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

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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.

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

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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.

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

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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 ...

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.

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

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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.

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

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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.

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

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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.

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

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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.

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

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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.

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)

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

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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)

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

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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.

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

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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.

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.

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

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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.

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

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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.

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

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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...

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

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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.

CATTLE FEEDER BEHAVIOR AND FEEDER CATTLE PLACEMENTS

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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...

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

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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.

Human embryonic stem cells and microenvironment

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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

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

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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)

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

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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.

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

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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

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

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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.

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

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

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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.

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)

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...

Colour preferences of UK garden birds at supplementary seed feeders.

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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.

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

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)

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

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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.

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

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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

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.

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

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

Derivation of transgene-free human induced pluripotent stem cells from human peripheral T cells in defined culture conditions.

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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.

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

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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.

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.

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.

Generation of Megakaryocytes and Platelets from Human Pluripotent Stem Cells.

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Pick, Marjorie

2016-01-01

Human pluripotent stem cells (hPSC) have the potential to produce any tissue type in the body and thus represent a source of cells for regenerative medicine. Here we have shown that human platelets can be produced from embryonic or induced pluripotent stem cells in a defined culture system. We describe a serum- and feeder-free culture system that enabled the generation of megakaryocyte (Mk) progenitors and functional platelets from hPSCs. After 13 days the differentiated population included precursor cells that formed colonies containing differentiated Mks, and after 20 days these Mks were able to fragment into platelet-like particles that were functional. This protocol represents an important step towards the generation of human platelets for therapeutic use.

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

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.

Generating a non-integrating human induced pluripotent stem cell bank from urine-derived cells.

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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.

Generation of Corneal Keratocytes from Human Embryonic Stem Cells.

Science.gov (United States)

Hertsenberg, Andrew J; Funderburgh, James L

2016-01-01

Human Embryonic Stem Cells (hESC) offer an important resource as a limitless supply of any differentiated cell type of the human body. Keratocytes, cells from the corneal stroma, may have the potential for restoration of vision in cell therapy and biomedical engineering applications, but these specialized cells are not readily expanded in vitro. Here we describe a two-part method to produce keratocytes from the H1 hESC cell line. The hESC cells, maintained and expanded in feeder-free culture medium are first differentiated to neural crest cells using the stromal-derived inducing activity (SDIA) of the PA6 mouse embryonic fibroblast cell line. The resulting neural crest cells are selected by their expression of cell-surface CD271 and subsequently cultured as 3D pellets in a defined differentiation medium to induce a keratocyte phenotype.

NCR1 Expression Identifies Canine Natural Killer Cell Subsets with Phenotypic Similarity to Human Natural Killer Cells

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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.

Derivation of Huntington Disease affected Genea046 human embryonic stem cell line

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Biljana Dumevska

2016-03-01

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

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.

Using Human Gestures and Generic Skills to Instruct a Mobile Robot Arm in a Feeder Filling Scenario

DEFF Research Database (Denmark)

Pedersen, Mikkel Rath; Høilund, Carsten; Krüger, Volker

2012-01-01

Mobile robots that have the ability to cooperate with humans are able to provide new possibilities to manufac- turing industries. In this paper, we discuss our mobile robot arm that a) can provide assistance at different locations in a factory and b) that can be programmed using complex human...... actions such as pointing in Take this object. In this paper, we discuss the use of the mobile robot for a feeding scenario where a human operator specifies the parts and the feeders through pointing gestures. The system is partially built using generic robotic skills. Through extensive experiments, we...

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.

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...

Differentiation of human pluripotent stem cells into highly functional classical brown adipocytes.

Science.gov (United States)

Nishio, Miwako; Saeki, Kumiko

2014-01-01

We describe a detailed method for directed differentiation of human pluripotent stem cells, including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), into functional classical brown adipocytes (BAs) under serum-free and feeder-free conditions. It is a two-tiered culture system, based on very simple techniques, a floating culture and a subsequent adherent culture. It does not require gene transfer. The entire process can be carried out in about 10 days. The key point is the usage of our special hematopoietic cytokine cocktail. Almost all the differentiated cells express uncoupling protein 1, a BA-selective marker, as determined by immunostaining. The differentiated cells show characteristics of classical BA as assessed by morphology and gene/protein expression. Moreover, the expression of myoblast marker genes is transiently induced during the floating culture step. hESC/hiPSC-derived BAs show significantly higher oxygen consumption rates (OCRs) than white adipocytes generated from human mesenchymal stem cell. They also show responsiveness to adrenergic stimuli, with about twofold upregulation in OCR by β-adrenergic receptor (β-AR) agonist treatments. hESC/hiPSC-derived BAs exert in vivo calorigenic activities in response to β-AR agonist treatments as assessed by thermography. Finally, lipid and glucose metabolisms are significantly improved in hESC/hiPSC-derived BA-transplanted mice. Our system provides a highly feasible way to produce functional classical BA bearing metabolism-improving capacities from hESC/hiPSC under a feeder-free and serum-free condition without gene transfer. © 2014 Elsevier Inc. All rights reserved.

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.

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)

Efficient Generation of Human Embryonic Stem Cell-Derived Corneal Endothelial Cells by Directed Differentiation.

Directory of Open Access Journals (Sweden)

Kathryn L McCabe

Full Text Available To generate human embryonic stem cell derived corneal endothelial cells (hESC-CECs for transplantation in patients with corneal endothelial dystrophies.Feeder-free hESC-CECs were generated by a directed differentiation protocol. hESC-CECs were characterized by morphology, expression of corneal endothelial markers, and microarray analysis of gene expression.hESC-CECs were nearly identical morphologically to primary human corneal endothelial cells, expressed Zona Occludens 1 (ZO-1 and Na+/K+ATPaseα1 (ATPA1 on the apical surface in monolayer culture, and produced the key proteins of Descemet's membrane, Collagen VIIIα1 and VIIIα2 (COL8A1 and 8A2. Quantitative PCR analysis revealed expression of all corneal endothelial pump transcripts. hESC-CECs were 96% similar to primary human adult CECs by microarray analysis.hESC-CECs are morphologically similar, express corneal endothelial cell markers and express a nearly identical complement of genes compared to human adult corneal endothelial cells. hESC-CECs may be a suitable alternative to donor-derived corneal endothelium.

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.

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.

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.

Derivation of NEM2 affected human embryonic stem cell line Genea079

Directory of Open Access Journals (Sweden)

Biljana Dumevska

2016-03-01

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

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.

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-

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.

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)

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.

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

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.

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.

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.

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

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

Epigenetic stability, adaptability, and reversibility in human embryonic stem cells

OpenAIRE

Tompkins, Joshua D.; Hall, Christine; Chen, Vincent Chang-yi; Li, Arthur Xuejun; Wu, Xiwei; Hsu, David; Couture, Larry A.; Riggs, Arthur D.

2012-01-01

The stability of human embryonic stem cells (hESCs) is of critical importance for both experimental and clinical applications. We find that as an initial response to altered culture conditions, hESCs change their transcription profile for hundreds of genes and their DNA methylation profiles for several genes outside the core pluripotency network. After adaption to conditions of feeder-free defined and/or xeno-free culture systems, expression and DNA methylation profiles are quite stable for a...

Directed Differentiation of Human-Induced Pluripotent Stem Cells to Mesenchymal Stem Cells.

Science.gov (United States)

Lian, Qizhou; Zhang, Yuelin; Liang, Xiaoting; Gao, Fei; Tse, Hung-Fat

2016-01-01

Multipotent stromal cells, also known as mesenchymal stem cells (MSCs), possess great potential to generate a wide range of cell types including endothelial cells, smooth muscle cells, bone, cartilage, and lipid cells. This protocol describes in detail how to perform highly efficient, lineage-specific differentiation of human-induced pluripotent stem cells (iPSCs) with an MSCs fate. The approach uses a clinically compliant protocol with chemically defined media, feeder-free conditions, and a CD105 positive and CD24 negative selection to achieve a single cell-based MSCs derivation from differentiating human pluripotent cells in approximately 20 days. Cells generated with this protocol express typical MSCs surface markers and undergo adipogenesis, osteogenesis, and chondrogenesis similar to adult bone marrow-derived MSCs (BM-MSCs). Nonetheless, compared with adult BM-MSCs, iPSC-MSCs display a higher proliferative capacity, up to 120 passages, without obvious loss of self-renewal potential and constitutively express MSCs surface antigens. MSCs generated with this protocol have numerous applications, including expansion to large scale cell numbers for tissue engineering and the development of cellular therapeutics. This approach has been used to rescue limb ischemia, allergic disorders, and cigarette smoke-induced lung damage and to model mesenchymal and vascular disorders of Hutchinson-Gilford progeria syndrome (HGPS).

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)

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)

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.

Derivation and characterisation of the human embryonic stem cell lines, NOTT1 and NOTT2.

Science.gov (United States)

Priddle, Helen; Allegrucci, Cinzia; Burridge, Paul; Munoz, Maria; Smith, Nigel M; Devlin, Lyndsey; Sjoblom, Cecilia; Chamberlain, Sarah; Watson, Sue; Young, Lorraine E; Denning, Chris

2010-04-01

The ability to maintain human embryonic stem cells (hESCs) during long-term culture and yet induce differentiation to multiple lineages potentially provides a novel approach to address various biomedical problems. Here, we describe derivation of hESC lines, NOTT1 and NOTT2, from human blastocysts graded as 3BC and 3CB, respectively. Both lines were successfully maintained as colonies by mechanical passaging on mouse embryonic feeder cells or as monolayers by trypsin-passaging in feeder-free conditions on Matrigel. Undifferentiated cells retained expression of pluripotency markers (OCT4, NANOG, SSEA-4, TRA-1-60 and TRA-1-81), a stable karyotype during long-term culture and could be transfected efficiently with plasmid DNA and short interfering RNA. Differentiation via formation of embryoid bodies resulted in expression of genes associated with early germ layers and terminal lineage specification. The electrophysiology of spontaneously beating NOTT1-derived cardiomyocytes was recorded and these cells were shown to be pharmacologically responsive. Histological examination of teratomas formed by in vivo differentiation of both lines in severe immunocompromised mice showed complex structures including cartilage or smooth muscle (mesoderm), luminal epithelium (endoderm) and neuroectoderm (ectoderm). These observations show that NOTT1 and NOTT2 display the accepted characteristics of hESC pluripotency.

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...

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).

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.

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

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

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)

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.

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 ...

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

Generation of clinical-grade human induced pluripotent stem cells in Xeno-free conditions.

Science.gov (United States)

Wang, Juan; Hao, Jie; Bai, Donghui; Gu, Qi; Han, Weifang; Wang, Lei; Tan, Yuanqing; Li, Xia; Xue, Ke; Han, Pencheng; Liu, Zhengxin; Jia, Yundan; Wu, Jun; Liu, Lei; Wang, Liu; Li, Wei; Liu, Zhonghua; Zhou, Qi

2015-11-12

Human induced pluripotent stem cells (hiPSCs) are considered as one of the most promising seed cell sources in regenerative medicine. Now hiPSC-based clinical trials are underway. To ensure clinical safety, cells used in clinical trials or therapies should be generated under GMP conditions, and with Xeno-free culture media to avoid possible side effects like immune rejection that induced by the Xeno reagents. However, up to now there are no reports for hiPSC lines developed completely under GMP conditions using Xeno-free reagents. Clinical-grade human foreskin fibroblast (HFF) cells used as feeder cells and parental cells of the clinical-grade hiPSCs were isolated from human foreskin tissues and cultured in Xeno-free media. Clinical-grade hiPSCs were derived by integration-free Sendai virus-based reprogramming kit in Xeno-free pluriton™ reprogramming medium or X medium. Neural cells and cardiomyocytes differentiation were conducted following a series of spatial and temporal specific signals induction according to the corresponding lineage development signals. Biological safety evaluation of the clinical-grade HFF cells and hiPSCs were conducted following the guidance of the "Pharmacopoeia of the People's Republic of China, Edition 2010, Volume III". We have successfully derived several integration-free clinical-grade hiPSC lines under GMP-controlled conditions and with Xeno-free reagents culture media in line with the current guidance of international and national evaluation criteria. As for the source of hiPSCs and feeder cells, biological safety evaluation of the HFF cells have been strictly reviewed by the National Institutes for Food and Drug Control (NIFDC). The hiPSC lines are pluripotent and have passed the safety evaluation. Moreover, one of the randomly selected hiPSC lines was capable of differentiating into functional neural cells and cardiomyocytes in Xeno-free culture media. The clinical-grade hiPSC lines therefore could be valuable sources for

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.

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)

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

Efficient and Cost-Effective Generation of Mature Neurons From Human Induced Pluripotent Stem Cells

OpenAIRE

Badja , Cherif; Maleeva , Galyna; El-Yazidi , Claire; Barruet , Emilie; Lasserre , Manon; Tropel , Philippe; Binetruy , Bernard; Bregestovski , Piotr; Magdinier , Frédérique

2014-01-01

The authors describe a feeder-free method of generating induced pluripotent stem cells by relying on the use of a chemically defined medium that overcomes the need for embryoid body formation and neuronal rosette isolation for neuronal precursors and terminally differentiated neuron production. This specific and efficient single-step strategy allows the production of mature neurons in 20–40 days with multiple applications, especially for modeling human pathologies.

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...

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...

Small-Molecule Induction Promotes Corneal Epithelial Cell Differentiation from Human Induced Pluripotent Stem Cells

Directory of Open Access Journals (Sweden)

Alexandra Mikhailova

2014-02-01

Full Text Available Human induced pluripotent stem cells (hiPSCs offer unique opportunities for developing novel cell-based therapies and disease modeling. In this study, we developed a directed differentiation method for hiPSCs toward corneal epithelial progenitor cells capable of terminal differentiation toward mature corneal epithelial-like cells. In order to improve the efficiency and reproducibility of our method, we replicated signaling cues active during ocular surface ectoderm development with the help of two small-molecule inhibitors in combination with basic fibroblast growth factor (bFGF in serum-free and feeder-free conditions. First, small-molecule induction downregulated the expression of pluripotency markers while upregulating several transcription factors essential for normal eye development. Second, protein expression of the corneal epithelial progenitor marker p63 was greatly enhanced, with up to 95% of cells being p63 positive after 5 weeks of differentiation. Third, corneal epithelial-like cells were obtained upon further maturation.

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.

Protein biosynthesis in cultured human hair follicle cells.

Science.gov (United States)

Weterings, P J; Vermorken, A J; Bloemendal, H

1980-10-31

A new technique has been used for culturing human keratinocytes. The cells grow on the basement membrane-like capsules of bovine lenses. Lens cells were removed from the capsules by rigid trypsinization. In order to exclude any contamination with remaining living cells the isolated capsules were irradiated with X-rays at a dose of 10,000 rad. In this way human epithelial cells can be brought in culture from individual hair follicles. Since feeder cells are not used in this culture technique, the biosynthesis of keratinocyte proteins can be studied in these cultures. The newly synthesized proteins can be separated into a water-soluble, a urea-soluble, and a urea-insoluble fraction. Product analysis has been performed on the first two fractions revealing protein patterns identical to those of intact hair follicles. Product analysis of the urea-soluble fractions of microdissected hair follicles shows that the protein pattern of the cultured keratinocytes resembles the protein pattern of the hair follicle sheath. Studies on the metabolism of benzo(a)pyrene revealed that the enzyme aryl hydrocarbon hydroxylase (AHH) is present in cultured hair follicle cells. A possible use of our culture system for eventual detection of inherited predisposition for smoking-dependent lung cancer is discussed.

Generation of human-induced pluripotent stem cells from burn patient-derived skin fibroblasts using a non-integrative method.

Science.gov (United States)

Fu, Shangfeng; Ding, Jianwu; Liu, Dewu; Huang, Heping; Li, Min; Liu, Yang; Tu, Longxiang; Liu, Deming

2018-01-01

Patient specific induced pluripotent stem cells (iPSCs) have been recognized as a possible source of cells for skin tissue engineering. They have the potential to greatly benefit patients with large areas of burned skin or skin defects. However, the integration virus-based reprogramming method is associated with a high risk of genetic mutation and mouse embryonic fibroblast feeder-cells may be a pollutant. In the present study, human skin fibroblasts (HSFs) were successfully harvested from patients with burns and patient-specific iPSCs were generated using a non-integration method with a feeder-free approach. The octamer-binding transcription factor 4 (OCT4), sex-determining region Y box 2 (SOX2) and NANOG transcription factors were delivered using Sendai virus vectors. iPSCs exhibited representative human embryonic stem cell-like morphology and proliferation characteristics. They also expressed pluripotent markers, including OCT4, NANOG, SOX2, TRA181, stage-specific embryonic antigen 4 and TRA-160, and exhibited a normal karyotype. Teratoma and embryoid body formation revealed that iPSCs were able to differentiate into cells of all three germ layers in vitro and in vivo. The results of the present study demonstrate that HSFs derived from patients with burns, may be reprogrammed into stem cells with pluripotency, which provides a basis for cell‑based skin tissue engineering in the future.

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.

Derivation and characterization of the NYSCFe003-A human embryonic stem cell line

Directory of Open Access Journals (Sweden)

Ana Sevilla

2017-12-01

Full Text Available The human embryonic stem cell line NYSCFe003-A was derived from a day 5 to 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 NYSCFe003-A line expresses all the pluripotency markers and has the potential to differentiate into all three germ layers in vitro. The line presents normal karyotype and is mycoplasma free.

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

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

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.

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.

Clonal proliferation of cultured nonmalignant and malignant human breast epithelia

International Nuclear Information System (INIS)

Smith, H.S.; Lan, S.; Ceriani, R.; Hackett, A.J.; Stampfer, M.R.

1981-01-01

We have developed a method for clonal growth of human mammary epithelial cells of both nonmalignant and malignant origin. Plating efficiencies of 1 to 50% were obtained by seeding second-passage mammary epithelial cells on fibroblast feeder layers in an enriched medium composed of various hormones and growth factors, as well as conditioned media from three specific human cell lines. Single mammary epithelial cells seeded sparsely onto the fibroblasts underwent at least eight population doublings to form large, readily visible colonies. Optimal colony formation required both feeder cells and the enriched medium. Epithelial colonies containing at least 16 cells were visible 5 days postseeding, and these colonies continued to grow progressively. Plating efficiency and colony size were similar on ultraviolet-irradiated or nonirradiated fibroblasts. The number of colonies formed was proportional to the number of epithelial cells plated. The colonies were identified as epithelial by the presence of human mammary epithelial antigens

Scalable Generation of Universal Platelets from Human Induced Pluripotent Stem Cells

Directory of Open Access Journals (Sweden)

Qiang Feng

2014-11-01

Full Text Available Human induced pluripotent stem cells (iPSCs provide a potentially replenishable source for the production of transfusable platelets. Here, we describe a method to generate megakaryocytes (MKs and functional platelets from iPSCs in a scalable manner under serum/feeder-free conditions. The method also permits the cryopreservation of MK progenitors, enabling a rapid “surge” capacity when large numbers of platelets are needed. Ultrastructural/morphological analyses show no major differences between iPSC platelets and human blood platelets. iPSC platelets form aggregates, lamellipodia, and filopodia after activation and circulate in macrophage-depleted animals and incorporate into developing mouse thrombi in a manner identical to human platelets. By knocking out the β2-microglobulin gene, we have generated platelets that are negative for the major histocompatibility antigens. The scalable generation of HLA-ABC-negative platelets from a renewable cell source represents an important step toward generating universal platelets for transfusion as well as a potential strategy for the management of platelet refractoriness.

