Derivation of cat embryonic stem-like cells from in vitro-produced blastocysts on homologous and heterologous feeder cells.

PubMed

Gómez, M C; Serrano, M A; Pope, C Earle; Jenkins, J A; Biancardi, M N; López, M; Dumas, C; Galiguis, J; Dresser, B L

2010-09-01

The domestic cat is a focal mammalian species that is used as a model for developing assisted reproductive technologies for preserving endangered cats and for studying human diseases. The generation of stable characterized cat embryonic stem cells (ESC) lines to use as donor nuclei may help to improve the efficiency of interspecies somatic cell nuclear transfer for preserving endangered cats and allow the creation of knockout cell lines to generate knockout cats for studying function of specific genes related to human diseases. It will also enable the possibility of producing gametes in vitro from ESC of endangered cats. In the present study, we report the generation of cat embryonic stem-like (cESL) cells from blastocysts derived entirely in vitro. We generated 32 cESL cell lines from 331 in vitro derived blastocysts from which inner cell masses were isolated by immunosurgery or by a mechanical method. Inhibition of cat dermal fibroblast (CDF) proliferation after exposure to mitomycin-C was both dose and time dependent, where doses of 30 to 40 microg/mL for 5 h were most efficient. These dosages were higher than that required to inhibit cell proliferation of mouse fetal fibroblasts (MFF; 10 microg/mL for 2.5 h). Mitomycin-C did not significantly increase necrosis of cells from either species, and had an anti-proliferative effect at concentrations below cytotoxicity. A clear species-specific relationship between feeder layers and derivation of cESL cell lines was observed, where higher numbers of cESL cell lines were generated on homologous cat feeder layers (n = 26) than from those derived on heterologous mouse feeder layers (n = 6). Three cESL cell lines generated from immunosurgery and cultured on CDF maintained self-renewal and were morphologically undifferentiated for nine and twelve passages (69-102 days). These lines showed a tightly packed dome shaped morphology, exhibited alkaline phosphatase activity and immuno-expression of the pluripotent marker OCT-4 and surface marker SSEA-1. Primary colonies at P0 to P3 and cat blastocysts expressed transcription factors OCT-4, NANOG and SOX-2 and the proto-oncogene C-MYC. However, expression was at levels significantly lower than in vitro produced blastocysts. During culture, cESL colonies spontaneously differentiated into fibroblasts, cardiomyocytes, and embryoid bodies. Development of techniques to prevent differentiation of cESL cells will be essential for maintaining defined cell lines. Copyright 2010 Elsevier Inc. All rights reserved.

Development and evaluation of a new composite Laserskin graft.

PubMed

Lam, P K; Chan, E S; To, E W; Lau, C H; Yen, S C; King, W W

1999-11-01

Tremendous effort has been made to improve the graft take rate of cultured epidermal autograph. The purpose of this study is to develop and evaluate a new composite Laserskin graft (CLSG) as a human skin substitute for wound resurfacing. The seeding efficacy of cultured keratinocytes on plain Laserskin was compared with the 3T3 cell-seeded Laserskin and allogenic fibroblast-populated Laserskin. Three different types of CLSG, 2 cm in diameter each, were prepared and tested in rats. Type A CLSG consisted of proliferative allogenic rat fibroblasts on both sides of the Laserskin with autologous keratinocytes also on the upper side. Fibroblasts and keratinocytes were seeded only on the upper side of the Laserskin in type B CLSG. Keratinocytes alone were seeded on plain Laserskin in type C CLSG. Type B CLSG consisting of autologous keratinocytes and autologous dermal fibroblasts was tested on five selected wounds (5x5 cm each) of a patient with full-thickness burn. In another burn patient, type B CLSG consisting of autologous keratinocytes and allogenic dermal fibroblasts was grafted onto three wounds (5x5 cm each). The seeding efficacy of human keratinocytes on plain Laserskin increased from 75% to 95% when proliferative allogenic fibroblasts were grown as a feeder layer on the Laserskin. The seeding efficacy of rat keratinocytes increased from 36% to 88% in the presence of a proliferative allogenic fibroblast feeder layer, whereas human/rat keratinocytes had respective seeding efficacy of 98%/91% on Laserskin preseeded with mitomycin C-treated 3T3 cells. Skin biopsies of grafted type A CLSG on day 14 after grafting showed complete epithelialization without severe inflammation in 16 of 20 (80%) grafted surgical wounds in rats. There were eight (40%) and seven (35%) "takes" of the CLSG in types B and C, respectively. The infection rate in type B CLSG was two (10%). There was one (5%) infection in types A and C. The respective take rates on the two patients grafted with type B CLSG were 60% and 100%. The animal experiment and the preliminary clinical data showed that the CSLGs consisting of autologous keratinocytes and of autologous/allogenic fibroblasts are good human skin substitutes in terms of durability, biocompatibility, high seeding efficacy for keratinocytes, high graft take rate, and low infection rate.

Effect of 1,25-dihydroxyvitamin D3 on human keratinocytes grown under different culture conditions.

PubMed

McLane, J A; Katz, M; Abdelkader, N

1990-04-01

1,25-Dihydroxyvitamin D3 (1,25-(OH)2-D3) is known to decrease the proliferation and increase the differentiation of different cell types including human keratinocytes. The growth and differentiation of keratinocytes in the presence of 1,25-(OH)2-D3 using serum-free media formulations has been described previously. This investigation extends these studies to describe various culture conditions with human foreskin keratinocytes to determine the optimal antiproliferative activity of 1,25-(OH)2-D3. Keratinocytes were plated onto tissue culture dishes using one of three basic serum-free media protocols; a) with no feeder layer in keratinocyte growth medium (KGM); b) onto mitomycin C-treated 3T3 mouse embryo fibroblasts; or c) onto mitomycin C-treated dermal human fibroblasts. The last two protocols utilized Dulbecco's modified Eagle's Medium (DMEM) supplemented with growth factors. Keratinocyte cell growth was greatest in the KGM medium. Although the growth of keratinocytes on either feeder layer was similar, there were differences in the ability of the cells to form envelopes in the presence of 1,25-(OH)2-D3. The addition of hydrocortisone and cholera toxin to the medium also affected the response of the keratinocytes to 1,25-(OH)2-D3. The antiproliferative effect of 1,25-(OH)2-D3 was not altered by varying the extracellular calcium levels from 0.25 to 3 mM. The antiproliferative activity of 1,25-(OH)2-D3 is attenuated in cells at low density. Our results suggest that an optimal condition to investigate the ability of 1,25-(OH)2-D3 to inhibit keratinocyte proliferation is at preconfluent cell density in the presence of KGM supplemented with 1.5 mM calcium without a feeder layer. These conditions are not appropriate for investigating the enhancement of differentiation by 1,25-(OH)2-D3, but can be used to assay other agents that modulate keratinocyte proliferation.

Post-mitotic human dermal fibroblasts efficiently support the growth of human follicular keratinocytes.

PubMed

Limat, A; Hunziker, T; Boillat, C; Bayreuther, K; Noser, F

1989-05-01

For growth at low seeding densities, keratinocytes isolated from human tissues like epidermis or hair follicles are dependent on mesenchyme-derived feeder cells such as the 3T3-cell employed so far. As an alternative method, the present study describes the use of post-mitotic human dermal fibroblasts sublethally irradiated or mitomycin C-treated. Special emphasis was put on efficient growth of primary keratinocyte cultures plated at very low seeding densities. Thus, outer root sheath cells isolated from two anagen human hair follicles and plated in a 35-mm culture dish (3 - 6 X 10(2) attached cells) grew to confluence within 3 weeks (6 - 8 X 10(5) cells). Similar results were obtained for interfollicular keratinocytes. A crucial point for the function of these fibroblast feeder cells is plating at appropriate densities, considering their tremendous increase in cell size at the post-mitotic state. Plating densities of 4 - 5 X 10(3/cm2 allow full spreading of the feeder cells and do not impede the settling and expansion of the keratinocytes. Major advantages of this system include easier handling and better reproducibility than using 3T3-cells. Moreover, homologous fibroblast feeders mimic more closely the physiologic situation and therefore might provide a valuable tool for studying interactions between human mesenchymal and epithelial cells. Finally, potential hazards of using transformed feeder cells from a different species in keratinocyte cultures raised for wound covering in humans could be thus avoided.

A new system for cultivation of human keratinocytes on acellular dermal matrix substitute with the use of human fibroblast feeder layer.

PubMed

Xiao, S; Zhu, S; Ma, B; Xia, Z-F; Yang, J; Wang, G

2008-01-01

To improve the proliferative potential of human keratinocytes (HK) cultured on acellular dermal matrix (ADM), HK and mitomycin C-treated human fibroblasts (MMC-HF) were seeded onto ADM to form four types of composite skin: type I, HK were seeded onto the epidermal side of ADM; type II, both HK and MMC-HF were seeded onto the epidermal side; type III, MMC-HF were preseeded onto the dermal side of ADM, and then HK were seeded onto the epidermal side, and type IV, where MMC-HF were preseeded onto both sides, and then HK were seeded onto the epidermal side. Compared with type I and III, the proliferative potential of HK of type II and IV was significantly higher on day 3, 5, 7 and 9 in vitro. In type I and III, HK grew into one layer on day 7-9, while in type II and IV keratinocytes grew into a confluent monolayer by day 4-6. The adherence to ADM of HK in types II and IV was stronger than that in type I and III. The take rate of type II and IV composite skin was also significantly higher. In conclusion, when MMC-HF and HK were cocultured on the epidermal side of ADM, MMC-HF could serve as excellent feeder cells. Copyright 2007 S. Karger AG, Basel.

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

PubMed

Sakai, Yusuke; Koike, Makiko; Hasegawa, Hideko; Yamanouchi, Kosho; Soyama, Akihiko; Takatsuki, Mitsuhisa; Kuroki, Tamotsu; Ohashi, Kazuo; Okano, Teruo; Eguchi, Susumu

2013-01-01

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.

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

PubMed

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.

Properties of murine embryonic stem cells maintained on human foreskin fibroblasts without LIF.

PubMed

Meng, G L; Zur Nieden, N I; Liu, S Y; Cormier, J T; Kallos, M S; Rancourt, D E

2008-04-01

In embryonic stem (ES) cells, leukemia inhibitory factor (LIF)/STAT3, wnt and nodal/activin signaling are mainly active to control pluripotency during expansion. To maintain pluripotency, ES cells are typically cultured on feeder cells of varying origins. Murine ES cells are commonly cultured on murine embryonic fibroblasts (MEFs), which senesce early and must be frequently prepared. This process is laborious and leads to batch variation presenting a challenge for high-throughput ES cell expansion. Although some cell lines can be sustained by exogenous LIF, this method is costly. We present here a novel and inexpensive culture method for expanding murine ES cells on human foreskin fibroblast (HFF) feeders. After 20 passages on HFFs without LIF, ES cell lines showed normal expression levels of pluripotency markers, maintained a normal karyotype and retained the ability to contribute to the germline. As HFFs do not senesce for at least 62 passages, they present a vast supply of feeders. Copyright 2007 Wiley-Liss, Inc.

Rapid Fabricating Technique for Multi-Layered Human Hepatic Cell Sheets by Forceful Contraction of the Fibroblast Monolayer

PubMed Central

Sakai, Yusuke; Koike, Makiko; Hasegawa, Hideko; Yamanouchi, Kosho; Soyama, Akihiko; Takatsuki, Mitsuhisa; Kuroki, Tamotsu; Ohashi, Kazuo; Okano, Teruo; Eguchi, Susumu

2013-01-01

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. PMID:23923035

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

PubMed

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.

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

PubMed

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.

Feeder-free reprogramming of human fibroblasts with messenger RNA.

PubMed

Warren, Luigi; Wang, Jiwu

2013-11-13

This unit describes a feeder-free protocol for deriving induced pluripotent stem cells (iPSCs) from human fibroblasts by transfection of synthetic mRNA. The reprogramming of somatic cells requires transient expression of a set of transcription factors that collectively activate an endogenous gene regulatory network specifying the pluripotent phenotype. The necessary ectopic factor expression was first effected using retroviruses; however, as viral integration into the genome is problematic for cell therapy applications, the use of footprint-free vectors such as mRNA is increasingly preferred. Strong points of the mRNA approach include high efficiency, rapid kinetics, and obviation of a clean-up phase to purge the vector. Still, the method is relatively laborious and has, up to now, involved the use of feeder cells, which brings drawbacks including poor applicability to clinically oriented iPSC derivation. Using the methods described here, mRNA reprogramming can be performed without feeders at much-reduced labor and material costs relative to established protocols. Copyright © 2013 John Wiley & Sons, Inc.

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

PubMed Central

Zhao, Zhenqiang; Ma, Yanlin; Chen, Zhibin; Liu, Qian; Li, Qi; Kong, Deyan; Yuan, Kunxiong; Hu, Lan; Wang, Tan; Chen, Xiaowu; Peng, Yanan; Jiang, Weimin; Yu, Yanhong; Liu, Xinfeng

2016-01-01

Mouse embryonic fibroblasts (MEFs) and human foreskin fibroblasts (HFFs) are used for the culture of human embryonic stem cells (hESCs). MEFs and HFFs differed in their capacity to support the proliferation and pluripotency of hESCs and could affect cardiac differentiation potential of hESCs. The aim of this study was to evaluate the effect of MEFs and HFFs feeders on dopaminergic differentiation of hESCs lines. To minimize the impact of culture condition variation, two hESCs lines were cultured on mixed feeder cells (MFCs, MEFs: HFFs = 1:1) and HFFs feeder, respectively, and then were differentiated into dopaminergic (DA) neurons under the identical protocol. Dopaminergic differentiation was evaluated by immunocytochemistry, quantitative fluorescent real-time PCR, transmission and scanning electron microscopy, and patch clamp. Our results demonstrated that these hESCs-derived neurons were genuine and functional DA neurons. However, compared to hESCs line on MFCs feeder, hESCs line on HFFs feeder had a higher proportion of tyrosine hydroxylase (TH) positive cells and expressed higher levels of FOXA2, PITX3, NURR1, and TH genes. In addition, the values of threshold intensity and threshold membrane potential of DA neurons from hESCs line on HFFs feeder were lower than those of DA neurons from hESCs line on the MFCs feeder. In conclusion, HFFs feeder not only facilitated the differentiation of hESCs cells into dopaminergic neurons, but also induced hESCs-derived DA neurons to express higher electrophysiological excitability. Therefore, feeder cells could affect not only dopaminergic differentiation potential of different hESCs lines, but also electrophysiological properties of hESCs-derived DA neurons. PMID:28066186

Mechanism of induction of fibroblast to corneal endothelial cell.

PubMed

Jiang, Yan; Fu, Wei-Cai; Zhang, Lin

2014-08-01

To explore mechanism of nduction of fibroblast to corneal endothelial cell. Rabbit conjunctiva fibroblasts were used as feeder cells, rabbit oral mucosa epithelial cells were used as seed cells, and human denuded amniotic membrane was used as carrier to establish tissue engineering corneal endothelium. The transformation effect was observed. As concentration of mitomycin C increased, cell survival rate gradually decreased, cell proliferation was obviously inhibited when concentration≥25 μg/mL; 5 days after being treated by 5 μg/mL mitomycin C, cell body was enlarged and extended without cell fusion, however after being treated by 0.5 μg/mL mitomycin C, cell body was significantly proliferated and gradually fused; after 3 weeks of culture, stratified epithelium appeared on rabbit oral mucosa epithelial cells, differentiation layers were 4-5 and were well differentiated, the morphology was similar to corneal endothelial cells; Under electron microscope, surface layer of cells were polygonal, tightly connected to another with microvilli on the border, there was hemidesmosome between basal cells and human denuded amniotic membrane. Fibroblast cells have the potential of multi-directional differentiation, effective induction can promote emergence of intercellular desmosomes between seed cells and emergence of epithelial surface microvilli, and differentiate to the corneal endothelial cell. However, clinical application still needs more research and safety evaluation. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Rapid fibroblast removal from high density human embryonic stem cell cultures.

PubMed

Turner, William S; McCloskey, Kara E

2012-10-28

Mouse embryonic fibroblasts (MEFs) were used to establish human embryonic stem cells (hESCs) cultures after blastocyst isolation(1). This feeder system maintains hESCs from undergoing spontaneous differentiation during cell expansion. However, this co-culture method is labor intensive, requires highly trained personnel, and yields low hESC purity(4). Many laboratories have attempted to minimize the number of feeder cells in hESC cultures (i.e. incorporating matrix-coated dishes or other feeder cell types(5-8)). These modified culture systems have shown some promise, but have not supplanted the standard method for culturing hESCs with mitomycin C-treated mouse embyronic fibroblasts in order to retard unwanted spontaneous differentiation of the hESC cultures. Therefore, the feeder cells used in hESC expansion should be removed during differentiation experiments. Although several techniques are available for purifying the hESC colonies (FACS, MACS, or use of drug resistant vectors) from feeders, these techniques are labor intensive, costly and/or destructive to the hESC. The aim of this project was to invent a method of purification that enables the harvesting of a purer population of hESCs. We have observed that in a confluent hESC culture, the MEF population can be removed using a simple and rapid aspiration of the MEF sheet. This removal is dependent on several factors, including lateral cell-to-cell binding of MEFs that have a lower binding affinity to the styrene culture dish, and the ability of the stem cell colonies to push the fibroblasts outward during the generation of their own "niche". The hESC were then examined for SSEA-4, Oct3/4 and Tra 1-81 expression up to 10 days after MEF removal to ensure maintenance of pluripotency. Moreover, hESC colonies were able to continue growing from into larger formations after MEF removal, providing an additional level of hESC expansion.

Epithelial-mesenchymal transition in colonies of rhesus monkey embryonic stem cells: a model for processes involved in gastrulation.

PubMed

Behr, Rüdiger; Heneweer, Carola; Viebahn, Christoph; Denker, Hans-Werner; Thie, Michael

2005-01-01

Rhesus monkey embryonic stem (rhES) cells were grown on mouse embryonic fibroblast (MEF) feeder layers for up to 10 days to form multilayered colonies. Within this period, stem cell colonies differentiated transiently into complex structures with a disc-like morphology. These complex colonies were characterized by morphology, immunohistochemistry, and marker mRNA expression to identify processes of epithelialization as well as epithelial-mesenchymal transition (EMT) and pattern formation. Typically, differentiated colonies were comprised of an upper and a lower ES cell layer, the former growing on top of the layer of MEF cells whereas the lower ES cell layer spread out underneath the MEF cells. Interestingly, in the central part of the colonies, a roundish pit developed. Here the feeder layer disappeared, and upper layer cells seemed to ingress and migrate through the pit downward to form the lower layer while undergoing a transition from the epithelial to the mesenchymal phenotype, which was indicated by the loss of the marker proteins E-cadherin and ZO-1 in the lower layer. In support of this, we found a concomitant 10-fold upregulation of the gene Snail2, which is a key regulator of the EMT process. Conversion of epiblast to mesoderm was also indicated by the regulated expression of the mesoderm marker Brachyury. An EMT is a characteristic process of vertebrate gastrulation. Thus, these rhES cell colonies may be an interesting model for studies on some basic processes involved in early primate embryogenesis and may open new ways to study the regulation of EMT in vitro.

Reprogramming of somatic cells induced by fusion of embryonic stem cells using hemagglutinating virus of Japan envelope (HVJ-E)

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

Yue, Xiao-shan; Department of Biomolecular Engineering, Graduate School of Bioscience and Technology, Tokyo Institute of Technology, Nagatsuta-cho, Midori-ku, Yokohama-shi, Kanagawa 226-8501; Fujishiro, Masako

In this research, hemagglutinating virus of Japan envelope (HVJ-E) was used to reprogram somatic cells by fusion with mouse embryonic stem (ES) cells. Neomycin-resistant mouse embryonic fibroblasts (MEFs) were used as somatic cells. Nanog-overexpressing puromycin-resistant EB3 cells were used as mouse ES cells. These two cells were fused by exposing to HVJ-E and the generated fusion cells were selected by puromycin and G418 to get the stable fusion cell line. The fusion cells form colonies in feeder-free culture system. Microsatellite analysis of the fusion cells showed that they possessed genes from both ES cells and fibroblasts. The fusion cells weremore » tetraploid, had alkali phosphatase activity, and expressed stem cell marker genes such as Pou5f1, Nanog, and Sox2, but not the fibroblast cell marker genes such as Col1a1 and Col1a2. The pluripotency of fusion cells was confirmed by their expression of marker genes for all the three germ layers after differentiation induction, and by their ability to form teratoma which contained all the three primary layers. Our results show that HVJ-E can be used as a fusion reagent for reprogramming of somatic cells.« less

Quantitative and semi-quantitative immunoassay of growth factors and cytokines in the conditioned medium of STO and CF-1 mouse feeder cells

USDA-ARS?s Scientific Manuscript database

Feeder-cells of irradiated mouse fibroblasts are commonly used for, and are generally necessary for, the in vitro maintenance and growth of many fastidious cell types, particularly embryonic stem cells or induced pluripotent stem cells. Quantitative and semi-quantitative immunoassays were performed...

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

PubMed

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.

Extending Human Hematopoietic Stem Cell Survival In Vitro with Adipocytes

PubMed Central

Glettig, Dean Liang

2013-01-01

Abstract Human hematopoietic stem cells (hHSCs) cannot be maintained in vitro for extended time periods because they rapidly differentiate or die. To extend in vitro culture time, researchers have made attempts to use human mesenchymal stem cells (hMSCs) to create feeder layers that mimic the stem cell niche. We have conducted an array of experiments including adipocytes in these feeder layers that inhibit hHSC differentiation and by that prolong stem cell survival in vitro. The amount of CD34+ cells was quantified using flow cytometry. In a first experiment, feeder layers of undifferentiated hMSCs were compared with feeder layers differentiated toward osteoblasts or adipocytes using minimal medium, showing the highest survival rate where adipocytes were included. The same conclusion was drawn in a second experiment in comparing hMSCs with adipogenic feeder cells, using a culture medium supplemented with a cocktail of hHSC growth factors. In a third experiment, it was shown that direct cell–cell contact is necessary for the supportive effect of the feeder layers. In a fourth and fifth experiment the amount of adipocytes in the feeder layers were varied, and in all experiments a higher amount of adipocytes in the feeder layers showed a less rapid decay of CD34+ cells at later time points. We therefore concluded that adipocytes assist in suppressing hHSC differentiation and aid in prolonging their survival in vitro. PMID:23741628

Cellular responses to disruption of the permeability barrier in a three-dimensional organotypic epidermal model

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

Ajani, Gati; Sato, Nobuyuki; Mack, Judith A.

2007-08-15

Repeated injury to the stratum corneum of mammalian skin (caused by friction, soaps, or organic solvents) elicits hyperkeratosis and epidermal thickening. Functionally, these changes serve to restore the cutaneous barrier and protect the organism. To better understand the molecular and cellular basis of this response, we have engineered an in vitro model of acetone-induced injury using organotypic epidermal cultures. Rat epidermal keratinocytes (REKs), grown on a collagen raft in the absence of any feeder fibroblasts, developed all the hallmarks of a true epidermis including a well-formed cornified layer. To induce barrier injury, REK cultures were treated with intermittent 30-s exposuresmore » to acetone then were fixed and paraffin-sectioned. After two exposures, increased proliferation (Ki67 and BrdU staining) was observed in basal and suprabasal layers. After three exposures, proliferation became confined to localized buds in the basal layer and increased terminal differentiation was observed (compact hyperkeratosis of the stratum corneum, elevated levels of K10 and filaggrin, and heightened transglutaminase activity). Thus, barrier disruption causes epidermal hyperplasia and/or enhances differentiation, depending upon the extent and duration of injury. Given that no fibroblasts are present in the model, the ability to mount a hyperplastic response to barrier injury is an inherent property of keratinocytes.« less

Increased Risk of Genetic and Epigenetic Instability in Human Embryonic Stem Cells Associated with Specific Culture Conditions

PubMed Central

Garitaonandia, Ibon; Amir, Hadar; Boscolo, Francesca Sesillo; Wambua, Gerald K.; Schultheisz, Heather L.; Sabatini, Karen; Morey, Robert; Waltz, Shannon; Wang, Yu-Chieh; Tran, Ha; Leonardo, Trevor R.; Nazor, Kristopher; Slavin, Ileana; Lynch, Candace; Li, Yingchun; Coleman, Ronald; Gallego Romero, Irene; Altun, Gulsah; Reynolds, David; Dalton, Stephen; Parast, Mana; Loring, Jeanne F.; Laurent, Louise C.

2015-01-01

The self-renewal and differentiation capacities of human pluripotent stem cells (hPSCs) make them a promising source of material for cell transplantation therapy, drug development, and studies of cellular differentiation and development. However, the large numbers of cells necessary for many of these applications require extensive expansion of hPSC cultures, a process that has been associated with genetic and epigenetic alterations. We have performed a combinatorial study on both hESCs and hiPSCs to compare the effects of enzymatic vs. mechanical passaging, and feeder-free vs. mouse embryonic fibroblast feeder substrate, on the genetic and epigenetic stability and the phenotypic characteristics of hPSCs. In extensive experiments involving over 100 continuous passages, we observed that both enzymatic passaging and feeder-free culture were associated with genetic instability, higher rates of cell proliferation, and persistence of OCT4/POU5F1-positive cells in teratomas, with enzymatic passaging having the stronger effect. In all combinations of culture conditions except for mechanical passaging on feeder layers, we noted recurrent deletions in the genomic region containing the tumor suppressor gene TP53, which was associated with decreased mRNA expression of TP53, as well as alterations in the expression of several downstream genes consistent with a decrease in the activity of the TP53 pathway. Among the hESC cultures, we also observed culture-associated variations in global gene expression and DNA methylation. The effects of enzymatic passaging and feeder-free conditions were also observed in hiPSC cultures. Our results highlight the need for careful assessment of the effects of culture conditions on cells intended for clinical therapies. PMID:25714340

Investigation of mitomycin-C-treated fibroblasts in 3-D collagen gel and conditioned medium for keratinocyte proliferation.

PubMed

Huang, Yi-Chau; Wang, Tzu-Wei; Sun, Jui-Sheng; Lin, Feng-Huei

2006-03-01

Fibroblasts produce a spectrum of necessary growth factors essential for growth and proliferation of a variety of cell types. In this study, the paracrine effect of mitomycin-C-treated fibroblasts with various densities in collagen gel for keratinocyte proliferation was investigated from which an optimum cell density and optimum conditioned medium would be determined to expand keratinocyte without further differentiation for skin equivalent tissue engineering. The optimum cell density in collagen feeder gel for optimum collected medium preparation will be determined by checking the level of keratinocyte growth factor and granulocyte macrophage colony-stimulating factor in conventional medium. The results showed that the cell density of 1 x 10(5) cells/gel in the feeder gel is better to produce optimum collected medium. The conditioned medium is prepared by mixing together the optimum collected medium and molecular cellular and developmental biology (MCDB) 153 medium in different ratios for keratinocyte growth. The keratinocyte viability will be measured by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to determine the optimum conditioned medium. From the study, 67% conditioned medium was supposed as the better medium for keratinocyte proliferation. In this experiment, the optimum cell density in feeder gel to coculture with keratinocytes is also determined as 1 x 10(5) cells/gel. Keratin 10 (K10) and Terminal Deoxynucleotidyl Transferase Mediated dUTP Nick End Labeling stain will be used to check the cell differentiation and apoptosis, respectively. The results suggest that keratinocytes should not be cultured in postconfluent conditions due to undesired apoptosis and differentiation. The result of cell viability from passages to passages shows that the optimum feeder gel plays a more important role to the keratinocyte proliferation than that of optimum conditioned medium. Keratinocytes cultured with optimum feeder gel in 67% conditioned medium could effectively promote proliferation, inhibit apoptosis, and prevent differentiation. The combination of conditioned media and feeder gel to culture keratinocytes without external supplements can provide an inexpensive way for keratinocyte proliferation and construct an environment for real-time communication between the two cells. The results conclude that keratinocyte cultivation in feeder gel with modified medium should be feasible in the production of high quality keratinocytes for skin equivalents preparation.

Basic FGF Support of Human Embryonic Stem Cell Self-Renewal

PubMed Central

Levenstein, Mark E.; Ludwig, Tenneille E.; Xu, Ren-He; Llanas, Rachel A.; VanDenHeuvel-Kramer, Kaitlyn; Manning, Daisy; Thomson, James A.

2015-01-01

Human embryonic stem (ES) cells have most commonly been cultured in the presence of basic FGF (FGF2) either on fibroblast feeder layers or in fibroblast-conditioned medium. Recently, it has been reported that elevated concentrations of FGF2 permit the culture of human ES cells in the absence of fibroblasts or fibroblast-conditioned medium. Here we compare the ability of unconditioned medium (UM) supplemented with 4, 24, 40, 80, 100 and 250 ng/ml FGF2 to sustain low-density human ES cell cultures through multiple passages. In these stringent culture conditions, 4, 24, and 40 ng/ml FGF2 failed to sustain human ES cells through three passages, but 100 ng/ml sustained human ES cells with an effectiveness comparable to conditioned medium (CM). Two human ES cell lines (H1 and H9) were maintained for up to 164 population doublings (7 and 4 months) in UM supplemented with 100 ng/ml FGF2. After prolonged culture the cells formed teratomas when injected into SCID-beige mice, and expressed markers characteristic of undifferentiated human ES cells. We also demonstrate that FGF2 is degraded more rapidly in UM than in CM, partly explaining the need for higher concentrations of FGF2 in UM. These results further facilitate the large-scale, routine culture of human ES cells, and suggest that fibroblasts and fibroblast-conditioned medium sustain human ES cells in part by stabilizing FGF signaling above a critical threshold. PMID:16282444

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

PubMed Central

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

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

PubMed Central

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

In vitro culture of functionally active buffalo hepatocytes isolated by using a simplified manual perfusion method.

PubMed

Panda, Santanu; Bisht, Sonu; Malakar, Dhruba; Mohanty, Ashok K; Kaushik, Jai K

2015-01-01

In farm animals, there is no suitable cell line available to understand liver-specific functions. This has limited our understanding of liver function and metabolism in farm animals. Culturing and maintenance of functionally active hepatocytes is difficult, since they survive no more than few days. Establishing primary culture of hepatocytes can help in studying cellular metabolism, drug toxicity, hepatocyte specific gene function and regulation. Here we provide a simple in vitro method for isolation and short-term culture of functionally active buffalo hepatocytes. Buffalo hepatocytes were isolated from caudate lobes by using manual enzymatic perfusion and mechanical disruption of liver tissue. Hepatocyte yield was (5.3 ± 0.66)×107 cells per gram of liver tissue with a viability of 82.3 ± 3.5%. Freshly isolated hepatocytes were spherical with well contrasted border. After 24 hours of seeding onto fibroblast feeder layer and different extracellular matrices like dry collagen, matrigel and sandwich collagen coated plates, hepatocytes formed confluent monolayer with frequent clusters. Cultured hepatocytes exhibited typical cuboidal and polygonal shape with restored cellular polarity. Cells expressed hepatocyte-specific marker genes or proteins like albumin, hepatocyte nuclear factor 4α, glucose-6-phosphatase, tyrosine aminotransferase, cytochromes, cytokeratin and α1-antitrypsin. Hepatocytes could be immunostained with anti-cytokeratins, anti-albumin and anti α1-antitrypsin antibodies. Abundant lipid droplets were detected in the cytosol of hepatocytes using oil red stain. In vitro cultured hepatocytes could be grown for five days and maintained for up to nine days on buffalo skin fibroblast feeder layer. Cultured hepatocytes were viable for functional studies. We developed a convenient and cost effective technique for hepatocytes isolation for short-term culture that exhibited morphological and functional characteristics of active hepatocytes for studying gene expression, regulation, hepatic genomics and proteomics in farm animals.

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

PubMed

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.

Characterization of vibrissa germinative cells: transition of cell types.

PubMed

Osada, A; Kobayashi, K

2001-12-01

Germinative cells, small cell masses attached to the stalks of dermal papillae that are able to differentiate into the hair shaft and inner root sheath, form follicular bulb-like structures when co-cultured with dermal papilla cells. We studied the growth characteristics of germinative cells to determine the cell types in the vibrissa germinative tissue. Germinative tissues, attaching to dermal papillae, were cultured on 3T3 feeder layers. The cultured keratinocytes were harvested and transferred, equally and for two passages, onto lined dermal papilla cells (LDPC) and/or 3T3 feeder layers. The resulting germinative cells were classified into three types in the present experimental condition. Type 1 cells grow very well on either feeder layer, whereas Type 3 cells scarcely grow on either feeder layer. Type 2 cells are very conspicuous and are reversible. They grow well on 3T3 but growth is suppressed on LDPC feeder layers. The Type 2 cells that grow well on 3T3 feeder layers, however, are suppressed when transferred onto LDPC and the Type 2 cells that are suppressed on LDPC begin to grow again on 3T3. The transition of one cell type to another in vitro and the cell types that these germinative cell types correspond to in vivo is discussed. It was concluded that stem cells or their close progenitors reside in the germinative tissues of the vibrissa bulb except at late anagen-early catagen.

