediator transcriptional coregulator complex. When normal mouse bone marrow (BM) cells were cocultured with Med1{sup +/+} MEFs or BM stromal cells in the presence of anti-FGF7 antibody, the growth of BM cells and the number of long-time culture-initiating cells (LTC-ICs) decreased significantly. Anti-FGF7 antibody also attenuated the proliferation and cobblestone formation of MB1 stromal cell-dependent myeloblastoma cells. The addition of recombinant FGF7 to the coculture of BM cells and Med1{sup −/−} MEFs increased BM cells and LTC-ICs. FGF7 and its cognate receptor, FGFR2IIIb, were undetectable in BM cells, but MEFs and BM stromal cells expressed both. FGF7 activated downstream targets of FGFR2IIIb in Med1{sup +/+} and Med1{sup −/−} MEFs and BM stromal cells. Taken together, we propose that FGF7 supports HSPCs and leukemia-initiating cells indirectly via FGFR2IIIb expressed on stromal cells.

  15. FGF7 supports hematopoietic stem and progenitor cells and niche-dependent myeloblastoma cells via autocrine action on bone marrow stromal cells in vitro

    International Nuclear Information System (INIS)

    Ishino, Ruri; Minami, Kaori; Tanaka, Satowa; Nagai, Mami; Matsui, Keiji; Hasegawa, Natsumi; Roeder, Robert G.; Asano, Shigetaka; Ito, Mitsuhiro

    2013-01-01

    Highlights: •FGF7 is downregulated in MED1-deficient mesenchymal cells. •FGF7 produced by mesenchymal stromal cells is a novel hematopoietic niche molecule. •FGF7 supports hematopoietic progenitor cells and niche-dependent leukemia cells. •FGF7 activates FGFR2IIIb of bone marrow stromal cells in an autocrine manner. •FGF7 indirectly acts on hematopoietic cells lacking FGFR2IIIb via stromal cells. -- Abstract: FGF1 and FGF2 support hematopoietic stem and progenitor cells (HSPCs) under stress conditions. In this study, we show that fibroblast growth factor (FGF7) may be a novel niche factor for HSPC support and leukemic growth. FGF7 expression was attenuated in mouse embryonic fibroblasts (MEFs) deficient for the MED1 subunit of the Mediator transcriptional coregulator complex. When normal mouse bone marrow (BM) cells were cocultured with Med1 +/+ MEFs or BM stromal cells in the presence of anti-FGF7 antibody, the growth of BM cells and the number of long-time culture-initiating cells (LTC-ICs) decreased significantly. Anti-FGF7 antibody also attenuated the proliferation and cobblestone formation of MB1 stromal cell-dependent myeloblastoma cells. The addition of recombinant FGF7 to the coculture of BM cells and Med1 −/− MEFs increased BM cells and LTC-ICs. FGF7 and its cognate receptor, FGFR2IIIb, were undetectable in BM cells, but MEFs and BM stromal cells expressed both. FGF7 activated downstream targets of FGFR2IIIb in Med1 +/+ and Med1 −/− MEFs and BM stromal cells. Taken together, we propose that FGF7 supports HSPCs and leukemia-initiating cells indirectly via FGFR2IIIb expressed on stromal cells

  16. Both canonical and non-canonical Wnt signaling independently promote stem cell growth in mammospheres.

    Directory of Open Access Journals (Sweden)

    Alexander M Many

    Full Text Available The characterization of mammary stem cells, and signals that regulate their behavior, is of central importance in understanding developmental changes in the mammary gland and possibly for targeting stem-like cells in breast cancer. The canonical Wnt/β-catenin pathway is a signaling mechanism associated with maintenance of self-renewing stem cells in many tissues, including mammary epithelium, and can be oncogenic when deregulated. Wnt1 and Wnt3a are examples of ligands that activate the canonical pathway. Other Wnt ligands, such as Wnt5a, typically signal via non-canonical, β-catenin-independent, pathways that in some cases can antagonize canonical signaling. Since the role of non-canonical Wnt signaling in stem cell regulation is not well characterized, we set out to investigate this using mammosphere formation assays that reflect and quantify stem cell properties. Ex vivo mammosphere cultures were established from both wild-type and Wnt1 transgenic mice and were analyzed in response to manipulation of both canonical and non-canonical Wnt signaling. An increased level of mammosphere formation was observed in cultures derived from MMTV-Wnt1 versus wild-type animals, and this was blocked by treatment with Dkk1, a selective inhibitor of canonical Wnt signaling. Consistent with this, we found that a single dose of recombinant Wnt3a was sufficient to increase mammosphere formation in wild-type cultures. Surprisingly, we found that Wnt5a also increased mammosphere formation in these assays. We confirmed that this was not caused by an increase in canonical Wnt/β-catenin signaling but was instead mediated by non-canonical Wnt signals requiring the receptor tyrosine kinase Ror2 and activity of the Jun N-terminal kinase, JNK. We conclude that both canonical and non-canonical Wnt signals have positive effects promoting stem cell activity in mammosphere assays and that they do so via independent signaling mechanisms.