Efficient generation of functional pancreatic β-cells from human induced pluripotent stem cells.

Science.gov (United States)

Yabe, Shigeharu G; Fukuda, Satsuki; Takeda, Fujie; Nashiro, Kiyoko; Shimoda, Masayuki; Okochi, Hitoshi

2017-02-01

Insulin-secreting cells have been generated from human embryonic or induced pluripotent stem cells (iPSCs) by mimicking developmental processes. However, these cells do not always secrete glucose-responsive insulin, one of the most important characteristics of pancreatic β-cells. We focused on the importance of endodermal differentiation from human iPSCs in order to obtain functional pancreatic β-cells. A six-stage protocol was established for the differentiation of human iPSCs to pancreatic β-cells using defined culture media without feeders or serum. The effects of CHIR99021, a selective glycogen synthase kinase-3β inhibitor, were examined in the presence of fibroblast growth factor 2, activin, and bone morphogenetic protein 4 (FAB) during definitive endodermal induction by immunostaining for SRY (sex determining region Y)-box 17 (SOX17) and Forkhead box protein A2 (FOXA2). Insulin secretion was compared between the last stage of monolayer culture and spheroid culture conditions. Cultured cells were transplanted under kidney capsules of streptozotocin-diabetic non-obese diabetic-severe combined immunodeficiency mice, and blood glucose levels were measured once a week. Immunohistochemical analyses were performed 4 and 12 weeks after transplantation. Addition of CHIR99021 (3 μmol/L) in the presence of FAB for 2 days improved endodermal cell viability, maintaining the high SOX17-positive rate. Spheroid formation after the endocrine progenitor stage showed more efficient insulin secretion than did monolayer culture. After cell transplantation, diabetic mice had lower blood glucose levels, and islet-like structures were detected in vivo. Functional pancreatic β-cells were generated from human iPSCs. Induction of definitive endoderm and spheroid formation may be key steps for producing these cells. © 2016 Ruijin Hospital, Shanghai Jiaotong University School of Medicine and John Wiley & Sons Australia, Ltd.

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.

YKL-40 is differentially expressed in human embryonic stem cells and in cell progeny of the three germ layers

DEFF Research Database (Denmark)

Brøchner, Christian B; Johansen, Julia S; Larsen, Lars A

2012-01-01

oxygen tension, in culture medium with or without basic fibroblast growth factor, and on feeder layers comprising mouse embryonic fibroblasts or human foreskin fibroblasts to evaluate whether hESCs and their progeny produced YKL-40 and to characterize YKL-40 expression during differentiation. Secreted......The secreted glycoprotein YKL-40 participates in cell differentiation, inflammation, and cancer progression. High YKL-40 expression is reported during early human development, but its functions are unknown. Six human embryonic stem cell (hESC) lines were cultured in an atmosphere of low or high...... YKL-40 protein and YKL-40 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative RT-PCR. Serial-sectioned colonies were stained for YKL-40 protein and for pluripotent hESC (OCT4, NANOG) and germ layer (HNF-3ß, PDX1, CD34, p63, nestin, PAX6) markers. Double...

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

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...

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

Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Including Primary Cilia Associated Signaling Pathways

DEFF Research Database (Denmark)

Vestergaard, Maj Linea; Awan, Aashir; Warzecha, Caroline Becker

2016-01-01

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

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

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)

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...

Chemosensitivity and radiosensitivity testing of freshly explanted human tumour cells in vitro

International Nuclear Information System (INIS)

Wells, J.

1977-10-01

In this thesis, in vitro testing for the chemosensitivity and radiosensitivity of freshly explanted human tumour cells is described. The cells were incubated with anti-tumour drugs and either a 6-day growth test performed or a clonal growth test as a measure of survival of cell reproductive capacity. It was shown that if one aims to develop a suitable in vitro method for predicting the subsequent response of human tumour cells in situ to cytotoxic chemotherapy, the test procedure must be initiated before the explanted cells have undergone significant growth in vitro. The survival of the reproductive capacity of tumour cell explants following X-radiation was also studied. Using a 'feeder' layer technique, values for the survival curve parameter Dsub(q) were in the range 400-610 rad and the values for D 0 were in the range 120-160 rad. The shape of the X-ray survival curves did not change when cells were retested after repeated subculturing in vitro. Therefore, unlike chemosensitivity measured by the same biological end-point, radiosensitivity apparently does not change once cells have reached their maximum growth potential. (UK)

Low calcium culture condition induces mesenchymal cell-like phenotype in normal human epidermal keratinocytes

International Nuclear Information System (INIS)

Takagi, Ryo; Yamato, Masayuki; Murakami, Daisuke; Sugiyama, Hiroaki; Okano, Teruo

2011-01-01

Highlights: → Normal human epidermal keratinocytes serially cultured under low calcium concentration were cytokeratin and vimentin double positive cells. → The human keratinocytes expressed some epithelial stem/progenitor cell makers, mesenchymal cell markers, and markers of epithelial-mesenchymal transition. → Mesenchymal cell-like phenotype in the keratinocytes was suppressed under high-calcium condition. -- Abstract: Epithelial-mesenchymal transition (EMT) is an important cellular phenomenon in organ developments, cancer invasions, and wound healing, and many types of transformed cell lines are used for investigating for molecular mechanisms of EMT. However, there are few reports for EMT in normal human epithelial cells, which are non-transformed or non-immortalized cells, in vitro. Therefore, normal human epidermal keratinocytes (NHEK) serially cultured in low-calcium concentration medium (LCM) were used for investigating relations between differentiation and proliferation and mesenchymal-like phenotype in the present study, since long-term cultivation of NHEK is achieved in LCM. Interestingly, NHEK serially cultured in LCM consisted essentially of cytokeratin-vimentin double positive cells (98%), although the NHEK exhibited differentiation under high-calcium culture condition with 3T3 feeder layer. The vimentin expression was suppressed under high-calcium condition. These results may indicate the importance of mesenchymal-like phenotype for serially cultivation of NHEK in vitro.

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.

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)

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

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)

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.

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.

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

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

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

Rapid fabricating technique for multi-layered human hepatic cell sheets by forceful contraction of the fibroblast monolayer.

Directory of Open Access Journals (Sweden)

Yusuke Sakai

Full Text Available Cell sheet engineering is attracting attention from investigators in various fields, from basic research scientists to clinicians focused on regenerative medicine. However, hepatocytes have a limited proliferation potential in vitro, and it generally takes a several days to form a sheet morphology and multi-layered sheets. We herein report our rapid and efficient technique for generating multi-layered human hepatic cell (HepaRG® cell sheets using pre-cultured fibroblast monolayers derived from human skin (TIG-118 cells as a feeder layer on a temperature-responsive culture dish. Multi-layered TIG-118/HepaRG cell sheets with a thick morphology were harvested on day 4 of culturing HepaRG cells by forceful contraction of the TIG-118 cells, and the resulting sheet could be easily handled. In addition, the human albumin and alpha 1-antitrypsin synthesis activities of TIG-118/HepaRG cells were approximately 1.2 and 1.3 times higher than those of HepaRG cells, respectively. Therefore, this technique is considered to be a promising modality for rapidly fabricating multi-layered human hepatocyte sheets from cells with limited proliferation potential, and the engineered cell sheet could be used for cell transplantation with highly specific functions.

The role of NF-κB signaling in the maintenance of pluripotency of human induced pluripotent stem cells.

Directory of Open Access Journals (Sweden)

Osamu Takase

Full Text Available NF-κB signaling plays an essential role in maintaining the undifferentiated state of embryonic stem (ES cells. However, opposing roles of NF-κB have been reported in mouse and human ES cells, and the role of NF-κB in human induced pluripotent stem (iPS cells has not yet been clarified. Here, we report the role of NF-κB signaling in maintaining the undifferentiated state of human iPS cells. Compared with differentiated cells, undifferentiated human iPS cells showed an augmentation of NF-κB activity. During differentiation induced by the removal of feeder cells and FGF2, we observed a reduction in NF-κB activity, the expression of the undifferentiation markers Oct3/4 and Nanog, and the up-regulation of the differentiated markers WT-1 and Pax-2. The specific knockdown of NF-κB signaling using p65 siRNA also reduced the expression of Oct3/4 and Nanog and up-regulated WT-1 and Pax-2 but did not change the ES-like colony formation. Our results show that the augmentation of NF-κB signaling maintains the undifferentiated state of human iPS and suggest the importance of this signaling pathway in maintenance of human iPS cells.

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.

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.

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

Generation of Hepatocyte-like Cells from Human Induced Pluripotent Stem (iPS) Cells By Co-culturing Embryoid Body Cells with Liver Non-parenchymal Cell Line TWNT-1

International Nuclear Information System (INIS)

Javed, M. S.; Yaqoob, N.; Iwamuro, M.; Kobayashi, N.; Fujiwara, T.

2014-01-01

Objective: To generate a homogeneous population of patient-specific hepatocyte-like cells (HLCs) from human iPS cells those show the morphologic and phenotypic properties of primary human hepatocytes. Study Design: An experimental study. Place and Duration of Study: Department of Surgery, Okayama University, Graduate School of Medicine, Japan, from April to December 2011. Methodology: Human iPS cells were generated and maintained on ES qualified matrigel coated plates supplemented with mTeSR medium or alternatively on mitotically inactivated MEF feeder layer in DMEM/F12 medium containing 20% KOSR, 4ng/ml bFGF-2, 1 x 10-4 M 2-mercaptoethanol, 1 mmol/L NEAA, 2mM L-glutamine and 1% penicillin-streptomycin. iPS cells were differentiated to HLCs by sequential culture using a four step differentiation protocol: (I) Generation of embryoid bodies (EBs) in suspension culture; (II) Induction of definitive endoderm (DE) from 2 days old EBs by growth in human activin-A (100 ng/ml) and basic fibroblasts growth factor (bFGF2) (100 ng/ml) on matrigel coated plates; (III) Induction of hepatic progenitors by co-culture with non-parenchymal human hepatic stellate cell line (TWNT-1); and (IV) Maturation by culture in dexamethasone. Characterization was performed by RT-PCR and functional assays. Results: The generated HLCs showed microscopically morphological phenotype of human hepatocytes, expressed liver specific genes (ASGPR, Albumin, AFP, Sox17, Fox A2), secreted human liver-specific proteins such as albumin, synthesized urea and metabolized ammonia. Conclusion: Functional HLCs were generated from human iPS cells, which could be used for autologus hepatocyte transplantation for liver failure and as in vitro model for determining the metabolic and toxicological properties of drug compounds. (author)

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.

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.

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

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...

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.

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)

Defined Conditions for the Isolation and Expansion of Basal Prostate Progenitor Cells of Mouse and Human Origin

Directory of Open Access Journals (Sweden)

Thomas Höfner

2015-03-01

Full Text Available Methods to isolate and culture primary prostate epithelial stem/progenitor cells (PESCs have proven difficult and ineffective. Here, we present a method to grow and expand both murine and human basal PESCs long term in serum- and feeder-free conditions. The method enriches for adherent mouse basal PESCs with a Lin−SCA-1+CD49f+TROP2high phenotype. Progesterone and sodium selenite are additionally required for the growth of human Lin−CD49f+TROP2high PESCs. The gene-expression profiles of expanded basal PESCs show similarities to ESCs, and NF-kB function is critical for epithelial differentiation of sphere-cultured PESCs. When transplanted in combination with urogenital sinus mesenchyme, expanded mouse and human PESCs generate ectopic prostatic tubules, demonstrating their stem cell activity in vivo. This novel method will facilitate the molecular, genomic, and functional characterization of normal and pathologic prostate glands of mouse and human origin.

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.

[The characters and specific features of new human embryonic stem cells lines].

Science.gov (United States)

Krylova, T A; Kol'tsova, A M; Zenin, V V; Gordeeva, O F; Musorina, A S; Goriachaia, T S; Shlykova, S A; Kamenetskaia, Iu K; Pinaev, G P; Polianskaia, G G

2009-01-01

Four continuous human embryonic stem cell lines (SC1, SC2, SC3 and SC4), derived from the blastocysts has been described. The cell lines were cultivated on mitotically inactivated human feeder cells. The cell lines SC1 and SC2 have passed through 150 population doublings and the cell lines SC3 and SC4 -- near 120 populations doublings, which exceeds Hayflick limit sufficiently. These cell lines maintain high activity of alkaline phosphatase, expression of transcription factor OCT-4 and cell surface antigens (SSEA-4, TRA-1-60 and TRA-1-81), confirming their ESC status and human specificity. Immunofluorescent detection of antigens, characteristic of ectoderm, endoderm and mesoderm confirms the ability of these cells to retain their pluripotency under in vitro condition. PCR analysis revealed expression of six genes specific for pluripotent cells (OCT-4, NANOG, DPPA3/STELLA, TDGF/CRIPTO and LEFTYA). Correlation between the level of proliferative activity and the character of DNA-bound fluorescent staining was found. Fluorescent dyes, Hoechst 33342 and PI, produced diffuse staining of the nuclei in slowly proliferating cells of the SC1 and SC2 lines. In contrast, in actively proliferating cells of the SC3 and SC4 lines, the clear staining of the nuclei was observed. Upon changing the cultivation condition, proliferative activity of SC3 and SC4 lines decreased and became similar to that of SC1 and SC2 lines. The character of the fluorescent staining of all these lines was also shown to be similar. These results show that quality of the fluorescent staining with Hoechst 33342 and PI reflects the level of proliferation. Possible causes and mechanisms of this feature of human ESC are discussed.

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).

Human corneal epithelial subpopulations

DEFF Research Database (Denmark)

Søndergaard, Chris Bath

2013-01-01

Corneal epithelium is being regenerated throughout life by limbal epithelial stem cells (LESCs) believed to be located in histologically defined stem cell niches in corneal limbus. Defective or dysfunctional LESCs result in limbal stem cell deficiency (LSCD) causing pain and decreased visual acuity...... subpopulations in human corneal epithelium using a combination of laser capture microdissection and RNA sequencing for global transcriptomic profiling. We compared dissociation cultures, using either expansion on γ-irradiated NIH/3T3 feeder cells in serum-rich medium or expansion directly on plastic in serum...

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.

Derivation of Stromal (Skeletal and Mesenchymal) Stem-Like Cells from Human Embryonic Stem Cells

Science.gov (United States)

Harkness, Linda; Abdallah, Basem M.; Elsafadi, Mona; Al-Nbaheen, May S.; Aldahmash, Abdullah; Kassem, Moustapha

2012-01-01

Derivation of bone forming cells (osteoblasts) from human embryonic stem cells (hESCs) is a prerequisite for their use in clinical applications. However, there is no standard protocol for differentiating hESCs into osteoblastic cells. The aim of this study was to identify the emergence of a human stromal (mesenchymal and 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 hESCs in a feeder-free environment using serum replacement and as suspension aggregates (embryoid bodies; hEBs). Over a 20 day developmental period, the hEBs demonstrated increasing enrichment for cells expressing hMSC markers: CD29, CD44, CD63, CD56, CD71, CD73, CD105, CD106, and CD166 as revealed by immunohistochemical staining and flow cytometry (fluorescence-activated cell sorting) analysis. Ex vivo differentiation of hEBs using bone morphogenic protein 2 (BMP2) 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, revealed bone and cartilage, and fibrous tissue elements after 8 weeks. These tissues were of human origin and there was no evidence of differentiation to nonmesodermal tissues. hEBs implanted in the absence of HA/TCP formed vacuolated tissue containing glandular, fibrous and muscle-like tissue elements. Conversely, implantation of undifferentiated hESCs resulted in the formation of a teratoma containing a mixture of endodermal, mesodermal, and ectodermal tissues. Our study demonstrates that hMSC-like cells can be obtained from hESCs and they can be induced to form skeletal tissues in vivo when combined with HA/TCP. These findings are relevant for tissue engineering and suggest that differentiated hEBs can provide an unlimited source for

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.

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.

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

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

YKL-40 is differentially expressed in human embryonic stem cells and in cell progeny of the three germ layers.

Science.gov (United States)

Brøchner, Christian B; Johansen, Julia S; Larsen, Lars A; Bak, Mads; Mikkelsen, Hanne B; Byskov, Anne Grete; Andersen, Claus Yding; Møllgård, Kjeld

2012-03-01

The secreted glycoprotein YKL-40 participates in cell differentiation, inflammation, and cancer progression. High YKL-40 expression is reported during early human development, but its functions are unknown. Six human embryonic stem cell (hESC) lines were cultured in an atmosphere of low or high oxygen tension, in culture medium with or without basic fibroblast growth factor, and on feeder layers comprising mouse embryonic fibroblasts or human foreskin fibroblasts to evaluate whether hESCs and their progeny produced YKL-40 and to characterize YKL-40 expression during differentiation. Secreted YKL-40 protein and YKL-40 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative RT-PCR. Serial-sectioned colonies were stained for YKL-40 protein and for pluripotent hESC (OCT4, NANOG) and germ layer (HNF-3β, PDX1, CD34, p63, nestin, PAX6) markers. Double-labeling showed YKL-40 expression in OCT4-positive hESCs, PAX6-positive neuroectodermal cells, and HNF-3β-positive endodermal cells. The differentiating progeny showed strong YKL-40 expression. Abrupt transition between YKL-40 and OCT4-positive hESCs and YKL-40-positive ecto- and neuroectodermal lineages was observed within the same epithelial-like layer. YKL-40-positive cells within deeper layers lacked contact with OCT4-positive cells. YKL-40 may be important in initial cell differentiation from hESCs toward ectoderm and neuroectoderm, with retained epithelial morphology, whereas later differentiation into endoderm and mesoderm involves a transition into the deeper layers of the colony.

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.

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)

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)

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)

Generation of a human induced pluripotent stem cell line (MUSIi001-A from caesarean section scar fibroblasts using Sendai viral vectors

Directory of Open Access Journals (Sweden)

Methichit Wattanapanitch

2018-03-01

Full Text Available We generated a human induced pluripotent stem cell (iPSC line from caesarean section scar fibroblasts of a 33-year-old healthy woman using transgene-free Sendai viral vectors under feeder-free condition. The established iPSC line, designated as MUSIi001-A, exhibited a normal karyotype, expressed pluripotent markers, differentiated into cells of three embryonic germ layers. Further analyses showed that the Sendai viral genome was absent at passage 25. The MUSIi001-A line can serve as a control for studying developmental biology and phenotypic comparison with disease-specific iPSCs.