Comparison of defined culture systems for feeder cell free propagation of human embryonic stem cells

PubMed Central

Akopian, Veronika; Beil, Stephen; Benvenisty, Nissim; Brehm, Jennifer; Christie, Megan; Ford, Angela; Fox, Victoria; Gokhale, Paul J.; Healy, Lyn; Holm, Frida; Hovatta, Outi; Knowles, Barbara B.; Ludwig, Tenneille E.; McKay, Ronald D. G.; Miyazaki, Takamichi; Nakatsuji, Norio; Oh, Steve K. W.; Pera, Martin F.; Rossant, Janet; Stacey, Glyn N.; Suemori, Hirofumi

2010-01-01

There are many reports of defined culture systems for the propagation of human embryonic stem cells in the absence of feeder cell support, but no previous study has undertaken a multi-laboratory comparison of these diverse methodologies. In this study, five separate laboratories, each with experience in human embryonic stem cell culture, used a panel of ten embryonic stem cell lines (including WA09 as an index cell line common to all laboratories) to assess eight cell culture methods, with propagation in the presence of Knockout Serum Replacer, FGF-2, and mouse embryonic fibroblast feeder cell layers serving as a positive control. The cultures were assessed for up to ten passages for attachment, death, and differentiated morphology by phase contrast microscopy, for growth by serial cell counts, and for maintenance of stem cell surface marker expression by flow cytometry. Of the eight culture systems, only the control and those based on two commercial media, mTeSR1 and STEMPRO, supported maintenance of most cell lines for ten passages. Cultures grown in the remaining media failed before this point due to lack of attachment, cell death, or overt cell differentiation. Possible explanations for relative success of the commercial formulations in this study, and the lack of success with other formulations from academic groups compared to previously published results, include: the complex combination of growth factors present in the commercial preparations; improved development, manufacture, and quality control in the commercial products; differences in epigenetic adaptation to culture in vitro between different ES cell lines grown in different laboratories. PMID:20186512

Establishment of goat embryonic stem cells from in vivo produced blastocyst-stage embryos.

PubMed

Behboodi, E; Bondareva, A; Begin, I; Rao, K; Neveu, N; Pierson, J T; Wylie, C; Piero, F D; Huang, Y J; Zeng, W; Tanco, V; Baldassarre, H; Karatzas, C N; Dobrinski, I

2011-03-01

Embryonic stem (ES) cells with the capacity for germ line transmission have only been verified in mouse and rat. Methods for derivation, propagation, and differentiation of ES cells from domestic animals have not been fully established. Here, we describe derivation of ES cells from goat embryos. In vivo-derived embryos were cultured on goat fetal fibroblast feeders. Embryos either attached to the feeder layer or remained floating and expanded in culture. Embryos that attached showed a prominent inner cell mass (ICM) and those that remained floating formed structures resembling ICM disks surrounded by trophectodermal cells. ICM cells and embryonic disks were isolated mechanically, cultured on feeder cells in the presence of hLIF, and outgrown into ES-like colonies. Two cell lines were cultured for 25 passages and stained positive for alkaline phosphatase, POU5F1, NANOG, SOX2, SSEA-1, and SSEA-4. Embryoid bodies formed in suspension culture without hLIF. One cell line was cultured for 2 years (over 120 passages). This cell line differentiated in vitro into epithelia and neuronal cells, and could be stably transfected and selected for expression of a fluorescent marker. When cells were injected into SCID mice, teratomas were identified 5-6 weeks after transplantation. Expression of known ES cell markers, maintenance in vitro for 2 years in an undifferentiated state, differentiation in vitro, and formation of teratomas in immunodeficient mice provide evidence that the established cell line represents goat ES cells. This also is the first report of teratoma formation from large animal ES cells. Copyright © 2011 Wiley-Liss, Inc.

Alveolar type II cell-fibroblast interactions, synthesis and secretion of surfactant and type I collagen.

PubMed

Griffin, M; Bhandari, R; Hamilton, G; Chan, Y C; Powell, J T

1993-06-01

During alveolar development and alveolar repair close contacts are established between fibroblasts and lung epithelial cells through gaps in the basement membrane. Using co-culture systems we have investigated whether these close contacts influence synthesis and secretion of the principal surfactant apoprotein (SP-A) by cultured rat lung alveolar type II cells and the synthesis and secretion of type I collagen by fibroblasts. The alveolar type II cells remained cuboidal and grew in colonies on fibroblast feeder layers and on Matrigel-coated cell culture inserts but were progressively more flattened on fixed fibroblast monolayers and plastic. Alveolar type II cells cultured on plastic released almost all their SP-A into the medium by 4 days. Alveolar type II cells cultured on viable fibroblasts or Matrigel-coated inserts above fibroblasts accumulated SP-A in the medium at a constant rate for the first 4 days, and probably recycle SP-A by endocytosis. The amount of mRNA for SP-A was very low after 4 days of culture of alveolar type II cells on plastic, Matrigel-coated inserts or fixed fibroblast monolayers: relatively, the amount of mRNA for SP-A was increased 4-fold after culture of alveolar type II cells on viable fibroblasts. Co-culture of alveolar type II cells with confluent human dermal fibroblasts stimulated by 2- to 3-fold the secretion of collagen type I into the culture medium, even after the fibroblasts' growth had been arrested with mitomycin C. Collagen secretion, by fibroblasts, also was stimulated 2-fold by conditioned medium from alveolar type II cells cultured on Matrigel. The amount of mRNA for type I collagen increased only modestly when fibroblasts were cultured in this conditioned medium. This stimulation of type I collagen secretion diminished as the conditioned medium was diluted out, but at high dilutions further stimulation occurred, indicating that a factor that inhibited collagen secretion also was being diluted out. The conditioned medium contained low levels of IGF-1 and the stimulation of type I collagen secretion was abolished when the conditioned medium was pre-incubated with antibodies to insulin-like growth factor 1 (IGF-1). There are important reciprocal interactions between alveolar type II cells and fibroblasts in co-culture. Direct contacts between alveolar type II cells and fibroblasts appear to have a trophic effect on cultured alveolar type II cells, increasing the levels of mRNA for SP-A. Rat lung alveolar type II cells appear to release a factor (possibly IGF-1) that stimulates type I collagen secretion by fibroblasts.

Successful Consecutive Expansion of Limbal Explants Using a Biosafe Culture Medium under Feeder Layer-Free Conditions.

PubMed

López-Paniagua, Marina; Nieto-Miguel, Teresa; de la Mata, Ana; Galindo, Sara; Herreras, José M; Corrales, Rosa M; Calonge, Margarita

2017-05-01

Transplantation of in vitro cultured limbal epithelial stem cells (LESCs) is a treatment widely used for LESC deficiency. However, the number of limbal tissue donors is limited, and protocols for LESC cultivation often include compounds and/or feeder layers that can induce side effects and/or increase the cost of the culture procedure. We investigated the feasibility of obtaining more than one limbal primary culture (LPC) from the same biopsy using a culture medium in which several potentially harmful compounds were replaced at the same time by biosafe supplements, allowing the LESC cultivation without feeder layers. We established feeder layer-free LPCs with three culture media: (1) a modified supplemental hormonal epithelial medium, containing potential harmful components (cholera toxin, dimethylsulfoxide, and fetal bovine serum [FBS]), (2) IOBA-FBS, a medium with FBS but with no other harmful supplements, and (3) IOBA-HS, similar to IOBA-FBS but with human serum instead of FBS. Additionally, the same limbal explant was consecutively cultured with IOBA-HS producing three cultures. LPCs were characterized by real-time reverse transcription polymerase chain reaction and/or immunofluorescence. LPCs cultured with the three media under feeder layer-free conditions showed cuboidal cells and no significant differences in the percentage of positive cells for limbal (ABCG2, p63, and K14) and corneal (K3, K12) proteins. Except for ABCG2, the relative mRNA expression of the LESC markers was significantly higher when IOBA-FBS or IOBA-HS was used. LPC1 showed characteristics similar to LPC0, while LPC2 cell morphology became elongated and the expression of some LESC markers was diminished. IOBA-HS enables the culturing of up to two biosafe homologous LPCs from one limbal tissue under feeder layer-free conditions. The routine use of this culture medium could improve both the biosafety and the number of available LPCs for potential clinical transplantation, as well as decrease the expense of the culture procedure.

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

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

Kamada, Mizuna; Mitsui, Youji, E-mail: y-mitsui8310@hb.tp1.jp; Kumazaki, Tsutomu

2014-10-24

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

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

PubMed

Lamb, Rebecca; Ambler, Carrie A

2013-01-01

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.

Keratinocytes Propagated in Serum-Free, Feeder-Free Culture Conditions Fail to Form Stratified Epidermis in a Reconstituted Skin Model

PubMed Central

Lamb, Rebecca; Ambler, Carrie A.

2013-01-01

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. PMID:23326335

Effect of mitomycin-C on human foreskin fibroblasts used as feeders in human embryonic stem cells: immunocytochemistry MIB1 score and DNA ploidy and apoptosis evaluated by flow cytometry.

PubMed

Nieto, A; Cabrera, C M; Catalina, P; Cobo, F; Barnie, A; Cortés, J L; Barroso del Jesus, A; Montes, R; Concha, A

2007-03-01

Mitomycin C (MMC) treatment has been used to arrest cell proliferation but not much is known about the effect of MMC on human foreskin fibroblasts (HFF) used as feeders for human embryonic stem cells (hESC). We tested the ability of MMC to stop the proliferation of HFF and to induce apoptosis. MMC inhibited the proliferation of HFF at 10 microg/ml over 2.5h of MMC treatment showing a decrease in the proliferation index measured by Ki-67 and S and G2/M phases related to active HFF. A low percentage of cells showed necrotic or apoptotic features using different lengths of incubation. Over time, the majority of cells remained in a mitotically inactive state. The percentage of apoptotic cells increased from day 2 to day 10, at the same time as the necrotic ones increased. The HS181 hESC line grew in an undifferentiated state on inactive HFF throughout the study.

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

PubMed

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 differentiation of hESCs on subsequent replicative life span and cellular senescence of the differentiated keratinocytes, with implications for use of these cells for applications in tissue engineering and regenerative medicine.

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

PubMed Central

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

2010-01-01

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

Multiphoton autofluorescence lifetime imaging of induced pluripotent stem cells

NASA Astrophysics Data System (ADS)

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.

Wharton's Jelly Mesenchymal Stromal Cells as a Feeder Layer for the Ex Vivo Expansion of Hematopoietic Stem and Progenitor Cells: a Review.

PubMed

Lo Iacono, Melania; Anzalone, Rita; La Rocca, Giampiero; Baiamonte, Elena; Maggio, Aurelio; Acuto, Santina

2017-02-01

In recent years, umbilical cord blood (UCB) has been widely used as an alternative source to bone marrow (BM) for transplantation of hematopoietic stem and progenitor cells (HSPCs) in a variety of hematological and non-hematological disorders. Nevertheless, the insufficient number of UCB-HSPCs for graft represents a major challenge. HSPCs ex vivo expansion prior to transplantation is a valid strategy to overcome this limit. Several attempts to optimize the expansion conditions have been reported, including the use of mesenchymal stromal cells (MSCs) as feeder layer. Wharton's Jelly (WJ), the main component of umbilical cord (UC) matrix, is especially rich in MSCs, which are considered ideal candidates for feeder layer in co-culture systems. In fact, they can be easily harvested and grow robustly in culture, producing a confluent monolayer in a short time. Similarly to bone marrow-mesenchymal stromal cells (BM-MSCs), WJ-derived MSCs (WJ-MSCs) have been used to support hematopoiesis in vitro and in vivo. Here, we review the rationale for using MSCs, particularly WJ-MSCs, as a feeder layer for UCB-HSPCs ex vivo expansion. In addition, we report the main findings attesting the use of these MSCs as a support in hematopoiesis.

Exposure cell number during feeder cell growth-arrest by Mitomycin C is a critical pharmacological aspect in stem cell culture system.

PubMed

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

2016-01-01

Growth-arrested feeder cells following Mitomycin C treatment are instrumental in stem cell culture allowing development of regenerative strategies and alternatives to animal testing in drug discovery. The concentration of Mitomycin C and feeder cell type was described to affect feeder performance but the criticality of feeder cell exposure density was not addressed. We hypothesize that the exposure cell density influences the effectiveness of Mitomycin C in an arithmetic manner. Three different exposure cell densities of Swiss 3T3 fibroblasts were treated with a range of Mitomycin C concentrations for 2h. The cells were replaced and the viable cells counted on 3, 6, 9, 12 and 20days. The cell extinctions were compared with doses per cell which were derived by dividing the product of concentration and volume of Mitomycin C solution with exposure cell number. The periodic post-treatment feeder cell extinctions were not just dependent on Mitomycin C concentration but also on dose per cell. Analysis of linearity between viable cell number and Mitomycin C dose per cell derived from the concentrations of 3 to 10μg/ml revealed four distinct categories of growth-arrest. Confluent cultures exposed to low concentration showed growth-arrest failure. The in vitro cell density titration can facilitate prediction of a compound's operational in vivo dosing. For containing the growth arrest failure, an arithmetic volume derivation strategy is proposed by fixing the exposure density to a safe limit. The feeder extinction characteristics are critical for streamlining the stem cell based pharmacological and toxicological assays. Copyright © 2016 Elsevier Inc. All rights reserved.

Human dermal papilla cells and outer root sheath cells: no follicular differentiation in nude mice and chicken embryos.

PubMed

Chiu, H C; Chang, C H; Jee, S H; Chang, C C; Wu, Y C

1994-09-01

Human scalp specimens were incubated in 5 U/ml dispase solution at 4 degrees C overnight before the isolation of dermal papillae and follicle epithelium. This pretreatment not only facilitated the attachment and cell outgrowth of dermal papillae but also made it easier to pluck out hairs with intact follicle epithelium. The outer root sheath cells were released from the follicle epithelium and grown on a feeder layer of mitomycin C-treated human dermal fibroblasts. The subcultured outer root sheath cells were grown in a serum-free medium. When the mixtures of early-passage dermal papilla cells and outer root sheath cells were injected into the subcutis of nude mice, an epidermal cyst surrounded by layers of fibrous tissue was found in three weeks. No hair follicles were found when the mixtures were implanted onto the chorioallantoic membrane of nine-day-old chicken embryos. A keratinized mass lying on the chorionic epithelium with or without smaller similar masses in the chorioallantoic membrane was found in eight days. No hair follicle-like structure could be found. Possible factors contributing to the failure to undergo follicular differentiation in this study are discussed.

Long-term xeno-free culture of human pluripotent stem cells on hydrogels with optimal elasticity.

PubMed

Higuchi, Akon; Kao, Shih-Hsuan; Ling, Qing-Dong; Chen, Yen-Ming; Li, Hsing-Fen; Alarfaj, Abdullah A; Munusamy, Murugan A; Murugan, Kadarkarai; Chang, Shih-Chang; Lee, Hsin-Chung; Hsu, Shih-Tien; Kumar, S Suresh; Umezawa, Akihiro

2015-12-14

The tentative clinical application of human pluripotent stem cells (hPSCs), such as human embryonic stem cells and human induced pluripotent stem cells, is restricted by the possibility of xenogenic contamination resulting from the use of mouse embryonic fibroblasts (MEFs) as a feeder layer. Therefore, we investigated hPSC cultures on biomaterials with different elasticities that were grafted with different nanosegments. We prepared dishes coated with polyvinylalcohol-co-itaconic acid hydrogels grafted with an oligopeptide derived from vitronectin (KGGPQVTRGDVFTMP) with elasticities ranging from 10.3 to 30.4 kPa storage moduli by controlling the crosslinking time. The hPSCs cultured on the stiffest substrates (30.4 kPa) tended to differentiate after five days of culture, whereas the hPSCs cultured on the optimal elastic substrates (25 kPa) maintained their pluripotency for over 20 passages under xeno-free conditions. These results indicate that cell culture matrices with optimal elasticity can maintain the pluripotency of hPSCs in culture.

Culture of porcine hepatocytes or bile duct epithelial cells by inductive serum-free media

USDA-ARS?s Scientific Manuscript database

A serum-free, feeder-cell-dependent, selective culture system for the long-term culture of porcine hepatocytes or cholangiocytes was developed. Liver cells were isolated from 1 wk old pigs or young adult pigs (25 and 63 kg live weight) and were placed in primary culture on feeder-cell layers of mit...

Reconstituted human gingival epithelium: nonsubmerged in vitro model.

PubMed

Delcourt-Huard, A; Corlu, A; Joffre, A; Magloire, H; Bonnaure-Mallet, M

1997-01-01

Many studies have shown that human gingival keratinocytes grown in submerged culture fail to attain optimal differentiation. This study reports an in vitro culture system for oral gingival epithelial cells, in which they are grown at the air-liquid interface, on polycarbonate inserts, in the presence of an NIH-3T3 feeder layer. This model was compared with two submerged culture methods for gingival keratinocytes, on type 1 collagen gel and on an NIH-3T3 feeder layer. Transmission electron microscopy showed an advanced level of stratification (over six layers of cells) for cultures grown at the air-liquid interface. Immunofluorescence and electrophoretic patterns showed the presence of cytokeratins 10 and 11 in cytoskeletal protein extracts of these cultured keratinocytes. In this air-liquid interface culture model, in the presence of NIH-3T3 feeder cells, keratinocytes can achieve an advanced level of stratification and differentiation and a resemblance to in vivo gingiva. The obtention of a highly differentiated epithelium will permit in vitro pharmacological studies and studies on the biocompatability of certain alloys with the superficial periodontium; it will also provide grafts for patients undergoing periodontal surgery.

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

NASA Astrophysics Data System (ADS)

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 pipe-like feeder and 2) a linear feeder representing a dike-like feeder. In the experiments with point inlet, circular saucer-shaped sills formed. In the experiments with linear feeder, elliptical saucer-shaped sills formed. In the latter experiments, the long axes of the saucers were parallel to, and located directly above, the linear feeder. The experiments show that the feeder geometry has an important influence on the geometry of the emplaced sills. There are close similarities between the geometry of the Golden Valley sill and the intrusions formed in the experiments. The elliptical shape of the Golden Valley sill suggests that it was fed by an elongated feeder, probably the D4 dike. In general, our results show that the three-dimensional geometry of saucer-shaped sills observed in sedimentary basins, may constrain the shape of their feeders, i.e. their emplacement mechanisms.

Plasma membrane characterization, by scanning electron microscopy, of multipotent myoblasts-derived populations sorted using dielectrophoresis.

PubMed

Muratore, Massimo; Mitchell, Steve; Waterfall, Martin

2013-09-06

Multipotent progenitor cells have shown promise for use in biomedical applications and regenerative medicine. The implementation of such cells for clinical application requires a synchronized, phenotypically and/or genotypically, homogenous cell population. Here we have demonstrated the implementation of a biological tag-free dielectrophoretic device used for discrimination of multipotent myoblastic C2C12 model. The multipotent capabilities in differentiation, for these cells, diminishes with higher passage number, so for cultures above 70 passages only a small percentage of cells is able to differentiate into terminal myotubes. In this work we demonstrated that we could recover, above 96% purity, specific cell types from a mixed population of cells at high passage number without any biological tag using dielectrophoresis. The purity of the samples was confirmed by cytometric analysis using the cell specific marker embryonic myosin. To further investigate the dielectric properties of the cell plasma membrane we co-culture C2C12 with similar size, when in suspension, GFP-positive fibroblast as feeder layer. The level of separation between the cell types was above 98% purity which was confirmed by flow cytometry. These levels of separation are assumed to account for cell size and for the plasma membrane morphological differences between C2C12 and fibroblast unrelated to the stages of the cell cycle which was assessed by immunofluorescence staining. Plasma membrane conformational differences were further confirmed by scanning electron microscopy. Copyright © 2013 Elsevier Inc. All rights reserved.

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

PubMed

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

2017-06-01

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.

In vitro culture of primary plasmacytomas requires stromal cell feeder layers.

PubMed Central

Degrassi, A; Hilbert, D M; Rudikoff, S; Anderson, A O; Potter, M; Coon, H G

1993-01-01

Attempts to grow primary murine plasmacytomas in vitro have, to date, been largely unsuccessful. In this study, we demonstrate that long-term in vitro growth of primary plasmacytomas is accomplished by using feeder layers composed of stromal cells from the initial site of plasmacytomagenesis. The early neoplastic lines established in this manner are dependent on physical contact with the stromal layer, which is mediated in part by CD44, for growth and survival. The stromal cells provide at least two stimuli for the plasma cells, one being interleukin 6 and the second, of unknown nature, resulting from direct physical interaction that cannot be replaced by soluble factors. These plasma cell lines have been passaged for as long as 20 months yet still maintain characteristics associated with primary plasmacytomas as they will grow in vivo only in pristane-primed animals, indicating a continued dependence on the pristane-induced microenvironment characteristic of early-stage tumors. The ability to grow primary plasmacytomas in culture and maintain their "primary" properties provides a model system for detailed analysis of early events in plasma cell tumor progression involving neoplastic cells completely dependent on physical contact with a stromal feeder layer for survival and expansion. Images Fig. 1 Fig. 2 PMID:8446628

Evidence of two distinct functionally specialized fibroblast lineages in breast stroma.

PubMed

Morsing, Mikkel; Klitgaard, Marie Christine; Jafari, Abbas; Villadsen, René; Kassem, Moustapha; Petersen, Ole William; Rønnov-Jessen, Lone

2016-11-03

The terminal duct lobular unit (TDLU) is the most dynamic structure in the human breast and the putative site of origin of human breast cancer. Although stromal cells contribute to a specialized microenvironment in many organs, this component remains largely understudied in the human breast. We here demonstrate the impact on epithelium of two lineages of breast stromal fibroblasts, one of which accumulates in the TDLU while the other resides outside the TDLU in the interlobular stroma. The two lineages are prospectively isolated by fluorescence activated cell sorting (FACS) based on different expression levels of CD105 and CD26. The characteristics of the two fibroblast lineages are assessed by immunocytochemical staining and gene expression analysis. The differentiation capacity of the two fibroblast populations is determined by exposure to specific differentiating conditions followed by analysis of adipogenic and osteogenic differentiation. To test whether the two fibroblast lineages are functionally imprinted by their site of origin, single cell sorted CD271 low /MUC1 high normal breast luminal epithelial cells are plated on fibroblast feeders for the observation of morphological development. Epithelial structure formation and polarization is shown by immunofluorescence and digitalized quantification of immunoperoxidase-stained cultures. Lobular fibroblasts are CD105 high /CD26 low while interlobular fibroblasts are CD105 low /CD26 high . Once isolated the two lineages remain phenotypically stable and functionally distinct in culture. Lobular fibroblasts have properties in common with bone marrow derived mesenchymal stem cells and they specifically convey growth and branching morphogenesis of epithelial progenitors. Two distinct functionally specialized fibroblast lineages exist in the normal human breast, of which the lobular fibroblasts have properties in common with mesenchymal stem cells and support epithelial growth and morphogenesis. We propose that lobular fibroblasts constitute a specialized microenvironment for human breast luminal epithelial progenitors, i.e. the putative precursors of breast cancer.

Hydrodynamic control of phytoplankton loss to the benthos in an estuarine environment

USGS Publications Warehouse

Jones, Nicole L.; Thompson, Janet K.; Arrigo, Kevin R.; Monismith, Stephen G.

2009-01-01

Field experiments were undertaken to measure the influence of hydrodynamics on the removal of phytoplankton by benthic grazers in Suisun Slough, North San Francisco Bay. Chlorophyll a concentration boundary layers were found over beds inhabited by the active suspension feeders Corbula amurensis and Corophium alienense and the passive suspension feeders Marenzellaria viridis and Laonome sp. Benthic losses of phytoplankton were estimated via both the control volume and the vertical flux approach, in which chlorophyll a concentration was used as a proxy for phytoplankton biomass. The rate of phytoplankton loss to the bed was positively correlated to the bed shear stress. The maximum rate of phytoplankton loss to the bed was five times larger than estimated by laboratory-derived pumping rates for the active suspension feeders. Reasons for this discrepancy are explored including a physical mechanism whereby phytoplankton is entrained in a near-bed fluff layer where aggregation is mediated by the presence of mucus produced by the infaunal community.

Hydrodynamic control of phytoplankton loss to the benthos in an estuarine environment

USGS Publications Warehouse

Jones, N.L.; Thompson, J.K.; Arrigo, K.R.; Monismith, Stephen G.

2009-01-01

Field experiments were undertaken to measure the influence of hydrodynamics on the removal of phytoplankton by benthic grazers in Suisun Slough, North San Francisco Bay. Chlorophyll a concentration boundary layers were found over beds inhabited by the active suspension feeders Corbula amurensis and Corophium alienense and the passive suspension feeders Marenzellaria viridis and Laonome sp. Benthic losses of phytoplankton were estimated via both the control volume and the vertical flux approach, in which chlorophyll a concentration was used as a proxy for phytoplankton biomass. The rate of phytoplankton loss to the bed was positively correlated to the bed shear stress. The maximum rate of phytoplankton loss to the bed was five times larger than estimated by laboratory-derived pumping rates for the active suspension feeders. Reasons for this discrepancy are explored including a physical mechanism whereby phytoplankton is entrained in a near-bed fluff layer where aggregation is mediated by the presence of mucus produced by the infaunal community. ?? 2009, by the American Society of Limnology and Oceanography, Inc.

Evaluation of the Fretting Resistance of the High Voltage Insulation on the ITER Magnet Feeder Busbars

NASA Astrophysics Data System (ADS)

Clayton, N.; Crouchen, M.; Evans, D.; Gung, C.-Y.; Su, M.; Devred, A.; Piccin, R.

2017-12-01

The high voltage (HV) insulation on the ITER magnet feeder superconducting busbars and current leads will be prepared from S-glass fabric, pre-impregnated with an epoxy resin, which is interleaved with polyimide film and wrapped onto the components and cured during feeder manufacture. The insulation architecture consists of nine half-lapped layers of glass/Kapton, which is then enveloped in a ground-screen, and two further half-lapped layers of glass pre-preg for mechanical protection. The integrity of the HV insulation is critical in order to inhibit electrical arcs within the feeders. The insulation over the entire length of the HV components (bus bar, current leads and joints) must provide a level of voltage isolation of 30 kV. In operation, the insulation on ITER busbars will be subjected to high mechanical loads, arising from Lorentz forces, and in addition will be subjected to fretting erosion against stainless steel clamps, as the pulsed nature of some magnets results in longitudinal movement of the busbar. This work was aimed at assessing the wear on, and the changes in, the electrical properties of the insulation when subjected to typical ITER operating conditions. High voltage tests demonstrated that the electrical isolation of the insulation was intact after the fretting test.

Transmissible Gastroenteritis in Feeder Pigs: Observations on the Jejunal Epithelium of Normal Feeder Pigs and Feeder Pigs Infected with TGE Virus

PubMed Central

Morin, M.; Morehouse, L. G.

1974-01-01

Light and electron microscopy findings in the jejunal mucosa of the normal feeder pig and feeder pigs infected with transmissible gastroenteritis (TGE) virus are reported. Villi in the mid jejunum of the normal feeder pig were elongated, finger shaped and covered with a layer of columnar absorptive cells with a well developed and regular brush border. Severe lesions of villous atrophy were present in all jejunal segments of feeder swine killed 96 hours post infection with TGE virus. Atrophic villi were covered by flat to cuboidal cells with a poorly developed brush border in some areas. In other segments, cells varied in appearance from sub-columnar to columnar type of near normal appearance. The ultrastructure of the jejunal absorptive cells in the normal feeder pig was found to be similar to that described for the jejunal cells of other adult mammals. There were no significant indications of high pinocytotic activity. The epithelial cells covering the atrophic villi of TGE infected pigs had a fine structure similar to that described for the crypt cells, ranging in appearance from very immature to moderately differentiated cells. Microvilli were very short, decreased markedly in number and irregular in arrangement. The terminal web was poorly developed. Strands of rough endoplasmic reticulum were markedly diminished and an increase in free ribosomes was noted. The significance of these observations in explaining pathogenesis of TGE in feeder pigs is discussed. ImagesFig. 1.Fig. 2.Fig. 3.Fig. 4.Fig. 5.Fig. 6.Fig. 7.Fig. 8. PMID:4277743

Papillary fibroblasts differentiate into reticular fibroblasts after prolonged in vitro culture.

PubMed

Janson, David; Saintigny, Gaëlle; Mahé, Christian; El Ghalbzouri, Abdoelwaheb

2013-01-01

The dermis can be divided into two morphologically different layers: the papillary and reticular dermis. Fibroblasts isolated from these layers behave differently when cultured in vitro. During skin ageing, the papillary dermis decreases in volume. Based on the functional differences in vitro, it is hypothesized that the loss of papillary fibroblasts contributes to skin ageing. In this study, we aimed to mimic certain aspects of skin ageing by using high-passage cultures of reticular and papillary fibroblasts and investigated the effect of these cells on skin morphogenesis in reconstructed human skin equivalents. Skin equivalents generated with reticular fibroblasts showed a reduced terminal differentiation and fewer proliferating basal keratinocytes. Aged in vitro papillary fibroblasts had increased expression of biomarkers specific to reticular fibroblasts. The phenotype and morphology of skin equivalents generated with high-passage papillary fibroblasts resembled that of reticular fibroblasts. This demonstrates that papillary fibroblasts can differentiate into reticular fibroblasts in vitro. Therefore, we hypothesize that papillary fibroblasts represent an undifferentiated phenotype, while reticular fibroblasts represent a more differentiated population. The differentiation process could be a new target for anti-skin-ageing strategies. © 2013 John Wiley & Sons A/S.

Defined Medium Conditions for the Induction and Expansion of Human Pluripotent Stem Cell-Derived Retinal Pigment Epithelium.

PubMed

Lidgerwood, Grace E; Lim, Shiang Y; Crombie, Duncan E; Ali, Ray; Gill, Katherine P; Hernández, Damián; Kie, Josh; Conquest, Alison; Waugh, Hayley S; Wong, Raymond C B; Liang, Helena H; Hewitt, Alex W; Davidson, Kathryn C; Pébay, Alice

2016-04-01

We demonstrate that a combination of Noggin, Dickkopf-1, Insulin Growth Factor 1 and basic Fibroblast Growth Factor, promotes the differentiation of human pluripotent stem cells into retinal pigment epithelium (RPE) cells. We describe an efficient one-step approach that allows the generation of RPE cells from both human embryonic stem cells and human induced pluripotent stem cells within 40-60 days without the need for manual excision, floating aggregates or imbedded cysts. Compared to methods that rely on spontaneous differentiation, our protocol results in faster differentiation into RPE cells. This pro-retinal culture medium promotes the growth of functional RPE cells that exhibit key characteristics of the RPE including pigmentation, polygonal morphology, expression of mature RPE markers, electrophysiological membrane potential and the ability to phagocytose photoreceptor outer segments. This protocol can be adapted for feeder, feeder-free and serum-free conditions. This method thereby provides a rapid and simplified production of RPE cells for downstream applications such as disease modelling and drug screening.

A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells

PubMed Central

Nakagawa, Masato; Taniguchi, Yukimasa; Senda, Sho; Takizawa, Nanako; Ichisaka, Tomoko; Asano, Kanako; Morizane, Asuka; Doi, Daisuke; Takahashi, Jun; Nishizawa, Masatoshi; Yoshida, Yoshinori; Toyoda, Taro; Osafune, Kenji; Sekiguchi, Kiyotoshi; Yamanaka, Shinya

2014-01-01

In order to apply human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) to regenerative medicine, the cells should be produced under restricted conditions conforming to GMP guidelines. Since the conventional culture system has some issues that need to be addressed to achieve this goal, we developed a novel culture system. We found that recombinant laminin-511 E8 fragments are useful matrices for maintaining hESCs and hiPSCs when used in combination with a completely xeno-free (Xf) medium, StemFit™. Using this system, hESCs and hiPSCs can be easily and stably passaged by dissociating the cells into single cells for long periods, without any karyotype abnormalities. Human iPSCs could be generated under feeder-free (Ff) and Xf culture systems from human primary fibroblasts and blood cells, and they possessed differentiation abilities. These results indicate that hiPSCs can be generated and maintained under this novel Ff and Xf culture system. PMID:24399248

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

PubMed

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.

Mapping the 3-D extent of the Northern Lobe of the Bushveld layered mafic intrusion from geophysical data

USGS Publications Warehouse

Finn, Carol A.; Bedrosian, Paul A.; Cole, Janine; Khoza, Tshepo David; Webb, Susan J.

2015-01-01

Geophysical models image the 3D geometry of the mafic portion of the Bushveld Complex north of the Thabazimbi-Murchison Lineament (TML), critical for understanding the origin of the world's largest layered mafic intrusion and platinum group element deposits. The combination of the gravity and magnetic data with recent seismic, MT, borehole and rock property measurements powerfully constrains the models. The intrusion north of the TML is generally shallowly buried (generally <1500 m) with a modeled area of ∼160 km × ∼125 km. The modeled thicknesses are not well constrained but vary from ∼<1000 to >12,000 m, averaging ∼4000 m. A feeder, suggested by a large modeled thickness (>10,000 m) and funnel shape, for Lower Zone magmas could have originated near the intersection of NS and NE trending TML faults under Mokopane. The TML has been thought to be the feeder zone for the entire Bushveld Complex but the identification of local feeders and/or dikes in the TML in the models is complicated by uncertainties on the syn- and post-Bushveld deformation history. However, modeled moderately thick high density material near the intersection of faults within the central and western TML may represent feeders for parts of the Bushveld Complex if deformation was minimal. The correspondence of flat, high resistivity and density regions reflect the sill-like geometry of the Bushveld Complex without evidence for feeders north of Mokopane. Magnetotelluric models indicate that the Transvaal sedimentary basin underlies much of the Bushveld Complex north of the TML, further than previously thought and important because the degree of reaction and assimilation of the Transvaal rocks with the mafic magmas resulted in a variety of mineralization zones.

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

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

Yin, Xiaohui; Peking University Stem Cell Research Center and Department of Cell Biology, School of Basic Medical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191; Li, Yang

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 maintainedmore » 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.« less

An Alternative Method for Long-Term Culture of Chicken Embryonic Stem Cell In Vitro.