  17. Fault Diagnosis for Distribution Networks Using Enhanced Support Vector Machine Classifier with Classical Multidimensional Scaling

    Directory of Open Access Journals (Sweden)

    Ming-Yuan Cho

    2017-09-01

    Full Text Available In this paper, a new fault diagnosis techniques based on time domain reflectometry (TDR method with pseudo-random binary sequence (PRBS stimulus and support vector machine (SVM classifier has been investigated to recognize the different types of fault in the radial distribution feeders. This novel technique has considered the amplitude of reflected signals and the peaks of cross-correlation (CCR between the reflected and incident wave for generating fault current dataset for SVM. Furthermore, this multi-layer enhanced SVM classifier is combined with classical multidimensional scaling (CMDS feature extraction algorithm and kernel parameter optimization to increase training speed and improve overall classification accuracy. The proposed technique has been tested on a radial distribution feeder to identify ten different types of fault considering 12 input features generated by using Simulink software and MATLAB Toolbox. The success rate of SVM classifier is over 95% which demonstrates the effectiveness and the high accuracy of proposed method.

  18. The impact of deprivation on youth violence: a comparison of cities and their feeder towns.

    Science.gov (United States)

    Jones, S J; Sivarajasingam, V; Shepherd, J

    2011-06-01

    66,000 children and adolescents are treated at emergency departments (EDs) in England and Wales each year for assault injury. The aim of this study was to compare adolescent assault injury rates in cities and towns and determine how assault injury varies with deprivation and gender. The study was set in three cities in Wales, UK, and their surrounding towns. Subjects were 11-17 year olds treated for assault injury at one of seven EDs from 1 October 2005 to 30 September 2006. Area of residence (electoral divisions, EDivs) was identified from patient postcode. EDivs were aggregated into deprivation fifths for males and females and cities and towns. Assault injury rates, rate ratios and 95% CIs were calculated. 1472 children and adolescents of 11-17 years old were treated for assault injury. Male city assault injury rates were 14.2/1000 11-17 year olds; and 13.1 in towns. Female city assault injury rates were 6.0; and 5.6 in towns. In the most deprived city areas males had assault injury rates 2.6 times (95% CI 1.85 to 3.59) that of the most affluent, compared with 2.0 times in towns (95% CI 1.39 to 2.86). For females, the most deprived city areas had assault injury rates 5.3 times that of the most affluent (95% CI 2.93 to 9.41), compared with 2.8 times in towns (95% CI 1.47 to 5.28). Injury in youth violence increased with increasing deprivation in cities and their feeder towns. This was true for boys and girls, though rates for boys were consistently higher. This link between assault injury and deprivation was stronger for girls in cities than in feeder towns. Strategies to prevent youth violence need to include improved safeguarding arrangements for girls living in the most deprived city areas.

  19. Support of enhanced oil recovery to independent producers in Texas. Quarterly technical progress report, July 1, 1995--September 30, 1995

    Energy Technology Data Exchange (ETDEWEB)

    Fotouh, K.H.

    1995-09-30

    The main objective of this project is to support independent oil producers in Texas and to improve the productivity of marginal wells utilizing enhanced oil recovery techniques. The main task carried out this quarter was the generation of an electronic data base.

  20. Met-Independent Hepatocyte Growth Factor-mediated regulation of cell adhesion in human prostate cancer cells

    Directory of Open Access Journals (Sweden)

    Davis Rodney

    2006-07-01

    Full Text Available Abstract Background Prostate cancer cells communicate reciprocally with the stromal cells surrounding them, inside the prostate, and after metastasis, within the bone. Each tissue secretes factors for interpretation by the other. One stromally-derived factor, Hepatocyte Growth Factor (HGF, was found twenty years ago to regulate invasion and growth of carcinoma cells. Working with the LNCaP prostate cancer progression model, we found that these cells could respond to HGF stimulation, even in the absence of Met, the only known HGF receptor. The new HGF binding partner we find on the cell surface may help to clarify conflicts in the past literature about Met expression and HGF response in cancer cells. Methods We searched for Met or any HGF binding partner on the cells of the PC3 and LNCaP prostate cancer cell models, using HGF immobilized on agarose beads. By using mass spectrometry analyses and sequencing we have identified nucleolin protein as a novel HGF binding partner. Antibodies against nucleolin (or HGF were able to ameliorate the stimulatory effects of HGF on met-negative prostate cancer cells. Western blots, RT-PCR, and immunohistochemistry were used to assess nucleolin levels during prostate cancer progression in both LNCaP and PC3 models. Results We have identified HGF as a major signaling component of prostate stromal-conditioned media (SCM and have implicated the protein nucleolin in HGF signal reception by the LNCaP model prostate cancer cells. Antibodies that silence either HGF (in SCM or nucleolin (on the cell surfaces eliminate the adhesion-stimulatory effects of the SCM. Likewise, addition of purified HGF to control media mimics the action of SCM. C4-2, an LNCaP lineage-derived, androgen-independent human prostate cancer cell line, responds to HGF in a concentration-dependent manner by increasing its adhesion and reducing its migration on laminin substratum. These HGF effects are not due to shifts in the expression levels of