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...

Effects of mechanical stimulation on the reprogramming of somatic cells into human-induced pluripotent stem cells.

Science.gov (United States)

Kim, Young Mi; Kang, Yun Gyeong; Park, So Hee; Han, Myung-Kwan; Kim, Jae Ho; Shin, Ji Won; Shin, Jung-Woog

2017-06-08

Mechanical stimuli play important roles in the proliferation and differentiation of adult stem cells. However, few studies on their effects on induced pluripotent stem cells (iPSCs) have been published. Human dermal fibroblasts were seeded onto flexible membrane-bottom plates, and infected with retrovirus expressing the four reprogramming factors OCT4, SOX2, KLF, and c-MYC (OSKM). The cells were subjected to equiaxial stretching (3% or 8% for 2, 4, or 7 days) and seeded on feeder cells (STO). The reprogramming into iPSCs was evaluated by the expression of pluripotent markers, in vitro differentiation into three germ layers, and teratoma formation. Equiaxial stretching enhanced reprogramming efficiency without affecting the viral transduction rate. iPSCs induced by transduction of four reprogramming factors and application of equiaxial stretching had characteristics typical of iPSCs in terms of pluripotency and differentiation potentials. This is the first study to show that mechanical stimuli can increase reprogramming efficiency. However, it did not enhance the infection rate, indicating that mechanical stimuli, defined as stretching in this study, have positive effects on reprogramming rather than on infection. Additional studies should evaluate the mechanism underlying the modulation of reprogramming of somatic cells into iPSCs.

Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) blocks differentiation and maintains the expression of pluripotency markers in human embryonic stem cells.

Science.gov (United States)

Burton, Peter; Adams, David R; Abraham, Achamma; Allcock, Robert W; Jiang, Zhong; McCahill, Angela; Gilmour, Jane; McAbney, John; Kaupisch, Alexandra; Kane, Nicole M; Baillie, George S; Baker, Andrew H; Milligan, Graeme; Houslay, Miles D; Mountford, Joanne C

2010-12-15

hESCs (human embryonic stem cells) have enormous potential for use in pharmaceutical development and therapeutics; however, to realize this potential, there is a requirement for simple and reproducible cell culture methods that provide adequate numbers of cells of suitable quality. We have discovered a novel way of blocking the spontaneous differentiation of hESCs in the absence of exogenous cytokines by supplementing feeder-free conditions with EHNA [erythro-9-(2-hydroxy-3-nonyl)adenine], an established inhibitor of ADA (adenosine deaminase) and cyclic nucleotide PDE2 (phosphodiesterase 2). hESCs maintained in feeder-free conditions with EHNA for more than ten passages showed no reduction in hESC-associated markers including NANOG, POU5F1 (POU domain class 5 transcription factor 1, also known as Oct-4) and SSEA4 (stage-specific embryonic antigen 4) compared with cells maintained in feeder-free conditions containing bFGF (basic fibroblast growth factor). Spontaneous differentiation was reversibly suppressed by the addition of EHNA, but, upon removing EHNA, hESC populations underwent efficient spontaneous, multi-lineage and directed differentiation. EHNA also acts as a strong blocker of directed neuronal differentiation. Chemically distinct inhibitors of ADA and PDE2 lacked the capacity of EHNA to suppress hESC differentiation, suggesting that the effect is not driven by inhibition of either ADA or PDE2. Preliminary structure-activity relationship analysis found the differentiation-blocking properties of EHNA to reside in a pharmacophore comprising a close adenine mimetic with an extended hydrophobic substituent in the 8- or 9-position. We conclude that EHNA and simple 9-alkyladenines can block directed neuronal and spontaneous differentiation in the absence of exogenous cytokine addition, and may provide a useful replacement for bFGF in large-scale or cGMP-compliant processes.

System-wide temporal characterization of the proteome and phosphoproteome of human embryonic stem cell differentiation

DEFF Research Database (Denmark)

Rigbolt, Kristoffer T.G.; Prokhorova, Tatyana; Akimov, Vyacheslav

2011-01-01

by feeder cells. We profiled 6521 proteins and 23,522 phosphorylation sites, of which almost 50% displayed dynamic changes in phosphorylation status during 24 hours of differentiation. These data are a resource for studies of the events associated with the maintenance of hESC pluripotency and those...... of the matching sequence motif. In addition to identifying previously unknown phosphorylation sites on factors associated with differentiation, such as kinases and transcription factors, we observed dynamic phosphorylation of DNA methyltransferases (DNMTs). We found a specific interaction of DNMTs during early......To elucidate cellular events underlying the pluripotency of human embryonic stem cells (hESCs), we performed parallel quantitative proteomic and phosphoproteomic analyses of hESCs during differentiation initiated by a diacylglycerol analog or transfer to media that had not been conditioned...

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.

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...

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.…

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...

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.

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

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.

An in vitro expansion system for generation of human iPS cell-derived hepatic progenitor-like cells exhibiting a bipotent differentiation potential.

Directory of Open Access Journals (Sweden)

Ayaka Yanagida

Full Text Available Hepatoblasts, hepatic stem/progenitor cells in liver development, have a high proliferative potential and the ability to differentiate into both hepatocytes and cholangiocytes. In regenerative medicine and drug screening for the treatment of severe liver diseases, human induced pluripotent stem (iPS cell-derived mature functional hepatocytes are considered to be a potentially good cell source. However, induction of proliferation of these cells is difficult ex vivo. To circumvent this problem, we generated hepatic progenitor-like cells from human iPS cells using serial cytokine treatments in vitro. Highly proliferative hepatic progenitor-like cells were purified by fluorescence-activated cell sorting using antibodies against CD13 and CD133 that are known cell surface markers of hepatic stem/progenitor cells in fetal and adult mouse livers. When the purified CD13(highCD133(+ cells were cultured at a low density with feeder cells in the presence of suitable growth factors and signaling inhibitors (ALK inhibitor A-83-01 and ROCK inhibitor Y-27632, individual cells gave rise to relatively large colonies. These colonies consisted of two types of cells expressing hepatocytic marker genes (hepatocyte nuclear factor 4α and α-fetoprotein and a cholangiocytic marker gene (cytokeratin 7, and continued to proliferate over long periods of time. In a spheroid formation assay, these cells were found to express genes required for mature liver function, such as cytochrome P450 enzymes, and secrete albumin. When these cells were cultured in a suitable extracellular matrix gel, they eventually formed a cholangiocytic cyst-like structure with epithelial polarity, suggesting that human iPS cell-derived hepatic progenitor-like cells have a bipotent differentiation ability. Collectively these data indicate that this novel procedure using an in vitro expansion system is useful for not only liver regeneration but also for the determination of molecular mechanisms that

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.

Directed differentiation of basal forebrain cholinergic neurons from human pluripotent stem cells.

Science.gov (United States)

Hu, Yao; Qu, Zhuang-Yin; Cao, Shi-Ying; Li, Qi; Ma, Lixiang; Krencik, Robert; Xu, Min; Liu, Yan

2016-06-15

Basal forebrain cholinergic neurons (BFCNs) play critical roles in learning, memory and cognition. Dysfunction or degeneration of BFCNs may connect to neuropathology, such as Alzheimer's disease, Down's syndrome and dementia. Generation of functional BFCNs may contribute to the studies of cell-based therapy and pathogenesis that is related to learning and memory deficits. Here we describe a detail method for robust generation of BFCNs from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). In this method, BFCN progenitors are patterned from hESC or hiPSC-derived primitive neuroepithelial cells, with the treatment of sonic hedgehog (SHH) or combination with its agonist Purmorphamine, and by co-culturing with human astrocytes. At day 20, ∼90% hPSC-derived progenitors expressed NKX2.1, which is a transcriptional marker for MGE. Moreover, around 40% of NKX2.1+ cells co-expressed OLIG2 and ∼15% of NKX2.1+ cells co-expressed ISLET1, which are ventral markers. At day 35, ∼40% neurons robustly express ChAT, most of which are co-labeled with NKX2.1, ISLET1 and FOXG1, indicating the basal forebrain-like identity. At day 45, these neurons express mature neuronal markers MAP2, Synapsin, and VAChT. In this method, undefined conditions including genetic modification or cell-sorting are avoided. As a choice, feeder free conditions are used to avoid ingredients of animal origin. Moreover, Purmorphamine can be substituted for SHH to induce ventral progenitors effectively and economically. We provide an efficient method to generate BFCNs from multiple hPSC lines, which offers the potential application for disease modeling and pharmacological studies. Copyright © 2016 Elsevier B.V. All rights reserved.

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

Human T cell colony formation in microculture: analysis of growth requirements and functional activities.

Science.gov (United States)

Gelfand, E W; Lee, J W; Dosch, H M; Price, G B

1981-03-01

A microculture method in methylcellulose has been developed for the study of human T cell colony formation. The technique is simple, reliable, does not require preincubation with lectin and requires small numbers of cells. Colony formation was dependent on the presence of phytohemagglutin-conditioned medium, a T colony precursor cell (TCPC), and a "helper" or accessory T cell. Plating efficiency was increased 10-fold in the presence of irradiated feeder cells. Progenitors of the T colony cells were identified in peripheral blood, tonsil, and spleen but not in thymus or thoracic duct. They were isolated in the E-rosetting, theophylline-resistant, Fc-IgG-negative cell populations. In peripheral blood the frequency of TCPC and accessory cells, the T colony forming unit, was estimated to be 8 X 10(-3). Colony cells proliferated in response to lectins and allogeneic cells. Forty to 80% of the cells were Ia-positive and stimulated both autologous and allogeneic mixed lymphocyte responses. They were incapable of mediating antibody-dependent cytotoxicity. In contrast, they were effective in assays of spontaneous cytotoxicity but only against certain target cells. This method for the analysis of T colony formation should prove valuable in the functional analysis of T cell subsets in immunodeficiency states or the transplant recipient.

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...

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

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.

Functional vascular smooth muscle cells derived from human induced pluripotent stem cells via mesenchymal stem cell intermediates

Science.gov (United States)

Bajpai, Vivek K.; Mistriotis, Panagiotis; Loh, Yuin-Han; Daley, George Q.; Andreadis, Stelios T.

2012-01-01

Aims Smooth muscle cells (SMC) play an important role in vascular homeostasis and disease. Although adult mesenchymal stem cells (MSC) have been used as a source of contractile SMC, they suffer from limited proliferation potential and culture senescence, particularly when originating from older donors. By comparison, human induced pluripotent stem cells (hiPSC) can provide an unlimited source of functional SMC for autologous cell-based therapies and for creating models of vascular disease. Our goal was to develop an efficient strategy to derive functional, contractile SMC from hiPSC. Methods and results We developed a robust, stage-wise, feeder-free strategy for hiPSC differentiation into functional SMC through an intermediate stage of multipotent MSC, which could be coaxed to differentiate into fat, bone, cartilage, and muscle. At this stage, the cells were highly proliferative and displayed higher clonogenic potential and reduced senescence when compared with parental hair follicle mesenchymal stem cells. In addition, when exposed to differentiation medium, the myogenic proteins such as α-smooth muscle actin, calponin, and myosin heavy chain were significantly upregulated and displayed robust fibrillar organization, suggesting the development of a contractile phenotype. Indeed, tissue constructs prepared from these cells exhibited high levels of contractility in response to receptor- and non-receptor-mediated agonists. Conclusion We developed an efficient stage-wise strategy that enabled hiPSC differentiation into contractile SMC through an intermediate population of clonogenic and multipotent MSC. The high yield of MSC and SMC derivation suggests that our strategy may facilitate an acquisition of the large numbers of cells required for regenerative medicine or for studying vascular disease pathophysiology. PMID:22941255

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.

Efficient, long term production of monocyte-derived macrophages from human pluripotent stem cells under partly-defined and fully-defined conditions.

Directory of Open Access Journals (Sweden)

Bonnie van Wilgenburg

Full Text Available Human macrophages are specialised hosts for HIV-1, dengue virus, Leishmania and Mycobacterium tuberculosis. Yet macrophage research is hampered by lack of appropriate cell models for modelling infection by these human pathogens, because available myeloid cell lines are, by definition, not terminally differentiated like tissue macrophages. We describe here a method for deriving monocytes and macrophages from human Pluripotent Stem Cells which improves on previously published protocols in that it uses entirely defined, feeder- and serum-free culture conditions and produces very consistent, pure, high yields across both human Embryonic Stem Cell (hESC and multiple human induced Pluripotent Stem Cell (hiPSC lines over time periods of up to one year. Cumulatively, up to ∼3×10(7 monocytes can be harvested per 6-well plate. The monocytes produced are most closely similar to the major blood monocyte (CD14(+, CD16(low, CD163(+. Differentiation with M-CSF produces macrophages that are highly phagocytic, HIV-1-infectable, and upon activation produce a pro-inflammatory cytokine profile similar to blood monocyte-derived macrophages. Macrophages are notoriously hard to genetically manipulate, as they recognise foreign nucleic acids; the lentivector system described here overcomes this, as pluripotent stem cells can be relatively simply genetically manipulated for efficient transgene expression in the differentiated cells, surmounting issues of transgene silencing. Overall, the method we describe here is an efficient, effective, scalable system for the reproducible production and genetic modification of human macrophages, facilitating the interrogation of human macrophage biology.

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.

Generation of hematopoietic stem cells from human embryonic stem cells using a defined, stepwise, serum-free, and serum replacement-free monolayer culture method.

Science.gov (United States)

Kim, So-Jung; Jung, Ji-Won; Ha, Hye-Yeong; Koo, Soo Kyung; Kim, Eung-Gook; Kim, Jung-Hyun

2017-03-01

Embryonic stem cells (ESCs) can be expanded infinitely in vitro and have the potential to differentiate into hematopoietic stem cells (HSCs); thus, they are considered a useful source of cells for HSC production. Although several technical in vitro methods for engineering HSCs from pluripotent stem cells have been developed, clinical application of HSCs engineered from pluripotent stem cells is restricted because of the possibility of xenogeneic contamination resulting from the use of murine materials. Human ESCs (CHA-hES15) were cultured on growth factor-reduced Matrigel-coated dishes in the mTeSR1 serum-free medium. When the cells were 70% confluent, we initiated HSC differentiation by three methods involving (1) knockout serum replacement (KSR), cytokines, TGFb1, EPO, and FLT3L; (2) KSR, cytokines, and bFGF; or (3) cytokines and bFGF. Among the three differentiation methods, the minimal number of cytokines without KSR resulted in the greatest production of HSCs. The optimized method resulted in a higher proportion of CD34 + CD43 + hematopoietic progenitor cells (HPCs) and CD34 + CD45 + HPCs compared to the other methods. In addition, the HSCs showed the potential to differentiate into multiple lineages of hematopoietic cells in vitro . In this study, we optimized a two-step, serum-free, animal protein-free, KSR-free, feeder-free, chemically defined monolayer culture method for generation of HSCs and hematopoietic stem and progenitor cells (HSPCs) from human ESCs.

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.

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

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....

GMP scale-up and banking of pluripotent stem cells for cellular therapy applications.

Science.gov (United States)

Ausubel, Lara J; Lopez, Patricia M; Couture, Larry A

2011-01-01

Human pluripotent stem cells (PSCs), which include human embryonic stem cells (ESCs) as well as induced pluripotent stem cells (iPSCs), represent an important source of cellular therapies in regenerative medicine and the study of early human development. As such, it is becoming increasingly important to develop methods for the large-scale banking of human PSC lines. There are several well-established methods for the propagation of human PSCs. The key to development of a good manufacturing practice (GMP) bank is to determine a manufacturing method that is amenable to large-scale production using materials that are fully documented. We have developed several banks of hESCs using animal feeder cells, animal-based matrices, or animal-free matrices. Protocols for growing hESCs on mouse embryonic fibroblasts (MEFs) are well established and are very helpful for producing research grade banks of cells. As most human ESCs cultured by research laboratories have been exposed to xenogeneic reagents, it is not imperative that all materials used in the production of a master cell bank be animal-free in origin. Nevertheless, as the field develops, it will no doubt become increasingly important to produce a bank of cells for clinical use without xenogeneic reagents, particularly nonhuman feeder cells which might harbor viruses with potential risk to human health or cell product integrity. Thus, even for cell lines previously exposed to xenogeneic reagents, it is important to minimize any subsequent exposure of the cell lines to additional adventitious agents. We have specifically described procedures for the growth of hESCs on Matrigel, an animal-matrix, and CELLstart, an animal-free matrix, and these can be used to produce hESCs as part of a clinical manufacturing process.

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.)

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...

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.

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...

Serial cultivation of human scalp hair follicle keratinocytes.

Science.gov (United States)

Weterings, P J; Roelofs, H M; Vermorken, A J; Bloemendal, H

1983-01-01

A method is described for the serial cultivation of adult human hair follicle keratinocytes. Plucked scalp hair follicles, placed on bovine eye lens capsules as a growth substrate, give rise to quickly expanding colonies within a few days. After trypsinization, the cells are replated with irradiated 3T3 cells as 'feeders'. Using this combination of techniques the keratinocytes can be subcultured up to four times. In this way about 10(7) keratinocytes can be generated from one single hair follicle. Moreover, the technique enables cryogenic storage of the cells, allowing for instance, convenient transportation. Subcultured hair follicle keratinocytes can be plated on glass coverslips. This allows immunofluorescence studies. The keratin cytoskeletons visualized using an antiserum against human keratin.

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

Energy Technology Data Exchange (ETDEWEB)

Stoupis, James; Mousavi, Mirrasoul

2014-09-30

New data collection system equipment was installed in Xcel Energy substations and data was collected from 6 substations and 20 feeders. During Phase I, ABB collected and analyzed 793 real-time events to date from 6 Xcel Energy substations and continues today. The development and integration of several applications was completed during the course of this project, including a model-based faulted segment identification algorithm, with very positive results validated with field-gathered data discussed and included in this report. For mostly underground feeders, the success rate is 90% and the overreach rate is 90%. For mostly overhead feeders, the success rate is 74% and the overreach rate is 50%. The developed method is producing very accurate results for mostly underground feeders. For mostly overhead feeders, due to the bad OMS data quality and varying fault resistance when arcing, the developed method is producing good results but with much room for improvement. One area where the algorithm can be improved is the accuracy for sub-cycle fault events. In these cases, the accuracy of the conventional signal processing methods suffers due to most of these methods being based on a one-cycle processing window. By improving the signal processing accuracy, the accuracy of the faulted segment identification algorithm will also improve significantly. ABB intends to devote research in this area in the near future to help solve this problem. Other new applications developed during the course of the project include volt/VAR monitoring, unbalanced capacitor switching detection, unbalanced feeder loading detection, and feeder overloading detection. An important aspect of the demonstration phase of the project is to show the ability to provide adequate “heads-up” time ahead of customer calls or AMI reports so that the operators are provided with the much needed time to collect information needed to address an outage. The advance notification feature of the demonstration system

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...