PubMed

Zhang, Li; Wu, Yenan; Li, Xiang; Wei, Shao; Xing, Yiming; Lian, Zhengxing; Han, Hongbing

2018-01-01

Chicken embryonic stem cells (cESCs) obtained from stage X embryos provide a novel model for the study of avian embryonic development. A new way to maintain cESCs for a long period in vitro still remains unexplored. We found that the cESCs showed stem cell-like properties in vitro for a long term with the support of DF-1 feeder and basic culture medium supplemented with human basic fibroblast growth factor (hbFGF), mouse stem cell factor (mSCF), and human leukemia inhibitory factor (hLIF). During the long culture period, the cESCs showed typical ES cell morphology and expressed primitive stem cell markers with a relatively stable proliferation rate and high telomerase activity. These cells also exhibited the capability to differentiate into cardiac myocytes, smooth muscle cells, neural cells, osteoblast, and adipocyte in vitro . Chimera chickens were produced by cESCs cultured for 25 passages with this new culture system. The experiments showed that DF-1 was the optimal feeder and hbFGF was an important factor for maintaining the pluripotency of cESCs in vitro .

Organogenesis from transformed tomato explants.

PubMed

Frary, Anne; Van Eck, Joyce

2005-01-01

Tomato was one of the first crops for which a genetic transformation system was reported involving regeneration by organogenesis from Agrobacterium-transformed explants. Since the initial reports, various factors have been studied that affect the efficiency of tomato transformation and the technique has been useful for the isolation and identification of many genes involved in plant disease resistance, morphology and development. In this method, cotyledon explants from in vitro-grown seedlings are precultured overnight on a tobacco suspension feeder layer. The explants are then inoculated with Agrobacterium and returned to the feeder layer for a 2-d period of cocultivation. After cocultivation, the explants are transferred to an MS-based selective regeneration medium containing zeatin. Regenerated shoots are then rooted on a separate selective medium. This protocol has been used with several tomato cultivars and routinely yields transformation efficiencies of 10-15%.

Stromalized microreactor supports murine hematopoietic progenitor enrichment.

PubMed

Khong, Danika; Li, Matthew; Singleton, Amy; Chin, Ling-Yee; Parekkadan, Biju

2018-01-20

There is an emerging need to process, expand, and even genetically engineer hematopoietic stem and progenitor cells (HSPCs) prior to administration for blood reconstitution therapy. A closed-system and automated solution for ex vivo HSC processing can improve adoption and standardize processing techniques. Here, we report a recirculating flow bioreactor where HSCs are stabilized and enriched for short-term processing by indirect fibroblast feeder coculture. Mouse 3 T3 fibroblasts were seeded on the extraluminal membrane surface of a hollow fiber micro-bioreactor and were found to support HSPC cell number compared to unsupported BMCs. CFSE analysis indicates that 3 T3-support was essential for the enhanced intrinsic cell cycling of HSPCs. This enhanced support was specific to the HSPC population with little to no effect seen with the Lineage positive and Lineage negative cells. Together, these data suggest that stromal-seeded hollow fiber micro-reactors represent a platform to screening various conditions that support the expansion and bioprocessing of HSPCs ex vivo.

Gastrointestinal Fibroblasts Have Specialized, Diverse Transcriptional Phenotypes: A Comprehensive Gene Expression Analysis of Human Fibroblasts

PubMed Central

Ishii, Genichiro; Aoyagi, Kazuhiko; Sasaki, Hiroki; Ochiai, Atsushi

2015-01-01

Background Fibroblasts are the principal stromal cells that exist in whole organs and play vital roles in many biological processes. Although the functional diversity of fibroblasts has been estimated, a comprehensive analysis of fibroblasts from the whole body has not been performed and their transcriptional diversity has not been sufficiently explored. The aim of this study was to elucidate the transcriptional diversity of human fibroblasts within the whole body. Methods Global gene expression analysis was performed on 63 human primary fibroblasts from 13 organs. Of these, 32 fibroblasts from gastrointestinal organs (gastrointestinal fibroblasts: GIFs) were obtained from a pair of 2 anatomical sites: the submucosal layer (submucosal fibroblasts: SMFs) and the subperitoneal layer (subperitoneal fibroblasts: SPFs). Using hierarchical clustering analysis, we elucidated identifiable subgroups of fibroblasts and analyzed the transcriptional character of each subgroup. Results In unsupervised clustering, 2 major clusters that separate GIFs and non-GIFs were observed. Organ- and anatomical site-dependent clusters within GIFs were also observed. The signature genes that discriminated GIFs from non-GIFs, SMFs from SPFs, and the fibroblasts of one organ from another organ consisted of genes associated with transcriptional regulation, signaling ligands, and extracellular matrix remodeling. Conclusions GIFs are characteristic fibroblasts with specific gene expressions from transcriptional regulation, signaling ligands, and extracellular matrix remodeling related genes. In addition, the anatomical site- and organ-dependent diversity of GIFs was also discovered. These features of GIFs contribute to their specific physiological function and homeostatic maintenance, and create a functional diversity of the gastrointestinal tract. PMID:26046848

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

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

Taru Sharma, G., E-mail: gts553@gmail.com; Dubey, Pawan K.; Verma, Om Prakash

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

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

PubMed

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.

An atomic-force-microscopy study of the structure of surface layers of intact fibroblasts

NASA Astrophysics Data System (ADS)

Khalisov, M. M.; Ankudinov, A. V.; Penniyaynen, V. A.; Nyapshaev, I. A.; Kipenko, A. V.; Timoshchuk, K. I.; Podzorova, S. A.; Krylov, B. V.

2017-02-01

Intact embryonic fibroblasts on a collagen-treated substrate have been studied by atomic-force microscopy (AFM) using probes of two types: (i) standard probes with tip curvature radii of 2-10 nm and (ii) special probes with a calibrated 325-nm SiO2 ball radius at the tip apex. It is established that, irrespective of probe type, the average maximum fibroblast height is on a level of 1.7 μm and the average stiffness of the probe-cell contact amounts to 16.5 mN/m. The obtained AFM data reveal a peculiarity of the fibroblast structure, whereby its external layers move as a rigid shell relative to the interior and can be pressed inside to a depth dependent on the load only.

Matrix remodeling maintains ESC self-renewal by activating Stat3

PubMed Central

Przybyla, Laralynne M.; Theunissen, Thorold W.; Jaenisch, Rudolf; Voldman, Joel

2013-01-01

While a variety of natural and synthetic matrices have been used to influence embryonic stem cell (ESC) self-renewal or differentiation, and ESCs also deposit a rich matrix of their own, the mechanisms behind how extracellular matrix affects cell fate are largely unexplored. The ESC matrix is continuously remodeled by matrix metalloproteinases (MMPs), a process that we find is enhanced by the presence of mouse embryonic fibroblast feeders in a paracrine manner. Matrix remodeling by MMPs aids in the self-renewal of ESCs, as inhibition of MMPs inhibits the ability of ESCs to self-renew. We also find that addition of the interstitial collagenase MMP1 is sufficient to maintain long-term LIF-independent mESC self-renewal in a dose-dependent manner. This remarkable ability is due to the presence of endogenously produced self-renewal-inducing signals, including the LIF-family ligand CNTF, that are normally trapped within the ECM and become exposed upon MMP-induced matrix remodeling to signal through JAK and Stat3. These results uncover a new role for feeder cells in maintaining self-renewal and show that mESCs normally produce sufficient levels of autocrine-acting pro-self-renewal ligands. PMID:23404867

CFD Application to Flow-Accelerated Corrosion in Feeder Bends

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

Pietralik, John M.; Smith, Bruce A.W.

2006-07-01

Feeder piping in CANDU{sup R} plants experiences a thinning degradation mechanism called Flow-Accelerated Corrosion (FAC). The piping is made of carbon steel and has high water flow speeds. Although the water chemistry is highly alkaline with room-temperature pH in a range of 10.0-10.5, the piping has FAC rates exceeding 0.1 mm/year in some locations, e.g., in bends. One of the most important parameters affecting the FAC rate is the mass transfer coefficient for convective mass transport of ferrous ions. The ions are created at the pipe wall as a result of corrosion, diffuse through the oxide layer, and are transportedmore » from the oxide-layer/water interface to the bulk water by mass transport. Consequently, the local flow characteristics contribute to the highly turbulent convective mass transfer. Plant data and laboratory experiments indicate that the mass transfer step dominates FAC under feeder conditions. In this study, the flow and mass transfer in a feeder bend under operating conditions were simulated using the Fluent{sup TM} computer code. Because the flow speed is very high, with the Reynolds numbers in a range of several millions, and because the geometry is complex, experiments in a 1:1 scale were conducted with the main objective to validate flow simulations. The experiments measured pressure at several key locations and visualized the flow. The flow and mass transfer models were validated using available friction-factor and mass transfer correlations and literature experiments on mass transfer in a bend. The validation showed that the turbulence model that best predicts the experiments is the realizable k-{epsilon} model. Other two-equation turbulence models, as well as one-equation models and Reynolds stress models were tried. The near-wall treatment used the non-equilibrium wall functions. The wall functions were modified for surface roughness when necessary. A comparison of the local mass transfer coefficient with measured FAC rate in plant specimens shows very good agreement. Visualization experiments indicate secondary flows in the bends. No boundary layer separation was observed in experiments or in simulations. (authors)« less

Stimulatory effect of fibroblast-derived prostaglandin E₂ on keratinocyte stratification in the skin equivalent.

PubMed

Arai, Koji Y; Fujioka, Atsuko; Okamura, Ryoko; Nishiyama, Toshio

2014-01-01

Epidermal-dermal interaction plays important roles in physiological events such as wound healing. In this study, we examined a double paracrine mechanism between keratinocytes and fibroblasts through interleukin-1 (IL-1) and an IL-1-induced inflammatory mediator prostaglandin E₂ (PGE₂) using the skin equivalent. The epidermal layer of the skin equivalent expressed high levels of IL-1α mRNA (IL1A mRNA) and relatively low levels of IL-1β mRNA (IL1B mRNA). IL1A mRNA was not detected in fibroblasts. Fibroblasts also expressed low but not negligible levels of IL1B mRNA only in the presence of keratinocytes. Expression of prostaglandin-endoperoxide synthase 2 mRNA (PTGS2 mRNA) and production of PGE₂ in three-dimensionally cultured fibroblasts were noticeably stimulated by co-culture with keratinocytes, whereas PTGS2 mRNA expression in the epidermal layer was very low. In addition, hydroxyprostaglandin dehydrogenase 15-(NAD) mRNA was highly expressed in keratinocytes but not in fibroblasts, and exogenous IL-1β stimulated PTGS2 mRNA expression in the dermal equivalent. The thickness of the epidermal layer and the number of MKI67-positive keratinocytes in the skin equivalent were decreased by treatment with indomethacin, and the decrease recovered when exogenous PGE₂ was added. These results indicate that keratinocytes stimulate their own proliferation through a double paracrine mechanism mediated by IL-1 and PGE₂. © 2014 by the Wound Healing Society.

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

PubMed

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.

Placental growth factor neutralising antibodies give limited anti-angiogenic effects in an in vitro organotypic angiogenesis model.

PubMed

Brave, Sandra R; Eberlein, Cath; Shibuya, Masabumi; Wedge, Stephen R; Barry, Simon T

2010-12-01

Vascular Endothelial Growth Factor Receptor (VEGFR) mediated signalling drives angiogenesis. This is predominantly attributed to the activity of VEGFR-2 following binding of VEGF-A. Whether other members of the VEGFR and ligand families such as VEGFR-1 and its ligand Placental Growth Factor (PlGF) can also contribute to developmental and pathological angiogenesis is less clear. We explored the function of PlGF in VEGF-A dependent angiogenesis using an in vitro co-culture assay in which endothelial cells are cultured on a fibroblast feeder layer. In the presence of 2% FS MCDB media (containing limited growth factors) in vitro endothelial tube formation is driven by endogenous angiogenic stimuli which are produced by the fibroblast and endothelial cells. Under these conditions independent sequestration of either free VEGF-A or PlGF with polyclonal and monoclonal antibodies inhibited tube formation suggesting that both ligands are required to drive an angiogenic response. Endothelial tube formation could only be driven within this assay by the addition of exogenous VEGF-A, VEGF-E or VEGF-A/PlGF heterodimer, but not by PlGF alone, implying that activation of either VEGFR-2/VEGFR-1 heterodimers or VEGFR-2 homodimers were responsible for eliciting an angiogenic response directly, but not VEGFR-1 homodimers. In contrast to results obtained with an endogenous angiogenic drive, sequestration of PlGF did not affect endothelial tube formation when the assay was driven by 1 ng/ml exogenous VEGF-A. These data suggest that although neutralising PlGF can be shown to reduce endothelial tube formation in vitro, this effect is only observed under restricted culture conditions and is influenced by VEGF-A. Such data questions whether neutralising PlGF would have a therapeutic benefit in vivo in the presence of pathological concentrations of VEGF-A.

Granular Material Scoop and Near-Vertical Lifting Feeder/Conveyor

NASA Technical Reports Server (NTRS)

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

2017-01-01

An integrated granular-material scoop and near-vertical lifting feeder/conveyor includes special connections and skirts between a bullnose rotating scoop and an open-helical screw that provides the rotations and material lift and evacuation. A conical working-face of the bullnose rotating scoop has symmetrically distributed graters and vents to break loose and force-in granular material from natural deposits and cargo holds. The bullnose rotating scoop and the open-helical screw its attached to move the material into a continuous layer on the inside surface of an outer stationary sheathing. A motor drive attached to the open-helical screw above at the delivery end provides the lifting force necessary.

Nanog is an essential factor for induction of pluripotency in somatic cells from endangered felids.

PubMed

Verma, Rajneesh; Liu, Jun; Holland, Michael Kenneth; Temple-Smith, Peter; Williamson, Mark; Verma, Paul John

2013-02-01

Nanog has an important role in pluripotency induction in bovines and snow leopards. To examine whether it was required for wild felids globally, we examined the induction of pluripotency in felids from Asia (Bengal tiger, Panthera tigris), Africa (serval, Leptailurus serval), and the Americas (jaguar, Panthera onca). Dermal fibroblasts were transduced with genes encoding the human transcription factors OCT4, SOX2, KLF4, and cMYC with or without NANOG. Both four- and five-factor induction resulted in colony formation at day 3 in all three species tested; however, we were not able to maintain colonies that were generated without NANOG beyond passage (P) 7. Five-factor induced pluripotent stem cell (iPSC) colonies from wild cats were expanded in vitro on feeder layers and were positive for alkaline phosphatase and protein expression of OCT-4, NANOG, and stage-specific embryonic antigen-4 at P4 and P14. Reverse-transcription polymerase chain reaction confirmed that all five human transgenes were transcribed at P4; however, OCT4, SOX2, and NANOG transgenes were silenced by P14. Endogenous OCT4 and NANOG transcripts were detected at P4 and P14 in all cell lines confirming successful reprogramming. At P14, the iPSCs from all three species remained euploid and differentiated in vivo and in vitro into derivatives of the three germ layers. This study describes an effective method for inducing pluripotency in three endangered wild cats from across the globe and confirms Nanog as an essential factor in the reprogramming event. Efficient production of iPSC from endangered felids creates a unique opportunity for species preservation through gamete production, nuclear transfer, embryo complementation, and future novel technologies.

Nanog Is an Essential Factor for Induction of Pluripotency in Somatic Cells from Endangered Felids

PubMed Central

Verma, Rajneesh; Liu, Jun; Holland, Michael Kenneth; Temple-Smith, Peter; Williamson, Mark

2013-01-01

Abstract Nanog has an important role in pluripotency induction in bovines and snow leopards. To examine whether it was required for wild felids globally, we examined the induction of pluripotency in felids from Asia (Bengal tiger, Panthera tigris), Africa (serval, Leptailurus serval), and the Americas (jaguar, Panthera onca). Dermal fibroblasts were transduced with genes encoding the human transcription factors OCT4, SOX2, KLF4, and cMYC with or without NANOG. Both four- and five-factor induction resulted in colony formation at day 3 in all three species tested; however, we were not able to maintain colonies that were generated without NANOG beyond passage (P) 7. Five-factor induced pluripotent stem cell (iPSC) colonies from wild cats were expanded in vitro on feeder layers and were positive for alkaline phosphatase and protein expression of OCT-4, NANOG, and stage-specific embryonic antigen-4 at P4 and P14. Reverse-transcription polymerase chain reaction confirmed that all five human transgenes were transcribed at P4; however, OCT4, SOX2, and NANOG transgenes were silenced by P14. Endogenous OCT4 and NANOG transcripts were detected at P4 and P14 in all cell lines confirming successful reprogramming. At P14, the iPSCs from all three species remained euploid and differentiated in vivo and in vitro into derivatives of the three germ layers. This study describes an effective method for inducing pluripotency in three endangered wild cats from across the globe and confirms Nanog as an essential factor in the reprogramming event. Efficient production of iPSC from endangered felids creates a unique opportunity for species preservation through gamete production, nuclear transfer, embryo complementation, and future novel technologies. PMID:23514873

Unsuccessful derivation of human embryonic stem cell lines from pairs of human blastomeres.

PubMed

Fong, Chui-Yee; Richards, Mark; Bongso, Ariff

2006-08-01

Human embryonic stem cells (hESC) that differentiate into all three primordial germ layers have been established. Differentiation of these cells into desirable lineages offers hope for future transplantation therapies. Currently, hESC lines are derived from the inner cell mass (ICM) of blastocysts, leading to destruction of the embryo, and thus the process is ethically controversial. Successful attempts at deriving hESC lines from blastomeres without destruction of the ensuing embryo have not been reported. One or two blastomeres are routinely biopsied from 8-cell embryos for preimplantation genetic diagnosis. In this study it was therefore attempted to derive hESC lines from paired blastomeres. Of 66 pairs of 8-cell stage blastomeres, four pairs produced two morula and two blastocyst-like structures. When plated on mitomycin-C-treated mouse embryonic fibroblasts, one morula and one blastocyst-like structure separately produced small colonies containing hESC-like cells with prominent nucleoli and high nuclear-cytoplasmic ratios. When these colonies were detached and plated onto fresh feeders, there was no further colony formation or ensuing hESC lines. The results showed that it might not be possible to derive hESC lines directly from paired blastomeres. A minimum number of blastomeres in close contact with one another may be required to successfully generate an hESC line as blastomeres, like ICM and hESC cells, may be 'social' cells.

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

PubMed

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 study shows that certain nonviral reprogramming methods may not necessarily be safer than viral approaches and that maximizing exogenous gene expression of reprogramming factors is not sufficient to ensure successful reprogramming.

Comparison of two different culture conditions for derivation of early hiPSC.

PubMed

Hey, Caroline A B; Saltõkova, Katarina B; Bisgaard, Hanne C; Møller, Lisbeth B

2018-03-30

Different culture-systems for derivation of induced pluripotent stem cells (iPSC) in vitro from human fibroblasts have been established. Here, we compared the efficacy of two different feeder-free culture-systems; Matrigel-coated surfaces in combination with mTeSR1 medium versus Vitronectin-coated surfaces in combination with Essential 8 (E8) medium. The comparison was performed by counting the number of emerging iPSC-looking colonies of re-programmed fibroblasts. The fibroblasts were re-programmed using episomal plasmids expressing OCT3/4, SOX2, KLF4, L-MYC, LIN28, and a p53 knock down shP53. Three different fibroblast lines, K40 and K48 from healthy controls and BBS1 from a patient with Bardet-Biedl syndrome, were used in two independent setups. The BBS1 line was used in both setups in combination with K40 and K48 respectively. In all four re-programming experiments, we observed a significantly higher number of emerging colonies with the combination Matrigel/mTeSR1 as compared to the combination Vitronectin/E8. The presence of iPSC was verified by alkaline phosphatase and Tra-1-60 staining. Furthermore, a higher expression of the pluripotency-associated markers NANOG and SOX2 in cells under Matrigel/mTeSR1 conditions compared with Vitronectin/E8 supported the higher proportion of iPSC on Matrigel/mTeSR1 plates. In conclusion, the combination Matrigel/mTeSR1 is more efficient for derivation of iPSC compared to the Vitronectin/E8 combination. © 2018 The Authors. Cell Biology International Published by John Wiley & Sons Ltd on behalf of International Federation of Cell Biology.

The influence of SrO and CaO in silicate and phosphate bioactive glasses on human gingival fibroblasts.

PubMed

Massera, J; Kokkari, A; Närhi, T; Hupa, L

2015-06-01

In this paper, we investigate the effect of substituting SrO for CaO in silicate and phosphate bioactive glasses on the human gingival fibroblast activity. In both materials the presence of SrO led to the formation of a CaP layer with partial Sr substitution for Ca. The layer at the surface of the silicate glass consisted of HAP whereas at the phosphate glasses it was close to the DCPD composition. In silicate glasses, SrO gave a faster initial dissolution and a thinner reaction layer probably allowing for a continuous ion release into the solution. In phosphate glasses, SrO decreased the dissolution process and gave a more strongly bonded reaction layer. Overall, the SrO-containing silicate glass led to a slight enhancement in the activity of the gingival fibroblasts cells when compared to the SrO-free reference glass, S53P4. The cell activity decreased up to 3 days of culturing for all phosphate glasses containing SrO. Whereas culturing together with the SrO-free phosphate glass led to complete cell death at 7 days. The glasses containing SrO showed rapid cell proliferation and growth between 7 and 14 days, reaching similar activity than glass S53P4. The addition of SrO in both silicate and phosphate glasses was assumed beneficial for proliferation and growth of human gingival fibroblasts due to Sr incorporation in the reaction layer at the glass surface and released in the cell culture medium.

Determination of body width in brown and white layer pullets by image analyses.

PubMed

Giersberg, M F; Kemper, N; Hartung, J; Schrader, L; Spindler, B

2017-06-01

1. Specific legal requirements for keeping pullets are not available in the European Union. However, two of the most important rearing factors for pullets are sufficient perching and feeder space. Both factors represent horizontal space dimensions which derive from the body width of the birds. 2. The body width of two strains of layer pullets (brown (BL) and white (WL) layer pullets) based on the measurement of distances in digital images was conducted on front-view digital photographs of BL and WL pullets taken at 8, 12 and 19 weeks of life. 3. Depending on live weight, age and body position, BL pullets measured an average body width between 10.70 ± 1.10 and 13.96 ± 1.11 cm. The width of WL pullets ranged from 10.30 ± 0.86 to 13.00 ± 1.14 cm. 4. Compared with WL, BL pullets occupied more horizontal space during rearing. Age influenced the body width of BL and WL pullets at the end of rearing. The tested body positions of the pullets did not affect the measured body width. 5. The biometric data obtained in this study are a useful basis for developing legal requirements for pullets, especially for defining minimum perch width and feeder space allowances.

Fabrication of In Vitro Cancer Microtissue Array on Fibroblast-Layered Nanofibrous Membrane by Inkjet Printing

PubMed Central

Park, Tae-Min; Kang, Donggu; Jang, Ilho; Yun, Won-Soo; Shim, Jin-Hyung; Jeong, Young Hun; Kwak, Jong-Young; Yoon, Sik; Jin, Songwan

2017-01-01

In general, a drug candidate is evaluated using 2D-cultured cancer cells followed by an animal model. Despite successful preclinical testing, however, most drugs that enter human clinical trials fail. The high failure rates are mainly caused by incompatibility between the responses of the current models and humans. Here, we fabricated a cancer microtissue array in a multi-well format that exhibits heterogeneous and batch-to-batch structure by continuous deposition of collagen-suspended Hela cells on a fibroblast-layered nanofibrous membrane via inkjet printing. Expression of both Matrix Metalloproteinase 2 (MMP2) and Matrix Metalloproteinase 9 (MMP9) was higher in cancer microtissues than in fibroblast-free microtissues. The fabricated microtissues were treated with an anticancer drug, and high drug resistance to doxorubicin occurred in cancer microtissues but not in fibroblast-free microtissues. These results introduce an inkjet printing fabrication method for cancer microtissue arrays, which can be used for various applications such as early drug screening and gradual 3D cancer studies. PMID:29112150

Substrate stiffness affects skeletal myoblast differentiation in vitro

NASA Astrophysics Data System (ADS)

Romanazzo, Sara; Forte, Giancarlo; Ebara, Mitsuhiro; Uto, Koichiro; Pagliari, Stefania; Aoyagi, Takao; Traversa, Enrico; Taniguchi, Akiyoshi

2012-12-01

To maximize the therapeutic efficacy of cardiac muscle constructs produced by stem cells and tissue engineering protocols, suitable scaffolds should be designed to recapitulate all the characteristics of native muscle and mimic the microenvironment encountered by cells in vivo. Moreover, so not to interfere with cardiac contractility, the scaffold should be deformable enough to withstand muscle contraction. Recently, it was suggested that the mechanical properties of scaffolds can interfere with stem/progenitor cell functions, and thus careful consideration is required when choosing polymers for targeted applications. In this study, cross-linked poly-ɛ-caprolactone membranes having similar chemical composition and controlled stiffness in a supra-physiological range were challenged with two sources of myoblasts to evaluate the suitability of substrates with different stiffness for cell adhesion, proliferation and differentiation. Furthermore, muscle-specific and non-related feeder layers were prepared on stiff surfaces to reveal the contribution of biological and mechanical cues to skeletal muscle progenitor differentiation. We demonstrated that substrate stiffness does affect myogenic differentiation, meaning that softer substrates can promote differentiation and that a muscle-specific feeder layer can improve the degree of maturation in skeletal muscle stem cells.

Dermis, acellular dermal matrix, and fibroblasts from different layers of pig skin exhibit different profibrotic characteristics: evidence from in vivo study

PubMed Central

Zuo, Yanhai; Lu, Shuliang

2017-01-01

To explore the profibrotic characteristics of the autografted dermis, acellular dermal matrix, and dermal fibroblasts from superficial/deep layers of pig skin, 93 wounds were established on the dorsa of 7 pigs. 72 wounds autografted with the superficial/deep dermis and acellular dermal matrix served as the superficial/deep dermis and acellular dermal matrix group, respectively, and were sampled at 2, 4, and 8 weeks post-wounding. 21 wounds autografted with/without superficial/deep dermal fibroblasts served as the superficial/deep dermal fibroblast group and the control group, respectively, and were sampled at 2 weeks post-wounding. The hematoxylin and eosin staining showed that the wounded skin thicknesses in the deep dermis group (superficial acellular dermal matrix group) were significantly greater than those in the superficial dermis group (deep acellular dermal matrix group) at each time point, the thickness of the cutting plane in the deep dermal fibroblast group was significantly greater than that in the superficial dermal fibroblast group and the control group. The western blots showed that the α-smooth muscle actin expression in the deep dermis group (superficial acellular dermal matrix group) was significantly greater than that in the superficial dermis group (deep acellular dermal matrix group) at each time point. In summary, the deep dermis and dermal fibroblasts exhibited more profibrotic characteristics than the superficial ones, on the contrary, the deep acellular dermal matrix exhibited less profibrotic characteristics than the superficial one. PMID:28423561

An in-vitro scaffold-free epithelial-fibroblast coculture model for the larynx

PubMed Central

Walimbe, Tanaya; Panitch, Alyssa; Sivasankar, M. Preeti

2017-01-01

Objective Physiologically relevant, well-characterized in vitro vocal fold coculture models are needed to test the effects of various challenges and therapeutics on vocal fold physiology. We characterize a healthy state coculture model, created by using bronchial/tracheal epithelial cells and immortalized vocal fold fibroblasts. We also demonstrate that this model can be induced into a fibroplastic state to overexpress stress fibers using TGFβ1. Method Cell metabolic activity of immortalized human vocal fold fibroblasts incubated in different media combinations were confirmed with MTT assay. Fibroblasts were grown to confluence and primary bronchial/tracheal epithelial cells suspended in coculture media were seeded directly over the base layer of the fibroblasts. Cells were treated with TGFβ1 to induce myofibroblast formation. Cell shape and position was confirmed by live cell tracking, fibrosis was confirmed by probing for α smooth muscle actin (α-SMA) and phenotype was confirmed by immunostaining for vimentin and E-cadherin. Results Fibroblasts retain metabolic activity in coculture epithelial media. Live cell imaging revealed a layer of epithelial cells atop fibroblasts. α-SMA expression was enhanced in TGFβ1 treated cells, confirming that both cell types maintained a healthy phenotype in coculture, and can be induced into overexpressing stress fibers. Vimentin and E-cadherin immunostaining show that cells retain phenotype in coculture. Conclusion These data lay effective groundwork for a functional coculture model that retains the reproducibility necessary to serve as a viable diagnostic and therapeutic screening platform. Level of Evidence NA PMID:27859361

Development of an ES-like cell culture system (RESC) from rohu, Labeo rohita (Ham.).

PubMed

Goswami, M; Lakra, W S; Yadav, Kamalendra; Jena, J K

2012-12-01

An embryonic stem (ES)-like cell culture system RESC from a commercially important freshwater carp, Labeo rohita, was developed using blastula stage embryos. The cells were cultured in Leibovitz-15 (L-15) medium in gelatin-coated cell culture flask supplemented with 15 % fetal bovine serum along with 10 ng ml(-1) basic fibroblast growth factor at 28 °C under feeder-free conditions. The ES-like cells were characterized by their unique morphology, alkaline phosphatase activity, embryoid body formation tendency, expression of transcription factor Oct4, and consistent chromosome count. The RESC cells when treated with retinoic acid differentiated into cells of different lineages. The RESC developed from mid-blastula embryos of L. rohita would be a useful tool for cellular differentiation and gene expression studies.

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

PubMed

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

2011-12-01

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

Efficient expansion of human keratinocyte stem/progenitor cells carrying a transgene with lentiviral vector

PubMed Central

2013-01-01

Introduction The development of an appropriate procedure for lentiviral gene transduction into keratinocyte stem cells is crucial for stem cell biology and regenerative medicine for genetic disorders of the skin. However, there is little information available on the efficiency of lentiviral transduction into human keratinocyte stem/progenitor cells and the effects of gene transduction procedures on growth potential of the stem cells by systematic assessment. Methods In this study, we explored the conditions for efficient expansion of human keratinocyte stem/progenitor cells carrying a transgene with a lentiviral vector, by using the culture of keratinocytes on a feeder layer of 3 T3 mouse fibroblasts. The gene transduction and expansion of keratinocytes carrying a transgene were analyzed by Western blotting, quantitative PCR, and flow cytometry. Results Polybrene (hexadiamine bromide) markedly enhanced the efficiency of lentiviral gene transduction, but negatively affected the maintenance of the keratinocyte stem/progenitor cells at a concentration higher than 5 μg/ml. Rho-assiciated kinase (ROCK) inhibitor Y-27632, a small molecule which enhanced keratinocyte proliferation, significantly interfered with the lentiviral transduction into cultured human keratinocytes. However, a suitable combination of polybrene and Y-27632 effectively expanded keratinocytes carrying a transgene. Conclusions This study provides information for effective expansion of cultured human keratinocyte stem/progenitor cells carrying a transgene. This point is particularly significant for the application of genetically modified keratinocyte stem/progenitor stem cells in regenerative medicine. PMID:24406242

Long-term Culture and Cloning of Primary Human Bronchial Basal Cells that Maintain Multipotent Differentiation Capacity and CFTR Channel Function.

PubMed

Peters-Hall, Jennifer Ruth; Coquelin, Melissa L; Torres, Michael J; LaRanger, Ryan; Alabi, Busola Ruth; Sho, Sei; Calva-Moreno, Jose Francisco; Thomas, Philip J; Shay, Jerry William

2018-05-03

While primary cystic fibrosis (CF) and non-CF human bronchial epithelial basal cells (HBECs) accurately represent in vivo phenotypes, one barrier to their wider use has been a limited ability to clone and expand cells in sufficient numbers to produce rare genotypes using genome editing tools. Recently, conditional reprogramming of cells (CRC) with a ROCK inhibitor and culture on an irradiated fibroblast feeder layer resulted in extension of the lifespan of HBECs, but differentiation capacity and CF transmembrane conductance regulator (CFTR) function decreased as a function of passage. This report details modifications to the standard HBEC CRC protocol (Mod CRC), including the use of bronchial epithelial growth medium instead of F-medium and 2% oxygen instead of 21% oxygen, that extend HBEC lifespan while preserving multipotent differentiation capacity and CFTR function. Critically, Mod CRC conditions support clonal growth of primary HBECs from a single cell and the resulting clonal HBEC population maintains multipotent differentiation capacity, including CFTR function, permitting gene editing of these cells. As a proof of concept, CRISPR/Cas9 genome editing and cloning was used to introduce insertions/deletions in CFTR exon 11. Mod CRC conditions overcome many barriers to the expanded use of HBECs for basic research and drug screens. Importantly, Mod CRC conditions support the creation of isogenic cell lines in which CFTR is mutant or wild-type in the same genetic background with no history of CF to enable determination of the primary defects of mutant CFTR.

Rational design of bottom blocks for development of ore deposits systems with caving of ore and enclosing rocks

NASA Astrophysics Data System (ADS)

Versilov, S. O.; Posylniy, Yu V.; Shurygin, D. N.; Tretyak, A. Ya

2017-10-01

The assessment of the geological conditions of development of existing ore deposits was made. For testing ore deposits in difficult mining and geological conditions, the authors proposed the system of development, accompanied by collapse of the mechanical ore with the use of feeders of active action that could be manufactured directly in the mine in accordance with the specific conditions of occurrence of minerals. The paper demonstrates the technology of manufacture of load-bearing structures of the feeder directly in the mine at the scene of the breaking of the first layer of ore, as well as the dynamics of the ore and the choice of parameters of concrete feeders. A new design of the bottom block was proposed, the idea of technical solution of which consists in the fact that it is offered to undergo the production of the smallest possible cross section, which is determined only by the dimensions of the conveyors to deliver ore. And before the explosion of fans of production wells, it is necessary to produce local collapse of the roof production to increase its height at the place of production of ore by blasting wellheads in two or three rows.