Higher-Density Culture in Human Embryonic Stem Cells Results in DNA Damage and Genome Instability

Directory of Open Access Journals (Sweden)

Kurt Jacobs

2016-03-01

Full Text Available Human embryonic stem cells (hESC show great promise for clinical and research applications, but their well-known proneness to genomic instability hampers the development to their full potential. Here, we demonstrate that medium acidification linked to culture density is the main cause of DNA damage and genomic alterations in hESC grown on feeder layers, and this even in the short time span of a single passage. In line with this, we show that increasing the frequency of the medium refreshments minimizes the levels of DNA damage and genetic instability. Also, we show that cells cultured on laminin-521 do not present this increase in DNA damage when grown at high density, although the (long-term impact on their genomic stability remains to be elucidated. Our results explain the high levels of genome instability observed over the years by many laboratories worldwide, and show that the development of optimal culture conditions is key to solving this problem.

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.

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.

The negative impact of Wnt signaling on megakaryocyte and primitive erythroid progenitors derived from human embryonic stem cells

Directory of Open Access Journals (Sweden)

Prasuna Paluru

2014-03-01

Full Text Available The Wnt gene family consists of structurally related genes encoding secreted signaling molecules that have been implicated in many developmental processes, including regulation of cell fate and patterning during embryogenesis. Previously, we found that Wnt signaling is required for primitive or yolk sac-derived-erythropoiesis using the murine embryonic stem cell (ESC system. Here, we examine the effect of Wnt signaling on the formation of early hematopoietic progenitors derived from human ESCs. The first hematopoietic progenitor cells in the human ESC system express the pan-hematopoietic marker CD41 and the erythrocyte marker, glycophorin A or CD235. We have developed a novel serum-free, feeder-free, adherent differentiation system that can efficiently generate large numbers of CD41 + CD235+ cells. We demonstrate that this cell population contains progenitors not just for primitive erythroid and megakaryocyte cells but for the myeloid lineage as well and term this population the primitive common myeloid progenitor (CMP. Treatment of mesoderm-specified cells with Wnt3a led to a loss of hematopoietic colony-forming ability while the inhibition of canonical Wnt signaling with DKK1 led to an increase in the number of primitive CMPs. Canonical Wnt signaling also inhibits the expansion and/or survival of primitive erythrocytes and megakaryocytes, but not myeloid cells, derived from this progenitor population. These findings are in contrast to the role of Wnt signaling during mouse ESC differentiation and demonstrate the importance of the human ESC system in studying species-specific differences in development.

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.

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.

Critical evaluation of gamma-irradiated serum used as feeder in the culture and demonstration of putative nanobacteria and calcifying nanoparticles.

Directory of Open Access Journals (Sweden)

Jan Martel

Full Text Available The culture and demonstration of putative nanobacteria (NB and calcifying nanoparticles (CNP from human and animal tissues has relied primarily on the use of a culture supplement consisting of FBS that had been gamma-irradiated at a dose of 30 kGy (gamma-FBS. The use of gamma-FBS is based on the assumption that this sterilized fluid has been rid entirely of any residual NB/CNP, while it continues to promote the slow growth in culture of NB/CNP from human/animal tissues. We show here that gamma-irradiation (5-50 kGy produces extensive dose-dependent serum protein breakdown as demonstrated through UV and visible light spectrophotometry, fluorometry, Fourier-transformed infrared spectroscopy, and gel electrophoresis. Yet, both gamma-FBS and gamma-irradiated human serum (gamma-HS produce NB/CNP in cell culture conditions that are morphologically and chemically indistinguishable from their normal serum counterparts. Contrary to earlier claims, gamma-FBS does not enhance the formation of NB/CNP from several human body fluids (saliva, urine, ascites, and synovial fluid tested. In the presence of additional precipitating ions, both gamma-irradiated serum (FBS and HS and gamma-irradiated proteins (albumin and fetuin-A retain the inherent dual NB inhibitory and seeding capabilities seen also with their untreated counterparts. By gel electrophoresis, the particles formed from both gamma-FBS and gamma-HS are seen to have assimilated into their scaffold the same smeared protein profiles found in the gamma-irradiated sera. However, their protein compositions as identified by proteomics are virtually identical to those seen with particles formed from untreated serum. Moreover, particles derived from human fluids and cultured in the presence of gamma-FBS contain proteins derived from both gamma-FBS and the human fluid under investigation-a confusing and unprecedented scenario indicating that these particles harbor proteins from both the host tissue and the FBS

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

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.

CD133-enriched Xeno-Free human embryonic-derived neural stem cells expand rapidly in culture and do not form teratomas in immunodeficient mice

Directory of Open Access Journals (Sweden)

Daniel L. Haus

2014-09-01

Full Text Available Common methods for the generation of human embryonic-derived neural stem cells (hNSCs result in cells with potentially compromised safety profiles due to maintenance of cells in conditions containing non-human proteins (e.g. in bovine serum or on mouse fibroblast feeders. Additionally, sufficient expansion of resulting hNSCs for scaling out or up in a clinically relevant time frame has proven to be difficult. Here, we report a strategy that produces hNSCs in completely “Xeno-Free” culture conditions. Furthermore, we have enriched the hNSCs for the cell surface marker CD133 via magnetic sorting, which has led to an increase in the expansion rate and neuronal fate specification of the hNSCs in vitro. Critically, we have also confirmed neural lineage specificity upon sorted hNSC transplantation into the immunodeficient NOD-scid mouse brain. The future use or adaptation of these protocols has the potential to better facilitate the advancement of pre-clinical strategies from the bench to the bedside.

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)

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.

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)

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.

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

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

Assessment of in vitro drug resistance of human breast cancer cells subcultured from biopsy specimens

Czech Academy of Sciences Publication Activity Database

Krásná, Luboslava; Netíková, I.; Chaloupková, Alena; Taišlová, Eva; Zimovjanová, M.; Veselý, Pavel; Daneš, J.; Petruželka, L.; Matoušková, Eva

2003-01-01

Roč. 23, 3B (2003), s. 2593-2600 ISSN 0250-7005 R&D Projects: GA MZd NC6734 Institutional research plan: CEZ:AV0Z5052915 Keywords : Breast cancer * 3T3 feeder-layer culture technique * MTT test Subject RIV: EA - Cell Biology Impact factor: 1.347, year: 2003

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

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

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

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

[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.

Inhibition of mitogen-activated protein kinase kinase, DNA methyltransferase, and transforming growth factor-β promotes differentiation of human induced pluripotent stem cells into enterocytes.

Science.gov (United States)

Kodama, Nao; Iwao, Takahiro; Kabeya, Tomoki; Horikawa, Takashi; Niwa, Takuro; Kondo, Yuki; Nakamura, Katsunori; Matsunaga, Tamihide

2016-06-01

We previously reported that small-molecule compounds were effective in generating pharmacokinetically functional enterocytes from human induced pluripotent stem (iPS) cells. In this study, to determine whether the compounds promote the differentiation of human iPS cells into enterocytes, we investigated the effects of a combination of mitogen-activated protein kinase kinase (MEK), DNA methyltransferase (DNMT), and transforming growth factor (TGF)-β inhibitors on intestinal differentiation. Human iPS cells cultured on feeder cells were differentiated into endodermal cells by activin A. These endodermal-like cells were then differentiated into intestinal stem cells by fibroblast growth factor 2. Finally, the cells were differentiated into enterocyte cells by epidermal growth factor and small-molecule compounds. After differentiation, mRNA expression levels and drug-metabolizing enzyme activities were measured. The mRNA expression levels of the enterocyte marker sucrase-isomaltase and the major drug-metabolizing enzyme cytochrome P450 (CYP) 3A4 were increased by a combination of MEK, DNMT, and TGF-β inhibitors. The mRNA expression of CYP3A4 was markedly induced by 1α,25-dihydroxyvitamin D3. Metabolic activities of CYP1A1/2, CYP2B6, CYP2C9, CYP2C19, CYP3A4/5, UDP-glucuronosyltransferase, and sulfotransferase were also observed in the differentiated cells. In conclusion, MEK, DNMT, and TGF-β inhibitors can be used to promote the differentiation of human iPS cells into pharmacokinetically functional enterocytes. Copyright © 2016 The Japanese Society for the Study of Xenobiotics. Published by Elsevier Ltd. All rights reserved.

Cytokine-free directed differentiation of human pluripotent stem cells efficiently produces hemogenic endothelium with lymphoid potential.

Science.gov (United States)

Galat, Yekaterina; Dambaeva, Svetlana; Elcheva, Irina; Khanolkar, Aaruni; Beaman, Kenneth; Iannaccone, Philip M; Galat, Vasiliy

2017-03-17

The robust generation of human hematopoietic progenitor cells from induced or embryonic pluripotent stem cells would be beneficial for multiple areas of research, including mechanistic studies of hematopoiesis, the development of cellular therapies for autoimmune diseases, induced transplant tolerance, anticancer immunotherapies, disease modeling, and drug/toxicity screening. Over the past years, significant progress has been made in identifying effective protocols for hematopoietic differentiation from pluripotent stem cells and understanding stages of mesodermal, endothelial, and hematopoietic specification. Thus, it has been shown that variations in cytokine and inhibitory molecule treatments in the first few days of hematopoietic differentiation define primitive versus definitive potential of produced hematopoietic progenitor cells. The majority of current feeder-free, defined systems for hematopoietic induction from pluripotent stem cells include prolonged incubations with various cytokines that make the differentiation process complex and time consuming. We established that the application of Wnt agonist CHIR99021 efficiently promotes differentiation of human pluripotent stem cells in the absence of any hematopoietic cytokines to the stage of hemogenic endothelium capable of definitive hematopoiesis. The hemogenic endothelium differentiation was accomplished in an adherent, serum-free culture system by applying CHIR99021. Hemogenic endothelium progenitor cells were isolated on day 5 of differentiation and evaluated for their endothelial, myeloid, and lymphoid potential. Monolayer induction based on GSK3 inhibition, described here, yielded a large number of CD31 + CD34 + hemogenic endothelium cells. When isolated and propagated in adherent conditions, these progenitors gave rise to mature endothelium. When further cocultured with OP9 mouse stromal cells, these progenitors gave rise to various cells of myeloid lineages as well as natural killer lymphoid, T

Scalable cultivation of human pluripotent stem cells on chemically-defined surfaces

Science.gov (United States)

Hsiung, Michael Chi-Wei

Human stem cells (SCs) are classified as self-renewing cells possessing great ability in therapeutic applications due of their ability to differentiate along any major cell lineage in the human body. Despite their restorative potential, widespread use of SCs is hampered by strenuous control issues. Along with the need for strict xeno-free environments to sustain growth in culture, current methods for growing human pluripotent stem cells (hPSCs) rely on platforms which impede large-scale cultivation and therapeutic delivery. Hence, any progress towards development of large-scale culture systems is severely hindered. In a concentrated effort to develop a scheme that can serve as a model precursor for large scale SC propagation in clinical use, we have explored methods for cultivating hPSCs on completely defined surfaces. We discuss novel approaches with the potential to go beyond the limitations presented by current methods. In particular, we studied the cultivation of human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) on surface which underwent synthetic or chemical modification. Current methods for hPSCs rely on animal-based extracellular matrices (ECMs) such as mouse embryonic fibroblasts (MEFs) or feeders and murine sacoma cell-derived substrates to facilitate their growth. While these layers or coatings can be used to maximize the output of hPSC production, they cannot be considered for clinical use because they risk introducing foreign pathogens into culture. We have identified and developed conditions for a completely defined xeno-free substrate used for culturing hPSCs. By utilizing coupling chemistry, we can functionalize ester groups on a given surface and conjugate synthetic peptides containing the arginine-glycine-aspartic acid (RGD) motif, known for their role in cell adhesion. This method offers advantages over traditional hPSC culture by keeping the modified substrata free of xenogenic response and can be scaled up in

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

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

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.

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.

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....

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.

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...

Human-induced pluripotent stem cell-derived macrophages and their immunological function in response to tuberculosis infection.

Science.gov (United States)

Hong, Danping; Ding, Jiongyan; Li, Ouyang; He, Quan; Ke, Minxia; Zhu, Mengyi; Liu, Lili; Ou, Wen-Bin; He, Yulong; Wu, Yuehong

2018-02-26

Induced pluripotent stem cells (iPS) represent an innovative source for the standardized in vitro generation of macrophages (Mφ). Mφ show great promise in disease pathogenesis, particularly tuberculosis. However, there is no information about human iPS-derived (hiPS) macrophages (hiPS-Mφ) in response to tuberculosis infection. In the present study, macrophages derived from hiPS were established via embryoid body (EB) formation by using feeder-free culture conditions, and the human monocyte cell line THP-1 (THP-1-Mφ) was used as control. iPS-Mφ were characterized by using morphology, Giemsa staining, nonspecific esterase staining (α-NAE), phagocytosis, and surface phenotype. Additionally, after treatment with Bacillus Calmette-Guérin (BCG) for 24 h, cell apoptosis was detected by using an Annexin V-FITC Apoptosis Detection assay. The production of nitric oxide (NO), expression of tumor necrosis factor alpha (TNF-α), activity of apoptosis-related protein cysteine-3 (Caspase-3) and expression of B-cell lymphoma-2 (Bcl-2) were analyzed. With respect to morphology, surface phenotype, and function, the iPS-Mφ closely resembled their counterparts generated in vitro from a human monocyte cell line. iPS-Mφ exhibited the typically morphological characteristics of macrophages, such as round, oval, fusiform and irregular characteristics. The cells were Giemsa-stained-positive, α-NAE-positive, and possessed phagocytic ability. iPS-Mφ express high levels of CD14, CD11b, CD40, CD68, and major histocompatibility complex II (MHC-II). Moreover, with regard to the apoptotic rate, the production of NO, expression of TNF-α, and activity of Caspase-3 and Bcl-2, iPS-Mφ closely resemble that of their counterparts generated in vitro from human monocyte cell line in response to BCG infection. The rate of apoptosis of BCG-treated iPS-Mφ was 37.77 ± 7.94% compared to that of the untreated group at 4.97 ± 1.60% (P immunological function in response to Bacillus Calmette

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.

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...

A Simple and Robust Method for Culturing Human-Induced Pluripotent Stem Cells in an Undifferentiated State Using Botulinum Hemagglutinin.

Science.gov (United States)

Kim, Mee-Hae; Matsubara, Yoshifumi; Fujinaga, Yukako; Kino-Oka, Masahiro

2018-02-01

Clinical and industrial applications of human-induced pluripotent stem cells (hiPSCs) is hindered by the lack of robust culture strategies capable of sustaining a culture in an undifferentiated state. Here, a simple and robust hiPSC-culture-propagation strategy incorporating botulinum hemagglutinin (HA)-mediated selective removal of cells deviating from an undifferentiated state is developed. After HA treatment, cell-cell adhesion is disrupted, and deviated cells detached from the central region of the colony to subsequently form tight monolayer colonies following prolonged incubation. The authors find that the temporal and dose-dependent activity of HA regulated deviated-cell removal and recoverability after disruption of cell-cell adhesion in hiPSC colonies. The effects of HA are confirmed under all culture conditions examined, regardless of hiPSC line and feeder-dependent or -free culture conditions. After routine application of our HA-treatment paradigm for serial passages, hiPSCs maintains expression of pluripotent markers and readily forms embryoid bodies expressing markers for all three germ-cell layers. This method enables highly efficient culturing of hiPSCs and use of entire undifferentiated portions without having to pick deviated cells manually. This simple and readily reproducible culture strategy is a potentially useful tool for improving the robust and scalable maintenance of undifferentiated hiPSC cultures. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Serially cultured keratinocytes from human scalp hair follicles: a tool for cytogenetic studies.

Science.gov (United States)

Weterings, P J; Roelofs, H M; Jansen, B A; Vermorken, A J

1983-01-01

Keratinocytes originating from adult human hair follicles, the most convenient biopsy tissue, can be serially cultured using a combination of two techniques. Primary cultures are established using plucked scalp hair follicles and the bovine eye lens capsule as a growth substrate. Subsequently, cells from these cultures are serially cultivated in the presence of irradiated 3T3 cells as feeders. By this combination of techniques many keratinocytes can be generated from one single hair follicle. These cultures, appropriately treated with colchicine, can provide an adequate number of metaphases suitable for chromosome studies.

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)

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.

A defined and xeno-free culture method enabling the establishment of clinical-grade human embryonic, induced pluripotent and adipose stem cells.

Directory of Open Access Journals (Sweden)

Kristiina Rajala

2010-04-01

Full Text Available The growth of stem cells in in vitro conditions requires optimal balance between signals mediating cell survival, proliferation, and self-renewal. For clinical application of stem cells, the use of completely defined conditions and elimination of all animal-derived materials from the establishment, culture, and differentiation processes is desirable.Here, we report the development of a fully defined xeno-free medium (RegES, capable of supporting the expansion of human embryonic stem cells (hESC, induced pluripotent stem cells (iPSC and adipose stem cells (ASC. We describe the use of the xeno-free medium in the derivation and long-term (>80 passages culture of three pluripotent karyotypically normal hESC lines: Regea 06/015, Regea 07/046, and Regea 08/013. Cardiomyocytes and neural cells differentiated from these cells exhibit features characteristic to these cell types. The same formulation of the xeno-free medium is capable of supporting the undifferentiated growth of iPSCs on human feeder cells. The characteristics of the pluripotent hESC and iPSC lines are comparable to lines derived and cultured in standard undefined culture conditions. In the culture of ASCs, the xeno-free medium provided significantly higher proliferation rates than ASCs cultured in medium containing allogeneic human serum (HS, while maintaining the differentiation potential and characteristic surface marker expression profile of ASCs, although significant differences in the surface marker expression of ASCs cultured in HS and RegES media were revealed.Our results demonstrate that human ESCs, iPSCs and ASCs can be maintained in the same defined xeno-free medium formulation for a prolonged period of time while maintaining their characteristics, demonstrating the applicability of the simplified xeno-free medium formulation for the production of clinical-grade stem cells. The basic xeno-free formulation described herein has the potential to be further optimized for specific

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

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...

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.

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 ...

Non-integrating episomal plasmid-based reprogramming of human amniotic fluid stem cells into induced pluripotent stem cells in chemically defined conditions.

Science.gov (United States)

Slamecka, Jaroslav; Salimova, Lilia; McClellan, Steven; van Kelle, Mathieu; Kehl, Debora; Laurini, Javier; Cinelli, Paolo; Owen, Laurie; Hoerstrup, Simon P; Weber, Benedikt

2016-01-01

Amniotic fluid stem cells (AFSC) represent an attractive potential cell source for fetal and pediatric cell-based therapies. However, upgrading them to pluripotency confers refractoriness toward senescence, higher proliferation rate and unlimited differentiation potential. AFSC were observed to rapidly and efficiently reacquire pluripotency which together with their easy recovery makes them an attractive cell source for reprogramming. The reprogramming process as well as the resulting iPSC epigenome could potentially benefit from the unspecialized nature of AFSC. iPSC derived from AFSC also have potential in disease modeling, such as Down syndrome or β-thalassemia. Previous experiments involving AFSC reprogramming have largely relied on integrative vector transgene delivery and undefined serum-containing, feeder-dependent culture. Here, we describe non-integrative oriP/EBNA-1 episomal plasmid-based reprogramming of AFSC into iPSC and culture in fully chemically defined xeno-free conditions represented by vitronectin coating and E8 medium, a system that we found uniquely suited for this purpose. The derived AF-iPSC lines uniformly expressed a set of pluripotency markers Oct3/4, Nanog, Sox2, SSEA-1, SSEA-4, TRA-1-60, TRA-1-81 in a pattern typical for human primed PSC. Additionally, the cells formed teratomas, and were deemed pluripotent by PluriTest, a global expression microarray-based in-silico pluripotency assay. However, we found that the PluriTest scores were borderline, indicating a unique pluripotent signature in the defined condition. In the light of potential future clinical translation of iPSC technology, non-integrating reprogramming and chemically defined culture are more acceptable.

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.

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.

The Human Cell Atlas.

Science.gov (United States)

Regev, Aviv; Teichmann, Sarah A; Lander, Eric S; Amit, Ido; Benoist, Christophe; Birney, Ewan; Bodenmiller, Bernd; Campbell, Peter; Carninci, Piero; Clatworthy, Menna; Clevers, Hans; Deplancke, Bart; Dunham, Ian; Eberwine, James; Eils, Roland; Enard, Wolfgang; Farmer, Andrew; Fugger, Lars; Göttgens, Berthold; Hacohen, Nir; Haniffa, Muzlifah; Hemberg, Martin; Kim, Seung; Klenerman, Paul; Kriegstein, Arnold; Lein, Ed; Linnarsson, Sten; Lundberg, Emma; Lundeberg, Joakim; Majumder, Partha; Marioni, John C; Merad, Miriam; Mhlanga, Musa; Nawijn, Martijn; Netea, Mihai; Nolan, Garry; Pe'er, Dana; Phillipakis, Anthony; Ponting, Chris P; Quake, Stephen; Reik, Wolf; Rozenblatt-Rosen, Orit; Sanes, Joshua; Satija, Rahul; Schumacher, Ton N; Shalek, Alex; Shapiro, Ehud; Sharma, Padmanee; Shin, Jay W; Stegle, Oliver; Stratton, Michael; Stubbington, Michael J T; Theis, Fabian J; Uhlen, Matthias; van Oudenaarden, Alexander; Wagner, Allon; Watt, Fiona; Weissman, Jonathan; Wold, Barbara; Xavier, Ramnik; Yosef, Nir

2017-12-05

The recent advent of methods for high-throughput single-cell molecular profiling has catalyzed a growing sense in the scientific community that the time is ripe to complete the 150-year-old effort to identify all cell types in the human body. The Human Cell Atlas Project is an international collaborative effort that aims to define all human cell types in terms of distinctive molecular profiles (such as gene expression profiles) and to connect this information with classical cellular descriptions (such as location and morphology). An open comprehensive reference map of the molecular state of cells in healthy human tissues would propel the systematic study of physiological states, developmental trajectories, regulatory circuitry and interactions of cells, and also provide a framework for understanding cellular dysregulation in human disease. Here we describe the idea, its potential utility, early proofs-of-concept, and some design considerations for the Human Cell Atlas, including a commitment to open data, code, and community.

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.

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.

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.

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.

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)

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

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.

Discarded human fetal tissue and cell cultures for transplantation research

International Nuclear Information System (INIS)

Hay, R.J.; Phillips, T.; Thompson, A.; Vilner, L.; Cleland, M.; Tchaw-ren Chen; Zabrenetzky, V.

1999-01-01

A feasibility study has been performed to explore the utility of various tissues from discarded human abortuses for transplantation and related research. Specifically, aborted fetuses plus parental blood samples and all relevant clinical data were obtained through a local hospital complex. Whenever possible, pancreas, skin and skeletal muscle, heart, liver, kidney, cartilage and lung tissues were removed, dissociated and subfractionated for cryopreservation, characterization and cultivation trials in vitro. Existing protocols for these manipulations were compared and improved upon as required. Clonal culture, cell aggregate maintenance techniques and use of feeder cell populations have been utilized where appropriate to develop quantitative comparative data. Histological and biochemical assays were applied both to evaluate separation/cultivation methods and to identify optimal culture conditions for maintaining functional cells. Immunochemical and molecular biological procedures were applied to study expression of Major Histocompatibility Vomplex (MHC) class 1 and 11 molecules on cell lines derived. Tissue and cell culture populations were examined for infections with bacteria, ftingi, mycoplasma, HIV, CMV, hepatitis B and other viruses. Only 1% of the abortuses tested were virally infected. Cytogenetic analyses confin-ned the normal diploid status in the vast majority (>98%) of lines tested. A total of over 250 abortuses have been obtained and processed. Only 25 were found to be contaminated with bacteria or fungi and unsuitable for further cultivation trials. A total of over 200 cell populations were isolated, characterized and cryopreserved for further study. Included were kidney, lung, liver and epidermal epithelia: cartilage-derived cells from the spine and epiphyses plus myogenic myoblasts. Selected lines have been immortalized using HPV I 6E6/E7 sequences. Epithelia from the liver and pancreas and cardiac myocytes were the most problematic in that initial

The human cell atlas

DEFF Research Database (Denmark)

Regev, Aviv; Teichmann, Sarah A.; Lander, Eric S.

2017-01-01

The recent advent of methods for high-throughput single-cell molecular profiling has catalyzed a growing sense in the scientific community that the time is ripe to complete the 150-year-old effort to identify all cell types in the human body. The Human Cell Atlas Project is an international...... collaborative effort that aims to define all human cell types in terms of distinctive molecular profiles (such as gene expression profiles) and to connect this information with classical cellular descriptions (such as location and morphology). An open comprehensive reference map of the molecular state of cells...... in healthy human tissues would propel the systematic study of physiological states, developmental trajectories, regulatory circuitry and interactions of cells, and also provide a framework for understanding cellular dysregulation in human disease. Here we describe the idea, its potential utility, early...

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.

Efficient generation of megakaryocytes from human induced pluripotent stem cells using food and drug administration-approved pharmacological reagents.

Science.gov (United States)

Liu, Yanfeng; Wang, Ying; Gao, Yongxing; Forbes, Jessica A; Qayyum, Rehan; Becker, Lewis; Cheng, Linzhao; Wang, Zack Z

2015-04-01

Megakaryocytes (MKs) are rare hematopoietic cells in the adult bone marrow and produce platelets that are critical to vascular hemostasis and wound healing. Ex vivo generation of MKs from human induced pluripotent stem cells (hiPSCs) provides a renewable cell source of platelets for treating thrombocytopenic patients and allows a better understanding of MK/platelet biology. The key requirements in this approach include developing a robust and consistent method to produce functional progeny cells, such as MKs from hiPSCs, and minimizing the risk and variation from the animal-derived products in cell cultures. In this study, we developed an efficient system to generate MKs from hiPSCs under a feeder-free and xeno-free condition, in which all animal-derived products were eliminated. Several crucial reagents were evaluated and replaced with Food and Drug Administration-approved pharmacological reagents, including romiplostim (Nplate, a thrombopoietin analog), oprelvekin (recombinant interleukin-11), and Plasbumin (human albumin). We used this method to induce MK generation from hiPSCs derived from 23 individuals in two steps: generation of CD34(+)CD45(+) hematopoietic progenitor cells (HPCs) for 14 days; and generation and expansion of CD41(+)CD42a(+) MKs from HPCs for an additional 5 days. After 19 days, we observed abundant CD41(+)CD42a(+) MKs that also expressed the MK markers CD42b and CD61 and displayed polyploidy (≥16% of derived cells with DNA contents >4N). Transcriptome analysis by RNA sequencing revealed that megakaryocytic-related genes were highly expressed. Additional maturation and investigation of hiPSC-derived MKs should provide insights into MK biology and lead to the generation of large numbers of platelets ex vivo. ©AlphaMed Press.

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)

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.

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.

Progression from productive infection to integration and oncogenic transformation in human papillomavirus type 59-immortalized foreskin keratinocytes.

Science.gov (United States)

Spartz, Helena; Lehr, Elizabeth; Zhang, Benyue; Roman, Ann; Brown, Darron R

2005-05-25

Studies of changes in the virus and host cell upon progression from human papillomavirus (HPV) episomal infection to integration are critical to understanding HPV-related malignant transformation. However, there exist only a few in vitro models of both productive HPV infection and neoplastic progression on the same host background. We recently described a unique foreskin keratinocyte cell line (ERIN 59) that contains HPV 59 (a close relative of HPV 18). Early passages of ERIN 59 cells (passages 9-13) contained approximately 50 copies of episomes/cell, were feeder cell-dependent, and could be induced to differentiate and produce infectious virus in a simple culture system. We now report that late passage cells (passages greater than 50) were morphologically different from early passage cells, were feeder cell independent, and did not differentiate or produce virus. These late passage cells contained HPV in an integrated form. An integration-derived oncogene transcript was expressed in late passage cells. The E2 open reading frame was interrupted in this transcript at nucleotide 3351. Despite a lower viral genome copy number in late passage ERIN 59 cells, expression of E6/E7 oncogene transcripts was similar to early passage cells. We conclude that ERIN 59 cells are a valuable cell line representing a model of progression from HPV 59 episomal infection and virus production to HPV 59 integration and associated oncogenic transformation on the same host background.

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

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...

Improved Survival and Initiation of Differentiation of Human Induced Pluripotent Stem Cells to Hepatocyte-Like Cells upon Culture in William's E Medium followed by Hepatocyte Differentiation Inducer Treatment.

Directory of Open Access Journals (Sweden)

Minoru Tomizawa

Full Text Available Hepatocyte differentiation inducer (HDI lacks both glucose and arginine, but is supplemented with galactose and ornithine, and is added together with other reagents such as apoptosis inhibitor and oncostatin M. Although human induced pluripotent stem (iPS cells initiate hepatocyte differentiation, most die within 7 days. In this study, we investigated both HDI and conventional media for their potential to improve cell survival.201B7 iPS cells were cultured in conventional media. This consisted of three cycles of 5-day culture in William's E (WE medium, followed by a 2-day culture in HDI.Expression levels of α-feto protein (AFP were higher in cells cultured in WE and in Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DF12. 201B7 cells expressed the highest AFP and albumin (ALB when cultured in HDI for 2 days following 7-day culture in WE. After three cycles of 5-day culture in WE followed by 2 days in HDI, 201B7 cells expressed AFP and ALB 54 ± 2.3 (average ± standard deviation and 73 ± 15.1 times higher, respectively, than those cultured in ReproFF (feeder-free condition.201B7 cells survived culture in WE for 7 days followed HDI for 2 days. After three cycles of culture under these conditions, hepatocyte differentiation was enhanced, as evidenced by increased AFP and ALB expression.

Vector-free and transgene-free human iPS cells differentiate into functional neurons and enhance functional recovery after ischemic stroke in mice.

Directory of Open Access Journals (Sweden)

Osama Mohamad

Full Text Available Stroke is a leading cause of human death and disability in the adult population in the United States and around the world. While stroke treatment is limited, stem cell transplantation has emerged as a promising regenerative therapy to replace or repair damaged tissues and enhance functional recovery after stroke. Recently, the creation of induced pluripotent stem (iPS cells through reprogramming of somatic cells has revolutionized cell therapy by providing an unlimited source of autologous cells for transplantation. In addition, the creation of vector-free and transgene-free human iPS (hiPS cells provides a new generation of stem cells with a reduced risk of tumor formation that was associated with the random integration of viral vectors seen with previous techniques. However, the potential use of these cells in the treatment of ischemic stroke has not been explored. In the present investigation, we examined the neuronal differentiation of vector-free and transgene-free hiPS cells and the transplantation of hiPS cell-derived neural progenitor cells (hiPS-NPCs in an ischemic stroke model in mice. Vector-free hiPS cells were maintained in feeder-free and serum-free conditions and differentiated into functional neurons in vitro using a newly developed differentiation protocol. Twenty eight days after transplantation in stroke mice, hiPS-NPCs showed mature neuronal markers in vivo. No tumor formation was seen up to 12 months after transplantation. Transplantation of hiPS-NPCs restored neurovascular coupling, increased trophic support and promoted behavioral recovery after stroke. These data suggest that using vector-free and transgene-free hiPS cells in stem cell therapy are safe and efficacious in enhancing recovery after focal ischemic stroke in mice.

Generation and periodontal differentiation of human gingival fibroblasts-derived integration-free induced pluripotent stem cells

Energy Technology Data Exchange (ETDEWEB)

Yin, Xiaohui [Department of Periodontology, School and Hospital of Stomatology, Peking University, 22 South Avenue Zhong-Guan-Cun, Beijing 100081 (China); Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191 (China); Li, Yang [Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191 (China); Li, Jingwen [Department of Periodontology, School and Hospital of Stomatology, Peking University, 22 South Avenue Zhong-Guan-Cun, Beijing 100081 (China); Li, Peng [Faculty of Dentistry, The University of Hong Kong, 34 Hospital Road, Hong Kong SAR (China); Liu, Yinan [Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191 (China); Wen, Jinhua, E-mail: jhwen@bjmu.edu.cn [Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191 (China); Luan, Qingxian, E-mail: kqluanqx@126.com [Department of Periodontology, School and Hospital of Stomatology, Peking University, 22 South Avenue Zhong-Guan-Cun, Beijing 100081 (China)

2016-05-06

Induced pluripotent stem cells (iPSCs) have been recognized as a promising cell source for periodontal tissue regeneration. However, the conventional virus-based reprogramming approach is associated with a high risk of genetic mutation and limits their therapeutic utility. Here, we successfully generated iPSCs from readily accessible human gingival fibroblasts (hGFs) through an integration-free and feeder-free approach via delivery of reprogramming factors of Oct4, Sox2, Klf4, L-myc, Lin28 and TP53 shRNA with episomal plasmid vectors. The iPSCs presented similar morphology and proliferation characteristics as embryonic stem cells (ESCs), and expressed pluripotent markers including Oct4, Tra181, Nanog and SSEA-4. Additionally, these cells maintained a normal karyotype and showed decreased CpG methylation ratio in the promoter regions of Oct4 and Nanog. In vivo teratoma formation assay revealed the development of tissues representative of three germ layers, confirming the acquisition of pluripotency. Furthermore, treatment of the iPSCs in vitro with enamel matrix derivative (EMD) or growth/differentiation factor-5 (GDF-5) significantly up-regulated the expression of periodontal tissue markers associated with bone, periodontal ligament and cementum respectively. Taken together, our data demonstrate that hGFs are a valuable cell source for generating integration-free iPSCs, which could be sequentially induced toward periodontal cells under the treatment of EMD and GDF-5. - Highlights: • Integration-free iPSCs are successfully generated from hGFs via an episomal approach. • EMD promotes differentiation of the hGFs-derived iPSCs toward periodontal cells. • GDF-5 promotes differentiation of the hGFs-derived iPSCs toward periodontal cells. • hGFs-derived iPSCs could be a promising cell source for periodontal regeneration.

Generation and periodontal differentiation of human gingival fibroblasts-derived integration-free induced pluripotent stem cells

International Nuclear Information System (INIS)

Yin, Xiaohui; Li, Yang; Li, Jingwen; Li, Peng; Liu, Yinan; Wen, Jinhua; Luan, Qingxian

2016-01-01

Induced pluripotent stem cells (iPSCs) have been recognized as a promising cell source for periodontal tissue regeneration. However, the conventional virus-based reprogramming approach is associated with a high risk of genetic mutation and limits their therapeutic utility. Here, we successfully generated iPSCs from readily accessible human gingival fibroblasts (hGFs) through an integration-free and feeder-free approach via delivery of reprogramming factors of Oct4, Sox2, Klf4, L-myc, Lin28 and TP53 shRNA with episomal plasmid vectors. The iPSCs presented similar morphology and proliferation characteristics as embryonic stem cells (ESCs), and expressed pluripotent markers including Oct4, Tra181, Nanog and SSEA-4. Additionally, these cells maintained a normal karyotype and showed decreased CpG methylation ratio in the promoter regions of Oct4 and Nanog. In vivo teratoma formation assay revealed the development of tissues representative of three germ layers, confirming the acquisition of pluripotency. Furthermore, treatment of the iPSCs in vitro with enamel matrix derivative (EMD) or growth/differentiation factor-5 (GDF-5) significantly up-regulated the expression of periodontal tissue markers associated with bone, periodontal ligament and cementum respectively. Taken together, our data demonstrate that hGFs are a valuable cell source for generating integration-free iPSCs, which could be sequentially induced toward periodontal cells under the treatment of EMD and GDF-5. - Highlights: • Integration-free iPSCs are successfully generated from hGFs via an episomal approach. • EMD promotes differentiation of the hGFs-derived iPSCs toward periodontal cells. • GDF-5 promotes differentiation of the hGFs-derived iPSCs toward periodontal cells. • hGFs-derived iPSCs could be a promising cell source for periodontal regeneration.

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.

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.

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.

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.

Coculture with BJ fibroblast cells inhibits the adipogenesis and lipogenesis in 3T3-L1 cells

International Nuclear Information System (INIS)

Jeong, Hyun Jeong; Park, Sahng Wook; Kim, Hojeong; Park, Sang-Kyu; Yoon, Dojun

2010-01-01

Mouse or human fibroblasts are commonly used as feeder cells to prevent differentiation in stem or primary cell culture. In the present study, we addressed whether fibroblasts can affect the differentiation of adipocytes. We found that the differentiation of 3T3-L1 preadipocytes was strongly suppressed when the cells were cocultured with human fibroblast (BJ) cells. BrdU incorporation analysis indicated that mitotic clonal expansion, an early event required for 3T3-L1 cell adipogenesis, was not affected by BJ cells. The 3T3-L1 cell expression levels of peroxisome proliferator-activated receptor γ2, CCAAT/enhancer-binding protein alpha (C/EBPα), sterol regulatory element binding protein-1c, and Krueppel-like factor 15, but not those of C/EBPβ or C/EBPδ, were decreased by coculture with BJ cells. When mature 3T3-L1 adipocytes were cocultured with BJ cells, their lipid contents were significantly reduced, with decreased fatty acid synthase expression and increased phosphorylated form of acetyl-CoA carboxylase 1. Our data indicate that coculture with BJ fibroblast cells inhibits the adipogenesis of 3T3-L1 preadipocytes and decreases the lipogenesis of mature 3T3-L1 adipocytes.