Binary colloidal crystals (BCCs) as a feeder-free system to generate human induced pluripotent stem cells (hiPSCs)

PubMed Central

Wang, Peng-Yuan; Hung, Sandy Shen-Chi; Thissen, Helmut; Kingshott, Peter; Wong, Raymond Ching-Bong

2016-01-01

Human induced pluripotent stem cells (hiPSCs) are capable of differentiating into any cell type and provide significant advances to cell therapy and regenerative medicine. However, the current protocol for hiPSC generation is relatively inefficient and often results in many partially reprogrammed colonies, which increases the cost and reduces the applicability of hiPSCs. Biophysical stimulation, in particular from tuning cell-surface interactions, can trigger specific cellular responses that could in turn promote the reprogramming process. In this study, human fibroblasts were reprogrammed into hiPSCs using a feeder-free system and episomal vectors using novel substrates based on binary colloidal crystals (BCCs). BCCs are made from two different spherical particle materials (Si and PMMA) ranging in size from nanometers to micrometers that self-assemble into hexagonal close-packed arrays. Our results show that the BCCs, particularly those made from a crystal of 2 μm Si and 0.11 μm PMMA particles (2SiPM) facilitate the reprogramming process and increase the proportion of fully reprogrammed hiPSC colonies, even without a vitronectin coating. Subsequent isolation of clonal hiPSC lines demonstrates that they express pluripotent markers (OCT4 and TRA-1-60). This proof-of-concept study demonstrates that cell reprogramming can be improved on substrates where surface properties are tailored to the application. PMID:27833126

Trace-fossil assemblages with a new ichnogenus in "spotted"

NASA Astrophysics Data System (ADS)

Šimo, Vladimír; Tomašových, Adam

2013-10-01

Highly-bioturbated "spotted" limestones and marls (Fleckenmergel-Fleckenkalk facies) of the Early Jurassic, which were deposited in broad and recurrent deep-shelf habitats of the Northern Tethys, are characterized by rare benthic carbonate-producing macroinvertebrates. To address this paradox, we analyse trace-fossil assemblages in a ~85 m-thick succession of Pliensbachian spotted deposits (Zliechov Basin, Western Carpathians). They are dominated by infaunal and semi-infaunal deposit-feeders, with 9 ichnogenera and pyritized tubes of the semi-infaunal foraminifer Bathysiphon, being dominated by Chondrites, Lamellaeichnus (new ichnogenus), and Teichichnus. Lamellaeichnus, represented by a horizontal basal cylindrical burrow and an upper row of stacked convex-up gutters, was produced by a mobile deposit-feeder inhabiting shallow tiers because it is crossed by most other trace fossils. We show that the spotty appearance of the deposits is generated by a mixture of (1) dark, organic-rich shallow- and deep-tier traces (TOC = 0.16-0.36), and (2) light grey, organic-poor mottled or structurless sediment (TOC = 0.09-0.22). The higher TOC in shallow-tier burrows of Lamellaeichnus demonstrates that uppermost sediment layers were affected by poor redox cycling. Such conditions imply a limited mixed-layer depth and inefficient nutrient recycling conditioned by hypoxic bottom-waters, allowed by poor circulation and high sedimentation rates in depocenters of the Zliechov Basin. Hypoxic conditions are further supported by (1) dominance of trace-fossils produced by infaunal deposit feeders, (2) high abundance of hypoxiatolerant agglutinated foraminifer Bathysiphon, and (3) high abundance of Chondrites with ~0.5 mm-sized branches. Oxygen-deficient bottom-conditions can thus simultaneously explain the rarity of benthic carbonate-producing macroinvertebrates and high standing abundance of tolerant soft-shell and agglutinated organisms in spotted deposits.

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

PubMed

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.

Investigating the feasibility of scale up and automation of human induced pluripotent stem cells cultured in aggregates in feeder free conditions☆

PubMed Central

Soares, Filipa A.C.; Chandra, Amit; Thomas, Robert J.; Pedersen, Roger A.; Vallier, Ludovic; Williams, David J.

2014-01-01

The transfer of a laboratory process into a manufacturing facility is one of the most critical steps required for the large scale production of cell-based therapy products. This study describes the first published protocol for scalable automated expansion of human induced pluripotent stem cell lines growing in aggregates in feeder-free and chemically defined medium. Cells were successfully transferred between different sites representative of research and manufacturing settings; and passaged manually and using the CompacT SelecT automation platform. Modified protocols were developed for the automated system and the management of cells aggregates (clumps) was identified as the critical step. Cellular morphology, pluripotency gene expression and differentiation into the three germ layers have been used compare the outcomes of manual and automated processes. PMID:24440272

The pulmonary mesenchymal tissue layer is defective in an in vitro recombinant model of nitrofen-induced lung hypoplasia.

PubMed

van Loenhout, Rhiannon B; Tseu, Irene; Fox, Emily K; Huang, Zhen; Tibboel, Dick; Post, Martin; Keijzer, Richard

2012-01-01

Despite modern treatments, congenital diaphragmatic hernia (CDH) remains associated with variable survival and significant morbidity. The associated pulmonary hypoplasia is a major determinant of outcome. To develop better treatments, improved comprehension of the pathogenesis of lung hypoplasia is warranted. We developed an in vitro cell recombinant model to mimic pulmonary hypoplasia and specifically to investigate epithelial-mesenchymal interactions and to decipher which tissue layer is primarily defective in nitrofen-induced CDH-associated lung hypoplasia. Epithelial cells (E) and fibroblasts (F) were isolated from E19 control ((C)) and nitrofen-induced hypoplastic rat lungs ((N)). Cells were recombined and cultured as either homotypic [(F(C))(E(C)) and (F(N))(E(N))] or heterotypic [(F(C))(E(N)) and (F(N))(E(C))] recombinants. Recombinants containing F(N) fibroblasts had a thickened fibroblast tissue layer and there were fewer organized alveolar-like epithelial structures compared with those in control (F(C))(E(C)) recombinants. These F(N) recombinants exhibited a decrease in terminal deoxynucleotidyl transferase dUTP nick end labeling and cleaved caspase-3 positive cells. Cell proliferation was arrested in recombinants containing F(N) fibroblasts, which also exhibited increased p27(Kip1) and p57(Kip2) expression. In conclusion, fibroblasts, and not epithelial cells, appear to be the defective cell type in nitrofen-induced hypoplastic lungs due to a decreased ability to undergo apoptosis and maintain overall proliferation. This may explain the characteristic pulmonary interstitial thickening and hypoplasia observed in both nitrofen-induced hypoplastic lungs as well as human hypoplastic CDH lungs. Copyright © 2012 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

Food web structure and seasonality of slope megafauna in the NW Mediterranean elucidated by stable isotopes: Relationship with available food sources

NASA Astrophysics Data System (ADS)

Papiol, V.; Cartes, J. E.; Fanelli, E.; Rumolo, P.

2013-03-01

The food-web structure and seasonality of the dominant taxa of benthopelagic megafauna (fishes and decapods) on the middle slope of the Catalan Sea (Balearic Basin, NW Mediterranean) were investigated using the carbon and nitrogen stable isotope ratios of 29 species. Macrofauna (infauna, suprabenthos and zooplankton) were also analysed as potential prey. Samples were collected on a seasonal basis from 600 to 1000 m depth between February 2007 and February 2008. The fishes and decapods were classified into feeding groups based on the literature: benthic feeders (including suprabenthos) and zooplankton feeders, the latter further separated into migratory and non-migratory species. Decapods exhibited depleted δ15N and enriched δ13C compared to fishes. Annual mean δ13C of fishes ranged from - 19.15‰ (Arctozenus risso) to - 16.65‰ (Phycis blennoides) and of δ15N from 7.27‰ (Lampanyctus crocodilus) to 11.31‰ (Nezumia aequalis). Annual mean values of δ13C of decapods were from - 18.94‰ (Sergestes arcticus) to - 14.78‰ (Pontophilus norvegicus), and of δ15N from 6.36‰ (Sergia robusta) to 9.72‰ (Paromola cuvieri). Stable isotopes distinguished well amongst the 3 feeding guilds established a priori, pointing to high levels of resource partitioning in deep-sea communities. The trophic structure of the community was a function of the position of predators along the benthic-pelagic gradient, with benthic feeders isotopically enriched relative to pelagic feeders. This difference allowed the identification of two food webs based on pelagic versus benthic consumption. Prey and predator sizes were also important in structuring the community. The most generalised seasonal pattern was δ13C depletion from winter to spring and summer, especially amongst migratory macroplankton feeders. This suggests greater consumption of pelagic prey, likely related with increases in pelagic production or with ontogenic migrations of organisms from mid-water to the Benthic Boundary Layer (BBL). δ15N enrichment was detected in periods of water column stratification, particularly amongst benthic feeder fishes. Megafauna relied on a single source of nutrition after peaks in surface production, presumably marine snow. Conversely, a larger array of food sources, probably from advection, sustained the community in periods of water column stratification. Benthic feeder δ13C values of both taxa were positively correlated with fluorescence measured 5 m above the seabed and negatively correlated with total organic carbon in the sediments, both being food sources for deposit feeding macroinfauna. Macroplankton feeder δ13C values were linked to environmental variables related to vertical transport from surface production, i.e. lipids and chlorophyll and their degradation products, likely due to their stronger reliance on sinking phytodetritus through consumption of planktonic prey.

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

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

Reno, Matthew J.; Coogan, Kyle; Seuss, John

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 canmore » 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.« less

Influence of human fibroblasts on development and quality of multilayered composite grafts in athymic nude mice.

PubMed

Cedidi, C Can; Wilkens, L; Berger, A; Ingianni, G

2007-11-05

In patients after extensive burn injury the lack of split thickness skin graft donor sites, and consecutive delay in wound closure are critical factors of morbidity and mortality. In addition limited functional and aesthetic results after transplantation of split thickness skin grafts present a socioeconomic problem. For improved wound closure the aim of this study was the development of a one stage technique for the establishment of a multi layer composite graft, existing of a collagen-GAG-matrix with silicon layer of a two layer synthetic dermal equivalent (DE) with integrated fibroblasts, and ceratinocytes. - In 64 athymic nude mice the evaluation of the multi layer skin grafts potential to re-establish a human epidermis, and high quality dermal structure was performed. In addition to clinical investigations we measured wound contraction, and analyzed histomorphologic, immunohistologic, "in situ hybridisation", and electro microscopic data. - Our results show, that the seeding of DE with human fibroblasts and ceratinocytes as a composite skin graft reproducible enabled a wound healing with an organised human dermis and epidermis within 10 - 15 days. The histological studies of the grafted composite skin grafts in this model showed morphologically a characteristic dermal-epidermal skin structure with a cornifying epithelium, being of human origin ("in situ hybridisation"). Through the co-cultivation of fibroblasts and ceratinocytes in the DE the generation and structural morphology of collagen fibres, and inflammatory reaction in the neodermis is positively influenced, and as a consequence wound contraction significantly reduced. In regard to the early preparation of composite grafts, and the minimal requirements for donor sites - with dependable stable reconstruction of the integument - this technique may present a step forward in the treatment of patients with extensive burns.

Generation of Corneal Keratocytes from Human Embryonic Stem Cells.

PubMed

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.

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

PubMed

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 < 0.001): Waste Less, 5%; Cinch Net, 6%; Hayhut, 9%; Covered Cradle, 11%; Tombstone Saver, 13%; Tombstone, Cone, and Ring, 19%; Hay Sleigh, 33%; and no-feeder control, 57%. Feeder design also affected payback (P < 0.01). The Cinch Net paid for itself in less than 1 mo; Tombstone and Ring, 2 mo; Hayhut and Tombstone Saver, 4 mo; Hay Sleigh, 5 mo; Waste Less, 8 mo; Cone, 9 mo; and Covered Cradle, 20 mo. Round-bale feeder design affected hay waste and payback, but not estimated hay intake or BW change during horse feeding.

Effects of feeder design and changing source of water to a location separate from the wet-dry feeder at 4 or 8 weeks before harvest on growth, feeding behavior, and carcass characteristics of finishing pigs.

PubMed

Bergstrom, J R; Nelssen, J L; Edwards, L N; Tokach, M D; Dritz, S S; Goodband, R D; DeRouchey, J M

2012-12-01

Our objectives were to compare a conventional dry (5-space, 152.4-cm-wide) and a wet-dry (double-sided, each side = 38.1-cm-wide single space) feeder and to determine if changing the source of water to a location separate from a wet-dry feeder would result in improved G:F and carcass characteristics. Water supply to the wet-dry feeder was shut off and the cup waterer was turned on in 8 pens at 8 (d 69) or 4 (d 97) wk prior to harvest. For the remaining 8 wet-dry feeder pens, the feeder provided the sole water source for the entire experiment. A total of 1,296 pigs (PIC, 337 × 1050; initially 19.4 kg BW) were used, with 27 pigs/pen (14 barrows and 13 gilts) and 24 pens/feeder design. From d 0 to 69, pigs fed with the wet-dry feeder had increased (P < 0.05) ADG, ADFI, G:F, and d 69 BW compared with those using the conventional dry feeder. Overall (d 0 to 124), pigs using fed with the water source in the wet-dry feeder the entire time had greater (P < 0.05) ADG, ADFI, final BW, and HCW the other treatments. The overall G:F was not different (P > 0.05) among pigs fed with the different feeder treatments. Pigs fed with the wet-dry feeder where water source was changed at 4 wk before harvest had greater (P < 0.05) ADG than pigs that used a conventional dry feeder. Pigs where the water source was changed at 4 wk had greater (P < 0.05) ADFI than those were the water source was changed 8 wk prior to harvest, and for pigs fed with the conventional dry feeder ADFI was intermediate. Back fat depth of pigs where the water source was changed at 8 wk before harvest was reduced (P < 0.05) compared with all other treatments and LM depth was greater (P < 0.05) than that of pigs using a conventional dry feeder and where the water source was changed at 4 week before harvest. Pigs fed using the wet-dry feeder visited the feeder less frequently (P < 0.05) and spent less total time at the feeder (P < 0.05) than those fed with the conventional dry feeder. The differences in feeding patterns remained even after the access to water was removed from the wet-dry feeder, with no change in the amount of aggressive behavior observed at the feeder. Pigs fed with a wet-dry feeder had an increased growth rate compared with those fed with a conventional dry feeder. Although measures of carcass leanness were improved by changing the location of the water, removing the water from the feeder also eliminated any net improvement in BW from using a wet-dry feeder.

95. VIEW OF ZINC FEEDER FROM SOUTHEAST. NOTE FEEDER CONE ...

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

95. VIEW OF ZINC FEEDER FROM SOUTHEAST. NOTE FEEDER CONE AND PIPING FROM VACUUM RECEIVER ON LEFT. PRECIPITATE PUMP MOTOR MOUNT VISIBLE BELOW FEEDER STAIRS, PUMP AND MOTOR MISSING. SUMPS ARE LOCATED UNDER THIS FLOOR, WITH ACCESS TO HATCH TO THE RIGHT OF FEEDER STAIR. - Bald Mountain Gold Mill, Nevada Gulch at head of False Bottom Creek, Lead, Lawrence County, SD

Hygiene at winter bird feeders in a southwestern Ontario city.

PubMed Central

Prescott, J F; Hunter, D B; Campbell, G D

2000-01-01

To further understand the source of the epidemic of salmonellosis in some species of birds using bird feeders in southern Ontario in the winter of 1997-1998, 124 bird feeder stations were examined for their state of hygiene and for Salmonella on 5 occasions during the winter of 1999 in a city of 100,000 people in southwestern Ontario. No Salmonella were isolated from feed contaminated with feces recovered from the feeders. Squirrel-proof feeders were significantly less contaminated with feces than were other feeder types (hopper, platform, silo), which did not differ significantly in their hygiene scores. Contamination of squirrel-proof feeders increased significantly through the course of the study, but other feeder types showed no significant change. Hygiene was poorer if feeders were maintained equally by both male and female household members, particularly as they grew older, but no age or gender effect was observed if only one person was largely responsible for maintaining the feeders. We concluded that winter bird feeder stations in a southern Ontario city were not contaminated with Salmonella but that bird feeder stations could be designed better to reduce fecal contamination of feed. PMID:10992987

Long-term culture of rat hippocampal neurons at low density in serum-free medium: combination of the sandwich culture technique with the three-dimensional nanofibrous hydrogel PuraMatrix.

PubMed

Kaneko, Ai; Sankai, Yoshiyuki

2014-01-01

The primary culture of neuronal cells plays an important role in neuroscience. There has long been a need for methods enabling the long-term culture of primary neurons at low density, in defined serum-free medium. However, the lower the cell density, the more difficult it is to maintain the cells in culture. Therefore, we aimed to develop a method for long-term culture of neurons at low density, in serum-free medium, without the need for a glial feeder layer. Here, we describe the work leading to our determination of a protocol for long-term (>2 months) primary culture of rat hippocampal neurons in serum-free medium at the low density of 3×10(4) cells/mL (8.9×10(3) cells/cm2) without a glial feeder layer. Neurons were cultured on a three-dimensional nanofibrous hydrogel, PuraMatrix, and sandwiched under a coverslip to reproduce the in vivo environment, including the three-dimensional extracellular matrix, low-oxygen conditions, and exposure to concentrated paracrine factors. We examined the effects of varying PuraMatrix concentrations, the timing and presence or absence of a coverslip, the timing of neuronal isolation from embryos, cell density at plating, medium components, and changing the medium or not on parameters such as developmental pattern, cell viability, neuronal ratio, and neurite length. Using our method of combining the sandwich culture technique with PuraMatrix in Neurobasal medium/B27/L-glutamine for primary neuron culture, we achieved longer neurites (≥3,000 µm), greater cell viability (≥30%) for 2 months, and uniform culture across the wells. We also achieved an average neuronal ratio of 97%, showing a nearly pure culture of neurons without astrocytes. Our method is considerably better than techniques for the primary culture of neurons, and eliminates the need for a glial feeder layer. It also exhibits continued support for axonal elongation and synaptic activity for long periods (>6 weeks).

Development of a Full-Thickness Human Skin Equivalent In Vitro Model Derived from TERT-Immortalized Keratinocytes and Fibroblasts

PubMed Central

Reijnders, Christianne M.A.; van Lier, Amanda; Roffel, Sanne; Kramer, Duco; Scheper, Rik J.

2015-01-01

Currently, human skin equivalents (HSEs) used for in vitro assays (e.g., for wound healing) make use of primary human skin cells. Limitations of primary keratinocytes and fibroblasts include availability of donor skin and donor variation. The use of physiologically relevant cell lines could solve these limitations. The aim was to develop a fully differentiated HSE constructed entirely from human skin cell lines, which could be applied for in vitro wound-healing assays. Skin equivalents were constructed from human TERT-immortalized keratinocytes and fibroblasts (TERT-HSE) and compared with native skin and primary HSEs. HSEs were characterized by hematoxylin–eosin and immunohistochemical stainings with markers for epidermal proliferation and differentiation, basement membrane (BM), fibroblasts, and the extracellular matrix (ECM). Ultrastructure was determined with electron microscopy. To test the functionality of the TERT-HSE, burn and cold injuries were applied, followed by immunohistochemical stainings, measurement of reepithelialization, and determination of secreted wound-healing mediators. The TERT-HSE was composed of a fully differentiated epidermis and a fibroblast-populated dermis comparable to native skin and primary HSE. The epidermis consisted of proliferating keratinocytes within the basal layer, followed by multiple spinous layers, a granular layer, and cornified layers. Within the TERT-HSE, the membrane junctions such as corneosomes, desmosomes, and hemidesmosomes were well developed as shown by ultrastructure pictures. Furthermore, the BM consisted of a lamina lucida and lamina densa comparable to native skin. The dermal matrix of the TERT-HSE was more similar to native skin than the primary construct, since collagen III, an ECM marker, was present in TERT-HSEs and absent in primary HSEs. After wounding, the TERT-HSE was able to reepithelialize and secrete inflammatory wound-healing mediators. In conclusion, the novel TERT-HSE, constructed entirely from human cell lines, provides an excellent opportunity to study in vitro skin biology and can also be used for drug targeting and testing new therapeutics, and ultimately, for incorporating into skin-on-a chip in the future. PMID:26135533

Development of a Full-Thickness Human Skin Equivalent In Vitro Model Derived from TERT-Immortalized Keratinocytes and Fibroblasts.

PubMed

Reijnders, Christianne M A; van Lier, Amanda; Roffel, Sanne; Kramer, Duco; Scheper, Rik J; Gibbs, Susan

2015-09-01

Currently, human skin equivalents (HSEs) used for in vitro assays (e.g., for wound healing) make use of primary human skin cells. Limitations of primary keratinocytes and fibroblasts include availability of donor skin and donor variation. The use of physiologically relevant cell lines could solve these limitations. The aim was to develop a fully differentiated HSE constructed entirely from human skin cell lines, which could be applied for in vitro wound-healing assays. Skin equivalents were constructed from human TERT-immortalized keratinocytes and fibroblasts (TERT-HSE) and compared with native skin and primary HSEs. HSEs were characterized by hematoxylin-eosin and immunohistochemical stainings with markers for epidermal proliferation and differentiation, basement membrane (BM), fibroblasts, and the extracellular matrix (ECM). Ultrastructure was determined with electron microscopy. To test the functionality of the TERT-HSE, burn and cold injuries were applied, followed by immunohistochemical stainings, measurement of reepithelialization, and determination of secreted wound-healing mediators. The TERT-HSE was composed of a fully differentiated epidermis and a fibroblast-populated dermis comparable to native skin and primary HSE. The epidermis consisted of proliferating keratinocytes within the basal layer, followed by multiple spinous layers, a granular layer, and cornified layers. Within the TERT-HSE, the membrane junctions such as corneosomes, desmosomes, and hemidesmosomes were well developed as shown by ultrastructure pictures. Furthermore, the BM consisted of a lamina lucida and lamina densa comparable to native skin. The dermal matrix of the TERT-HSE was more similar to native skin than the primary construct, since collagen III, an ECM marker, was present in TERT-HSEs and absent in primary HSEs. After wounding, the TERT-HSE was able to reepithelialize and secrete inflammatory wound-healing mediators. In conclusion, the novel TERT-HSE, constructed entirely from human cell lines, provides an excellent opportunity to study in vitro skin biology and can also be used for drug targeting and testing new therapeutics, and ultimately, for incorporating into skin-on-a chip in the future.

Isolation and culture of rabbit embryonic stem cells.

PubMed

Honda, Arata

2013-01-01

Mammalian stem cells are invaluable research resources for the study of cell and embryonic development as well as practical tools for use in the production of genetically engineered animals and further therapeutics. It is important that we further our knowledge and understanding of a variety of stem cells from several different animal species before trials in humans commence. Here we describe methods for establishing rabbit embryonic stem (rES) cell lines with indefinite proliferation potential. rES cells attain maximum proliferation potential when cultured at a feeder cell density of one-sixth of that of full confluency. Higher and lower densities of feeder cells induced ES cell differentiation or division arrest. Fibroblast growth factor (FGF)2 can maintain the undifferentiated status of rES cells; however leukemia inhibitory factor (LIF) is dispensable. Under optimized conditions, rES cells could be passaged by trypsinization 50 times. This culture system enabled efficient gene transduction and clonal expansion from single cells. rES cells grew as flat monolayer cell colonies, as reported for monkey and human ES cells, and expressed pluripotency markers. Embryoid bodies and teratomas formed readily in vitro and in vivo, respectively. Characterization of ES cells from different species is important for establishing common features of pluripotency. We have demonstrated the similarity of ES cells between rabbit and humans. These cell lines could be applied directly using gene-targeting techniques, or in combination with induced pluripotent stem cells. Thus, rES cells are a suitable model for studying human transplantation therapy and disease treatments.

Elimination of remaining undifferentiated induced pluripotent stem cells in the process of human cardiac cell sheet fabrication using a methionine-free culture condition.

PubMed

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.

Establishment of Trophectoderm Cell Lines from Buffalo (Bubalus bubalis) Embryos of Different Sources and Examination of In Vitro Developmental Competence, Quality, Epigenetic Status and Gene Expression in Cloned Embryos Derived from Them

PubMed Central

Mohapatra, Sushil Kumar; Sandhu, Anjit; Singh, Karn Pratap; Singla, Suresh Kumar; Chauhan, Manmohan Singh; Manik, Radheysham; Palta, Prabhat

2015-01-01

Despite being successfully used to produce live offspring in many species, somatic cell nuclear transfer (NT) has had a limited applicability due to very low (>1%) live birth rate because of a high incidence of pregnancy failure, which is mainly due to placental dysfunction. Since this may be due to abnormalities in the trophectoderm (TE) cell lineage, TE cells can be a model to understand the placental growth disorders seen after NT. We isolated and characterized buffalo TE cells from blastocysts produced by in vitro fertilization (TE-IVF) and Hand-made cloning (TE-HMC), and compared their growth characteristics and gene expression, and developed a feeder-free culture system for their long-term culture. The TE-IVF cells were then used as donor cells to produce HMC embryos following which their developmental competence, quality, epigenetic status and gene expression were compared with those of HMC embryos produced using fetal or adult fibroblasts as donor cells. We found that although TE-HMC and TE-IVF cells have a similar capability to grow in culture, significant differences exist in gene expression levels between them and between IVF and HMC embryos from which they are derived, which may have a role in the placental abnormalities associated with NT pregnancies. Although TE cells can be used as donor cells for producing HMC blastocysts, their developmental competence and quality is lower than that of blastocysts produced from fetal or adult fibroblasts. The epigenetic status and expression level of many important genes is different in HMC blastocysts produced using TE cells or fetal or adult fibroblasts or those produced by IVF. PMID:26053554

Establishment of Trophectoderm Cell Lines from Buffalo (Bubalus bubalis) Embryos of Different Sources and Examination of In Vitro Developmental Competence, Quality, Epigenetic Status and Gene Expression in Cloned Embryos Derived from Them.

PubMed

Mohapatra, Sushil Kumar; Sandhu, Anjit; Singh, Karn Pratap; Singla, Suresh Kumar; Chauhan, Manmohan Singh; Manik, Radheysham; Palta, Prabhat

2015-01-01

Despite being successfully used to produce live offspring in many species, somatic cell nuclear transfer (NT) has had a limited applicability due to very low (>1%) live birth rate because of a high incidence of pregnancy failure, which is mainly due to placental dysfunction. Since this may be due to abnormalities in the trophectoderm (TE) cell lineage, TE cells can be a model to understand the placental growth disorders seen after NT. We isolated and characterized buffalo TE cells from blastocysts produced by in vitro fertilization (TE-IVF) and Hand-made cloning (TE-HMC), and compared their growth characteristics and gene expression, and developed a feeder-free culture system for their long-term culture. The TE-IVF cells were then used as donor cells to produce HMC embryos following which their developmental competence, quality, epigenetic status and gene expression were compared with those of HMC embryos produced using fetal or adult fibroblasts as donor cells. We found that although TE-HMC and TE-IVF cells have a similar capability to grow in culture, significant differences exist in gene expression levels between them and between IVF and HMC embryos from which they are derived, which may have a role in the placental abnormalities associated with NT pregnancies. Although TE cells can be used as donor cells for producing HMC blastocysts, their developmental competence and quality is lower than that of blastocysts produced from fetal or adult fibroblasts. The epigenetic status and expression level of many important genes is different in HMC blastocysts produced using TE cells or fetal or adult fibroblasts or those produced by IVF.

Human fibroblast matrices bio-assembled under macromolecular crowding support stable propagation of human embryonic stem cells.

PubMed

Peng, Yanxian; Bocker, Michael Thomas; Holm, Jennifer; Toh, Wei Seong; Hughes, Christopher Stephen; Kidwai, Fahad; Lajoie, Gilles Andre; Cao, Tong; Lyko, Frank; Raghunath, Michael

2012-11-01

Stable pluripotent feeder-free propagation of human embryonic stem cells (hESCs) prior to their therapeutic applications remains a major challenge. Matrigel™ (BD Singapore) is a murine sarcoma-derived extracellular matrix (ECM) widely used as a cell-free support combined with conditioned or chemically defined media; however, inherent xenogenic and immunological threats invalidate it for clinical applications. Using human fibrogenic cells to generate ECM is promising but currently suffers from inefficient and time-consuming deposition in vitro. We recently showed that macromolecular crowding (MMC) accelerated ECM deposition substantially in vitro. In the current study, we used dextran sulfate 500 kDa as a macromolecular crowder to induce WI-38 fetal human lung fibroblasts at 0.5% serum condition to deposit human ECM in three days. After decellularization, the generated ECMs allowed stable propagation of H9 hESCs over 20 passages in chemically-defined medium (mTEsR1) with an overall improved outcome compared to Matrigel in terms of population doubling while retaining teratoma formation and differentiation capacity. Of significance, only ECMs generated by MMC allowed the successful propagation of hESCs. ECMs were highly complex and in contrast to Matrigel, contained no vitronectin but did contain collagen XII, ig-h3 and novel for hESC-supporting human matrices, substantial amounts of transglutaminase 2. Genome-wide analysis of promoter DNA methylation states revealed high overall similarity between human ECM- and Matrigel-cultured hESCs; however, distinct differences were observed with 49 genes associated with a variety of cellular functions. Thus, human ECMs deposited by MMC by selected fibroblast lines are a suitable human microenvironment for stable hESC propagation and clinically translational settings. Copyright © 2012 John Wiley & Sons, Ltd.

Selection of appropriate isolation method based on morphology of blastocyst for efficient derivation of buffalo embryonic stem cells.

PubMed

Kumar, R; Ahlawat, S P S; Sharma, M; Verma, O P; Sai Kumar, G; Taru Sharma, G

2014-03-01

The efficiency of embryonic stem cell (ESC) derivation from all species except for rodents and primates is very low. There are however, multiple interests in obtaining pluripotent cells from these animals with main expectations in the fields of transgenesis, cloning, regenerative medicine and tissue engineering. Researches are being carried out in laboratories throughout the world to increase the efficiency of ESC isolation for their downstream applications. Thus, the present study was undertaken to study the effect of different isolation methods based on the morphology of blastocyst for efficient derivation of buffalo ESCs. Embryos were produced in vitro through the procedures of maturation, fertilization and culture. Hatched blastocysts or isolated inner cell masses (ICMs) were seeded on mitomycin-C inactivated buffalo fetal fibroblast monolayer for the development of ESC colonies. The ESCs were analyzed for alkaline phosphatase activity, expression of pluripotency markers and karyotypic stability. Primary ESC colonies were obtained after 2-5 days of seeding hatched blastocysts or isolated ICMs on mitomycin-C inactivated feeder layer. Mechanically isolated ICMs attached and formed primary cell colonies more efficiently than ICMs isolated enzymatically. For derivation of ESCs from poorly defined ICMs intact hatched blastocyst culture was the most successful method. Results of this study implied that although ESCs can be obtained using all three methods used in this study, efficiency varies depending upon the morphology of blastocyst and isolation method used. So, appropriate isolation method must be selected depending on the quality of blastocyst for efficient derivation of ESCs.

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

PubMed

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.

Regulation by basic fibroblast growth factor of glycosaminoglycan biosynthesis in cultured vascular endothelial cells.

PubMed

Kaji, T; Hiraga, S; Ohkawara, S; Inada, M; Yamamoto, C; Kozuka, H; Koizumi, F

1995-05-01

The alteration of glycosaminoglycans (GAGs) in cultured bovine aortic endothelial cells after exposure to basic fibroblast growth factor (bFGF) was investigated. It was found that the incorporation of [3H]glucosamine into GAGs was markedly increased by bFGF in both the cell layer and the conditioned medium; however, that of [35S]sulfate was not changed by the growth factor. These results indicated that bFGF enhanced the sugar-chain formation but did not affect their sulfation in endothelial GAG production. Similar changes were observed in either bovine aortic smooth-muscle cells and human fibroblastic IMR-90 cells to greater and lesser degrees, respectively. Characterization of GAGs in the endothelial cell layer and the conditioned medium revealed that bFGF enhanced both heparan sulfate and the other GAGs to a similar degree. The present data suggest that bFGF may be involved in the regulation of the blood coagulation system via altering GAGs of the vascular tissue when the endothelium was damaged.

Evaluation of ERDA-sponsored coal feed system development

NASA Technical Reports Server (NTRS)

Phen, R. L.; Luckow, W. K.; Mattson, L.; Otth, D.; Tsou, P.

1977-01-01

Coal feeders were evaluated based upon criteria such as technical feasibility, performance (i.e. ability to meet process requirements), projected life cycle costs, and projected development cost. An initial set of feeders was selected based on the feeders' cost savings potential compared with baseline lockhopper systems. Additional feeders were considered for selection based on: (1) increasing the probability of successful feeder development; (2) application to specific processes; and (3) technical merit. A coal feeder development program is outlined.

Skin Equivalent Tissue-Engineered Construct: Co-Cultured Fibroblasts/ Keratinocytes on 3D Matrices of Sericin Hope Cocoons

PubMed Central

Nayak, Sunita; Dey, Sancharika; Kundu, Subhas C.

2013-01-01

The development of effective and alternative tissue-engineered skin replacements to autografts, allografts and xenografts has became a clinical requirement due to the problems related to source of donor tissue and the perceived risk of disease transmission. In the present study 3D tissue engineered construct of sericin is developed using co-culture of keratinocytes on the upper surface of the fabricated matrices and with fibroblasts on lower surface. Sericin is obtained from “Sericin Hope” silkworm of Bombyx mori mutant and is extracted from cocoons by autoclave. Porous sericin matrices are prepared by freeze dried method using genipin as crosslinker. The matrices are characterized biochemically and biophysically. The cell proliferation and viability of co-cultured fibroblasts and keratinocytes on matrices for at least 28 days are observed by live/dead assay, Alamar blue assay, and by dual fluorescent staining. The growth of the fibroblasts and keratinocytes in co-culture is correlated with the expression level of TGF-β, b-FGF and IL-8 in the cultured supernatants by enzyme-linked immunosorbent assay. The histological analysis further demonstrates a multi-layered stratified epidermal layer of uninhibited keratinocytes in co-cultured constructs. Presence of involucrin, collagen IV and the fibroblast surface protein in immuno-histochemical stained sections of co-cultured matrices indicates the significance of paracrine signaling between keratinocytes and fibroblasts in the expression of extracellular matrix protein for dermal repair. No significant amount of pro inflammatory cytokines (TNF-α, IL-1β and nitric oxide) production are evidenced when macrophages grown on the sericin matrices. The results all together depict the potentiality of sericin 3D matrices as skin equivalent tissue engineered construct in wound repair. PMID:24058626

Skin equivalent tissue-engineered construct: co-cultured fibroblasts/ keratinocytes on 3D matrices of sericin hope cocoons.