A chemically defined culture medium containing Rho kinase inhibitor Y-27632 for the fabrication of stratified squamous epithelial cell grafts

International Nuclear Information System (INIS)

Aslanova, Afag; Takagi, Ryo; Yamato, Masayuki; Okano, Teruo; Yamamoto, Masakazu

2015-01-01

With the development of a culture method for stratified squamous epithelial cells, tissue-engineered epithelial cell sheets have been successfully applied as clinical cell grafts. However, the implementation of these cell sheets without the use of any animal-derived materials is highly desirable. In this study, Rho-associated protein kinase inhibitor Y-27632 was used to develop a chemically defined culture medium for the fabrication of stratified epithelial cell grafts consisting of human epidermal and oral keratinocytes, and the proliferation activity, cell morphology, and gene expressions of the keratinocytes were analyzed. The results of a colorimetric assay indicated that Y-27632 significantly promoted the proliferation of the keratinocytes in culture media both with and without fetal bovine serum (FBS), although there were no indications of Y-27632 efficacy on cell morphology and stratification of the keratinocytes in culture medium without any animal-derived materials. The results of quantitative RT-PCR revealed that gene expressions correlated with cell adhesion, cell–cell junction, proliferation markers, and stem/progenitor markers in cultured keratinocytes were not strongly affected by the addition of Y-27632 to the culture medium. Moreover, gene expressions of differentiation markers in stratified keratinocytes cultured in medium without FBS were nearly identical to those of keratinocytes co-cultured with 3T3 feeder cells. Interestingly, the expressions of differentiation markers in cultured stratified keratinocytes were suppressed by FBS, whereas they were reconstructed by either co-culture of a 3T3 feeder layer or addition of Y-27632 into the culture medium containing FBS. These findings indicate that Y-27632 is a useful supplement for the development of a chemically defined culture medium for fabrication of stratified epithelial cell grafts for clinical applications for the purpose of developing the culture medium with a lower risk of pathogen

A chemically defined culture medium containing Rho kinase inhibitor Y-27632 for the fabrication of stratified squamous epithelial cell grafts

Energy Technology Data Exchange (ETDEWEB)

Aslanova, Afag [Department of Surgery, Institute of Gastroenterology, Tokyo Women' s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Institute of Advanced Biomedical Engineering and Science, Tokyo Women' s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Takagi, Ryo; Yamato, Masayuki; Okano, Teruo [Institute of Advanced Biomedical Engineering and Science, Tokyo Women' s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Yamamoto, Masakazu, E-mail: yamamoto.ige@twmu.ac.jp [Department of Surgery, Institute of Gastroenterology, Tokyo Women' s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan)

2015-05-01

With the development of a culture method for stratified squamous epithelial cells, tissue-engineered epithelial cell sheets have been successfully applied as clinical cell grafts. However, the implementation of these cell sheets without the use of any animal-derived materials is highly desirable. In this study, Rho-associated protein kinase inhibitor Y-27632 was used to develop a chemically defined culture medium for the fabrication of stratified epithelial cell grafts consisting of human epidermal and oral keratinocytes, and the proliferation activity, cell morphology, and gene expressions of the keratinocytes were analyzed. The results of a colorimetric assay indicated that Y-27632 significantly promoted the proliferation of the keratinocytes in culture media both with and without fetal bovine serum (FBS), although there were no indications of Y-27632 efficacy on cell morphology and stratification of the keratinocytes in culture medium without any animal-derived materials. The results of quantitative RT-PCR revealed that gene expressions correlated with cell adhesion, cell–cell junction, proliferation markers, and stem/progenitor markers in cultured keratinocytes were not strongly affected by the addition of Y-27632 to the culture medium. Moreover, gene expressions of differentiation markers in stratified keratinocytes cultured in medium without FBS were nearly identical to those of keratinocytes co-cultured with 3T3 feeder cells. Interestingly, the expressions of differentiation markers in cultured stratified keratinocytes were suppressed by FBS, whereas they were reconstructed by either co-culture of a 3T3 feeder layer or addition of Y-27632 into the culture medium containing FBS. These findings indicate that Y-27632 is a useful supplement for the development of a chemically defined culture medium for fabrication of stratified epithelial cell grafts for clinical applications for the purpose of developing the culture medium with a lower risk of pathogen

Generation and characterization of induced pluripotent stem cells from guinea pig fetal fibroblasts

Science.gov (United States)

Wu, Yuehong; Li, Ouyang; He, Chengwen; Li, Yong; Li, Min; Liu, Xiaoming; Wang, Yujiong; He, Yulong

2017-01-01

Induced pluripotent stem cells (iPS) represent an important tool to develop disease-modeling assays, drug testing assays and cell-based replacement therapies. The application of iPS in these fields requires the development of suitable animal models. Of the suitable species, guinea pigs are particularly important and offer significant advantages. Successful iPS generation has been accomplished in a number of species; however, it has not been reported in the guinea pig. The present study successfully generated iPS from guinea pigs (giPS) using single polycistronic virus transduction with mouse octamer-binding transcription factor 4 (Oct4), sex determining region Y-box 2 (Sox2), Kruppel-like factor 4 and c-Myc. The giPS cell lines were cultured in media containing leukemia inhibitory factor and guinea pig fibroblast cells were used as feeder cells. These cultures were expanded under feeder-free culture conditions using ESGRO Complete Plus Clonal Grade medium containing 15% fetal bovine serum on gelatin-coated dishes. The resultant cells had a normal karyotype, exhibited alkaline phosphatase activity and expressed the pluripotency markers Oct4, Sox2 and Nanog. The cells differentiated in vivo to form teratomas that contained all three germ layers of the tissue cells. The generation of giPS may facilitate future studies investigating the mechanisms underlying innate immunity, particularly for tuberculosis. These experiments provide proof of principle that iPS technology may be adapted to use the guinea pig as a model of human diseases. PMID:28393187

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.

Derivation of xeno-free and GMP-grade human embryonic stem cells--platforms for future clinical applications.

Directory of Open Access Journals (Sweden)

Shelly E Tannenbaum

Full Text Available Clinically compliant human embryonic stem cells (hESCs should be developed in adherence to ethical standards, without risk of contamination by adventitious agents. Here we developed for the first time animal-component free and good manufacturing practice (GMP-compliant hESCs. After vendor and raw material qualification, we derived xeno-free, GMP-grade feeders from umbilical cord tissue, and utilized them within a novel, xeno-free hESC culture system. We derived and characterized three hESC lines in adherence to regulations for embryo procurement, and good tissue, manufacturing and laboratory practices. To minimize freezing and thawing, we continuously expanded the lines from initial outgrowths and samples were cryopreserved as early stocks and banks. Batch release criteria included DNA-fingerprinting and HLA-typing for identity, characterization of pluripotency-associated marker expression, proliferation, karyotyping and differentiation in-vitro and in-vivo. These hESCs may be valuable for regenerative therapy. The ethical, scientific and regulatory methodology presented here may serve for development of additional clinical-grade hESCs.

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

Evaluating the potential of poly(beta-amino ester) nanoparticles for reprogramming human fibroblasts to become induced pluripotent stem cells.

Science.gov (United States)

Bhise, Nupura S; Wahlin, Karl J; Zack, Donald J; Green, Jordan J

2013-01-01

Gene delivery can potentially be used as a therapeutic for treating genetic diseases, including neurodegenerative diseases, as well as an enabling technology for regenerative medicine. A central challenge in many gene delivery applications is having a safe and effective delivery method. We evaluated the use of a biodegradable poly(beta-amino ester) nanoparticle-based nonviral protocol and compared this with an electroporation-based approach to deliver episomal plasmids encoding reprogramming factors for generation of human induced pluripotent stem cells (hiPSCs) from human fibroblasts. A polymer library was screened to identify the polymers most promising for gene delivery to human fibroblasts. Feeder-independent culturing protocols were developed for nanoparticle-based and electroporation-based reprogramming. The cells reprogrammed by both polymeric nanoparticle-based and electroporation-based nonviral methods were characterized by analysis of pluripotency markers and karyotypic stability. The hiPSC-like cells were further differentiated toward the neural lineage to test their potential for neurodegenerative retinal disease modeling. 1-(3-aminopropyl)-4-methylpiperazine end-terminated poly(1,4-butanediol diacry-late-co-4-amino-1-butanol) polymer (B4S4E7) self-assembled with plasmid DNA to form nanoparticles that were more effective than leading commercially available reagents, including Lipofectamine® 2000, FuGENE® HD, and 25 kDa branched polyethylenimine, for nonviral gene transfer. B4S4E7 nanoparticles showed effective gene delivery to IMR-90 human primary fibroblasts and to dermal fibroblasts derived from a patient with retinitis pigmentosa, and enabled coexpression of exogenously delivered genes, as is needed for reprogramming. The karyotypically normal hiPSC-like cells generated by conventional electroporation, but not by poly(beta-amino ester) reprogramming, could be differentiated toward the neuronal lineage, specifically pseudostratified optic cups. This

Defective repair of UV-damaged DNA in human tumor and SV40-transformed human cells but not in adenovirus-transformed human cells

International Nuclear Information System (INIS)

Rainbow, A.J.

1989-01-01

The DNA repair capacities of five human tumor cell lines, one SV40-transformed human cell line and one adenovirus-transformed human cell line were compared with that of normal human fibroblasts using a sensitive host cell reactivation (HCR) technique. Unirradiated and UV-irradiated suspensions of adenovirus type 2 (Ad 2) were assayed for their ability to form viral structural antigens (Vag) in the various cell types using immunofluorescent staining. The survival of Vag formation for UV-irradiated Ad 2 was significantly reduced in all the human tumor cell lines and the SV40-transformed human line compared to the normal human fibroblasts, but was apparently normal in the adenovirus-transformed human cells. D 0 values for the UV survival of Ad 2 Vag synthesis in the tumor and virally transformed lines expressed as a percentage of that obtained on normal fibroblast strains were used as a measure of DNA repair capacity. Percent HCR values ranged from 26 to 53% in the tumor cells. These results indicate a deficiency in the repair of UV-induced DNA damage associated with human tumorigenesis and the transformation of human cells by SV40 but not the transformation of human cells by adenovirus. (author)

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...

Genome engineering in human cells.

Science.gov (United States)

Song, Minjung; Kim, Young-Hoon; Kim, Jin-Soo; Kim, Hyongbum

2014-01-01

Genome editing in human cells is of great value in research, medicine, and biotechnology. Programmable nucleases including zinc-finger nucleases, transcription activator-like effector nucleases, and RNA-guided engineered nucleases recognize a specific target sequence and make a double-strand break at that site, which can result in gene disruption, gene insertion, gene correction, or chromosomal rearrangements. The target sequence complexities of these programmable nucleases are higher than 3.2 mega base pairs, the size of the haploid human genome. Here, we briefly introduce the structure of the human genome and the characteristics of each programmable nuclease, and review their applications in human cells including pluripotent stem cells. In addition, we discuss various delivery methods for nucleases, programmable nickases, and enrichment of gene-edited human cells, all of which facilitate efficient and precise genome editing in human cells.

Plasma membrane proteomics of human embryonic stem cells and human embryonal carcinoma cells.

NARCIS (Netherlands)

Dormeyer, W.; van Hoof, D.; Braam, S.R.; Heck, A.J.R.; Mummery, C.L.; Krijgsveld, J.

2008-01-01

Human embryonic stem cells (hESCs) are of immense interest in regenerative medicine as they can self-renew indefinitely and can give rise to any adult cell type. Human embryonal carcinoma cells (hECCs) are the malignant counterparts of hESCs found in testis tumors. hESCs that have acquired

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

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

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

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

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...

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.

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

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.

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.

Immunosuppressive Mesenchymal Stromal Cells Derived from Human-Induced Pluripotent Stem Cells Induce Human Regulatory T Cells In Vitro and In Vivo.

Science.gov (United States)

Roux, Clémence; Saviane, Gaëlle; Pini, Jonathan; Belaïd, Nourhène; Dhib, Gihen; Voha, Christine; Ibáñez, Lidia; Boutin, Antoine; Mazure, Nathalie M; Wakkach, Abdelilah; Blin-Wakkach, Claudine; Rouleau, Matthieu

2017-01-01

Despite mesenchymal stromal cells (MSCs) are considered as a promising source of cells to modulate immune functions on cells from innate and adaptive immune systems, their clinical use remains restricted (few number, limited in vitro expansion, absence of a full phenotypic characterization, few insights on their in vivo fate). Standardized MSCs derived in vitro from human-induced pluripotent stem (huIPS) cells, remediating part of these issues, are considered as well as a valuable tool for therapeutic approaches, but their functions remained to be fully characterized. We generated multipotent MSCs derived from huiPS cells (huiPS-MSCs), and focusing on their immunosuppressive activity, we showed that human T-cell activation in coculture with huiPS-MSCs was significantly reduced. We also observed the generation of functional CD4 + FoxP3 + regulatory T (Treg) cells. Further tested in vivo in a model of human T-cell expansion in immune-deficient NSG mice, huiPS-MSCs immunosuppressive activity prevented the circulation and the accumulation of activated human T cells. Intracytoplasmic labeling of cytokines produced by the recovered T cells showed reduced percentages of human-differentiated T cells producing Th1 inflammatory cytokines. By contrast, T cells producing IL-10 and FoxP3 + -Treg cells, absent in non-treated animals, were detected in huiPS-MSCs treated mice. For the first time, these results highlight the immunosuppressive activity of the huiPS-MSCs on human T-cell stimulation with a concomitant generation of human Treg cells in vivo . They may favor the development of new tools and strategies based on the use of huiPS cells and their derivatives for the induction of immune tolerance.

Immunosuppressive Mesenchymal Stromal Cells Derived from Human-Induced Pluripotent Stem Cells Induce Human Regulatory T Cells In Vitro and In Vivo

Directory of Open Access Journals (Sweden)

Clémence Roux

2018-01-01

Full Text Available Despite mesenchymal stromal cells (MSCs are considered as a promising source of cells to modulate immune functions on cells from innate and adaptive immune systems, their clinical use remains restricted (few number, limited in vitro expansion, absence of a full phenotypic characterization, few insights on their in vivo fate. Standardized MSCs derived in vitro from human-induced pluripotent stem (huIPS cells, remediating part of these issues, are considered as well as a valuable tool for therapeutic approaches, but their functions remained to be fully characterized. We generated multipotent MSCs derived from huiPS cells (huiPS-MSCs, and focusing on their immunosuppressive activity, we showed that human T-cell activation in coculture with huiPS-MSCs was significantly reduced. We also observed the generation of functional CD4+ FoxP3+ regulatory T (Treg cells. Further tested in vivo in a model of human T-cell expansion in immune-deficient NSG mice, huiPS-MSCs immunosuppressive activity prevented the circulation and the accumulation of activated human T cells. Intracytoplasmic labeling of cytokines produced by the recovered T cells showed reduced percentages of human-differentiated T cells producing Th1 inflammatory cytokines. By contrast, T cells producing IL-10 and FoxP3+-Treg cells, absent in non-treated animals, were detected in huiPS-MSCs treated mice. For the first time, these results highlight the immunosuppressive activity of the huiPS-MSCs on human T-cell stimulation with a concomitant generation of human Treg cells in vivo. They may favor the development of new tools and strategies based on the use of huiPS cells and their derivatives for the induction of immune tolerance.

Alloimmune Responses of Humanized Mice to Human Pluripotent Stem Cell Therapeutics

Directory of Open Access Journals (Sweden)

Nigel G. Kooreman

2017-08-01

Full Text Available There is growing interest in using embryonic stem cell (ESC and induced pluripotent stem cell (iPSC derivatives for tissue regeneration. However, an increased understanding of human immune responses to stem cell-derived allografts is necessary for maintaining long-term graft persistence. To model this alloimmunity, humanized mice engrafted with human hematopoietic and immune cells could prove to be useful. In this study, an in-depth analysis of graft-infiltrating human lymphocytes and splenocytes revealed that humanized mice incompletely model human immune responses toward allogeneic stem cells and their derivatives. Furthermore, using an “allogenized” mouse model, we show the feasibility of reconstituting immunodeficient mice with a functional mouse immune system and describe a key role of innate immune cells in the rejection of mouse stem cell allografts.

Trophoblast lineage cells derived from human induced pluripotent stem cells

International Nuclear Information System (INIS)

Chen, Ying; Wang, Kai; Chandramouli, Gadisetti V.R.; Knott, Jason G.; Leach, Richard

2013-01-01

Highlights: •Epithelial-like phenotype of trophoblast lineage cells derived from human iPS cells. •Trophoblast lineage cells derived from human iPS cells exhibit trophoblast function. •Trophoblasts from iPS cells provides a proof-of-concept in regenerative medicine. -- Abstract: Background: During implantation, the blastocyst trophectoderm attaches to the endometrial epithelium and continues to differentiate into all trophoblast subtypes, which are the major components of a placenta. Aberrant trophoblast proliferation and differentiation are associated with placental diseases. However, due to ethical and practical issues, there is almost no available cell or tissue source to study the molecular mechanism of human trophoblast differentiation, which further becomes a barrier to the study of the pathogenesis of trophoblast-associated diseases of pregnancy. In this study, our goal was to generate a proof-of-concept model for deriving trophoblast lineage cells from induced pluripotency stem (iPS) cells from human fibroblasts. In future studies the generation of trophoblast lineage cells from iPS cells established from patient’s placenta will be extremely useful for studying the pathogenesis of individual trophoblast-associated diseases and for drug testing. Methods and results: Combining iPS cell technology with BMP4 induction, we derived trophoblast lineage cells from human iPS cells. The gene expression profile of these trophoblast lineage cells was distinct from fibroblasts and iPS cells. These cells expressed markers of human trophoblasts. Furthermore, when these cells were differentiated they exhibited invasive capacity and placental hormone secretive capacity, suggesting extravillous trophoblasts and syncytiotrophoblasts. Conclusion: Trophoblast lineage cells can be successfully derived from human iPS cells, which provide a proof-of-concept tool to recapitulate pathogenesis of patient placental trophoblasts in vitro

Trophoblast lineage cells derived from human induced pluripotent stem cells

Energy Technology Data Exchange (ETDEWEB)

Chen, Ying, E-mail: ying.chen@hc.msu.edu [Department of Obstetrics, Gynecology and Reproductive Biology, Michigan State University, 333 Bostwick NE, Grand Rapids, MI 49503 (United States); Wang, Kai; Chandramouli, Gadisetti V.R. [Department of Obstetrics, Gynecology and Reproductive Biology, Michigan State University, 333 Bostwick NE, Grand Rapids, MI 49503 (United States); Knott, Jason G. [Developmental Epigenetics Laboratory, Department of Animal Science, Michigan State University (United States); Leach, Richard, E-mail: Richard.leach@hc.msu.edu [Department of Obstetrics, Gynecology and Reproductive Biology, Michigan State University, 333 Bostwick NE, Grand Rapids, MI 49503 (United States); Department of Obstetrics, Gynecology and Women’s Health, Spectrum Health Medical Group (United States)

2013-07-12

Highlights: •Epithelial-like phenotype of trophoblast lineage cells derived from human iPS cells. •Trophoblast lineage cells derived from human iPS cells exhibit trophoblast function. •Trophoblasts from iPS cells provides a proof-of-concept in regenerative medicine. -- Abstract: Background: During implantation, the blastocyst trophectoderm attaches to the endometrial epithelium and continues to differentiate into all trophoblast subtypes, which are the major components of a placenta. Aberrant trophoblast proliferation and differentiation are associated with placental diseases. However, due to ethical and practical issues, there is almost no available cell or tissue source to study the molecular mechanism of human trophoblast differentiation, which further becomes a barrier to the study of the pathogenesis of trophoblast-associated diseases of pregnancy. In this study, our goal was to generate a proof-of-concept model for deriving trophoblast lineage cells from induced pluripotency stem (iPS) cells from human fibroblasts. In future studies the generation of trophoblast lineage cells from iPS cells established from patient’s placenta will be extremely useful for studying the pathogenesis of individual trophoblast-associated diseases and for drug testing. Methods and results: Combining iPS cell technology with BMP4 induction, we derived trophoblast lineage cells from human iPS cells. The gene expression profile of these trophoblast lineage cells was distinct from fibroblasts and iPS cells. These cells expressed markers of human trophoblasts. Furthermore, when these cells were differentiated they exhibited invasive capacity and placental hormone secretive capacity, suggesting extravillous trophoblasts and syncytiotrophoblasts. Conclusion: Trophoblast lineage cells can be successfully derived from human iPS cells, which provide a proof-of-concept tool to recapitulate pathogenesis of patient placental trophoblasts in vitro.