PubMed

Nayak, Sunita; Dey, Sancharika; Kundu, Subhas C

2013-01-01

The development of effective and alternative tissue-engineered skin replacements to autografts, allografts and xenografts has became a clinical requirement due to the problems related to source of donor tissue and the perceived risk of disease transmission. In the present study 3D tissue engineered construct of sericin is developed using co-culture of keratinocytes on the upper surface of the fabricated matrices and with fibroblasts on lower surface. Sericin is obtained from "Sericin Hope" silkworm of Bombyx mori mutant and is extracted from cocoons by autoclave. Porous sericin matrices are prepared by freeze dried method using genipin as crosslinker. The matrices are characterized biochemically and biophysically. The cell proliferation and viability of co-cultured fibroblasts and keratinocytes on matrices for at least 28 days are observed by live/dead assay, Alamar blue assay, and by dual fluorescent staining. The growth of the fibroblasts and keratinocytes in co-culture is correlated with the expression level of TGF-β, b-FGF and IL-8 in the cultured supernatants by enzyme-linked immunosorbent assay. The histological analysis further demonstrates a multi-layered stratified epidermal layer of uninhibited keratinocytes in co-cultured constructs. Presence of involucrin, collagen IV and the fibroblast surface protein in immuno-histochemical stained sections of co-cultured matrices indicates the significance of paracrine signaling between keratinocytes and fibroblasts in the expression of extracellular matrix protein for dermal repair. No significant amount of pro inflammatory cytokines (TNF-α, IL-1β and nitric oxide) production are evidenced when macrophages grown on the sericin matrices. The results all together depict the potentiality of sericin 3D matrices as skin equivalent tissue engineered construct in wound repair.

Pre-treatment of synthetic elastomeric scaffolds by cardiac fibroblasts improves engineered heart tissue

PubMed Central

Radisic, Milica; Park, Hyoungshin; Martens, Timothy P.; Salazar-Lazaro, Johanna E.; Geng, Wenliang; Wang, Yadong; Langer, Robert; Freed, Lisa E.; Vunjak-Novakovic, Gordana

2009-01-01

Native myocardium consists of several cell types, of which approximately one-third are myocytes and most of the nonmyocytes are fibroblasts. By analogy with monolayer culture in which fibroblasts were removed to prevent overgrowth, early attempts to engineer myocardium utilized cell populations enriched for cardiac myocytes (CMs; ~80–90% of total cells). We hypothesized that the pre-treatment of synthetic elastomeric scaffolds with cardiac fibroblasts (CFs) will enhance the functional assembly of the engineered cardiac constructs by creating an environment supportive of cardiomyocyte attachment and function. Cells isolated from neonatal rat ventricles were prepared to form three distinct populations: rapidly plating cells identified as CFs, slowly plating cells identified as CMs, and unseparated initial population of cells (US). The cell fractions (3 × 106 cells total) were seeded into poly(glycerol sebacate) scaffolds (highly porous discs, 5 mm in diameter × 2-mm thick) using Matrigel™, either separately (CM or CF), concurrently (US), or sequentially (CF pre-treatment followed by CM culture, CF + CM), and cultured in spinner flasks. The CF + CM group had the highest amplitude of contraction and the lowest excitation threshold, superior DNA content, and higher glucose consumption rate. The CF + CM group exhibited compact 100- to 200-μm thick layers of elongated myocytes aligned in parallel over layers of collagen-producing fibroblasts, while US and CM groups exhibited scattered and poorly elongated myocytes. The sequential co-culture of CF and CM on a synthetic elastomer scaffold thus created an environment supportive of cardiomyocyte attachment, differentiation, and contractile function, presumably due to scaffold conditioning by cultured fibroblasts. When implanted over the infarcted myocardium in a nude rat model, cell-free poly(glycerol sebacate) remained at the ventricular wall after 2 weeks of in vivo, and was vascularized. PMID:18041719

Colour preferences of UK garden birds at supplementary seed feeders.

PubMed

Rothery, Luke; Scott, Graham W; Morrell, Lesley J

2017-01-01

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.

Precision powder feeder

DOEpatents

Schlienger, M. Eric; Schmale, David T.; Oliver, Michael S.

2001-07-10

A new class of precision powder feeders is disclosed. These feeders provide a precision flow of a wide range of powdered materials, while remaining robust against jamming or damage. These feeders can be precisely controlled by feedback mechanisms.

Substrate effect modulates adhesion and proliferation of fibroblast on graphene layer.

PubMed

Lin, Feng; Du, Feng; Huang, Jianyong; Chau, Alicia; Zhou, Yongsheng; Duan, Huiling; Wang, Jianxiang; Xiong, Chunyang

2016-10-01

Graphene is an emerging candidate for biomedical applications, including biosensor, drug delivery and scaffold biomaterials. Cellular functions and behaviors on different graphene-coated substrates, however, still remain elusive to a great extent. This paper explored the functional responses of cells such as adhesion and proliferation, to different kinds of substrates including coverslips, silicone, polydimethylsiloxane (PDMS) with different curing ratios, PDMS treated with oxygen plasma, and their counterparts coated with single layer graphene (SLG). Specifically, adherent cell number, spreading area and cytoskeleton configuration were exploited to characterize cell-substrate adhesion ability, while MTT assay was employed to test the proliferation capability of fibroblasts. Experimental outcome demonstrated graphene coating had excellent cytocompatibility, which could lead to an increase in early adhesion, spreading, proliferation, and remodeling of cytoskeletons of fibroblast cells. Notably, it was found that the underlying substrate effect, e.g., stiffness of substrate materials, could essentially regulate the adhesion and proliferation of cells cultured on graphene. The stiffer the substrates were, the stronger the abilities of adhesion and proliferation of fibroblasts were. This study not only deepens our understanding of substrate-modulated interfacial interactions between live cells and graphene, but also provides a valuable guidance for the design and application of graphene-based biomaterials in biomedical engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

Atmospheric Characteristics of Cool Season Intermittent Precipitation Near Portland, Oregon

NASA Astrophysics Data System (ADS)

Cunningham, Jeffrey Glenn

Pacific Northwest cool season precipitation is often described as mostly stratiform (i.e. steady and continuous). While most regional precipitation is stratiform in terms of area and duration, embedded convective cells within stratiform precipitation occur frequently enough to warrant study. Embedded cells locally increase rain rate, total precipitation, and streamflow discharge and hence raise the risk of flooding, landslides, and debris flows. Analysis of vertically pointing radar data near Portland, Oregon for three cool seasons (2005 to 2008) indicates that fallstreaks in the snow layer, locally enhanced precipitation regions a few kilometers in size indicated in radar reflectivity data above the 0° C altitude, are nearly ubiquitous on days with significant rainfall accumulation and large areas of precipitation. The observed fallstreaks in snow enhance rainfall immediately below the snow fallstreak. Compared to stratiform periods, embedded convective periods include higher Doppler vertical velocity values and higher variability in velocities especially in the snow layer. The combination of these findings points to generating cells within the snow layer and the seeder-feeder mechanism as important sources of surface precipitation variability for periods of embedded convective cells within stratiform precipitation. The primary goal of this study was to determine the sources of instability typically associated with convective cells embedded within stratiform precipitation for Pacific Northwest cool season storms. Storm periods occurring over six cool seasons (2002 to 2008, totaling 1923 hours) of operational radar data (KRTX) and 166 upper air soundings (KSLE) are analyzed. A new method was employed to objectively determine the degree of precipitation intermittency in sequences of radar scans. The resulting continuum of intermittency values was grouped into four categories: mostly convective precipitation, mostly stratiform precipitation, embedded convective cells within stratiform precipitation, and other. Atmospheric soundings during periods with embedded convective cells within stratiform precipitation are more likely to have convective available potential energy (CAPE) than soundings during periods of mostly stratiform precipitation. Specifically, most unstable parcel CAPE (MUCAPE) > 0 J kg-1 occurs 2.8 more frequently during embedded periods than for mostly stratiform periods. Over 90% of embedded periods have MUCAPE > 0 J kg-1 or at least two 500 meter layers of potential instability. In contrast to the near surface based instability most commonly associated with the mostly convective precipitation, embedded convection is elevated. The median most unstable parcel height of origin for embedded convective periods is 2.5 km compared to 0.5 km for mostly convective periods. Although this present research did not deal directly with orographic precipitation enhancement, it does address synoptic and mesoscale precipitation processes that frequently occur near terrain in the Pacific Northwest. The exclusion of the seeder-feeder mechanism as a mode of cellularity for orographic precipitation in recent work is inconsistent with the observations presented here and inconsistent with much of the pre-2000 literature, which show the seeder-feeder mechanism directly modulating surface rain rate with or without terrain present. Numerical models, whether operational or idealized, need to represent the seeder-feeder process in order to accurately simulate precipitation variability at small spatial scales (less than < 5-10 km) and temporal scales (< 3 hours) within the warm sector of Pacific Northwest extratropical cyclones.

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.

PubMed

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 modulated the effect of the number of feeder places/pig on FCR (P < 0.05), with a gradual reduction of FCR in entire males and females when increasing feeder space allowance. Genetics tended to modulate the effect of diets' CP content on FCR (P < 0.10). Overall, these results may contribute to the improvement of the welfare and performance of growing-finishing pigs by a better knowledge of the influence of the rearing environment and may help optimize the feeding strategies in current production systems.

Encapsulation of basic fibroblast growth factor by polyelectrolyte multilayer microcapsules and its controlled release for enhancing cell proliferation.

PubMed

She, Zhen; Wang, Chunxia; Li, Jun; Sukhorukov, Gleb B; Antipina, Maria N

2012-07-09

Basic fibroblast growth factor (FGF2) is an important protein for cellular activity and highly vulnerable to environmental conditions. FGF2 protected by heparin and bovine serum albumin was loaded into the microcapsules by a coprecipitation-based layer-by-layer encapsulation method. Low cytotoxic and biodegradable polyelectrolytes dextran sulfate and poly-L-arginine were used for capsule shell assembly. The shell thickness-dependent encapsulation efficiency was measured by enzyme-linked immunosorbent assay. A maximum encapsulation efficiency of 42% could be achieved by microcapsules with a shell thickness of 14 layers. The effects of microcapsule concentration and shell thickness on cytotoxicity, FGF2 release kinetics, and L929 cell proliferation were evaluated in vitro. The advantage of using microcapsules as the carrier for FGF2 controlled release for enhancing L929 cell proliferation was analyzed.

30 CFR 77.1802 - Insulation of trolley wires, trolley feeder wires and bare signal wires; guarding of trolley...

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Insulation of trolley wires, trolley feeder wires and bare signal wires; guarding of trolley wires and trolley feeder wires. 77.1802 Section 77.1802... Wires and Trolley Feeder Wires § 77.1802 Insulation of trolley wires, trolley feeder wires and bare...

30 CFR 77.1802 - Insulation of trolley wires, trolley feeder wires and bare signal wires; guarding of trolley...

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Insulation of trolley wires, trolley feeder wires and bare signal wires; guarding of trolley wires and trolley feeder wires. 77.1802 Section 77.1802... Wires and Trolley Feeder Wires § 77.1802 Insulation of trolley wires, trolley feeder wires and bare...

Improved Small-Particle Powders for Plasma Spraying

NASA Technical Reports Server (NTRS)

Nguyen, QuynhGiao, N.; Miller, Robert A.; Leissler, George W.

2005-01-01

Improved small-particle powders and powder-processing conditions have been developed for use in plasma spray deposition of thermal-barrier and environmental barrier coatings. Heretofore, plasma-sprayed coatings have typically ranged in thickness from 125 to 1,800 micrometers. As explained below, the improved powders make it possible to ensure complete coverage of substrates at unprecedently small thicknesses of the order of 25 micrometers. Plasma spraying involves feeding a powder into a hot, high-velocity plasma jet. The individual powder particles melt in the plasma jet as they are propelled towards a substrate, upon which they splat to build up a coating. In some cases, multiple coating layers are required. The size range of the powder particles necessarily dictates the minimum thickness of a coating layer needed to obtain uniform or complete coverage. Heretofore, powder particle sizes have typically ranged from 40 to 70 micrometers; as a result, the minimum thickness of a coating layer for complete coverage has been about 75 micrometers. In some applications, thinner coatings or thinner coating layers are desirable. In principle, one can reduce the minimum complete-coverage thickness of a layer by using smaller powder particles. However, until now, when powder particle sizes have been reduced, the powders have exhibited a tendency to cake, clogging powder feeder mechanisms and feed lines. Hence, the main problem is one of synthesizing smaller-particle powders having desirable flow properties. The problem is solved by use of a process that begins with a spray-drying subprocess to produce spherical powder particles having diameters of less than 30 micrometers. (Spherical-particle powders have the best flow properties.) The powder is then passed several times through a commercial sifter with a mesh to separate particles having diameters less than 15 micrometers. The resulting fine, flowable powder is passed through a commercial fluidized bed powder feeder into a plasma spray jet.

Feeder density enhances house finch disease transmission in experimental epidemics.

PubMed

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).

Colour preferences of UK garden birds at supplementary seed feeders

PubMed Central

Rothery, Luke; Scott, Graham W.

2017-01-01

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. PMID:28212435

Feed intake and behavior of dairy goats when offered an elevated feed bunk.

PubMed

Neave, Heather W; von Keyserlingk, Marina A G; Weary, Daniel M; Zobel, Gosia

2018-04-01

Goats are browsers and select vegetation at various heights when foraging. On commercial farms, dairy goats are typically fed from low-level feed bunks. The objective of this study was to determine how feed intake and feeding behavior vary when goats are offered feed at variable heights, with the potential of evaluating the benefits of offering an elevated feeder to dairy goats. Thirteen Saanen X dairy goats were housed in a home pen with a lying area of wood shavings, where they were pre-exposed for 24 d to 3 feeder heights designed to result in differences in head height while feeding: floor level (head lowered relative to body), head level (head level relative to body), and elevated level (head and neck angled upward). Nine groups of 3 goats each were randomly selected and housed for 24 h in a test pen identical to the home pen except that it contained 1 of each of the 3 feeder heights. Each feeder contained ad libitum chopped alfalfa silage and a top-dressed corn-based supplement, refreshed twice daily. Refusals from inside and under each feeder were weighed to calculate intake. Feed intake increased with increasing feeder height (mean ± SE; 0.18, 0.29, and 0.34 ± 0.04 kg of DM/goat for floor-level, head-level, and elevated-level feeders, respectively). Total feeding time did not vary with feeder height, but feeding rate tended to be faster at the elevated-level feeder (14.5 ± 2.1 g of DM/min) compared with head-level (9.2 ± 2.3 g of DM/min) and floor-level (8.9 ± 2.1 g of DM/min) feeders. Goats visited the floor-level feeder (36.4 ± 8.4 visits/goat) less than the head-level (79.4 ± 8.4 visits/goat) and elevated-level (74.8 ± 8.4 visits/goat) feeders. The number of displacements per minute of feeding time (physical removal of another goat from the feeding place) was greater at the elevated-level feeder (0.46 ± 0.06 displacements/min) compared with the floor-level feeder (0.23 ± 0.06 displacements/min) and tended to differ from the head-level feeder (0.27 ± 0.06 displacements/min). We conclude that goats eat more from an elevated feeder and compete more to access this feeder. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Secretion of wound healing mediators by single and bi-layer skin substitutes.

PubMed

Maarof, Manira; Law, Jia Xian; Chowdhury, Shiplu Roy; Khairoji, Khairul Anuar; Saim, Aminuddin Bin; Idrus, Ruszymah Bt Hj

2016-10-01

Limitations of current treatments for skin loss caused by major injuries leads to the use of skin substitutes. It is assumed that secretion of wound healing mediators by these skin substitutes plays a role in treating skin loss. In our previous study, single layer keratinocytes (SK), single layer fibroblast (SF) and bilayer (BL; containing keratinocytes and fibroblasts layers) skin substitutes were fabricated using fibrin that had shown potential to heal wounds in preclinical studies. This study aimed to quantify the secretion of wound healing mediators, and compare between single and bi-layer skin substitutes. Skin samples were digested to harvest fibroblasts and keratinocytes, and expanded to obtain sufficient cells for the construction of skin substitutes. Acellular fibrin (AF) construct was used as control. Substitutes i.e. AF, SK, SF and BL were cultured for 2 days, and culture supernatant was collected to analyze secretion of wound healing mediators via multiplex ELISA. Among 19 wound healing mediators tested, BL substitute secreted significantly higher amounts of CXCL1 and GCSF compared to SF and AF substitute but this was not significant with respect to SK substitute. The BL substitute also secreted significantly higher amounts of CXCL5 and IL-6 compared to other substitutes. In contrast, the SK substitute secreted significantly higher amounts of VCAM-1 compared to other substitutes. However, all three skin substitutes also secreted CCL2, CCL5, CCL11, GM-CSF, IL8, IL-1α, TNF-α, ICAM-1, FGF-β, TGF-β, HGF, VEGF-α and PDGF-BB factors, but no significant difference was seen. Secretion of these mediators after transplantation may play a significant role in promoting wound healing process for the treatment of skin loss.

Use of versican variant V3 and versican antisense expression to engineer cultured human skin containing increased content of insoluble elastin.

PubMed

Merrilees, Mervyn J; Falk, Ben A; Zuo, Ning; Dickinson, Michelle E; May, Barnaby C H; Wight, Thomas N

2017-01-01

Skin substitutes for repair of dermal wounds are deficient in functional elastic fibres. We report that the content of insoluble elastin in the dermis of cultured human skin can be increased though the use of two approaches that enhance elastogenesis by dermal fibroblasts, forced expression of versican variant V3, which lacks glycosaminoglycan (GAG) chains, and forced expression of versican antisense to decrease levels of versican variant V1 with GAG chains. Human dermal fibroblasts transduced with V3 or anti-versican were cultured under standard conditions over a period of 4 weeks to produce dermal sheets, with growth enhanced though multiple seedings for the first 3 weeks. Human keratinocytes, cultured in supplemented media, were added to the 4-week dermal sheets and the skin layer cultured for a further week. At 5 weeks, keratinocytes were multilayered and differentiated, with desmosome junctions thoughout and keratin deposits in the upper squamous layers. The dermal layer was composed of layered fibroblasts surrounded by extracellular matrix of collagen bundles and, in control cultures, small scattered elastin deposits. Forced expression of V3 and versican antisense slowed growth, decreased versican V1 expression, increased tropoelastin expression and/or the deposition of large aggregates of insoluble elastin in the dermal layer, and increased tissue stiffness, as measured by nano-indentation. Skin sheets were also cultured on Endoform Dermal Template™, the biodegradable wound dressing made from the lamina propria of sheep foregut. Skin structure and the enhanced deposition of elastin by forced expression of V3 and anti-versican were preserved on this supportive substrate. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

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

PubMed

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 feeders improved the germ cell/somatic cell ratio. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Thermal imaging for assessment of electron-beam freeform fabrication (EBF3) additive manufacturing deposits

NASA Astrophysics Data System (ADS)

Zalameda, Joseph N.; Burke, Eric R.; Hafley, Robert A.; Taminger, Karen M.; Domack, Christopher S.; Brewer, Amy; Martin, Richard E.

2013-05-01

Additive manufacturing is a rapidly growing field where 3-dimensional parts can be produced layer by layer. NASA's electron beam freeform fabrication (EBF3) technology is being evaluated to manufacture metallic parts in a space environment. The benefits of EBF3 technology are weight savings to support space missions, rapid prototyping in a zero gravity environment, and improved vehicle readiness. The EBF3 system is composed of 3 main components: electron beam gun, multi-axis position system, and metallic wire feeder. The electron beam is used to melt the wire and the multi-axis positioning system is used to build the part layer by layer. To insure a quality deposit, a near infrared (NIR) camera is used to image the melt pool and solidification areas. This paper describes the calibration and application of a NIR camera for temperature measurement. In addition, image processing techniques are presented for deposit assessment metrics.

Ascorbate-apatite composite and ascorbate-FGF-2-apatite composite layers formed on external fixation rods and their effects on cell activity in vitro.

PubMed

Wang, Xiupeng; Ito, Atsuo; Sogo, Yu; Li, Xia; Tsurushima, Hideo; Oyane, Ayako

2009-09-01

Ascorbate-apatite and ascorbate-fibroblast growth factor-2 (FGF-2)-apatite composite layers were successfully formed on anodically oxidized Ti rods clinically used for external fixation by a one-step procedure at 25 degrees C, using a metastable supersaturated calcium phosphate solution supplemented with l-ascorbic acid phosphate magnesium salt n-hydrate (AsMg) and FGF-2. The AsMg-apatite and AsMg-FGF-2-apatite composite layers were evaluated in vitro using fibroblastic NIH3T3 and osteoblastic MC3T3-E1 cells. The AsMg-FGF-2-apatite composite layer markedly enhanced the NIH3T3 cell proliferation and procollagen type capital I, Ukrainian gene expression. Without FGF-2, the AsMg-apatite composite layer whose ascorbate content was 3.64+/-1.27microgcm(-2) obviously enhanced osteoblastic proliferation and differentiation. However, the AsMg-FGF-2-apatite composite layers whose FGF-2 contents were from 0.15+/-0.03 to 0.31+/-0.04microgcm(-2) inhibited osteoblastic differentiation in vitro. Thus, the AsMg-FGF-2-apatite composite layer should be precipitated on the surface of external fixators attached to skin and soft tissue. On the other hand, the AsMg-apatite composite layer should be precipitated at the part attached to bone tissue.

35. INTERIOR VIEW, SAME AS ABOVE WITH THE FEEDER ROD ...

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

35. INTERIOR VIEW, SAME AS ABOVE WITH THE FEEDER ROD INSERTED INTO THE MACHINE; NOTE THE WOOD FEEDER ROD AND FEEDER ROD REST ON THE BACK OF THE STOOL - LaBelle Iron Works, Thirtieth & Wood Streets, Wheeling, Ohio County, WV

Analytic Considerations and Design Basis for the IEEE Distribution Test Feeders

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

Schneider, K. P.; Mather, B. A.; Pal, B. C.

For nearly 20 years the Test Feeder Working Group of the Distribution System Analysis Subcommittee has been developing openly available distribution test feeders for use by researchers. The purpose of these test feeders is to provide models of distribution systems that reflect the wide diversity in design and their various analytic challenges. Because of their utility and accessibility, the test feeders have been used for a wide range of research, some of which has been outside the original scope of intended uses. This paper provides an overview of the existing distribution feeder models and clarifies the specific analytic challenges thatmore » they were originally designed to examine. Additionally, the paper will provide guidance on which feeders are best suited for various types of analysis. The purpose of this paper is to provide the original intent of the Working Group and to provide the information necessary so that researchers may make an informed decision on which of the test feeders are most appropriate for their work.« less

Analytic Considerations and Design Basis for the IEEE Distribution Test Feeders

DOE PAGES

Schneider, K. P.; Mather, B. A.; Pal, B. C.; ...

2017-10-10

For nearly 20 years the Test Feeder Working Group of the Distribution System Analysis Subcommittee has been developing openly available distribution test feeders for use by researchers. The purpose of these test feeders is to provide models of distribution systems that reflect the wide diversity in design and their various analytic challenges. Because of their utility and accessibility, the test feeders have been used for a wide range of research, some of which has been outside the original scope of intended uses. This paper provides an overview of the existing distribution feeder models and clarifies the specific analytic challenges thatmore » they were originally designed to examine. Additionally, the paper will provide guidance on which feeders are best suited for various types of analysis. The purpose of this paper is to provide the original intent of the Working Group and to provide the information necessary so that researchers may make an informed decision on which of the test feeders are most appropriate for their work.« less

Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions.

PubMed

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.

Lipid profiling of parkin-mutant human skin fibroblasts.

PubMed

Lobasso, Simona; Tanzarella, Paola; Vergara, Daniele; Maffia, Michele; Cocco, Tiziana; Corcelli, Angela

2017-12-01

Parkin mutations are a major cause of early-onset Parkinson's disease (PD). The impairment of protein quality control system together with defects in mitochondria and autophagy process are consequences of the lack of parkin, which leads to neurodegeneration. Little is known about the role of lipids in these alterations of cell functions. In the present study, parkin-mutant human skin primary fibroblasts have been considered as cellular model of PD to investigate on possible lipid alterations associated with the lack of parkin protein. Dermal fibroblasts were obtained from two unrelated PD patients with different parkin mutations and their lipid compositions were compared with that of two control fibroblasts. The lipid extracts of fibroblasts have been analyzed by combined matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) and thin-layer chromatography (TLC). In parallel, we have performed direct MALDI-TOF/MS lipid analyses of intact fibroblasts by skipping lipid extraction steps. Results show that the proportions of some phospholipids and glycosphingolipids were altered in the lipid profiles of parkin-mutant fibroblasts. The detected higher level of gangliosides, phosphatidylinositol, and phosphatidylserine could be linked to dysfunction of autophagy and mitochondrial turnover; in addition, the lysophosphatidylcholine increase could represent the marker of neuroinflammatory state, a well-known component of PD. © 2017 Wiley Periodicals, Inc.

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

PubMed

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 hummingbirds carried no pollen. Mist nets located 3km from La Georgina returned few captures (one-to-three specimens) per sampling date, contrasting with observations made before feeders were present. These results suggest that sugar-water feeders gather hummingbirds in over considerable distances drawing them away from flowers. The competitive and antagonistic pattern shown between feeders and flowers indicate that natural pollination system could be significantly altered. Supplementing hummingbirds with food seems likely to interfere with pollination networks already stressed by many anthropogenic effects.

Production of Zebrafish Offspring from Cultured Female Germline Stem Cells

PubMed Central

Wong, Ten-Tsao; Tesfamichael, Abraham; Collodi, Paul

2013-01-01

Zebrafish female germline stem cell (FGSC) cultures were generated from a transgenic line of fish that expresses Neo and DsRed under the control of the germ cell specific promoter, ziwi [Tg(ziwi:neo);Tg(ziwi:DsRed)]. Homogeneous FGSC cultures were established by G418 selection and continued to express ziwi for more than 6 weeks along with the germ cell markers nanos3, dnd, dazl and vasa. A key component of the cell culture system was the use of a feeder cell line that was initiated from ovaries of a transgenic line of fish [Tg(gsdf:neo)] that expresses Neo controlled by the zebrafish gonadal soma derived factor (gsdf) promoter. The feeder cell line was selected in G418 and engineered to express zebrafish leukemia inhibitory factor (Lif), basic fibroblast growth factor (Fgf2) and glial-cell-line derived neurotrophic factor (Gdnf). These factors were shown to significantly enhance FGSC growth, survival and germline competency in culture. Results from cell transplantation experiments revealed that the cultured FGSCs were able to successfully colonize the gonad of sterile recipient fish and generate functional gametes. Up to 20% of surviving recipient fish that were injected with the cultured FGSCs were fertile and generated multiple batches of normal offspring for at least 6 months. The FGSC cultures will provide an in vitro system for studies of zebrafish germ cell growth and differentiation and their high frequency of germline transmission following transplantation could form the basis of a stem cell-mediated strategy for gene transfer and manipulation of the zebrafish genome. PMID:23671620

A composite chitosan-gelatin bi-layered, biomimetic macroporous scaffold for blood vessel tissue engineering.

PubMed

Badhe, Ravindra V; Bijukumar, Divya; Chejara, Dharmesh R; Mabrouk, Mostafa; Choonara, Yahya E; Kumar, Pradeep; du Toit, Lisa C; Kondiah, Pierre P D; Pillay, Viness

2017-02-10

A composite chitosan-gelatin macroporous hydrogel-based scaffold with bi-layered tubular architecture was engineered by solvent casting-co-particulate leaching. The scaffold constituted an inner macroporous layer concealed by a non-porous outer layer mimicking the 3D matrix of blood vessels with cellular adhesion and proliferation. The scaffold was evaluated for its morphological, physicochemical, physicomechanical and biodurability properties employing SEM, FTIR, DSC, XRD, porositometry, rheology and texture analysis. The fluid uptake and biodegradation in the presence of lysozymes was also investigated. Cellular attachment and proliferation was analysed using human dermal fibroblasts (HDF-a) seeded onto the scaffold and evaluated by MTT assay, SEM, and confocal microscopy. Results demonstrated that the scaffold had a desirable tensile strength=95.81±11kPa, elongation at break 112.5±13%, porosity 82% and pores between 100 and 230μm, 50% in vitro biodegradation at day 16 and proliferated fibroblasts over 20 days. These results demonstrate that scaffold may be an excellent tubular archetype for blood vessel tissue engineering. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

PubMed Central

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

Enhanced cell adhesion on severe peened-plasma nitrided 316L stainless steel

NASA Astrophysics Data System (ADS)

Jayalakshmi, M.; Bhat, Badekai Ramachandra; Bhat, K. Udaya

2018-04-01

Plasma nitriding is an effective technique to enhance the wear resistance of austenitic stainless steels. Recently, severe surface deformation techniques are extensively used prior to nitriding to enhance diffusion kinetics. In the present study, AISI 316L austenitic stainless steel is subjected to peening-nitriding duplex treatment and biocompatibility of treated surfaces is assessed through adhesion of the fibroblast cells. Three-fold increase in the surface microhardness is observed from the un-peened sample to the peened-nitrided sample; with severe peened sample showing intermediate hardness. Similar trend is observed in the number of the fibroblast cells attached to the sample surface. Spreading of some of the fibroblast cells is observed on the sample subjected to duplex treatment; while the other two samples showed only the spindle shaped fibroblasts. Combined influence of surface nanocrystallization and presence of nitride layer is responsible for the improved biocompatibility.

Analysis of material particle motion and optimizing parameters of vibration of two-mass GZS vibratory feeder

NASA Astrophysics Data System (ADS)

Nguyen, Van Xo; Golikov, N. S.

2018-05-01

The structure and kinematics of the two-mass GZS vibratory feeder operation are considered. It is established that the movement of the material's particles on the feeder surface determines its capacity. The development and analysis of the mathematical model of material's particle movement on the two-mass GZS vibratory feeder surface are shown. The results of Matlab optimization of material particles velocity function are given that allows setting rational kinematics of the feeder.

ITER Magnet Feeder: Design, Manufacturing and Integration

NASA Astrophysics Data System (ADS)

CHEN, Yonghua; ILIN, Y.; M., SU; C., NICHOLAS; BAUER, P.; JAROMIR, F.; LU, Kun; CHENG, Yong; SONG, Yuntao; LIU, Chen; HUANG, Xiongyi; ZHOU, Tingzhi; SHEN, Guang; WANG, Zhongwei; FENG, Hansheng; SHEN, Junsong

2015-03-01

The International Thermonuclear Experimental Reactor (ITER) feeder procurement is now well underway. The feeder design has been improved by the feeder teams at the ITER Organization (IO) and the Institute of Plasma Physics, Chinese Academy of Sciences (ASIPP) in the last 2 years along with analyses and qualification activities. The feeder design is being progressively finalized. In addition, the preparation of qualification and manufacturing are well scheduled at ASIPP. This paper mainly presents the design, the overview of manufacturing and the status of integration on the ITER magnet feeders. supported by the National Special Support for R&D on Science and Technology for ITER (Ministry of Public Security of the People's Republic of China-MPS) (No. 2008GB102000)

Dry coal feeder development program at Ingersoll-Rand Research, Incorporated. [for coal gasification systems

NASA Technical Reports Server (NTRS)

Mistry, D. K.; Chen, T. N.

1977-01-01

A dry coal screw feeder for feeding coal into coal gasification reactors operating at pressures up to 1500 psig is described. Results on the feeder under several different modes of operation are presented. In addition, three piston feeder concepts and their technical and economical merits are discussed.

Using supplemental food and its influence on survival of northern bobwhite (Colinus virginianus)

USGS Publications Warehouse

Townsend, D.E.; Lochmiller, R.L.; DeMaso, S.J.; Leslie, David M.; Peoples, A.D.; Scott, A C.; Parry, E.S.

2000-01-01

Biologists have debated the effectiveness of supplemental feeders as a management tool for the northern bobwhite (Colinus virginianus), but few extensive evaluations have been conducted. We examined 783 crops from harvested bobwhites during 1992-1996 to determine effects of climatic stress in winter on use of supplemental feeders and their impact on survival rate in winter. Crops of bobwhites harvested from areas with supplemental feeders contained 28.2% supplemental food compared with 5.5% (P<0.001) for those from areas without supplemental feeders. Winter climate was not a significant predictor of the proportional use of supplemental feeders. Rates of winter survival were greater on areas with supplemental feeders compared with non-supplemented areas in winters 1992-1993 (P=0.001) and 1993-1994 (P=0.002), but in 1994-1995, rates were greater on nonsupplemented areas (P=0.032). Cause-specific mortality rates indicated that supplemental feeders did not predispose bobwhites to hunter harvest or predators. Results suggested that bobwhites can gain nutritional benefits from supplemental feeders during times of severe winter stress.

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

PubMed

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.

Exclusion of broiler breeder females from male feeders with a male only grill. 1. Behavioral and technical aspects of design and development.

PubMed

Brake, J; Khamidullin, T N; Fisinin, V I

1993-03-01

A series of experiments was conducted in order to develop a feeder grill that would allow broiler breeder males, but not females, to eat. Females are typically excluded from male feeders by increasing the height of the feeder. However, male feeder heights that exclude most females increase the time required for males to consume a given amount of feed 19 to 60%. Typical male feeder height from floor to feeder pan lip in commercial practice is 55 cm, which is about the distance from floor to the head of the male in an erect posture. Females measure about 40 cm in an erect posture. Because the necks of males (8.0 cm) are longer than those of females (6.3 cm), the lip of male feeder pans was extended horizontally 12.7 cm with a wire mesh on the assumption that females could not stretch their necks enough to reach the feed. However, the females learned quickly to perch on the extended lip and eat. Total exclusion of females from the male feeder, regardless of feeder height, was achieved by placing a horizontal upper mesh 5.6 to 10.2 cm above the extended lip, and connecting the upper mesh to the lower extended lip of the feeder with vertical bars spaced 5.1 cm apart. The horizontal upper mesh prevented perching by females and the 5.1-cm spacing of the vertical bars allowed males but not females to reach the feed by inserting their heads and necks up to their shoulders.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of long-term culture of mouse embryonic stem cells under low oxygen concentration as well as on glycosaminoglycan hyaluronan on cell proliferation and differentiation.

PubMed

Ramírez, M Á; Pericuesta, E; Yáñez-Mó, M; Palasz, A; Gutiérrez-Adán, A

2011-02-01

Maintaining undifferentiated stem cells in defined conditions is of critical importance to improve their in vitro culture. We have evaluated the effects of culturing mouse stem (mES) cells under physiological oxygen concentration as well as by replacing fibroblast feeder layer (mEF) with gelatin or glycosaminoglycan hyaluronan (HA), on cell proliferation and differentiation. After 3 days culture or after long-term cell culture under different conditions, levels of apoptotic cell death were determined by cell cycle and TUNEL (TdT-mediated dUTP nick end labelling) assays and levels of cell proliferation by CFSE (5-(and-6)-carboxyfluorescein diacetate succinimidyl ester) labelling. We assessed spontaneous differentiation into cardiomyocytes and mRNA expression of pluripotency and differentiation biomarkers. After 3 days culture under hypoxic conditions, levels of proliferation and apoptosis of mES cells were higher, in correlation with increase in intracellular reactive oxygen species. However, when cells were continuously grown for 1 month under those conditions, the level of apoptosis was, in all cases, under 4%. Hypoxia reduced spontaneous differentiation of mES into cardiomyocytes. Long-term culture on HA was more effective in maintaining the pluripotent state of the mES cells when compared to that on gelatin. Level of terminal differentiation was highest on mEF, intermediate on HA and lowest on gelatin. Our data suggest that hypoxia is not necessary for maintaining pluripotency of mES cells and appeared to be detrimental during ES differentiation. Moreover, HA may offer a valuable alternative for long-term culture of mES cells in vitro. © 2010 Blackwell Publishing Ltd.

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.

PubMed

Javed, M Shahid; Yaqoob, Naeem; Iwamuro, Masaya; Kobayashi, Naoya; Fujiwara, Toshiyoshi

2014-02-01

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. An experimental study. Department of Surgery, Okayama University, Graduate School of Medicine, Japan, from April to December 2011. 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. 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. 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.

Periodic harvesting of embryonic stem cells from a hollow-fiber membrane based four-compartment bioreactor.

PubMed

Knöspel, Fanny; Freyer, Nora; Stecklum, Maria; Gerlach, Jörg C; Zeilinger, Katrin

2016-01-01

Different types of stem cells have been investigated for applications in drug screening and toxicity testing. In order to provide sufficient numbers of cells for such in vitro applications a scale-up of stem cell culture is necessary. Bioreactors for dynamic three-dimensional (3D) culture of growing cells offer the option for culturing large amounts of stem cells at high densities in a closed system. We describe a method for periodic harvesting of pluripotent stem cells (PSC) during expansion in a perfused 3D hollow-fiber membrane bioreactor, using mouse embryonic stem cells (mESC) as a model cell line. A number of 100 × 10(6) mESC were seeded in bioreactors in the presence of mouse embryonic fibroblasts (MEF) as feeder cells. Over a cultivation interval of nine days cells were harvested by trypsin perfusion and mechanical agitation every second to third culture day. A mean of 380 × 10(6) mESC could be removed with every harvest. Subsequent to harvesting, cells continued growing in the bioreactor, as determined by increasing glucose consumption and lactate production. Immunocytochemical staining and mRNA expression analysis of markers for pluripotency and the three germ layers showed a similar expression of most markers in the harvested cells and in mESC control cultures. In conclusion, successful expansion and harvesting of viable mESC from bioreactor cultures with preservation of sterility was shown. The present study is the first one showing the feasibility of periodic harvesting of adherent cells from a continuously perfused four-compartment bioreactor including further cultivation of remaining cells. © 2015 American Institute of Chemical Engineers.

Derivation and characterization of novel nonhuman primate embryonic stem cell lines from in vitro-fertilized baboon preimplantation embryos.

PubMed

Chang, Tien-Cheng; Liu, Ya-Guang; Eddy, Carlton A; Jacoby, Ethan S; Binkley, Peter A; Brzyski, Robert G; Schenken, Robert S

2011-06-01

The development of nonhuman primate (NHP) embryonic stem cell (ESC) models holds great promise for cell-mediated treatment of debilitating diseases and to address numerous unanswered questions regarding the therapeutic efficacy of ESCs while supplanting ethical considerations involved with human studies. Here we report successful establishment and characterization of 3 novel baboon (Papio cynocephalus) ESC lines from the inner cell mass of intracytoplasmic sperm injection-derived blastocysts. Embryos were cultured in an improved baboon embryo in vitro culture protocol. The inner cell mass of blastocyst was laser-dissected and plated on mouse embryonic fibroblast feeder cell monolayer in the NHP ESC culture medium. Three cell lines with characteristic ESC morphology have been cultured through an extended period (>14 months), with 2 male cell lines (UT-1 and -2) and 1 female cell line (UT-3) displaying normal baboon karyotypes. Reverse transcription-polymerase chain reaction analysis confirmed that all 3 lines express primate ESC pluripotency markers, including OCT-4, NANOG, SOX-2, TERT, TDGF, LEFTYA, and REX-1. All 3 lines demonstrated positive immunocytochemical staining for OCT-4, stage-specific embryonic antigen-3, stage-specific embryonic antigen-4, TRA-1-60, and TRA-1-81. Baboon ESCs injected into NOD/SCID mice formed teratomas with all 3 germ layers. In addition, embryoid body-like spherical structures were derived and initial outgrowth was observed when embedded into extracellular matrix Matrigel. The ESC lines established in this NHP model have the potential to extend our knowledge in the fields of developmental biology, regenerative medicine, and future applications, including preclinical safety assessment of in vivo stem cell therapy.

Down-regulation of Pax6 is associated with abnormal differentiation of corneal epithelial cells in severe ocular surface diseases

PubMed Central

Li, W; Chen, Y-T; Hayashida, Y; Blanco, G; Kheirkah, A; He, H; Chen, S-Y; Liu, C-Y; Tseng, SCG

2010-01-01

Pax6 is the universal master control gene for eye morphogenesis. Other than retina and lens, Pax6 also expressed in the ocular surface epithelium from early gestation until the postnatal stage, in which little is known about the function of Pax6. In this study, corneal pannus tissues from patients with ocular surface diseases such as Stevens–Johnson syndrome (SJS), chemical burn, aniridia and recurrent pterygium were investigated. Our results showed that normal ocular surface epithelial cells expressed Pax6. However, corneal pannus epithelial cells from the above patients showed a decline or absence of Pax6 expression, accompanied by a decline or absence of K12 keratin but an increase of K10 keratin and filaggrin expression. Pannus basal epithelial cells maintained nuclear p63 expression and showed activated proliferation, evidenced by positive Ki67 and K16 keratin staining. On 3T3 fibroblast feeder layers, Pax6 immunostaining was negative in clones generated from epithelial cells harvested from corneal pannus from SJS or aniridia, but positive in those from the normal limbal epithelium; whereas western blots showed that some epithelial clones expanded from pannus retained Pax6 expression. Transient transfection of an adenoviral vector carrying EGFP–Pax6 transgenes into these Pax6− clones increased both Pax6 and K12 keratin expression. These results indicate that Pax6 helps to maintain the normal corneal epithelial phenotype postnatally, and that down-regulation of Pax6 is associated with abnormal epidermal differentiation in severe ocular surface diseases. Reintroduction of activation of the Pax6 gene might be useful in treating squamous metaplasia of the ocular surface epithelium. PMID:18027901

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

PubMed

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 significantly and negatively associated with neutering rate. These findings are in accordance with the suggestion that neutering may potentially improve cat welfare by reducing morbidity. Collaboration by the authorities with these heavy feeders, who represent a small number of FRC feeders and feed substantial FRC numbers, may be significant for the control and monitoring of FRC populations and their resources.

Preclinical study of mouse pluripotent parthenogenetic embryonic stem cell derivatives for the construction of tissue-engineered skin equivalent.

PubMed

Rao, Yang; Cui, Jihong; Yin, Lu; Liu, Wei; Liu, Wenguang; Sun, Mei; Yan, Xingrong; Wang, Ling; Chen, Fulin

2016-10-22

Embryonic stem cell (ESC) derivatives hold great promise for the construction of tissue-engineered skin equivalents (TESE). However, harvesting of ESCs destroys viable embryos and may lead to political and ethical concerns over their application. In the current study, we directed mouse parthenogenetic embryonic stem cells (pESCs) to differentiate into fibroblasts, constructed TESE, and evaluated its function in vivo. The stemness marker expression and the pluripotent differentiation ability of pESCs were tested. After embryoid body (EB) formation and adherence culture, mesenchymal stem cells (MSCs) were enriched and directed to differentiate into fibroblastic lineage. Characteristics of derived fibroblasts were assessed by quantitative real-time PCR and ELISA. Functional ability of the constructed TESE was tested by a mouse skin defects repair model. Mouse pESCs expressed stemness marker and could form teratoma containing three germ layers. MSCs could be enriched from outgrowths of EBs and directed to differentiate into fibroblastic lineage. These cells express a high level of growth factors including FGF, EGF, VEGF, TGF, PDGF, and IGF1, similar to those of ESC-derived fibroblasts and mouse fibroblasts. Seeded into collagen gels, the fibroblasts derived from pESCs could form TESE. Mouse skin defects could be successfully repaired 15 days after transplantation of TESE constructed by fibroblasts derived from pESCs. pESCs could be induced to differentiate into fibroblastic lineage, which could be applied to the construction of TESE and skin defect repair. Particularly, pESC derivatives avoid the limitations of political and ethical concerns, and provide a promising source for regenerative medicine.

30 CFR 75.1003-1 - Other requirements for guarding of trolley wires and trolley feeder wires.

Code of Federal Regulations, 2010 CFR

2010-07-01

... wires and trolley feeder wires. 75.1003-1 Section 75.1003-1 Mineral Resources MINE SAFETY AND HEALTH... Trolley Wires and Trolley Feeder Wires § 75.1003-1 Other requirements for guarding of trolley wires and trolley feeder wires. Adequate precaution shall be taken to insure that equipment being moved along...

30 CFR 75.1003-1 - Other requirements for guarding of trolley wires and trolley feeder wires.

Code of Federal Regulations, 2011 CFR

2011-07-01

... wires and trolley feeder wires. 75.1003-1 Section 75.1003-1 Mineral Resources MINE SAFETY AND HEALTH... Trolley Wires and Trolley Feeder Wires § 75.1003-1 Other requirements for guarding of trolley wires and trolley feeder wires. Adequate precaution shall be taken to insure that equipment being moved along...

Polymeric nanofibrous substrates stimulate pluripotent stem cells to form three-dimensional multilayered patty-like spheroids in feeder-free culture and maintain their pluripotency.

PubMed

Alamein, Mohammad A; Wolvetang, Ernst J; Ovchinnikov, Dmitry A; Stephens, Sebastien; Sanders, Katherine; Warnke, Patrick H

2015-09-01

Expansion of pluripotent stem cells in defined media devoid of animal-derived feeder cells to generate multilayered three-dimensional (3D) bulk preparations or spheroids, rather than two-dimensional (2D) monolayers, is advantageous for many regenerative, biological or disease-modelling studies. Here we show that electrospun polymer matrices comprised of nanofibres that mimic the architecture of the natural fibrous extracellular matrix allow for feeder-free expansion of pluripotent human induced pluripotent stem cells (IPSCs) and human embryonic stem cells (HESCs) into multilayered 3D 'patty-like' spheroid structures in defined xeno-free culture medium. The observation that IPSCs and HESCs readily revert to 2D growth in the absence of the synthetic nanofibre membranes suggests that this 3D expansion behaviour is mediated by the physical microenvironment and artificial niche provided by the nanofibres only. Importantly, we could show that such 3D growth as patties maintained the pluripotency of cells as long as they were kept on nanofibres. The generation of complex multilayered 3D structures consisting of only pluripotent cells on biodegradable nanofibre matrices of the desired shape and size will enable both industrial-scale expansion and intricate organ-tissue engineering applications with human pluripotent stem cells, where simultaneous coupling of differentiation pathways of all germ layers from one stem cell source may be required for organ formation. Copyright © 2014 John Wiley & Sons, Ltd.

Higher trophic level affects nutrient, silicon, metal(loid), and radionuclide mobilization from freshwater sediments

NASA Astrophysics Data System (ADS)

Schaller, Jörg; Planer-Friedrich, Britta

2017-04-01

Organic sediments in aquatic ecosystems are well known sinks for nutrients, silicon, and metal(loid)s. Organic matter-decomposing organisms like invertebrate shredders, grazers, bioturbators, and filter feeder are key-species for the carbon and energy turnover within the decomposer community. We could show that invertebrate shredders and grazer affect element fixation or remobilization by changing binding properties of organic sediments and the attached biofilm. Bioturbators affect element fixation or remobilization by changing redox conditions within the uppermost sediment layer. Last but not least filter feeders, like the zebra mussel Dreissena polymorpha, an invasive organism in North American and European freshwater ecosystems significantly contributed to element mobilization of silicon, iron, phosphorus, arsenic, and copper and to immobilization of uranium (p<0.001), probably driven by redox conditions, microbial activity within the gut system, or active control of element homeostasis. Except of the filter feeder D. polymorpha, the invertebrates are able to minimize the accumulation of non-nutrient elements due to specific strategies, which is an important strategy for species living in systems tending to element accumulation. However, D. polymorpha revealed a significant uptake and accumulation of arsenic, copper, iron, and especially uranium both into the soft body tissues and the seashell. This accumulation by D. polymorpha is in line with previous observations of metal(loid) accumulation from biomonitoring studies. In summary, higher trophic level strongly contributes to element fixation or remobilization in aquatic systems.

[Effects of canopy density on the functional group of soil macro fauna in Pinus massoniana plantations].

PubMed

Zhou, Hong Yang; Zhang, Dan Ju; Zhang, Jie; Zhao, Yan Bo; Zhao, Bo; Wei, Da Ping; Zhang, Jian

2017-06-18

In order to understand the effects of canopy density on the functional group characteristics of soil macrofauna in Pinus massoniana plantations, we divided the captured soil fauna into five types including xylophages, predators, saprophages, omnivores and fungal feeders. The results showed that 1) Saprozoic feeders had the highest percentage of total individuals, and the omnivores and xylophages occupied higher percentages of total taxa. 2) The individual and group number of the predators, and the group number of xylophages did not change significantly under 0.5-0.6 and then decreased significantly under 0.6-0.9 canopy density. 3) With the increasing canopy density, the individual an dgroup number of predators in litter layer decreased significantly, the saprozoic individual number in 5-10 cm soil layer represented irregular trends. The individual number of xylophage increased with the depth of soil, and the group number in litter layer, the individual and group number in 5-10 cm soil layer decreased significantly. 4) Pielou evenness of xylophage had no significant changes with the canopy density, all the other diversity index of xylophage and saprophage were various with the increasing canopy density. The predatory Simpson index was stable under 0.5-0.8, and then decreased significantly under 0.8-0.9 canopy density. 5) The CCA (canonical correlation analysis) indicated that soil bulk density and moisture content were the main environmental factors affecting functional groups of soil macro fauna. Moisture content greatly impacted on the number of saprophagous individuals. But xylophage and predators were mostly affected by soil bulk density, and the predatory Simpson index was mainly affected by soil pH value and total phosphorus. Our research indicated that the structure of soil macro faunal functional group under 0.7 canopy density was comparatively stable, which would facilitate the maintenance of soil fertility and ecological function in Pinus massoniana plantation.

Effect of in vitro gingival fibroblast seeding on the in vivo incorporation of acellular dermal matrix allografts in dogs.

PubMed

Novaes, Arthur B; Marchesan, Julie Teresa; Macedo, Guilherme O; Palioto, Daniela B

2007-02-01

Acellular dermal matrix allograft (ADMA) has been used in various periodontal procedures with successful results. Because ADMA has no blood vessels or cells, slower healing and incorporation are observed compared to a subepithelial connective tissue graft. Fibroblasts accelerate the healing process by regulation of matrix deposition and synthesis of a variety of growth factors. Thus, the objective of this study was to evaluate histologically if gingival fibroblasts affect healing and incorporation of ADMA in dogs when used as a subepithelial allograft. Gingival fibroblasts were established from explant culture from the connective tissue of keratinized gingiva collected from the maxilla of seven mongrel dogs. ADMA was seeded with gingival fibroblasts and transferred to dogs. Surgery was performed bilaterally, and the regions were divided into two groups: ADMA+F (ADMA containing fibroblasts) and ADMA (ADMA only). Biopsies were performed after 2, 4, and 8 weeks of healing. The quantity of blood vessels was significantly higher in the ADMA+F group at 2 weeks of healing (Kruskal-Wallis; P <0.05). There was no statistical difference (P >0.05) in the number of cell layers, epithelial area, or inflammatory infiltrate between the two groups at any stage of healing. The enhanced vascularization in vivo in early stages supports the important role of fibroblasts in improving graft performance and wound healing of cultured graft substitutes.

Increased efficiency of gamma-irradiated versus mitomycin C-treated feeder cells for the expansion of normal human cells in long-term cultures.

PubMed

Roy, A; Krzykwa, E; Lemieux, R; Néron, S

2001-12-01

Several normal human cells, such as hematopoietic stem cells, dendritic cells, and B cells, can be cultured in vitro in defined optimal conditions. Several ex vivo culture systems require the use of feeder cells to support the growth of target cells. In such systems, proliferation of feeder cells has to be stopped, so that they can be used as nonreplicating viable support cells. Because feeder cells need to provide one or few active signals, it is important to maintain them in an metabolically active state, allowing continued expression of specific ligands or cytokines. Mitomycin C and gamma-irradiation treatments are commonly used to prepare nonproliferating feeder cells and are usually considered to be equivalent. Normal human B lymphocytes can be expanded in vitro in the presence of feeder cells expressing the CD40 ligand CD154. Here we compared the ability of gamma-irradiation- and mitomycin C-treated feeder cells to support the expansion of normal human B lymphocytes. The results indicate that expansion of B cells during a long-term culture was 100 times more potent using gamma-irradiated feeder cells compared to mitomycin C-treated cells. This difference could be related to a significant reduction in both cellular metabolism and level of CD154 expression observed in mitomycin C-treated feeder cells, but not in gamma-irradiated cells nor in control untreated cells. These results indicate that mitomycin C-treated feeder cells are metabolically altered, and consequently less efficient at maintaining cell expansion in the long-term cell culture system used.

30 CFR 75.1003 - Insulation of trolley wires, trolley feeder wires and bare signal wires; guarding of trolley...

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Insulation of trolley wires, trolley feeder wires and bare signal wires; guarding of trolley wires and trolley feeder wires. 75.1003 Section 75.1003... MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Trolley Wires and Trolley Feeder Wires § 75.1003...

30 CFR 75.1003 - Insulation of trolley wires, trolley feeder wires and bare signal wires; guarding of trolley...

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Insulation of trolley wires, trolley feeder wires and bare signal wires; guarding of trolley wires and trolley feeder wires. 75.1003 Section 75.1003... MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Trolley Wires and Trolley Feeder Wires § 75.1003...

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

PubMed

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.

MAME Models for 4D Live-cell Imaging of Tumor: Microenvironment Interactions that Impact Malignant Progression

PubMed Central

Sameni, Mansoureh; Anbalagan, Arulselvi; Olive, Mary B.; Moin, Kamiar; Mattingly, Raymond R.; Sloane, Bonnie F.

2012-01-01

We have developed 3D coculture models, which we term MAME (mammary architecture and microenvironment engineering), and used them for live-cell imaging in real-time of cell:cell interactions. Our overall goal was to develop models that recapitulate the architecture of preinvasive breast lesions to study their progression to an invasive phenotype. Specifically, we developed models to analyze interactions among pre-malignant breast epithelial cell variants and other cell types of the tumor microenvironment that have been implicated in enhancing or reducing the progression of preinvasive breast epithelial cells to invasive ductal carcinomas. Other cell types studied to date are myoepithelial cells, fibroblasts, macrophages and blood and lymphatic microvascular endothelial cells. In addition to the MAME models, which are designed to recapitulate the cellular interactions within the breast during cancer progression, we have developed comparable models for the progression of prostate cancers. Here we illustrate the procedures for establishing the 3D cocultures along with the use of live-cell imaging and a functional proteolysis assay to follow the transition of cocultures of breast ductal carcinoma in situ (DCIS) cells and fibroblasts to an invasive phenotype over time, in this case over twenty-three days in culture. The MAME cocultures consist of multiple layers. Fibroblasts are embedded in the bottom layer of type I collagen. On that is placed a layer of reconstituted basement membrane (rBM) on which DCIS cells are seeded. A final top layer of 2% rBM is included and replenished with every change of media. To image proteolysis associated with the progression to an invasive phenotype, we use dye-quenched (DQ) fluorescent matrix proteins (DQ-collagen I mixed with the layer of collagen I and DQ-collagen IV mixed with the middle layer of rBM) and observe live cultures using confocal microscopy. Optical sections are captured, processed and reconstructed in 3D with Volocity visualization software. Over the course of 23 days in MAME cocultures, the DCIS cells proliferate and coalesce into large invasive structures. Fibroblasts migrate and become incorporated into these invasive structures. Fluorescent proteolytic fragments of the collagens are found in association with the surface of DCIS structures, intracellularly, and also dispersed throughout the surrounding matrix. Drugs that target proteolytic, chemokine/cytokine and kinase pathways or modifications in the cellular composition of the cocultures can reduce the invasiveness, suggesting that MAME models can be used as preclinical screens for novel therapeutic approaches. PMID:22371028

MAME models for 4D live-cell imaging of tumor: microenvironment interactions that impact malignant progression.

PubMed

Sameni, Mansoureh; Anbalagan, Arulselvi; Olive, Mary B; Moin, Kamiar; Mattingly, Raymond R; Sloane, Bonnie F

2012-02-17

We have developed 3D coculture models, which we term MAME (mammary architecture and microenvironment engineering), and used them for live-cell imaging in real-time of cell:cell interactions. Our overall goal was to develop models that recapitulate the architecture of preinvasive breast lesions to study their progression to an invasive phenotype. Specifically, we developed models to analyze interactions among pre-malignant breast epithelial cell variants and other cell types of the tumor microenvironment that have been implicated in enhancing or reducing the progression of preinvasive breast epithelial cells to invasive ductal carcinomas. Other cell types studied to date are myoepithelial cells, fibroblasts, macrophages and blood and lymphatic microvascular endothelial cells. In addition to the MAME models, which are designed to recapitulate the cellular interactions within the breast during cancer progression, we have developed comparable models for the progression of prostate cancers. Here we illustrate the procedures for establishing the 3D cocultures along with the use of live-cell imaging and a functional proteolysis assay to follow the transition of cocultures of breast ductal carcinoma in situ (DCIS) cells and fibroblasts to an invasive phenotype over time, in this case over twenty-three days in culture. The MAME cocultures consist of multiple layers. Fibroblasts are embedded in the bottom layer of type I collagen. On that is placed a layer of reconstituted basement membrane (rBM) on which DCIS cells are seeded. A final top layer of 2% rBM is included and replenished with every change of media. To image proteolysis associated with the progression to an invasive phenotype, we use dye-quenched (DQ) fluorescent matrix proteins (DQ-collagen I mixed with the layer of collagen I and DQ-collagen IV mixed with the middle layer of rBM) and observe live cultures using confocal microscopy. Optical sections are captured, processed and reconstructed in 3D with Volocity visualization software. Over the course of 23 days in MAME cocultures, the DCIS cells proliferate and coalesce into large invasive structures. Fibroblasts migrate and become incorporated into these invasive structures. Fluorescent proteolytic fragments of the collagens are found in association with the surface of DCIS structures, intracellularly, and also dispersed throughout the surrounding matrix. Drugs that target proteolytic, chemokine/cytokine and kinase pathways or modifications in the cellular composition of the cocultures can reduce the invasiveness, suggesting that MAME models can be used as preclinical screens for novel therapeutic approaches.

Emplacement of Zebín Hill, Jičín Volcanic Field, Bohemian Paradise, Czech Republic: Anisotropy of Magnetic Susceptibility, Ground Magnetometry, Electric Resistivity Tomography, and Paleomagnetic Data

NASA Astrophysics Data System (ADS)

Petronis, M. S.; Rapprich, , V.; Valenta, J.; Leman, J.; Brister, A. R.; van Wyk de Vries, B.

2014-12-01

A well-preserved set of mid-Miocene tuff-cones and their feeders outcrop in the Jičín Volcanic Field, Czech Republic. Zebín Hill is a tuff cone that has been quarried to reveal the volcanoes feeder system. This edifice offers the opportunity to understand how magma is transported through a monogenetic pyroclastic cone. Rock types include a coarse-grained basal phreatomagmatic layer and a stratified upper wall facies both of which are penetrated by feeder dikes. Anisotropy of magnetic susceptibility (AMS) and paleomagnetic data were collected at twenty-one sites from feeder dikes and the main conduit of the volcano. A high-resolution ground magnetometry survey, electric resistivity tomography and seismic tomography were also conducted. Magnetic susceptibility intensity indicates that the dominant magnetic mineral is a ferromagnetic phase with little contribution from paramagnetic minerals. AMS ellipsoids shapes are both oblate and prolate and inferred magma flow directions indicate magma flow away from the central vent area and subhorizontal flow towards and away from the axial conduit; both upward and downward magma flow is evident at some sites. Curie point estimates yield a spectrum of results indicating a mixture of high-Ti titanomagnetite, iron sulfide, and low-Ti titanomagnetite. Ground magnetometry data indicate that both normal and reverse polarity rocks are present at Zebín Hill. Paleomagnetic data confirm the ground magnetic data in that both normal and reverse polarity rocks are present. Most sites yield a single component magnetization that is well grouped at the site level and carried by pseudosingle domain titanomagnetite. The presence of both normal and reverse polarity magnetizations from the volcano indicate that significant time passed during the growth of this monogenic system. Complex system of branching dikes has been also observed from electric resistivity tomography. The simple external structure of monogenetic volcanoes hides a rather complex magmatic plumbing system that dynamically evolves during the life of the volcano. As we show, the well-exposed roots of Zebín Hill reveals that the growth of a volcano occurs not due to simple central axis feeder systems but rather through interplay of local structures, magmatic effects, and construct evolution during the life of the volcano

Generation of induced pluripotent stem cells as a potential source of hematopoietic stem cells for transplant in PNH patients.

PubMed

Phondeechareon, Tanapol; Wattanapanitch, Methichit; U-Pratya, Yaowalak; Damkham, Chanapa; Klincumhom, Nuttha; Lorthongpanich, Chanchao; Kheolamai, Pakpoom; Laowtammathron, Chuti; Issaragrisil, Surapol

2016-10-01

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemolytic anemia caused by lack of CD55 and CD59 on blood cell membrane leading to increased sensitivity of blood cells to complement. Hematopoietic stem cell transplantation (HSCT) is the only curative therapy for PNH, however, lack of HLA-matched donors and post-transplant complications are major concerns. Induced pluripotent stem cells (iPSCs) derived from patients are an attractive source for generating autologous HSCs to avoid adverse effects resulting from allogeneic HSCT. The disease involves only HSCs and their progeny; therefore, other tissues are not affected by the mutation and may be used to produce disease-free autologous HSCs. This study aimed to derive PNH patient-specific iPSCs from human dermal fibroblasts (HDFs), characterize and differentiate to hematopoietic cells using a feeder-free protocol. Analysis of CD55 and CD59 expression was performed before and after reprogramming, and hematopoietic differentiation. Patients' dermal fibroblasts expressed CD55 and CD59 at normal levels and the normal expression remained after reprogramming. The iPSCs derived from PNH patients had typical pluripotent properties and differentiation capacities with normal karyotype. After hematopoietic differentiation, the differentiated cells expressed early hematopoietic markers (CD34 and CD43) with normal CD59 expression. The iPSCs derived from HDFs of PNH patients have normal levels of CD55 and CD59 expression and hold promise as a potential source of HSCs for autologous transplantation to cure PNH patients.

Cytokeratin expression of engrafted three-dimensional culture tissues using epithelial cells derived from porcine periodontal ligaments.

PubMed

Yamada, Rie; Kitajima, Kayoko; Arai, Kyoko; Igarashi, Masaru

2014-09-01

This study investigated the differentiation and proliferation of epithelial cells derived from periodontal ligaments after three-dimensional culture using collagen gel with fibroblasts in vitro and in vivo. Epithelial cells and fibroblasts were derived from porcine periodontal ligaments. Epithelial cells were labeled using a fluorescent red membrane marker (PKH-26GL) and were seeded onto collagen gel with fibroblasts, followed by incubation in an air-liquid interface for 7 days. Three-dimensional cultures were grafted onto the backs of nude mice and removed at 1, 7, and 14 days after surgery (in vivo model). Unfixed sections (5 μm) were used to detect the presence of red fluorescent cells. Paraffin sections were analyzed histologically and immunohistochemically. Specimens were compared with three-dimensional culture tissues at 8, 14 and 21 days (in vitro model). Grafted three-dimensional cultures formed a stratified epithelial structure similar to skin in vivo. Epithelial cells were sequenced in basal-layer-like structures at 14 days in vivo. Immunohistochemical findings showed that the expression of cytokeratin was detected in the epithelial layer in in vitro and in vivo models. Ck8 + 18 + 19 was expressed in the upper epithelial layer in the in vitro model at 14 and 21 days, but not in vivo. Involucrin was expressed in the certified layers in vitro at 14 days, but not in vivo. Laminin was detected at the dermo-epidermal junction in vivo at 7 and 14 days, but not in vitro. These results suggest that differentiation of three-dimensional culture tissues differs in vivo and in vitro. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Ultrastructural characteristics of the cranial dura mater-arachnoid interface layer.

PubMed

Angelov, D N

1990-01-01

The ultrastructural features of the encephalic dura mater-arachnoid borderline (interface) layer (zone) of rats, rabbits, cats and humans were studied. The rat's interface zone included the electron-lucent epithelium-like arranged fibroblasts of the inner dural layer, the rich in filaments cells of the dural neurothelium, a 20 nm wide intercellular cleft filled with electron-dense material and the dark mitochondria-rich cells of the outer arachnoidal layer; in rabbits and cats, this laminar distinction was less prominent, while in man, it was almost absent.

Functional Investigation of NCI-H460-Inducible Myofibroblasts on the Chemoresistance to VP-16 with a Microfluidic 3D Co-Culture Device

PubMed Central

He, Jiarui; Guo, Zhe; Ying, Li; Xu, Zhiyun; Zhang, Jianing; Lu, Jianxin; Wang, Qi

2013-01-01

Fibroblasts, the major cell type in tumor stroma, are essential for tumor growth and survival, and represent an important therapeutic target for cancers. Here we presented a microfluidic co-culture device in which the three-dimensional (3D) matrix was employed to reconstruct an in vivo-like fibroblast-tumor cell microenvironment for investigation of the role of myofibroblasts induced by lung cancer cells in the chemoresistance to VP-16. Composed of a double-layer chip and an injection pump, the device houses fibroblasts and lung cancer cells co-cultured in 3D matrix and 2D mode to induce fibroblasts to become myofibroblasts with the supplement of the medium continuously. With this device, we verified that the cytokines secreted by lung cancer cells could effectively transform the fibroblasts into myofibroblasts. Moreover, compared to fibroblasts, the myofibroblasts showed higher resistance to anticancer drug VP-16. We also demonstrated that this kind of acquired resistance in myofibroblasts was associated with the expression of Glucose-regulated protein 78 (GP78). We concluded that this device allows for the assay to characterize various cellular events in a single device sequentially, facilitating a better understanding of the interactions among heterotypic cells in a sophisticated microenvironment. PMID:23613925

Reprogramming fibroblasts into induced pluripotent stem cells with Bmi1

PubMed Central

Moon, Jai-Hee; Heo, June Seok; Kim, Jun Sung; Jun, Eun Kyoung; Lee, Jung Han; Kim, Aeree; Kim, Jonggun; Whang, Kwang Youn; Kang, Yong-Kook; Yeo, Seungeun; Lim, Hee-Joung; Han, Dong Wook; Kim, Dong-Wook; Oh, Sejong; Yoon, Byung Sun; Schöler, Hans R; You, Seungkwon

2011-01-01

Somatic cells can be reprogrammed into induced pluripotent stem (iPS) cells by the transcription factors Oct4, Sox2, and Klf4 in combination with c-Myc. Recently, Sox2 plus Oct4 was shown to reprogram fibroblasts and Oct4 alone was able to reprogram mouse and human neural stem cells (NSCs) into iPS cells. Here, we report that Bmi1 leads to the transdifferentiation of mouse fibroblasts into NSC-like cells, and, in combination with Oct4, can replace Sox2, Klf4 and c-Myc during the reprogramming of fibroblasts into iPS cells. Furthermore, activation of sonic hedgehog signaling (by Shh, purmorphamine, or oxysterol) compensates for the effects of Bmi1, and, in combination with Oct4, reprograms mouse embryonic and adult fibroblasts into iPS cells. One- and two-factor iPS cells are similar to mouse embryonic stem cells in their global gene expression profile, epigenetic status, and in vitro and in vivo differentiation into all three germ layers, as well as teratoma formation and germline transmission in vivo. These data support that converting fibroblasts with Bmi1 or activation of the sonic hedgehog pathway to an intermediate cell type that expresses Sox2, Klf4, and N-Myc allows iPS generation via the addition of Oct4. PMID:21709693

Synthesis, characterization, and efficacy of antituberculosis isoniazid zinc aluminum-layered double hydroxide based nanocomposites

PubMed Central

Saifullah, Bullo; El Zowalaty, Mohamed Ezzat; Arulselvan, Palanisamy; Fakurazi, Sharida; Webster, Thomas J; Geilich, Benjamin Mahler; Hussein, Mohd Zobir

2016-01-01

The chemotherapy for tuberculosis (TB) is complicated by its long-term treatment, its frequent drug dosing, and the adverse effects of anti-TB drugs. In this study, we have developed two nanocomposites (A and B) by intercalating the anti-TB drug isoniazid (INH) into Zn/Al-layered double hydroxides. The average size of the nanocomposites was found to bê164 nm. The efficacy of the Zn/Al-layered double hydroxides intercalated INH against Mycobacterium tuberculosis was increased by approximately three times more than free INH. The nanocomposites were also found to be active against Gram-positive and -negative bacteria. Compared to the free INH, the nanodelivery formulation was determined to be three times more biocompatible with human normal lung fibroblast MRC-5 cells and 3T3 fibroblast cells at a very high concentration of 50 µg/mL for up to 72 hours. The in vitro release of INH from the Zn/Al-layered double hydroxides was found to be sustained in human body-simulated buffer solutions of pH 4.8 and 7.4. This research is a step forward in making the TB chemotherapy patient friendly. PMID:27486322

Continuation Power Flow Analysis for PV Integration Studies at Distribution Feeders

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

Wang, Jiyu; Zhu, Xiangqi; Lubkeman, David L.