Formation of human hepatocyte-like cells with different cellular phenotypes by human umbilical cord blood-derived cells in the human-rat chimeras

International Nuclear Information System (INIS)

Sun, Yan; Xiao, Dong; Zhang, Ruo-Shuang; Cui, Guang-Hui; Wang, Xin-Hua; Chen, Xi-Gu

2007-01-01

We took advantage of the proliferative and permissive environment of the developing pre-immune fetus to develop a noninjury human-rat xenograft small animal model, in which the in utero transplantation of low-density mononuclear cells (MNCs) from human umbilical cord blood (hUCB) into fetal rats at 9-11 days of gestation led to the formation of human hepatocyte-like cells (hHLCs) with different cellular phenotypes, as revealed by positive immunostaining for human-specific alpha-fetoprotein (AFP), cytokeratin 19 (CK19), cytokeratin 8 (CK8), cytokeratin 18 (CK18), and albumin (Alb), and with some animals exhibiting levels as high as 10.7% of donor-derived human cells in the recipient liver. More interestingly, donor-derived human cells stained positively for CD34 and CD45 in the liver of 2-month-old rat. Human hepatic differentiation appeared to partially follow the process of hepatic ontogeny, as evidenced by the expression of AFP gene at an early stage and albumin gene at a later stage. Human hepatocytes generated in this model retained functional properties of normal hepatocytes. In this xenogeneic system, the engrafted donor-derived human cells persisted in the recipient liver for at least 6 months after birth. Taken together, these findings suggest that the donor-derived human cells with different cellular phenotypes are found in the recipient liver and hHLCs hold biological activity. This humanized small animal model, which offers an in vivo environment more closely resembling the situations in human, provides an invaluable approach for in vivo investigating human stem cell behaviors, and further in vivo examining fundamental mechanisms controlling human stem cell fates in the future

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

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

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.

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.

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

c-Myc-Dependent Cell Competition in Human Cancer Cells.

Science.gov (United States)

Patel, Manish S; Shah, Heta S; Shrivastava, Neeta

2017-07-01

Cell Competition is an interaction between cells for existence in heterogeneous cell populations of multicellular organisms. This phenomenon is involved in initiation and progression of cancer where heterogeneous cell populations compete directly or indirectly for the survival of the fittest based on differential gene expression. In Drosophila, cells having lower dMyc expression are eliminated by cell competition through apoptosis when present in the milieu of cells having higher dMyc expression. Thus, we designed a study to develop c-Myc (human homolog) dependent in vitro cell competition model of human cancer cells. Cells with higher c-Myc were transfected with c-myc shRNA to prepare cells with lower c-Myc and then co-cultured with the same type of cells having a higher c-Myc in equal ratio. Cells with lower c-Myc showed a significant decrease in numbers when compared with higher c-Myc cells, suggesting "loser" and "winner" status of cells, respectively. During microscopy, engulfment of loser cells by winner cells was observed with higher expression of JNK in loser cells. Furthermore, elimination of loser cells was prevented significantly, when co-cultured cells were treated with the JNK (apoptosis) inhibitor. Above results indicate elimination of loser cells in the presence of winner cells by c-Myc-dependent mechanisms of cell competition in human cancer cells. This could be an important mechanism in human tumors where normal cells are eliminated by c-Myc-overexpressed tumor cells. J. Cell. Biochem. 118: 1782-1791, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Human induced pluripotent stem cell-derived models to investigate human cytomegalovirus infection in neural cells.

Directory of Open Access Journals (Sweden)

Leonardo D'Aiuto

Full Text Available Human cytomegalovirus (HCMV infection is one of the leading prenatal causes of congenital mental retardation and deformities world-wide. Access to cultured human neuronal lineages, necessary to understand the species specific pathogenic effects of HCMV, has been limited by difficulties in sustaining primary human neuronal cultures. Human induced pluripotent stem (iPS cells now provide an opportunity for such research. We derived iPS cells from human adult fibroblasts and induced neural lineages to investigate their susceptibility to infection with HCMV strain Ad169. Analysis of iPS cells, iPS-derived neural stem cells (NSCs, neural progenitor cells (NPCs and neurons suggests that (i iPS cells are not permissive to HCMV infection, i.e., they do not permit a full viral replication cycle; (ii Neural stem cells have impaired differentiation when infected by HCMV; (iii NPCs are fully permissive for HCMV infection; altered expression of genes related to neural metabolism or neuronal differentiation is also observed; (iv most iPS-derived neurons are not permissive to HCMV infection; and (v infected neurons have impaired calcium influx in response to glutamate.

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.

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.

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

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

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...

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

Establishment of Human Neural Progenitor Cells from Human Induced Pluripotent Stem Cells with Diverse Tissue Origins

OpenAIRE

Hayato Fukusumi; Tomoko Shofuda; Yohei Bamba; Atsuyo Yamamoto; Daisuke Kanematsu; Yukako Handa; Keisuke Okita; Masaya Nakamura; Shinya Yamanaka; Hideyuki Okano; Yonehiro Kanemura

2016-01-01

Human neural progenitor cells (hNPCs) have previously been generated from limited numbers of human induced pluripotent stem cell (hiPSC) clones. Here, 21 hiPSC clones derived from human dermal fibroblasts, cord blood cells, and peripheral blood mononuclear cells were differentiated using two neural induction methods, an embryoid body (EB) formation-based method and an EB formation method using dual SMAD inhibitors (dSMADi). Our results showed that expandable hNPCs could be generated from hiPS...

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.

Incorporation of human factors into ship collision risk models focusing on human centred design aspects

International Nuclear Information System (INIS)

Sotiralis, P.; Ventikos, N.P.; Hamann, R.; Golyshev, P.; Teixeira, A.P.

2016-01-01

This paper presents an approach that more adequately incorporates human factor considerations into quantitative risk analysis of ship operation. The focus is on the collision accident category, which is one of the main risk contributors in ship operation. The approach is based on the development of a Bayesian Network (BN) model that integrates elements from the Technique for Retrospective and Predictive Analysis of Cognitive Errors (TRACEr) and focuses on the calculation of the collision accident probability due to human error. The model takes into account the human performance in normal, abnormal and critical operational conditions and implements specific tasks derived from the analysis of the task errors leading to the collision accident category. A sensitivity analysis is performed to identify the most important contributors to human performance and ship collision. Finally, the model developed is applied to assess the collision risk of a feeder operating in Dover strait using the collision probability estimated by the developed BN model and an Event tree model for calculation of human, economic and environmental risks. - Highlights: • A collision risk model for the incorporation of human factors into quantitative risk analysis is proposed. • The model takes into account the human performance in different operational conditions leading to the collision. • The most important contributors to human performance and ship collision are identified. • The model developed is applied to assess the collision risk of a feeder operating in Dover strait.

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....

The human airway epithelial basal cell transcriptome.

Directory of Open Access Journals (Sweden)

Neil R Hackett

2011-05-01

Full Text Available The human airway epithelium consists of 4 major cell types: ciliated, secretory, columnar and basal cells. During natural turnover and in response to injury, the airway basal cells function as stem/progenitor cells for the other airway cell types. The objective of this study is to better understand human airway epithelial basal cell biology by defining the gene expression signature of this cell population.Bronchial brushing was used to obtain airway epithelium from healthy nonsmokers. Microarrays were used to assess the transcriptome of basal cells purified from the airway epithelium in comparison to the transcriptome of the differentiated airway epithelium. This analysis identified the "human airway basal cell signature" as 1,161 unique genes with >5-fold higher expression level in basal cells compared to differentiated epithelium. The basal cell signature was suppressed when the basal cells differentiated into a ciliated airway epithelium in vitro. The basal cell signature displayed overlap with genes expressed in basal-like cells from other human tissues and with that of murine airway basal cells. Consistent with self-modulation as well as signaling to other airway cell types, the human airway basal cell signature was characterized by genes encoding extracellular matrix components, growth factors and growth factor receptors, including genes related to the EGF and VEGF pathways. Interestingly, while the basal cell signature overlaps that of basal-like cells of other organs, the human airway basal cell signature has features not previously associated with this cell type, including a unique pattern of genes encoding extracellular matrix components, G protein-coupled receptors, neuroactive ligands and receptors, and ion channels.The human airway epithelial basal cell signature identified in the present study provides novel insights into the molecular phenotype and biology of the stem/progenitor cells of the human airway epithelium.

Development and function of human innate immune cells in a humanized mouse model.

Science.gov (United States)

Rongvaux, Anthony; Willinger, Tim; Martinek, Jan; Strowig, Till; Gearty, Sofia V; Teichmann, Lino L; Saito, Yasuyuki; Marches, Florentina; Halene, Stephanie; Palucka, A Karolina; Manz, Markus G; Flavell, Richard A

2014-04-01

Mice repopulated with human hematopoietic cells are a powerful tool for the study of human hematopoiesis and immune function in vivo. However, existing humanized mouse models cannot support development of human innate immune cells, including myeloid cells and natural killer (NK) cells. Here we describe two mouse strains called MITRG and MISTRG, in which human versions of four genes encoding cytokines important for innate immune cell development are knocked into their respective mouse loci. The human cytokines support the development and function of monocytes, macrophages and NK cells derived from human fetal liver or adult CD34(+) progenitor cells injected into the mice. Human macrophages infiltrated a human tumor xenograft in MITRG and MISTRG mice in a manner resembling that observed in tumors obtained from human patients. This humanized mouse model may be used to model the human immune system in scenarios of health and pathology, and may enable evaluation of therapeutic candidates in an in vivo setting relevant to human physiology.

The comparison of two methods to obtain human oral keratinocytes in primary culture

International Nuclear Information System (INIS)

Klingbeil, Maria Fatima Guarizo

2006-01-01

The therapeutic procedures frequently used in oral treatments for the pathological diseases are surgical, resulting in failures of the mucosal continuity.The possibility to obtain transplantable oral epithelia from an in vitro cell culture opens new utilization perspectives not only to where it comes from, but also as a reconstructive material for other parts of the human body, such as: urethra, epithelia corneo-limbal, cornea, ocular surface. Many researchers still use controversial methods for obtaining cells. It was therefore evaluated and compared the efficiency in both methods: enzymatic and direct explant to obtain oral keratinocytes from human oral mucosa. Fragments of intra oral epithelial tissues from healthy human subjects, undergoing dental surgeries, were donated to the research project. The keratinocytes were cultivated over a feeder-layer from a previously irradiated 3T3 Swiss albino fibroblasts. In this study it was compared the time needed in the cell obtention, the best cell amount between both methods, the life-span, the cell capacity to form an in vitro epithelia and its morphologic structure. The results in the assessment of both methods have shown the possibility to obtain keratinocytes from a small oral fragment, but at the same time we may verify the advantages and peculiar restrictions for each one of both analyzed methods. (author)

Identification and cloning of a prethymic precursor T lymphocyte from a population of common acute lymphoblastic leukemia antigen (CALLA)-positive fetal bone marrow cells

DEFF Research Database (Denmark)

Hokland, P; Hokland, M; Daley, J

1987-01-01

We have cloned common acute lymphoblastic leukemia (CALLA)-positive cells from human fetal bone marrow containing less than 1 in 10,000 E-RFC in round-bottomed microtiter wells (one cell per well) using the autocloning unit of an EPICS-V cell sorter. Expansion of such cells (with IL-2 and heavily...... irradiated autologous thymocytes as feeder cells) resulted in growth in 6-14% of the wells (mean, 11%) with cells with mature T lymphocyte phenotype. Two-color fluorescence analysis of outgrowing cultures furthermore ascertained that these cells had differentiated through a phase of simultaneous expression...... of T4 and T8 antigens and at the same time expression of the thymocyte-associated T6 antigens. Thus, given the fact that 10-20% of T cell acute lymphoblastic leukemia (T-ALLs) are CALLA+, we have been able to identify a human prethymic T lymphocyte population that might be the normal counterpart...

Human monoclonal antibodies reactive with human myelomonocytic leukemia cells.

Science.gov (United States)

Posner, M R; Santos, D J; Elboim, H S; Tumber, M B; Frackelton, A R

1989-04-01

Peripheral blood mononuclear cells from a patient with chronic myelogenous leukemia (CML), in remission, were depleted of CD8-positive T-cells and cultured with Epstein-Barr virus. Four of 20 cultures (20%) secreted human IgG antibodies selectively reactive with the cell surfaces of certain human leukemia cell lines. Three polyclonal, Epstein-Barr virus-transformed, B-cell lines were expanded and fused with the human-mouse myeloma analogue HMMA2.11TG/O. Antibody from secreting clones HL 1.2 (IgG1), HL 2.1 (IgG3), and HL 3.1 (IgG1) have been characterized. All three react with HL-60 (promyelocytic), RWLeu4 (CML promyelocytic), and U937 (monocytic), but not with KG-1 (myeloblastic) or K562 (CML erythroid). There is no reactivity with T-cell lines, Burkitt's cell lines, pre-B-leukemia cell lines, or an undifferentiated CML cell line, BV173. Leukemic cells from two of seven patients with acute myelogenous leukemia and one of five with acute lymphocytic leukemia react with all three antibodies. Normal lymphocytes, monocytes, polymorphonuclear cells, red blood cells, bone marrow cells, and platelets do not react. Samples from patients with other diverse hematopoietic malignancies showed no reactivity. Immunoprecipitations suggest that the reactive antigen(s) is a lactoperoxidase iodinatable series of cell surface proteins with molecular weights of 42,000-54,000 and a noniodinatable protein with a molecular weight of 82,000. Based on these data these human monoclonal antibodies appear to react with myelomonocytic leukemic cells and may detect a leukemia-specific antigen or a highly restricted differentiation antigen.

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)

Signaling hierarchy regulating human endothelial cell development.

Science.gov (United States)

Kelly, Melissa A; Hirschi, Karen K

2009-05-01

Our present knowledge of the regulation of mammalian endothelial cell differentiation has been largely derived from studies of mouse embryonic development. However, unique mechanisms and hierarchy of signals that govern human endothelial cell development are unknown and, thus, explored in these studies. Using human embryonic stem cells as a model system, we were able to reproducibly and robustly generate differentiated endothelial cells via coculture on OP9 marrow stromal cells. We found that, in contrast to studies in the mouse, bFGF and VEGF had no specific effects on the initiation of human vasculogenesis. However, exogenous Ihh promoted endothelial cell differentiation, as evidenced by increased production of cells with cobblestone morphology that coexpress multiple endothelial-specific genes and proteins, form lumens, and exhibit DiI-AcLDL uptake. Inhibition of BMP signaling using Noggin or BMP4, specifically, using neutralizing antibodies suppressed endothelial cell formation; whereas, addition of rhBMP4 to cells treated with the hedgehog inhibitor cyclopamine rescued endothelial cell development. Our studies revealed that Ihh promoted human endothelial cell differentiation from pluripotent hES cells via BMP signaling, providing novel insights applicable to modulating human endothelial cell formation and vascular regeneration for human clinical therapies.

Stemcell Information: SKIP000533 [SKIP Stemcell Database[Archive

Lifescience Database Archive (English)

Full Text Available aucasian/Japanese -- No hiPSC derived from primary human dermal fibroblasts.(Control line with a wildtype ACVR1 gene.) ヒト線維芽細胞...T4, SOX2, KLF4, c-MYC ... Yes SNL feeder cells primate ES cell medium(ReproCELL) ... ...8-190 Induced pluripotent stem cells from patients with human fibrodysplasia ossi

Human regulatory B cells control the TFH cell response.

Science.gov (United States)

Achour, Achouak; Simon, Quentin; Mohr, Audrey; Séité, Jean-François; Youinou, Pierre; Bendaoud, Boutahar; Ghedira, Ibtissem; Pers, Jacques-Olivier; Jamin, Christophe

2017-07-01

Follicular helper T (T FH ) cells support terminal B-cell differentiation. Human regulatory B (Breg) cells modulate cellular responses, but their control of T FH cell-dependent humoral immune responses is unknown. We sought to assess the role of Breg cells on T FH cell development and function. Human T cells were polyclonally stimulated in the presence of IL-12 and IL-21 to generate T FH cells. They were cocultured with B cells to induce their terminal differentiation. Breg cells were included in these cultures, and their effects were evaluated by using flow cytometry and ELISA. B-cell lymphoma 6, IL-21, inducible costimulator, CXCR5, and programmed cell death protein 1 (PD-1) expressions increased on stimulated human T cells, characterizing T FH cell maturation. In cocultures they differentiated B cells into CD138 + plasma and IgD - CD27 + memory cells and triggered immunoglobulin secretions. Breg cells obtained by Toll-like receptor 9 and CD40 activation of B cells prevented T FH cell development. Added to T FH cell and B-cell cocultures, they inhibited B-cell differentiation, impeded immunoglobulin secretions, and expanded Foxp3 + CXCR5 + PD-1 + follicular regulatory T cells. Breg cells modulated IL-21 receptor expressions on T FH cells and B cells, and their suppressive activities involved CD40, CD80, CD86, and intercellular adhesion molecule interactions and required production of IL-10 and TGF-β. Human Breg cells control T FH cell maturation, expand follicular regulatory T cells, and inhibit the T FH cell-mediated antibody secretion. These novel observations demonstrate a role for the Breg cell in germinal center reactions and suggest that deficient activities might impair the T FH cell-dependent control of humoral immunity and might lead to the development of aberrant autoimmune responses. Copyright © 2016 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Hybrid clone cells derived from human breast epithelial cells and human breast cancer cells exhibit properties of cancer stem/initiating cells.