2017-10-30

This paper presents a method for conducting continuation power flow simulation on high-solar penetration distribution feeders. A load disaggregation method is developed to disaggregate the daily feeder load profiles collected in substations down to each load node, where the electricity consumption of residential houses and commercial buildings are modeled using actual data collected from single family houses and commercial buildings. This allows the modeling of power flow and voltage profile along a distribution feeder on a continuing fashion for a 24- hour period at minute-by-minute resolution. By separating the feeder into load zones based on the distance between the loadmore » node and the feeder head, we studied the impact of PV penetration on distribution grid operation in different seasons and under different weather conditions for different PV placements.« less

Metastable primordial germ cell-like state induced from mouse embryonic stem cells by Akt activation

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

Yamano, Noriko; Kimura, Tohru, E-mail: tkimura@patho.med.osaka-u.ac.jp; Watanabe-Kushima, Shoko

Specification to primordial germ cells (PGCs) is mediated by mesoderm-induction signals during gastrulation. We found that Akt activation during in vitro mesodermal differentiation of embryonic stem cells (ESCs) generated self-renewing spheres with differentiation states between those of ESCs and PGCs. Essential regulators for PGC specification and their downstream germ cell-specific genes were expressed in the spheres, indicating that the sphere cells had commenced differentiation to the germ lineage. However, the spheres did not proceed to spermatogenesis after transplantation into testes. Sphere cell transfer to the original feeder-free ESC cultures resulted in chaotic differentiation. In contrast, when the spheres were culturedmore » on mouse embryonic fibroblasts or in the presence of ERK-cascade and GSK3 inhibitors, reversion to the ESC-like state was observed. These results indicate that Akt signaling promotes a novel metastable and pluripotent state that is intermediate to those of ESCs and PGCs.« less

Conditionally reprogrammed primary airway epithelial cells maintain morphology, lineage and disease specific functional characteristics.

PubMed

Martinovich, Kelly M; Iosifidis, Thomas; Buckley, Alysia G; Looi, Kevin; Ling, Kak-Ming; Sutanto, Erika N; Kicic-Starcevich, Elizabeth; Garratt, Luke W; Shaw, Nicole C; Montgomery, Samuel; Lannigan, Francis J; Knight, Darryl A; Kicic, Anthony; Stick, Stephen M

2017-12-21

Current limitations to primary cell expansion led us to test whether airway epithelial cells derived from healthy children and those with asthma and cystic fibrosis (CF), co-cultured with an irradiated fibroblast feeder cell in F-medium containing 10 µM ROCK inhibitor could maintain their lineage during expansion and whether this is influenced by underlying disease status. Here, we show that conditionally reprogrammed airway epithelial cells (CRAECs) can be established from both healthy and diseased phenotypes. CRAECs can be expanded, cryopreserved and maintain phenotypes over at least 5 passages. Population doublings of CRAEC cultures were significantly greater than standard cultures, but maintained their lineage characteristics. CRAECs from all phenotypes were also capable of fully differentiating at air-liquid interface (ALI) and maintained disease specific characteristics including; defective CFTR channel function cultures and the inability to repair wounds. Our findings indicate that CRAECs derived from children maintain lineage, phenotypic and importantly disease-specific functional characteristics over a specified passage range.

Thermal Imaging for Assessment of Electron-Beam Free Form Fabrication (EBF(sup 3)) Additive Manufacturing Welds

NASA Technical Reports Server (NTRS)

Zalameda, Joseph N.; Burke, Eric R.; Hafley, Robert A.; Taminger, Karen M.; Domack, Christopher S.; Brewer, Amy R.; Martin, Richard E.

2013-01-01

Additive manufacturing is a rapidly growing field where 3-dimensional parts can be produced layer by layer. NASA s electron beam free-form fabrication (EBF(sup 3)) technology is being evaluated to manufacture metallic parts in a space environment. The benefits of EBF(sup 3) technology are weight savings to support space missions, rapid prototyping in a zero gravity environment, and improved vehicle readiness. The EBF(sup 3) system is composed of 3 main components: electron beam gun, multi-axis position system, and metallic wire feeder. The electron beam is used to melt the wire and the multi-axis positioning system is used to build the part layer by layer. To insure a quality weld, a near infrared (NIR) camera is used to image the melt pool and solidification areas. This paper describes the calibration and application of a NIR camera for temperature measurement. In addition, image processing techniques are presented for weld assessment metrics.

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

PubMed

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.

A Paracrine Mechanism Accelerating Expansion of Human Induced Pluripotent Stem Cell-Derived Hepatic Progenitor-Like Cells

PubMed Central

Tsuruya, Kota; Chikada, Hiromi; Ida, Kinuyo; Anzai, Kazuya; Kagawa, Tatehiro; Inagaki, Yutaka; Mine, Tetsuya

2015-01-01

Hepatic stem/progenitor cells in liver development have a high proliferative potential and the ability to differentiate into both hepatocytes and cholangiocytes. In this study, we focused on the cell surface molecules of human induced pluripotent stem (iPS) cell-derived hepatic progenitor-like cells (HPCs) and analyzed how these molecules modulate expansion of these cells. Human iPS cells were differentiated into immature hepatic lineage cells by cytokines. In addition to hepatic progenitor markers (CD13 and CD133), the cells were coimmunostained for various cell surface markers (116 types). The cells were analyzed by flow cytometry and in vitro colony formation culture with feeder cells. Twenty types of cell surface molecules were highly expressed in CD13+CD133+ cells derived from human iPS cells. Of these molecules, CD221 (insulin-like growth factor receptor), which was expressed in CD13+CD133+ cells, was quickly downregulated after in vitro expansion. The proliferative ability was suppressed by a neutralizing antibody and specific inhibitor of CD221. Overexpression of CD221 increased colony-forming ability. We also found that inhibition of CD340 (erbB2) and CD266 (fibroblast growth factor-inducible 14) signals suppressed proliferation. In addition, both insulin-like growth factor (a ligand of CD221) and tumor necrosis factor-like weak inducer of apoptosis (a ligand of CD266) were provided by feeder cells in our culture system. This study revealed the expression profiles of cell surface molecules in human iPS cell-derived HPCs and that the paracrine interactions between HPCs and other cells through specific receptors are important for proliferation. PMID:25808356

A Paracrine Mechanism Accelerating Expansion of Human Induced Pluripotent Stem Cell-Derived Hepatic Progenitor-Like Cells.

PubMed

Tsuruya, Kota; Chikada, Hiromi; Ida, Kinuyo; Anzai, Kazuya; Kagawa, Tatehiro; Inagaki, Yutaka; Mine, Tetsuya; Kamiya, Akihide

2015-07-15

Hepatic stem/progenitor cells in liver development have a high proliferative potential and the ability to differentiate into both hepatocytes and cholangiocytes. In this study, we focused on the cell surface molecules of human induced pluripotent stem (iPS) cell-derived hepatic progenitor-like cells (HPCs) and analyzed how these molecules modulate expansion of these cells. Human iPS cells were differentiated into immature hepatic lineage cells by cytokines. In addition to hepatic progenitor markers (CD13 and CD133), the cells were coimmunostained for various cell surface markers (116 types). The cells were analyzed by flow cytometry and in vitro colony formation culture with feeder cells. Twenty types of cell surface molecules were highly expressed in CD13(+)CD133(+) cells derived from human iPS cells. Of these molecules, CD221 (insulin-like growth factor receptor), which was expressed in CD13(+)CD133(+) cells, was quickly downregulated after in vitro expansion. The proliferative ability was suppressed by a neutralizing antibody and specific inhibitor of CD221. Overexpression of CD221 increased colony-forming ability. We also found that inhibition of CD340 (erbB2) and CD266 (fibroblast growth factor-inducible 14) signals suppressed proliferation. In addition, both insulin-like growth factor (a ligand of CD221) and tumor necrosis factor-like weak inducer of apoptosis (a ligand of CD266) were provided by feeder cells in our culture system. This study revealed the expression profiles of cell surface molecules in human iPS cell-derived HPCs and that the paracrine interactions between HPCs and other cells through specific receptors are important for proliferation.

A Batch Feeder for Inhomogeneous Bulk Materials

NASA Astrophysics Data System (ADS)

Vislov, I. S.; Kladiev, S. N.; Slobodyan, S. M.; Bogdan, A. M.

2016-04-01

The work includes the mechanical analysis of mechanical feeders and batchers that find application in various technological processes and industrial fields. Feeders are usually classified according to their design features into two groups: conveyor-type feeders and non-conveyor feeders. Batchers are used to batch solid bulk materials. Less frequently, they are used for liquids. In terms of a batching method, they are divided into volumetric and weighting batchers. Weighting batchers do not provide for sufficient batching accuracy. Automatic weighting batchers include a mass controlling sensor and systems for automatic material feed and automatic mass discharge control. In terms of operating principle, batchers are divided into gravitational batchers and batchers with forced feed of material using conveyors and pumps. Improved consumption of raw materials, decreased loss of materials, ease of use in automatic control systems of industrial facilities allows increasing the quality of technological processes and improve labor conditions. The batch feeder suggested by the authors is a volumetric batcher that has no comparable counterparts among conveyor-type feeders and allows solving the problem of targeted feeding of bulk material batches increasing reliability and hermeticity of the device.

Functional and aesthetic approach to design of bird feeders

NASA Astrophysics Data System (ADS)

Kukhta, A.; Kukhta, M.

2015-10-01

Anthropogenic objects which load the urban environment negatively affects the human psyche. The alternative is attracting elements of the natural environment into urban environment, of which some of the most frequently identified are birds. Attracting birds in the city is possible by means of feeders and artificial nests, however, both must be harmonious. The aim of this study is to analyze the essential functions of the feeders, and their integration into the environmental design and development of the city. On this basis an original feeder which is convenient for use by birds and attracts people's attention is developed. In this paper we apply comparative analysis of different types of feeders encountered in Tomsk, bird watching, and evaluate usability of different types of feeders from the position of their convenience both for birds and human beings. Historical-cultural analysis for determining features of the architectural and environmental design of Tomsk is carried out, the method allows us to solve engineering problems. In this study the feeder convenient for bird use is designed which blends harmoniously with the architectural design of Tomsk.

Construction and histological analysis of a 3D human arterial wall model containing vasa vasorum using a layer-by-layer technique.

PubMed

Shima, Fumiaki; Narita, Hirokazu; Hiura, Ayami; Shimoda, Hiroshi; Akashi, Mitsuru

2017-03-01

There is considerable global demand for three-dimensional (3D) functional tissues which mimic our native organs and tissues for use as in vitro drug screening systems and in regenerative medicine. In particular, there has been an increasing number of patients who suffer from arterial diseases such as arteriosclerosis. As such, in vitro 3D arterial wall models that can evaluate the effects of novel medicines and a novel artificial graft for the treatment are required. In our previous study, we reported the rapid construction of 3D tissues by employing a layer-by-layer (LbL) technique and revealed their potential applications in the pharmaceutical fields and tissue engineering. In this study, we successfully constructed a 3D arterial wall model containing vasa vasorum by employing a LbL technique for the first time. The cells were coated with extracellular matrix nanofilms and seeded into a culture insert using a cell accumulation method. This model had a three-layered hierarchical structure: a fibroblast layer, a smooth muscle layer, and an endothelial layer, which resembled the native arterial wall. Our method could introduce vasa vasorum into a fibroblast layer in vitro and the 3D arterial wall model showed barrier function which was evaluated by immunostaining and transendothelial electrical resistance measurement. Furthermore, electron microscopy observations revealed that the vasa vasorum was composed of single-layered endothelial cells, and the endothelial tubes were surrounded by the basal lamina, which are known to promote maturation and stabilization in native blood capillaries. These models should be useful for tissue engineering, regenerative medicine, and pharmaceutical applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 814-823, 2017. © 2016 Wiley Periodicals, Inc.

Stem Cell-Soluble Signals Enhance Multilumen Formation in SMG Cell Clusters.

PubMed

Maruyama, C L M; Leigh, N J; Nelson, J W; McCall, A D; Mellas, R E; Lei, P; Andreadis, S T; Baker, O J

2015-11-01

Saliva plays a major role in maintaining oral health. Patients with salivary hypofunction exhibit difficulty in chewing and swallowing foods, tooth decay, periodontal disease, and microbial infections. At this time, treatments for hyposalivation are limited to medications (e.g., muscarinic receptor agonists: pilocarpine and cevimeline) that induce saliva secretion from residual acinar cells as well as artificial salivary substitutes. Therefore, advancement of restorative treatments is necessary to improve the quality of life in these patients. Our previous studies indicated that salivary cells are able to form polarized 3-dimensional structures when grown on growth factor-reduced Matrigel. This basement membrane is rich in laminin-III (L1), which plays a critical role in salivary gland formation. Mitotically inactive feeder layers have been used previously to support the growth of many different cell types, as they provide factors necessary for cell growth and organization. The goal of this study was to improve salivary gland cell differentiation in primary cultures by using a combination of L1 and a feeder layer of human hair follicle-derived mesenchymal stem cells (hHF-MSCs). Our results indicated that the direct contact of mouse submandibular (mSMG) cell clusters and hHF-MSCs was not required for mSMG cells to form acinar and ductal structures. However, the hHF-MSC conditioned medium enhanced cell organization and multilumen formation, indicating that soluble signals secreted by hHF-MSCs play a role in promoting these features. © International & American Associations for Dental Research 2015.

Modeling the Formation of Transverse Weld during Billet-on-Billet Extrusion

PubMed Central

Mahmoodkhani, Yahya; Wells, Mary; Parson, Nick; Jowett, Chris; Poole, Warren

2014-01-01

A comprehensive mathematical model of the hot extrusion process for aluminum alloys has been developed and validated. The plasticity module was developed using a commercial finite element package, DEFORM-2D, a transient Lagrangian model which couples the thermal and deformation phenomena. Validation of the model against industrial data indicated that it gave excellent predictions of the pressure during extrusion. The finite element predictions of the velocity fields were post-processed to calculate the thickness of the surface cladding as one billet is fed in after another through the die (i.e., the transverse weld). The mathematical model was then used to assess the effect a change in feeder dimensions would have on the shape, thickness and extent of the transverse weld during extrusion. Experimental measurements for different combinations of billet materials show that the model is able to accurately predict the transverse weld shape as well as the clad surface layer to thicknesses of 50 μm. The transverse weld is significantly affected by the feeder geometry shape, but the effects of ram speed, billet material and temperature on the transverse weld dimensions are negligible. PMID:28788629

Shared Ride Taxi Feeder Service in Memphis, TN

DOT National Transportation Integrated Search

1988-03-01

From May 1983 through October 1984, the Memphis Area Transit Authority (MATA) conducted the Taxi Feeder Demonstration Project. It entailed the operation of fixed-route feeder services through three low-density neighborhoods and one industrial park, c...

In vitro co-culture of human skin keratinocytes and fibroblasts on a biocompatible and biodegradable scaffold.

PubMed

Pajoum Shariati, Seyed Ramin; Shokrgozar, Mohammad Ali; Vossoughi, Manouchehr; Eslamifar, Ali

2009-07-01

Extensive full-thickness burns require replacement of both epidermis and dermis. In designing skin replacements, the goal has been to re-create this model and make a product which has both essential components. In the present study, we developed procedures for establishing confluent, stratified layers of cultured human keratinocytes on the surface of modified collagen-chitosan scaffold that contains fibroblasts. The culture methods for propagation of keratinocytes and fibroblasts isolated from human neonatal foreskin were developed. The growth and proliferation of normal human keratinocytes were evaluated in serum-free (keratinocyte growth medium) and our modified medium. Characterization of human keratinocytes was determined by using pan-keratin and anti-involucrin monoclonal antibodies. For fabrication of relevant biodegradable and biocompatible collagen-chitosan porous scaffold with improved biostability, modified method of freeze-gelation was used. In generating organotypic co-cultures, epidermal keratinocytes were plated onto the upper surface of scaffold containing embedded fibroblasts. The results showed that the growth of isolated human skin fibroblasts and keratinocytes in our modified medium was more than that in the serum-free medium. The different evaluations of collagen-chitosan scaffold showed that it is relevant to growth of cells (fibroblast and keratinocyte) and has a good flexibility in manipulation of the living skin equivalents. These findings indicate that the integration of collagen-chitosan scaffold with co-cultured keratinocyte and fibroblast in vitro provides a potential source of living skin for grafting in vivo.

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

PubMed

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 < 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. © The Author 2017. Published by Oxford University Press on behalf of Poultry Science Association.

39. CLOSE UP DETAIL OF THE FEEDER AND STAMP CONNECTION. ...

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

39. CLOSE UP DETAIL OF THE FEEDER AND STAMP CONNECTION. THE STAMP AN MORTAR BOX ARE ON THE LEFT AND THE FEEDER WITH ITS FEEDER DISK IS ON THE RIGHT. NOTE THE COLLAR ON THE CENTER STAMP STEM (UPPER LEFT CORNER OF THE IMAGE) THAT ACTIVATES THE LEVER IN THE CENTER OF THE PHOTO. THE COLLAR IS POSITIONED SUCH THAT WHEN THE LEVEL OF THE MATERIAL REACHES A LOW POINT IN THE MORTAR BOX IT PUSHES DOWN ON THE LEVER WHICH IN TURN ACTIVATES THE AUTOMATIC FEEDER DRIVE MECHANISM WHICH THEM DELIVERS ORE INTO THE BACKSIDE OF THE MORTAR BOX. - Standard Gold Mill, East of Bodie Creek, Northeast of Bodie, Bodie, Mono County, CA

A pellet feeder for the birds

PubMed Central

Millard, W. J.

1979-01-01

An alternative means of delivering food to pigeons in operant conditioning research is described. The feeder allows greater control of the amount of food delivered and reduces the amount of time necessary for the pigeon to collect the food. It is possible to extend the length of experimental sessions due to the reduction of food intake. Data obtained using the pellet feeder indicated that the control of responding is comparable to that observed with the standard grain-magazine feeder. PMID:16812118

Microscopic suspension feeders near boundaries: Effects of external water flow

NASA Astrophysics Data System (ADS)

Pepper, Rachel; Koehl, M. A. R.

2015-11-01

Microscopic sessile suspension feeders are an important part of aquatic ecosystems and form a vital link in the transfer of carbon in aquatic food webs. These suspension feeders live attached to boundaries, consume bacteria and small detritus, and are in turn eaten by larger organisms. Many create a feeding current that draws fluid towards them, and from which they filter their food. In still water, the feeding current consists of recirculating eddies which form as a result of fluid forcing near a boundary. These recirculating eddies can be depleted of food and significantly decrease nutrient uptake; a variety of strategies have been proposed for how attached feeders increase their access to undepleted water. We investigate the interaction of the flow produced by a microscopic suspension feeder with external environmental flow, such as the current in a stream or ocean. We show through calculations that even very slow flow (on the order of microns per second) is sufficient to provide a constant supply of undepleted water to suspension feeders when the feeders are modeled with perfect nutrient capture efficiency and in the absence of diffusion. We also discuss which natural flow environments exceed the threshold to supply undepleted water and which do not, and we examine how characteristics of the suspension feeders themselves, such as stalk length and feeding disk size, influence feeding currents and their interactions with external flows.

Immobilization of type-I collagen and basic fibroblast growth factor (bFGF) onto poly (HEMA-co-MMA) hydrogel surface and its cytotoxicity study.

PubMed

Yan, Tuo; Sun, Rong; Li, Chun; Tan, Baihua; Mao, Xuan; Ao, Ningjian

2010-08-01

Type-I collagen and bFGF were immobilized onto the surface of poly (HEMA-co-MMA) hydrogel by grafting and coating methods to improve its cytotoxicity. The multi-layered structure of the biocompatible layer was confirmed by FTIR, AFM and static water contact angles. The layers were stable in body-like environment (pH 7.4). Human skin fibroblast cells (HSFC) were seeded onto Col/bFGF-poly (HEMA-co-MMA), Col-poly (HEMA-co-MMA) and poly (HEMA-co-MMA) films for 1, 3 and 5 day. MTT assay was performed to evaluate the extraction toxicity of the materials. Results showed that the cell attachment, proliferation and differentiation on Col/bFGF-poly (HEMA-co-MMA) film were higher than those of the control group, which indicated the improvement of cell-material interaction. The extraction toxicity of the modified materials was also lower than that of the unmodified group. The protein and bFGF immobilized poly (HEMA-co-MMA) hydrogel might hold great promise to be a biocompatible material.

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

PubMed

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

2009-11-01

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

Flow accelerated corrosion of carbon steel feeder pipes from pressurized heavy water reactors

NASA Astrophysics Data System (ADS)

Singh, J. L.; Kumar, Umesh; Kumawat, N.; Kumar, Sunil; Kain, Vivekanand; Anantharaman, S.; Sinha, A. K.

2012-10-01

Detailed investigation of a number of feeder pipes received from Rajasthan Atomic Power Station Unit 2 (RAPS#2) after en-masse feeder pipe replacement after 15.67 Effective Full Power Years (EFPYs) was carried out. Investigations included ultrasonic thickness measurement by ultrasonic testing, optical microscopy, scanning electron microscopy, chemical analysis and X-ray Diffraction (XRD). Results showed that maximum thickness reduction of the feeder had occurred downstream and close to the weld in 32 NB (1.25″/32.75 mm ID) elbows. Rate of Flow Accelerated Corrosion (FAC) was measured to be higher in the lower diameter feeder pipes due to high flow velocity and turbulence. Weld regions had thinned to a lower extent than the parent material due to higher chromium content in the weld. A weld protrusion has been shown to add to the thinning due to FAC and lead to faster thinning rate at localized regions. Surface morphology of inner surface of feeder had shown different size scallop pattern over the weld and parent material. Inter-granular cracks were also observed along the weld fusion line and in the parent material in 32 NB outlet feeder elbow.

7 CFR 1280.107 - Feeder.

Code of Federal Regulations, 2010 CFR

2010-01-01

... AND ORDERS; MISCELLANEOUS COMMODITIES), DEPARTMENT OF AGRICULTURE LAMB PROMOTION, RESEARCH, AND INFORMATION ORDER Lamb Promotion, Research, and Information Order Definitions § 1280.107 Feeder. Feeder means any person who acquires ownership of lambs and feeds such lambs in the U.S. until they reach slaughter...

Apparatus and method for optimal phase balancing using dynamic programming with spatial consideration

DOEpatents

Robertazzi, Thomas G.; Skiena, Steven; Wang, Kai

2017-08-08

Provided are an apparatus and method for load-balancing of a three-phase electric power distribution system having a multi-phase feeder, including obtaining topology information of the feeder identifying supply points for customer loads and feeder sections between the supply points, obtaining customer information that includes peak customer load at each of the points between each of the feeder sections, performing a phase balancing analysis, and recommending phase assignment at the customer load supply points.

Relative Preference and Localized Food Affect Predator Space Use and Consumption of Incidental Prey

PubMed Central

Schartel, Tyler E.; Schauber, Eric M.

2016-01-01

Abundant, localized foods can concentrate predators and their foraging efforts, thus altering both the spatial distribution of predation risk and predator preferences for prey that are encountered incidentally. However, few investigations have quantified the spatial scale over which localized foods affect predator foraging behavior and consumption of incidental prey. In spring 2010, we experimentally tested how point-source foods altered how generalist predators (white-footed mice, Peromyscus leucopus) utilized space and depredated two incidental prey items: almonds (Prunus dulcis; highly profitable) and maple seeds (Acer saccharum; less profitable). We estimated mouse population densities with trapping webs, quantified mouse consumption rates of these incidental prey items, and measured local mouse activity with track plates. We predicted that 1) mouse activity would be elevated near full feeders, but depressed at intermediate distances from the feeder, 2) consumption of both incidental prey would be high near feeders providing less-preferred food and, 3) consumption of incidental prey would be contingent on predator preference for prey relative to feeders providing more-preferred food. Mouse densities increased significantly from pre- to post-experiment. Mean mouse activity was unexpectedly greatest in control treatments, particularly <15 m from the control (empty) feeder. Feeders with highly preferred food (sunflower seeds) created localized refuges for incidental prey at intermediate distances (15 to 25m) from the feeder. Feeders with less-preferred food (corn) generated localized high risk for highly preferred almonds <10 m of the feeder. Our findings highlight the contingent but predictable effects of locally abundant food on risk experienced by incidental prey, which can be positive or negative depending on both spatial proximity and relative preference. PMID:26978659

Aflatoxin contamination in corn sold for wildlife feed in texas.

PubMed

Dunham, Nicholas R; Peper, Steven T; Downing, Carson D; Kendall, Ronald J

2017-05-01

Supplemental feeding with corn to attract and manage deer is a common practice throughout Texas. Other species, including northern bobwhites (Colinus virginianus), are commonly seen feeding around supplemental deer feeders. In many cases, supplemental feeding continues year-round so feed supply stores always have supplemental corn in stock. Fluctuating weather and improper storage of corn can lead to and/or amplify aflatoxin contamination. Due to the recent decline of bobwhites throughout the Rolling Plains ecoregion of Texas, there has been interest in finding factors such as toxins that could be linked to their decline. In this study, we purchased and sampled supplemental corn from 19 locations throughout this ecoregion to determine if aflatoxin contamination was present in individual bags prior to being dispersed to wildlife. Of the 57 bags sampled, 33 bags (approximately 58%) contained aflatoxin with a bag range between 0.0-19.91 parts per billion (ppb). Additionally, three metal and three polypropylene supplemental feeders were each filled with 45.4 kg of triple cleaned corn and placed in an open field to study long-term aflatoxin buildup. Feeders were sampled every 3 months from November 2013-November 2014. Average concentration of aflatoxin over the year was 4.08 ± 2.53 ppb (±SE) in metal feeders, and 1.43 ± 0.89 ppb (±SE) in polypropylene feeders. The concentration of aflatoxins is not affected by the type of feeder (metal vs polypropylene), the season corn was sampled, and the location in the feeder (top, middle, bottom) where corn is sampled. It is unlikely that corn used in supplemental feeders is contributing to the bobwhite decline due to the low levels of aflatoxin found in purchased corn and long-term storage of corn used in supplemental feeders.

Relative Preference and Localized Food Affect Predator Space Use and Consumption of Incidental Prey.

PubMed

Schartel, Tyler E; Schauber, Eric M

2016-01-01

Abundant, localized foods can concentrate predators and their foraging efforts, thus altering both the spatial distribution of predation risk and predator preferences for prey that are encountered incidentally. However, few investigations have quantified the spatial scale over which localized foods affect predator foraging behavior and consumption of incidental prey. In spring 2010, we experimentally tested how point-source foods altered how generalist predators (white-footed mice, Peromyscus leucopus) utilized space and depredated two incidental prey items: almonds (Prunus dulcis; highly profitable) and maple seeds (Acer saccharum; less profitable). We estimated mouse population densities with trapping webs, quantified mouse consumption rates of these incidental prey items, and measured local mouse activity with track plates. We predicted that 1) mouse activity would be elevated near full feeders, but depressed at intermediate distances from the feeder, 2) consumption of both incidental prey would be high near feeders providing less-preferred food and, 3) consumption of incidental prey would be contingent on predator preference for prey relative to feeders providing more-preferred food. Mouse densities increased significantly from pre- to post-experiment. Mean mouse activity was unexpectedly greatest in control treatments, particularly <15 m from the control (empty) feeder. Feeders with highly preferred food (sunflower seeds) created localized refuges for incidental prey at intermediate distances (15 to 25m) from the feeder. Feeders with less-preferred food (corn) generated localized high risk for highly preferred almonds <10 m of the feeder. Our findings highlight the contingent but predictable effects of locally abundant food on risk experienced by incidental prey, which can be positive or negative depending on both spatial proximity and relative preference.

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

NASA Astrophysics Data System (ADS)

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.

Laboratory procedure for estimating residue dynamics of xenobiotic contaminants in a freshwater food chain

USGS Publications Warehouse

Johnson, B. Thomas

1980-01-01

A laboratory method of measuring the accumulation, transfer, elimination, and degradation of xenobiotic contaminants is described for organisms in a freshwater food chain (microorganisms, filter-feeder, and fish). A flow-through diluter-system, 14C-labeled contaminants, gas and thin-layer chromatography, autoradiography, and liquid scintillation spectrometry are used in making residue determinations. Accumulation factors and various index values are developed for measuring and estimating potential accumulation of xenobiotic contaminants by aquatic organisms. The laboratory procedure is economical, simple, reproducible, and ecologically relevant.

Voltage Support Study of Smart PV Inverters on a High-Photovoltaic Penetration Utility Distribution Feeder

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

Ding, Fei; Pratt, Annabelle; Bialek, Tom

2016-11-21

This paper reports on tools and methodologies developed to study the impact of adding rooftop photovoltaic (PV) systems, with and without the ability to provide voltage support, on the voltage profile of distribution feeders. Simulation results are provided from a study of a specific utility feeder. The simulation model of the utility distribution feeder was built in OpenDSS and verified by comparing the simulated voltages to field measurements. First, we set all PV systems to operate at unity power factor and analyzed the impact on feeder voltages. Then we conducted multiple simulations with voltage support activated for all the smartmore » PV inverters. These included different constant power factor settings and volt/VAR controls.« less

Analysis of broadcasting satellite service feeder link power control and polarization

NASA Technical Reports Server (NTRS)

Sullivan, T. M.

1982-01-01

Statistical analyses of carrier to interference power ratios (C/Is) were performed in assessing 17.5 GHz feeder links using (1) fixed power and power control, and (2) orthogonal linear and orthogonal circular polarizations. The analysis methods and attenuation/depolarization data base were based on CCIR findings to the greatest possible extent. Feeder links using adaptive power control were found to neither cause or suffer significant C/I degradation relative to that for fixed power feeder links having similar or less stringent availability objectives. The C/Is for sharing between orthogonal linearly polarized feeder links were found to be significantly higher than those for circular polarization only in links to nominally colocated satellites from nominally colocated Earth stations in high attenuation environments.

Network Reduction Algorithm for Developing Distribution Feeders for Real-Time Simulators

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

Nagarajan, Adarsh; Nelson, Austin A; Prabakar, Kumaraguru

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 simulators and test PV inverters using power hardware-in-the-loop (PHIL) techniques. This paper presents a novel feeder model reduction algorithm using a ruin & reconstruct methodology 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-timemore » digital testing platform. Smart PV inverters were added to the realtime 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 feeders could be analyzed.« less

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

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

Nagarajan, Adarsh; Nelson, Austin; Prabakar, Kumaraguru

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 digitalmore » 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.« less

Economic feeder for recharging and ``topping off''

NASA Astrophysics Data System (ADS)

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.

Comparing the 2000 and 2005 factors affecting the selling price of feeder cattle sold at Arkansas livestock auctions.

PubMed

Troxel, T R; Barham, B L

2007-12-01

The objectives of the study were to determine how factors affecting the selling price of feeder calves changed from 2000 to 2005 and to examine the perception that discounts narrow or even disappear as calf supplies decrease and selling prices increase. Data from weekly Arkansas livestock auctions were collected from January 1 to December 31 in 2000 and 2005. Data included calf sex, breed type, color, muscle score, horn status, frame score, fill, condition, health, and BW. Mean selling prices for 2000 and 2005 were $92.91 +/- 15.05 and $118.32 +/- 15.13 (mean +/- SD; $/45.45 kg), respectively. Individual price observations were subtracted from the respective annual means and became the dependent variable. The selling prices for feeder calves sold in groups of 2 to 5 calves and in groups of >/= 6 calves were greater in 2005 than 2000 (P < 0.001). Steers received a greater premium ($6.48 +/- 0.09 vs. $6.02 +/- 0.08; mean +/- SE) and bull calves received greater discounts ($0.30 +/- 0.14 vs. $1.68 +/- 0.09) in 2005 than in 2000. Breeds types that increased in value from 2000 to 2005 were Angus x Hereford, Angus, Angus x Charolais, and Brahman (P < 0.001). Breed types that received a reduced selling price in 2005 compared with 2000 (P < 0.001) were one-fourth Brahman Cross, Charolais, Charolais x Limousin, Hereford x Limousin, Limousin, Limousin x one-fourth Brahman, Longhorn, Saler and Simmental. Yellow-white face, black-white face, black, and gray feeder calves received an increase in selling price from 2000 to 2005 (P < 0.001). Although fewer horned feeder calves were sold in 2005 (P < 0.01), they received greater discounts in 2005 than 2000 (-$2.86 +/- 0.16 and -$0.51 +/- 0.09; P < 0.001). In 2005, large-framed feeder calves did not receive the premium detected in 2000, but medium-framed feeder calves in 2005 received a greater selling price compared with 2000. Feeder calves with a muscle score of 1 received a greater premium in 2005 compared with 2000 ($2.58 +/- 0.06 and $0.02 +/- 0.09, respectively; P < 0.001). Feeder calves with a muscle score of 2 were discounted in both years, but the discount in 2005 was not as great as in 2000 (P < 0.001). Full and tanked feeder calves received greater discounts in 2005 than in 2000 (P < 0.001). Discounts for fleshy and fat feeder calves were greater in 2005 than in 2000. Most factors affecting the selling price of Arkansas feeder calves in 2000 affected the selling price in 2005. Although feeder calf supplies were smaller in 2005 than 2000, many discounts increased.