Science.gov (United States)

Gauck, Daria; Keil, Silvia; Niggemann, Bernd; Zänker, Kurt S; Dittmar, Thomas

2017-08-02

The biological phenomenon of cell fusion has been associated with cancer progression since it was determined that normal cell × tumor cell fusion-derived hybrid cells could exhibit novel properties, such as enhanced metastatogenic capacity or increased drug resistance, and even as a mechanism that could give rise to cancer stem/initiating cells (CS/ICs). CS/ICs have been proposed as cancer cells that exhibit stem cell properties, including the ability to (re)initiate tumor growth. Five M13HS hybrid clone cells, which originated from spontaneous cell fusion events between M13SV1-EGFP-Neo human breast epithelial cells and HS578T-Hyg human breast cancer cells, and their parental cells were analyzed for expression of stemness and EMT-related marker proteins by Western blot analysis and confocal laser scanning microscopy. The frequency of ALDH1-positive cells was determined by flow cytometry using AldeRed fluorescent dye. Concurrently, the cells' colony forming capabilities as well as the cells' abilities to form mammospheres were investigated. The migratory activity of the cells was analyzed using a 3D collagen matrix migration assay. M13HS hybrid clone cells co-expressed SOX9, SLUG, CK8 and CK14, which were differently expressed in parental cells. A variation in the ALDH1-positive putative stem cell population was observed among the five hybrids ranging from 1.44% (M13HS-7) to 13.68% (M13HS-2). In comparison to the parental cells, all five hybrid clone cells possessed increased but also unique colony formation and mammosphere formation capabilities. M13HS-4 hybrid clone cells exhibited the highest colony formation capacity and second highest mammosphere formation capacity of all hybrids, whereby the mean diameter of the mammospheres was comparable to the parental cells. In contrast, the largest mammospheres originated from the M13HS-2 hybrid clone cells, whereas these cells' mammosphere formation capacity was comparable to the parental breast cancer cells. All M13HS

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

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

Second generation codon optimized minicircle (CoMiC) for nonviral reprogramming of human adult fibroblasts.

Science.gov (United States)

Diecke, Sebastian; Lisowski, Leszek; Kooreman, Nigel G; Wu, Joseph C

2014-01-01

The ability to induce pluripotency in somatic cells is one of the most important scientific achievements in the fields of stem cell research and regenerative medicine. This technique allows researchers to obtain pluripotent stem cells without the controversial use of embryos, providing a novel and powerful tool for disease modeling and drug screening approaches. However, using viruses for the delivery of reprogramming genes and transcription factors may result in integration into the host genome and cause random mutations within the target cell, thus limiting the use of these cells for downstream applications. To overcome this limitation, various non-integrating techniques, including Sendai virus, mRNA, minicircle, and plasmid-based methods, have recently been developed. Utilizing a newly developed codon optimized 4-in-1 minicircle (CoMiC), we were able to reprogram human adult fibroblasts using chemically defined media and without the need for feeder cells.

Stem cells in the human breast

DEFF Research Database (Denmark)

Petersen, Ole William; Polyak, Kornelia

2010-01-01

The origins of the epithelial cells participating in the development, tissue homeostasis, and cancer of the human breast are poorly understood. However, emerging evidence suggests a role for adult tissue-specific stem cells in these processes. In a hierarchical manner, these generate the two main...... mammary cell lineages, producing an increasing number of cells with distinct properties. Understanding the biological characteristics of human breast stem cells and their progeny is crucial in attempts to compare the features of normal stem cells and cancer precursor cells and distinguish these from...... nonprecursor cells and cells from the bulk of a tumor. A historical overview of research on human breast stem cells in primary tissue and in culture reveals the progress that has been made in this area, whereas a focus on the cell-of-origin and reprogramming that occurs during neoplastic conversion provides...

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

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.

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.

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.

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.

Human prostatic cancer cells, PC3, elaborate mitogenic activity which selectively stimulates human bone cells

International Nuclear Information System (INIS)

Perkel, V.S.; Mohan, S.; Herring, S.J.; Baylink, D.J.; Linkhart, T.A.

1990-01-01

Prostatic cancer typically produces osteoblastic metastases which are not attended by marrow fibrosis. In the present study we sought to test the hypothesis that prostatic cancer cells produce factor(s) which act selectively on human osteoblasts. Such a paracrine mechanism would explain the observed increase in osteoblasts, unaccompanied by an increase in marrow fibroblasts. To test this hypothesis we investigated the mitogenic activity released by the human prostatic tumor cell line, PC3. PC3 cells have been reported previously to produce mitogenic activity for cells that was relatively specific for rat osteoblasts compared to rat fibroblasts. However, the effects of this activity on human cells has not been examined previously. PC3-conditioned medium (CM) (5-50 micrograms CM protein/ml) stimulated human osteoblast proliferation by 200-950% yet did not stimulate human fibroblast proliferation ([3H]thymidine incorporation). PC3 CM also increased cell numbers in human osteoblast but not fibroblast cell cultures. To determine whether the osteoblast-specific mitogenic activity could be attributed to known bone growth factors, specific assays for these growth factors were performed. PC3 CM contained 10 pg insulin-like growth factor (IGF) I, less than 2 pg IGF II, 54 pg basic fibroblast growth factor, and 16 pg transforming growth factor beta/microgram CM protein. None of these growth factors alone or in combination could account for the observed osteoblast-specific PC3 cell-derived mitogenic activity. Furthermore, when 5 micrograms/ml PC3 CM was tested in combination with maximally effective concentrations of either basic fibroblast growth factor, IGF I, IGF II, or transforming growth factor beta, it produced an additive effect suggesting that PC3 CM stimulates osteoblast proliferation by a mechanism independent of these bone mitogens

Tuft (caveolated) cells in two human colon carcinoma cell lines.

OpenAIRE

Barkla, D. H.; Whitehead, R. H.; Foster, H.; Tutton, P. J.

1988-01-01

The presence of an unusual cell type in two human colon carcinoma cell lines is reported. The cells show the same morphology as "tuft" (caveolated) cells present in normal gastrointestinal epithelium. Tuft cells were seen in cell line LIM 1863 growing in vitro and in human colon carcinoma cell line LIM 2210 growing as subcutaneous solid tumour xenografts in nude mice. Characteristic morphologic features of tuft cells included a wide base, narrow apex and a tuft of long microvilli projecting f...

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...

Modeling human infertility with pluripotent stem cells

Directory of Open Access Journals (Sweden)

Di Chen

2017-05-01

Full Text Available Human fertility is dependent upon the correct establishment and differentiation of the germline. This is because no other cell type in the body is capable of passing a genome and epigenome from parent to child. Terminally differentiated germline cells in the adult testis and ovary are called gametes. However, the initial specification of germline cells occurs in the embryo around the time of gastrulation. Most of our knowledge regarding the cell and molecular events that govern human germline specification involves extrapolating scientific principles from model organisms, most notably the mouse. However, recent work using next generation sequencing, gene editing and differentiation of germline cells from pluripotent stem cells has revealed that the core molecular mechanisms that regulate human germline development are different from rodents. Here, we will discuss the major molecular pathways required for human germline differentiation and how pluripotent stem cells have revolutionized our ability to study the earliest steps in human embryonic lineage specification in order to understand human fertility.

Primary chondrocytes enhance cartilage tissue formation upon co-culture with expanded chondrocytes, dermal fibroblasts, 3T3 feeder cells and embryonic stem cells

NARCIS (Netherlands)

Hendriks, J.A.A.; Miclea, Razvan L.; Schotel, Roka; de Bruijn, Ewart; Moroni, Lorenzo; Karperien, Hermanus Bernardus Johannes; Riesle, J.U.; van Blitterswijk, Clemens

2010-01-01

Co-culture models have been increasingly used in tissue engineering applications to understand cell–cell interactions and consequently improve regenerative medicine strategies. Aiming at further elucidating cartilage tissue formation, we co-cultured bovine primary chondrocytes (BPCs) with human

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.

Inhibition of JAK3 and PKC via Immunosuppressive Drugs Tofacitinib and Sotrastaurin Inhibits Proliferation of Human B Lymphocytes In Vitro.

Science.gov (United States)

Martina, M N; Ramirez Bajo, M J; Bañon-Maneus, E; Moya Rull, D; Hierro-Garcia, N; Revuelta, I; Campistol, J M; Rovira, J; Diekmann, F

2016-11-01

Antibody-mediated response in solid organ transplantation is critical for graft dysfunction and loss. The use of immunosuppressive agents partially inhibits the B-lymphocyte response leading to a risk of acute and chronic antibody-mediated rejection. This study evaluated the impact of JAK3 and PKC inhibitors tofacitinib (Tofa) and sotrastaurin (STN), respectively, on B-cell proliferation, apoptosis, and activation in vitro. Human B cells isolated from peripheral blood of healthy volunteers were cocultured with CD40 ligand-transfected fibroblasts as feeder cells in the presence of interleukin (IL) 2, IL-10, and IL-21. The cocultures were treated with immunosuppressants Tofa, STN, and rapamycin (as a control), to analyze the proliferation and apoptosis of B cells by means of Cyquant and flow cytometry, respectively. CD27 and IgG staining were applied to evaluate whether treatments modified the activation of B cells. Tofa and STN were able to inhibit B-cell proliferation to the same extent as rapamycin, without inducing cell apoptosis. After 6 days in coculture with feeder cells, all B cells showed CD27 memory B-cell phenotype. None of the immunosuppressive treatments modified the proportion between class-switched and non-class-switched memory B cells observed in nontreated cultures. The high predominance of CD27 + CD24 + phenotype was not modified by any immunosuppressive treatment. Our results show that Tofa and STN can suppress B-cell antibody responses to an extent similar to rapamycin, in vitro; therefore these compounds may be a useful therapy against antibody-mediated rejection in transplantation. Copyright © 2016. Published by Elsevier Inc.

Establishment of Human Neural Progenitor Cells from Human Induced Pluripotent Stem Cells with Diverse Tissue Origins

Directory of Open Access Journals (Sweden)

Hayato Fukusumi

2016-01-01

Full Text Available Human neural progenitor cells (hNPCs have previously been generated from limited numbers of human induced pluripotent stem cell (hiPSC clones. Here, 21 hiPSC clones derived from human dermal fibroblasts, cord blood cells, and peripheral blood mononuclear cells were differentiated using two neural induction methods, an embryoid body (EB formation-based method and an EB formation method using dual SMAD inhibitors (dSMADi. Our results showed that expandable hNPCs could be generated from hiPSC clones with diverse somatic tissue origins. The established hNPCs exhibited a mid/hindbrain-type neural identity and uniform expression of neural progenitor genes.

3 CFR - Guidelines for Human Stem Cell Research

Science.gov (United States)

2010-01-01

... 3 The President 1 2010-01-01 2010-01-01 false Guidelines for Human Stem Cell Research Presidential Documents Other Presidential Documents Memorandum of July 30, 2009 Guidelines for Human Stem Cell Research..., scientifically worthy human stem cell research, including human embryonic stem cell research, to the extent...

In vitro proliferation of adult human beta-cells.

Directory of Open Access Journals (Sweden)

Sabine Rutti

Full Text Available A decrease in functional beta-cell mass is a key feature of type 2 diabetes. Glucagon-like peptide 1 (GLP-1 analogues induce proliferation of rodent beta-cells. However, the proliferative capacity of human beta-cells and its modulation by GLP-1 analogues remain to be fully investigated. We therefore sought to quantify adult human beta-cell proliferation in vitro and whether this is affected by the GLP-1 analogue liraglutide.Human islets from 7 adult cadaveric organ donors were dispersed into single cells. Beta-cells were purified by FACS. Non-sorted cells and the beta-cell enriched ("beta-cells" population were plated on extracellular matrix from rat (804G and human bladder carcinoma cells (HTB9 or bovine corneal endothelial ECM (BCEC. Cells were maintained in culture+/-liraglutide for 4 days in the presence of BrdU.Rare human beta-cell proliferation could be observed either in the purified beta-cell population (0.051±0.020%; 22 beta-cells proliferating out of 84'283 beta-cells counted or in the non-sorted cell population (0.055±0.011%; 104 proliferating beta-cells out of 232'826 beta-cells counted, independently of the matrix or the culture conditions. Liraglutide increased human beta-cell proliferation on BCEC in the non-sorted cell population (0.082±0.034% proliferating beta-cells vs. 0.017±0.008% in control, p<0.05.These results indicate that adult human beta-cell proliferation can occur in vitro but remains an extremely rare event with these donors and particular culture conditions. Liraglutide increases beta-cell proliferation only in the non-sorted cell population and only on BCEC. However, it cannot be excluded that human beta-cells may proliferate to a greater extent in situ in response to natural stimuli.

Glycogen synthase kinase 3 (GSK3) inhibitor 6-bromoindirubin-3 ...

African Journals Online (AJOL)

... male germline stem cells (mGSCs) under serum- and feeder-free conditions. ... to maintain the pluripotency of human and mouse embryonic stem cells (ESCs). ... capacity of mGSC-induced embryoid bodies (EBs) were examined as well.

Human innate lymphoid cells.

Science.gov (United States)

Mjösberg, Jenny; Spits, Hergen

2016-11-01

Innate lymphoid cells (ILCs) are increasingly acknowledged as important mediators of immune homeostasis and pathology. ILCs act as early orchestrators of immunity, responding to epithelium-derived signals by expressing an array of cytokines and cell-surface receptors, which shape subsequent immune responses. As such, ILCs make up interesting therapeutic targets for several diseases. In patients with allergy and asthma, group 2 innate lymphoid cells produce high amounts of IL-5 and IL-13, thereby contributing to type 2-mediated inflammation. Group 3 innate lymphoid cells are implicated in intestinal homeostasis and psoriasis pathology through abundant IL-22 production, whereas group 1 innate lymphoid cells are accumulated in chronic inflammation of the gut (inflammatory bowel disease) and lung (chronic obstructive pulmonary disease), where they contribute to IFN-γ-mediated inflammation. Although the ontogeny of mouse ILCs is slowly unraveling, the development of human ILCs is far from understood. In addition, the growing complexity of the human ILC family in terms of previously unrecognized functional heterogeneity and plasticity has generated confusion within the field. Here we provide an updated view on the function and plasticity of human ILCs in tissue homeostasis and disease. Copyright © 2016 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Evaluation of umbilical cord blood CD34+ hematopoietic stem cells expansion with inhibition of TGF-β receptorII in co-culture with bone marrow mesenchymal stromal cells.

Science.gov (United States)

Sohrabi Akhkand, Saman; Amirizadeh, Naser; Nikougoftar, Mahin; Alizadeh, Javad; Zaker, Farhad; Sarveazad, Arash; Joghataei, Mohammad Taghi; Faramarzi, Mahmood

2016-08-01

Umbilical cord blood (UCB) is an important source of hematopoietic stem cells (HSCs). However, low number of HSCs in UCB has been an obstacle for adult hematopoietic stem cell transplantation. The expansion of HSCs in culture is one approach to overcome this problem. In this study, we investigated the expansion of UCB-HSCs by using human bone marrow mesenchymal stromal cells (MSCs) as feeder layer as well as inhibiting the TGF-β signaling pathway through reduction of TGF-βRII expression. CD34(+) cells were isolated from UCB and transfected by SiRNA targeting TGF-βRII mRNA. CD34(+) cells were expanded in four culture media with different conditions, including 1) expansion of CD34(+) cells in serum free medium containing growth factors, 2) expansion of cells transfected with SiRNA targeting TGF-βRII in medium containing growth factors, 3) expansion of cells in presence of growth factors and MSCs, 4) expansion of cells transfected with SiRNA targeting TGF-βRII on MSCs feeder layer in medium containing growth factors. These culture conditions were evaluated for the number of total nucleated cells (TNCs), CD34 surface marker as well as using CFU assay on 8th day after culture. The fold increase in CD34(+) cells, TNCs, and colony numbers (71.8±6.9, 93.2±10.2 and 128±10, respectively) was observed to be highest in fourth culture medium compared to other culture conditions. The difference between number of cells in four culture media in 8th day compared to unexpanded cells (0day) before expansion was statistically significant (P<0.05). The results showed that transfection of CD34(+) cells with SiRNA targeting TGF-βRII and their co-culture with MSCs could considerably increase the number of progenitors. Therefore, this method could be useful for UCB-HSCs expansion. Copyright © 2016 Elsevier Ltd. All rights reserved.

Some Ethical Concerns About Human Induced Pluripotent Stem Cells.

Science.gov (United States)

Zheng, Yue Liang

2016-10-01

Human induced pluripotent stem cells can be obtained from somatic cells, and their derivation does not require destruction of embryos, thus avoiding ethical problems arising from the destruction of human embryos. This type of stem cell may provide an important tool for stem cell therapy, but it also results in some ethical concerns. It is likely that abnormal reprogramming occurs in the induction of human induced pluripotent stem cells, and that the stem cells generate tumors in the process of stem cell therapy. Human induced pluripotent stem cells should not be used to clone human beings, to produce human germ cells, nor to make human embryos. Informed consent should be obtained from patients in stem cell therapy.

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...

Comprehensive evaluation of leukocyte lineage derived from human hematopoietic cells in humanized mice.

Science.gov (United States)

Takahashi, Masayuki; Tsujimura, Noriyuki; Otsuka, Kensuke; Yoshino, Tomoko; Mori, Tetsushi; Matsunaga, Tadashi; Nakasono, Satoshi

2012-04-01

Recently, humanized animals whereby a part of the animal is biologically engineered using human genes or cells have been utilized to overcome interspecific differences. Herein, we analyzed the detail of the differentiation states of various human leukocyte subpopulations in humanized mouse and evaluated comprehensively the similarity of the leukocyte lineage between humanized mice and humans. Humanized mice were established by transplanting human CD34(+) cord blood cells into irradiated severely immunodeficient NOD/Shi-scid/IL2Rγ(null) (NOG) mice, and the phenotypes of human cells contained in bone marrow, thymus, spleen and peripheral blood from the mice were analyzed at monthly intervals until 4 months after cell transplantation. The analysis revealed that transplanted human hematopoietic stem cells via the caudal vein homed and engrafted themselves successfully at the mouse bone marrow. Subsequently, the differentiated leukocytes migrated to the various tissues. Almost all of the leukocytes within the thymus were human cells. Furthermore, analysis of the differentiation states of human leukocytes in various tissues and organs indicated that it is highly likely that the human-like leukocyte lineage can be developed in mice. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.