On the influence of various physicochemical properties of the CNTs based implantable devices on the fibroblasts' reaction in vitro.

PubMed

Benko, Aleksandra; Frączek-Szczypta, Aneta; Menaszek, Elżbieta; Wyrwa, Jan; Nocuń, Marek; Błażewicz, Marta

2015-11-01

Coating the material with a layer of carbon nanotubes (CNTs) has been a subject of particular interest for the development of new biomaterials. Such coatings, made of properly selected CNTs, may constitute an implantable electronic device that facilitates tissue regeneration both by specific surface properties and an ability to electrically stimulate the cells. The goal of the presented study was to produce, evaluate physicochemical properties and test the applicability of highly conductible material designed as an implantable electronic device. Two types of CNTs with varying level of oxidation were chosen. The process of coating involved suspension of the material of choice in the diluent followed by the electrophoretic deposition to fabricate layers on the surface of a highly biocompatible metal-titanium. Presented study includes an assessment of the physicochemical properties of the material's surface along with an electrochemical evaluation and in vitro biocompatibility, cytotoxicity and apoptosis studies in contact with the murine fibroblasts (L929) in attempt to answer the question how the chemical composition and CNTs distribution in the layer alters the electrical properties of the sample and whether any of these properties have influenced the overall biocompatibility and stimulated adhesion of fibroblasts. The results indicate that higher level of oxidation of CNTs yielded materials more conductive than the metal they are deposited on. In vitro study revealed that both materials were biocompatible and that the cells were not affected by the amount of the functional group and the morphology of the surface they adhered to.

Stabilisation of cables of fibronectin with micromolar concentrations of copper: in vitro cell substrate properties.

PubMed

Ahmed, Zubair; Briden, Anita; Hall, Susan; Brown, Robert A

2004-02-01

We have previously described the production of large cables of fibronectin, a large extracellular matrix cell adhesion glycoprotein, which has a potential application in tissue engineering. Here we have stabilised these cables for longer survival and looked at their ultrastructural cell-substrate behaviour in vitro. Dissolution experiments showed that low concentrations of copper not only caused significant material stabilisation but left pores which could promote cell ingrowth, as we have previously reported with Fn-mats. Indeed, the greatest amount of cell ingrowth was observed for copper treated cables. Immunostaining showed S-100(+) multi-layers of cells around the edge of cables while ultrastructural analysis confirmed the presence of a mixture of fibroblasts and bipolar cells associated with fragments of basal lamina, which is a Schwann cell phenotype. Interestingly, the outermost layers of cells consisted of S-100(-) cells, presumed fibroblasts, apparently 'capping' the Schwann cells. Toxicity tests revealed that Schwann cells were only able to grow at the lowest concentration of copper used (1microM) while fibroblasts grew at all concentrations tested. These results could be used to design biomaterials with optimum properties for promoting cellular ingrowth and survival in tissue engineered grafts which may be used to improve peripheral nerve repair.

Interactions of 1D- and 2D-layered inorganic nanoparticles with fibroblasts and human mesenchymal stem cells

PubMed Central

Rashkow, Jason Thomas; Talukdar, Yahfi; Lalwani, Gaurav; Sitharaman, Balaji

2015-01-01

Aim This study investigates the effects of tungsten disulfide nanotubes (WSNTs) and molybdenum disulfide nanoplatelets (MSNPs) on fibroblasts (NIH-3T3) and mesenchymal stem cells (MSCs) to determine safe dosages for potential biomedical applications. Materials & methods Cytotoxicity of MSNPs and WSNTs (5–300 µg/ml) on NIH-3T3 and MSCs was assessed at 6, 12 or 24 h. MSC differentiation to adipocytes and osteoblasts was assessed following treatment for 24 h. Results Only NIH-3T3 cells treated with MSNPs showed dose or time dependent increase in cytotoxicity. Differentiation markers of MSCs in treated groups were unaffected compared with untreated controls. Conclusion MSNPs and WSNTs at concentrations less than 50 µg/ml are potentially safe for treatment of fibroblasts or MSCs for up to 24 h. PMID:26080694

Effects of protein-coated nanofibers on conformation of gingival fibroblast spheroids: potential utility for connective tissue regeneration.

PubMed

Kaufman, Gili; Whitescarver, Ryan A; Nunes, Laiz; Palmer, Xavier-Lewis; Skrtic, Drago; Tutak, Wojtek

2018-01-24

Deep wounds in the gingiva caused by trauma or surgery require a rapid and robust healing of connective tissues. We propose utilizing gas-brushed nanofibers coated with collagen and fibrin for that purpose. Our hypotheses are that protein-coated nanofibers will: (i) attract and mobilize cells in various spatial orientations, and (ii) regulate the expression levels of specific extracellular matrix (ECM)-associated proteins, determining the initial conformational nature of dense and soft connective tissues. Gingival fibroblast monolayers and 3D spheroids were cultured on ECM substrate and covered with gas-blown poly-(DL-lactide-co-glycolide) (PLGA) nanofibers (uncoated/coated with collagen and fibrin). Cell attraction and rearrangement was followed by F-actin staining and confocal microscopy. Thicknesses of the cell layers, developed within the nanofibers, were quantified by ImageJ software. The expression of collagen1α1 chain (Col1α1), fibronectin, and metalloproteinase 2 (MMP2) encoding genes was determined by quantitative reverse transcription analysis. Collagen- and fibrin- coated nanofibers induced cell migration toward fibers and supported cellular growth within the scaffolds. Both proteins affected the spatial rearrangement of fibroblasts by favoring packed cell clusters or intermittent cell spreading. These cell arrangements resembled the structural characteristic of dense and soft connective tissues, respectively. Within three days of incubation, fibroblast spheroids interacted with the fibers, and grew robustly by increasing their thickness compared to monolayers. While the ECM key components, such as fibronectin and MMP2 encoding genes, were expressed in both protein groups, Col1α1 was predominantly expressed in bundled fibroblasts grown on collagen fibers. This enhanced expression of collagen1 is typical for dense connective tissue. Based on results of this study, our gas-blown, collagen- and fibrin-coated PLGA nanofibers are viable candidates for engineering soft and dense connective tissues with the required structural characteristics and functions needed for wound healing applications. Rapid regeneration of these layers should enhance healing of open wounds in a harsh oral environment.

An RFID Based Smart Feeder for Hummingbirds.

PubMed

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

PubMed Central

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

2015-01-01

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. PMID:26694402

46 CFR 111.75-1 - Lighting feeders.

Code of Federal Regulations, 2010 CFR

2010-10-01

... COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) ELECTRICAL ENGINEERING ELECTRIC SYSTEMS-GENERAL REQUIREMENTS Lighting Circuits and Protection § 111.75-1 Lighting feeders. (a) Passenger vessels. On a... lighting, feeders, and branch circuits are in subpart 112.43 of this chapter. [CGD 74-125A, 47 FR 15236...

Endothelial network formed with human dermal microvascular endothelial cells in autologous multicellular skin substitutes.

PubMed

Ponec, Maria; El Ghalbzouri, Abdoelwaheb; Dijkman, Remco; Kempenaar, Johanna; van der Pluijm, Gabri; Koolwijk, Pieter

2004-01-01

A human skin equivalent from a single skin biopsy harboring keratinocytes and melanocytes in the epidermal compartment, and fibroblasts and microvascular dermal endothelial cells in the dermal compartment was developed. The results of the study revealed that the nature of the extracellular matrix of the dermal compartments plays an important role in establishment of endothelial network in vitro. With rat-tail type I collagen matrices only lateral but not vertical expansion of endothelial networks was observed. In contrast, the presence of extracellular matrix of entirely human origin facilitated proper spatial organization of the endothelial network. Namely, when human dermal fibroblasts and microvascular endothelial cells were seeded on the bottom of an inert filter and subsequently epidermal cells were seeded on top of it, fibroblasts produced extracellular matrix throughout which numerous branched tubes were spreading three-dimensionally. Fibroblasts also facilitated the formation of basement membrane at the epidermal/matrix interface. Under all culture conditions, fully differentiated epidermis was formed with numerous melanocytes present in the basal epidermal cell layer. The results of the competitive RT-PCR revealed that both keratinocytes and fibroblasts expressed VEGF-A, -B, -C, aFGF and bFGF mRNA, whereas fibroblasts also expressed VEGF-D mRNA. At protein level, keratinocytes produced 10 times higher amounts of VEGF-A than fibroblasts did. The generation of multicellular skin equivalent from a single human skin biopsy will stimulate further developments for its application in the treatment of full-thickness skin defects. The potential development of biodegradable, biocompatible material suitable for these purposes is a great challenge for future research.

Can the KTP laser change the cementum surface of healthy and diseased teeth providing an acceptable root surface for fibroblast attachment?

NASA Astrophysics Data System (ADS)

Mailhot, Jason M.; Garnick, Jerry J.

1996-04-01

The purpose of our research is to determine the effects of KTP laser on root cementum and fibroblast attachment. Initial work has been completed in testing the effect of different energy levels on root surfaces. From these studies optimal energy levels were determined. In subsequent studies the working distance and exposure time required to obtain significant fibroblast attachment to healthy cementum surfaces were investigated. Results showed that lased cemental surfaces exhibited changes in surface topography which ranged from a melted surface to an apparent slight fusion of the surface of the covering smear layer. When the optimal energy level was used, fibroblasts demonstrate attachment on the specimens, resulting in the presence of a monolayer of cells on the control surfaces as well as on the surfaces lased with this energy level. The present study investigates the treatment of pathological root surfaces and calculus with a KTP laser utilizing these optimal parameters determine previously. Thirty single rooted teeth with advanced periodontal disease and ten healthy teeth were obtained, crowns were sectioned and roots split longitudinally. Forty test specimens were assigned into 1 of 4 groups; pathologic root--not lased, pathologic root--lased, root planed root and health root planed root. Human gingival fibroblasts were seeded on specimens and cultured for 24 hours. Specimens were processed for SEM. The findings suggest that with the KTP laser using a predetermined energy level applied to pathological root surfaces, the lased surfaces provided an unacceptable surface for fibroblast attachment. However, the procedural control using healthy root planed surfaces did demonstrate fibroblast attachment.

46 CFR 112.43-15 - Emergency lighting feeders.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 4 2011-10-01 2011-10-01 false Emergency lighting feeders. 112.43-15 Section 112.43-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) ELECTRICAL ENGINEERING EMERGENCY LIGHTING AND POWER SYSTEMS Emergency Lighting Systems § 112.43-15 Emergency lighting feeders. For a vessel with...

46 CFR 112.43-15 - Emergency lighting feeders.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 4 2010-10-01 2010-10-01 false Emergency lighting feeders. 112.43-15 Section 112.43-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) ELECTRICAL ENGINEERING EMERGENCY LIGHTING AND POWER SYSTEMS Emergency Lighting Systems § 112.43-15 Emergency lighting feeders. For a vessel with...

46 CFR 112.43-15 - Emergency lighting feeders.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 46 Shipping 4 2014-10-01 2014-10-01 false Emergency lighting feeders. 112.43-15 Section 112.43-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) ELECTRICAL ENGINEERING EMERGENCY LIGHTING AND POWER SYSTEMS Emergency Lighting Systems § 112.43-15 Emergency lighting feeders. For a vessel with...

46 CFR 112.43-15 - Emergency lighting feeders.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 46 Shipping 4 2012-10-01 2012-10-01 false Emergency lighting feeders. 112.43-15 Section 112.43-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) ELECTRICAL ENGINEERING EMERGENCY LIGHTING AND POWER SYSTEMS Emergency Lighting Systems § 112.43-15 Emergency lighting feeders. For a vessel with...

46 CFR 112.43-15 - Emergency lighting feeders.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 46 Shipping 4 2013-10-01 2013-10-01 false Emergency lighting feeders. 112.43-15 Section 112.43-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) ELECTRICAL ENGINEERING EMERGENCY LIGHTING AND POWER SYSTEMS Emergency Lighting Systems § 112.43-15 Emergency lighting feeders. For a vessel with...

77 FR 34876 - Airworthiness Directives; The Boeing Company

Federal Register 2010, 2011, 2012, 2013, 2014

2012-06-12

... (a flammable fluid leakage zone) or heat damage to the APU power feeder cable, insulation blankets... heat damage to the APU power feeder cable, insulation blankets, or pressure bulkhead. Relevant Service... feeder cable and heat damage of the insulation blanket adjacent to the clamp, a detailed inspection for...

Feederism in a woman.

PubMed

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.

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

PubMed

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.

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

PubMed

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.

Central Role for Dermal Fibroblasts in Skin Model Protection against Candida albicans.

PubMed

Kühbacher, Andreas; Henkel, Helena; Stevens, Philip; Grumaz, Christian; Finkelmeier, Doris; Burger-Kentischer, Anke; Sohn, Kai; Rupp, Steffen

2017-06-01

The fungal pathogen Candida albicans colonizes basically all human epithelial surfaces, including the skin. Under certain conditions, such as immunosuppression, invasion of the epithelia occurs. Not much is known about defense mechanisms against C. albicans in subepithelial layers such as the dermis. Using immune cell-supplemented 3D skin models we defined a new role for fibroblasts in the dermis and identified a minimal set of cell types for skin protection against C. albicans invasion. Dual RNA sequencing of individual host cell populations and C. albicans revealed that dermal invasion is directly impeded by dermal fibroblasts. They are able to integrate signals from the pathogen and CD4+ T cells and shift toward an antimicrobial phenotype with broad specificity that is dependent on Toll-like receptor 2 and interleukin 1β. These results highlight a central function of dermal fibroblasts for skin protection, opening new possibilities for treatment of infectious diseases. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

The innate responses of bumble bees to flower patterns: separating the nectar guide from the nectary changes bee movements and search time

NASA Astrophysics Data System (ADS)

Goodale, Eben; Kim, Edward; Nabors, Annika; Henrichon, Sara; Nieh, James C.

2014-06-01

Nectar guides can enhance pollinator efficiency and plant fitness by allowing pollinators to more rapidly find and remember the location of floral nectar. We tested if a radiating nectar guide around a nectary would enhance the ability of naïve bumble bee foragers to find nectar. Most experiments that test nectar guide efficacy, specifically radiating linear guides, have used guides positioned around the center of a radially symmetric flower, where nectaries are often found. However, the flower center may be intrinsically attractive. We therefore used an off-center guide and nectary and compared "conjunct" feeders with a nectar guide surrounding the nectary to "disjunct" feeders with a nectar guide separated from the nectary. We focused on the innate response of novice bee foragers that had never previously visited such feeders. We hypothesized that a disjunct nectar guide would conflict with the visual information provided by the nectary and negatively affect foraging. Approximately, equal numbers of bumble bees ( Bombus impatiens) found nectar on both feeder types. On disjunct feeders, however, unsuccessful foragers spent significantly more time (on average 1.6-fold longer) searching for nectar than any other forager group. Successful foragers on disjunct feeders approached these feeders from random directions unlike successful foragers on conjunct feeders, which preferentially approached the combined nectary and nectar guide. Thus, the nectary and a surrounding nectar guide can be considered a combination of two signals that attract naïve foragers even when not in the floral center.

The innate responses of bumble bees to flower patterns: separating the nectar guide from the nectary changes bee movements and search time.

PubMed

Goodale, Eben; Kim, Edward; Nabors, Annika; Henrichon, Sara; Nieh, James C

2014-06-01

Nectar guides can enhance pollinator efficiency and plant fitness by allowing pollinators to more rapidly find and remember the location of floral nectar. We tested if a radiating nectar guide around a nectary would enhance the ability of naïve bumble bee foragers to find nectar. Most experiments that test nectar guide efficacy, specifically radiating linear guides, have used guides positioned around the center of a radially symmetric flower, where nectaries are often found. However, the flower center may be intrinsically attractive. We therefore used an off-center guide and nectary and compared "conjunct" feeders with a nectar guide surrounding the nectary to "disjunct" feeders with a nectar guide separated from the nectary. We focused on the innate response of novice bee foragers that had never previously visited such feeders. We hypothesized that a disjunct nectar guide would conflict with the visual information provided by the nectary and negatively affect foraging. Approximately, equal numbers of bumble bees (Bombus impatiens) found nectar on both feeder types. On disjunct feeders, however, unsuccessful foragers spent significantly more time (on average 1.6-fold longer) searching for nectar than any other forager group. Successful foragers on disjunct feeders approached these feeders from random directions unlike successful foragers on conjunct feeders, which preferentially approached the combined nectary and nectar guide. Thus, the nectary and a surrounding nectar guide can be considered a combination of two signals that attract naïve foragers even when not in the floral center.

26 CFR 1.502-1 - Feeder organizations.

Code of Federal Regulations, 2010 CFR

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...

75 FR 8395 - Bunker Hill Groundwater Basin, Riverside-Corona Feeder Project, San Bernardino and Riverside...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-24

... DEPARTMENT OF THE INTERIOR Bureau of Reclamation Bunker Hill Groundwater Basin, Riverside-Corona...) will prepare a joint EIS/EIR for the proposed Riverside-Corona Feeder Project. The public and agencies... participate in the planning, design, and construction of the Riverside-Corona Feeder Project including: (i) 20...

7 CFR 1230.71 - Assessments.

Code of Federal Regulations, 2011 CFR

2011-01-01

... feeder pig that is sold shall pay an assessment on that animal, unless such producer demonstrates to the Board by appropriate documentation that an assessment was previously paid on that animal as a feeder pig... purchaser of a porcine animal raised by a producer as a feeder pig or market hog shall collect an assessment...

7 CFR 1230.71 - Assessments.

Code of Federal Regulations, 2010 CFR

2010-01-01

... feeder pig that is sold shall pay an assessment on that animal, unless such producer demonstrates to the Board by appropriate documentation that an assessment was previously paid on that animal as a feeder pig... purchaser of a porcine animal raised by a producer as a feeder pig or market hog shall collect an assessment...

7 CFR 1230.71 - Assessments.

Code of Federal Regulations, 2012 CFR

2012-01-01

... feeder pig that is sold shall pay an assessment on that animal, unless such producer demonstrates to the Board by appropriate documentation that an assessment was previously paid on that animal as a feeder pig... purchaser of a porcine animal raised by a producer as a feeder pig or market hog shall collect an assessment...

7 CFR 1230.71 - Assessments.

Code of Federal Regulations, 2014 CFR

2014-01-01

... feeder pig that is sold shall pay an assessment on that animal, unless such producer demonstrates to the Board by appropriate documentation that an assessment was previously paid on that animal as a feeder pig... purchaser of a porcine animal raised by a producer as a feeder pig or market hog shall collect an assessment...

7 CFR 1230.71 - Assessments.

Code of Federal Regulations, 2013 CFR

2013-01-01

... feeder pig that is sold shall pay an assessment on that animal, unless such producer demonstrates to the Board by appropriate documentation that an assessment was previously paid on that animal as a feeder pig... purchaser of a porcine animal raised by a producer as a feeder pig or market hog shall collect an assessment...

A fundamental dispute: A discussion of "On some fundamentals of igneous petrology" by Bruce D. Marsh, Contributions to Mineralogy and Petrology (2013) 166: 665-690

NASA Astrophysics Data System (ADS)

Latypov, Rais; Morse, Tony; Robins, Brian; Wilson, Richard; Cawthorn, Grant; Tegner, Christian; Holness, Marian; Lesher, Charles; Barnes, Steve; O'Driscoll, Brian; Veksler, Ilya; Higgins, Michael; Wilson, Allan; Namur, Olivier; Chistyakova, Sofya; Naslund, Richard; Thy, Peter

2015-02-01

Marsh (Contrib Miner Petrol 166:665-690, 2013) again claims that crystal-free basalt magmas are unable to differentiate in crustal magma chambers and regards layered intrusions as primarily due to the repeated emplacement of crystal suspensions. He ignores an earlier critique of his unconventional inferences (Latypov, J Petrol 50:1047-1069, 2009) as well as a wealth of petrographic, geochemical and experimental evidence supporting the dominant role of fractional crystallization in the solidification of layered intrusions. Most tellingly, the cryptic variations preserved in the Skaergaard and many other basaltic layered intrusions would require an exceedingly implausible sequence of phenocrystic magmas but are wholly consistent with in situ fractional crystallization. A major flaw in Marsh's hypothesis is that it dismisses progressive fractional crystallization within any magma chamber and hence prohibits the formation of crystal slurries with phenocrysts and melts that change systematically in composition in any feeder system. This inherent attribute of the hypothesis excludes the formation of layered intrusions anywhere.

Efficient delivery to human lung fibroblasts (WI-38) of pirfenidone incorporated into liposomes modified with truncated basic fibroblast growth factor and its inhibitory effect on collagen synthesis in idiopathic pulmonary fibrosis.

PubMed

Togami, Kohei; Miyao, Aki; Miyakoshi, Kei; Kanehira, Yukimune; Tada, Hitoshi; Chono, Sumio

2015-01-01

In the present in vitro study, we assessed the delivery of pirfenidone incorporated into liposomes modified with truncated basic fibroblast growth factor (tbFGF) to lung fibroblasts and investigated the anti-fibrotic effect of the drug. The tbFGF peptide, KRTGQYKLC, was used to modify the surface of liposomes (tbFGF-liposomes). We used the thin-layer evaporation method, followed by sonication, to prepare tbFGF-liposomes containing pirfenidone. The cellular accumulation of tbFGF-liposomes was 1.7-fold greater than that of non-modified liposomes in WI-38 cells used as a model of lung fibroblasts. Confocal laser scanning microscopy showed that tbFGF-liposomes were widely localized in WI-38 cells. The inhibitory effects of pirfenidone incorporated into tbFGF-liposomes on transforming growth factor-β1 (TGF-β1)-induced collagen synthesis in WI-38 cells were evaluated by measuring the level of intracellular hydroxyproline, a major component of the protein collagen. Pirfenidone incorporated into tbFGF-liposomes at concentrations of 10, 30, and 100 µM significantly decreased the TGF-β1-induced hydroxyproline content in WI-38 cells. The anti-fibrotic effect of pirfenidone incorporated into tbFGF-liposomes was enhanced compared with that of pirfenidone solution. These results indicate that tbFGF-liposomes are a useful drug delivery system of anti-fibrotic drugs to lung fibroblasts for the treatment of idiopathic pulmonary fibrosis.

A Multistate Investigation of Antibiotic-Resistant Salmonella enterica Serotype I 4,[5],12:i:- Infections as Part of an International Outbreak Associated with Frozen Feeder Rodents

PubMed Central

Cartwright, E. J.; Nguyen, T.; Melluso, C.; Ayers, T.; Lane, C.; Hodges, A.; Li, X.; Quammen, J.; Yendell, S. J.; Adams, J.; Mitchell, J.; Rickert, R.; Klos, R.; Williams, I. T.; Behravesh, C. Barton; Wright, J.

2015-01-01

While most human Salmonella infections result from exposure to contaminated foods, an estimated 11% of all Salmonella infections are attributed to animal exposures, including both direct animal handling and indirect exposures such as cleaning cages and handling contaminated pet food. This report describes the epidemiologic, environmental and laboratory investigations conducted in the United States as part of the response to an international outbreak of tetracycline-resistant Salmonella enterica serotype I 4,[5],12:i:- infections with over 500 illnesses occurring from 2008 to 2010. This investigation found that illness due to the outbreak strain was significantly associated with exposure to pet reptiles and frozen feeder rodents used as food for pet reptiles. Salmonella isolates indistinguishable from the outbreak strain were isolated from a frozen feeder mice-fed reptile owned by a case patient, as well as from frozen feeder mice and environmental samples collected from a rodent producing facility (Company A). An international voluntary recall of all Company A produced frozen feeder animals sold between May 2009 and July 2010 occurred. Only 13% of cases in our investigation were aware of the association between Salmonella infection and mice or rats. Consumers, the pet industry, healthcare providers and veterinarians need to be aware of the potential health risk posed by feeder rodents, whether live or frozen. Frozen feeder rodent producers, suppliers and distributors should follow the animal food labelling requirements as described in 21 CFR §501.5, and all packages of frozen feeder rodents should include safe handling instructions. Persons should wash their hands thoroughly with soap and water after handling live or frozen feeder rodents, as well as reptiles or anything in the area where the animals live. Continued opportunities exist for public health officials, the pet industry, veterinarians and consumers to work together to prevent salmonellosis associated with pet food, pets and other animals. PMID:25996458

A Multistate Investigation of Antibiotic-Resistant Salmonella enterica Serotype I 4,[5],12:i:- Infections as Part of an International Outbreak Associated with Frozen Feeder Rodents.

PubMed

Cartwright, E J; Nguyen, T; Melluso, C; Ayers, T; Lane, C; Hodges, A; Li, X; Quammen, J; Yendell, S J; Adams, J; Mitchell, J; Rickert, R; Klos, R; Williams, I T; Barton Behravesh, C; Wright, J

2016-02-01

While most human Salmonella infections result from exposure to contaminated foods, an estimated 11% of all Salmonella infections are attributed to animal exposures, including both direct animal handling and indirect exposures such as cleaning cages and handling contaminated pet food. This report describes the epidemiologic, environmental and laboratory investigations conducted in the United States as part of the response to an international outbreak of tetracycline-resistant Salmonella enterica serotype I 4,[5],12:i:- infections with over 500 illnesses occurring from 2008 to 2010. This investigation found that illness due to the outbreak strain was significantly associated with exposure to pet reptiles and frozen feeder rodents used as food for pet reptiles. Salmonella isolates indistinguishable from the outbreak strain were isolated from a frozen feeder mice-fed reptile owned by a case patient, as well as from frozen feeder mice and environmental samples collected from a rodent producing facility (Company A). An international voluntary recall of all Company A produced frozen feeder animals sold between May 2009 and July 2010 occurred. Only 13% of cases in our investigation were aware of the association between Salmonella infection and mice or rats. Consumers, the pet industry, healthcare providers and veterinarians need to be aware of the potential health risk posed by feeder rodents, whether live or frozen. Frozen feeder rodent producers, suppliers and distributors should follow the animal food labelling requirements as described in 21 CFR §501.5, and all packages of frozen feeder rodents should include safe handling instructions. Persons should wash their hands thoroughly with soap and water after handling live or frozen feeder rodents, as well as reptiles or anything in the area where the animals live. Continued opportunities exist for public health officials, the pet industry, veterinarians and consumers to work together to prevent salmonellosis associated with pet food, pets and other animals. © 2015 Blackwell Verlag GmbH.

Interactions of 1D- and 2D-layered inorganic nanoparticles with fibroblasts and human mesenchymal stem cells

DOE PAGES

Rashkow, Jason Thomas; Talukdar, Yahfi; Lalwani, Gaurav; ...

2015-06-01

Here, this study investigates the effects of tungsten disulfide nanotubes (WSNTs) and molybdenum disulfide nanoplatelets (MSNPs) on fibroblasts (NIH-3T3) and mesenchymal stem cells (MSCs) to determine safe dosages for potential biomedical applications. Cytotoxicity of MSNPs and WSNTs (5–300 μg/ml) on NIH-3T3 and MSCs was assessed at 6, 12 or 24 h. MSC differentiation to adipocytes and osteoblasts was assessed following treatment for 24 h. Only NIH-3T3 cells treated with MSNPs showed dose or time dependent increase in cytotoxicity. Differentiation markers of MSCs in treated groups were unaffected compared with untreated controls. In conclusion, MSNPs and WSNTs at concentrations less thanmore » 50 μg/ml are potentially safe for treatment of fibroblasts or MSCs for up to 24 h.« less

Transgene-free human induced pluripotent stem cell line (HS5-SV.hiPS) generated from cesarean scar-derived fibroblasts.

PubMed

Rungsiwiwut, Ruttachuk; Pavarajarn, Wipawee; Numchaisrika, Pranee; Virutamasen, Pramuan; Pruksananonda, Kamthorn

2016-01-01

Transgene-free human HS5-SV.hiPS line was generated from human cesarean scar-derived fibroblasts using temperature-sensitive Sendai virus vectors carrying Oct4, Sox2, cMyc and Klf4 exogenous transcriptional factors. The viral constructs were eliminated from HS5-SV.hiPS line through heat treatment. Transgene-free HS5-SV.hiPS cells expressed pluripotent associated transcription factors Oct4, Nanog, Sox2, Rex1 and surface markers SSEA-4, TRA-1-60 and OCT4. HS5-SV.hiPS cells formed embryoid bodies and differentiated into three embryonic germ layers in vivo. HS5-SV.hiPS cells maintained their normal karyotype (46, XX) after culture for extended period. HS5-SV.hiPS displayed the similar pattern of DNA fingerprinting to the parenteral scar-derived fibroblasts. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Derivation of an induced pluripotent stem cell line (MUSIi004-A) from dermal fibroblasts of a 48-year-old spinocerebellar ataxia type 3 patient.

PubMed

Ritthaphai, Alisa; Wattanapanitch, Methichit; Pithukpakorn, Manop; Heepchantree, Worapa; Soi-Ampornkul, Rungtip; Mahaisavariya, Panchalee; Triwongwaranat, Daranporn; Pattanapanyasat, Kovit; Vatanashevanopakorn, Chinnavuth

2018-05-21

Dermal fibroblasts were obtained from a 48-year-old female patient with spinocerebellar ataxia type 3 (SCA3). Fibroblasts were reprogrammed by nucleofection with episomal plasmids, carrying L-MYC, LIN28, OCT4, SOX2, KLF4, EBNA-1 and shRNA against p53. The SCA3 patient-specific iPSC line, MUSIi004-A, was characterized by immunofluorescence staining to verify the expression of pluripotent markers. The iPSC line exhibited an ability to differentiate into three germ layers by embryoid body (EB) formation. Karyotypic analysis of the MUSIi004-A line was normal. The mutant allele was still present in the iPSC line. This iPSC line represents a useful tool for studying neurodegeneration in SCA3. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Properties of dehydrated human amnion/chorion composite grafts: Implications for wound repair and soft tissue regeneration.

PubMed

Koob, Thomas J; Lim, Jeremy J; Massee, Michelle; Zabek, Nicole; Denozière, Guilhem

2014-08-01

PURION(®) processed dehydrated human amnion/chorion membrane (dHACM; MiMedx Group, Marietta, GA) tissue products were analyzed for the effectiveness of the PURION(®) process in retaining the native composition of the amniotic membrane and preserving bioactivity in the resulting products. dHACM was analyzed for extracellular matrix (ECM) composition through histological staining and for growth factor content via multiplex ELISA arrays. Bioactivity was assessed by evaluating endogenous growth factor production by human dermal fibroblasts in response to dHACM and for thermal stability by mechanical tests and in vitro cell proliferation assays. Histology of dHACM demonstrated preservation of the native amnion and chorion layers with intact, nonviable cells, collagen, proteoglycan, and elastic fibers distributed in the individual layers. An array of 36 cytokines known to regulate processes involved in inflammation and wound healing were identified in dHACM. When treated with dHACM extracts, bioactivity was demonstrated through an upregulation of basic fibroblast growth factor, granulocyte colony-stimulating factor, and placental growth factor biosynthesis, three growth factors involved in wound healing, by dermal fibroblasts in vitro. After conditioning at temperatures ranging from -78.7 to +73.5°C, dHACM retained its tensile strength and ability to promote proliferation of dermal fibroblasts in vitro. Elution experiments demonstrated a soluble fraction of growth factors that eluted from the tissue and another fraction sequestered within the matrix. The PURION(®) process retains the native composition of ECM and signaling molecules and preserves bioactivity. The array of cytokines preserved in dHACM are in part responsible for its therapeutic efficacy in treating chronic wounds by orchestrating a "symphony of signals" to promote healing. © 2014 Wiley Periodicals, Inc.

Establishment of human induced pluripotent stem cell lines from normal fibroblast TIG-1.

PubMed

Kumazaki, Tsutomu; Kurata, Sayaka; Matsuo, Taira; Mitsui, Youji; Takahashi, Tomoko

2011-06-01

Normal human cells have a replicative life span and therefore senesce. Usually, normal human cell strains are differentiated cells and reach a terminally differentiated state after a number of cell divisions. At present, definitive differences are not known between replicative senescence and terminal differentiation. TIG-1 is a human fibroblast strain established from fetal lung and has been used extensively in studies of cellular senescence, and numerous data were accumulated at the molecular level. Recently, a method for generating induced pluripotent stem cells (iPSCs) was developed. Using the method, we introduced four reprogramming genes to TIG-1 fibroblasts and succeeded in isolating colonies that had embryonic stem cell (ESC)-like morphologies. They showed alkaline phosphatase activity and expressed ESC markers, as shown by immunostaining of OCT4, SOX2, SSEA4, and TRA-1-81 as well as reverse-transcription polymerase chain reaction (RT-PCR) for OCT4 and NANOG transcripts. Thus, we succeeded in establishing iPSC clones from TIG-1. The iPSC clones could differentiate to cells originated from all three germ-cell layers, as shown by RT-PCR, for messenger RNA (mRNA) expression of α-fetoprotein (endoderm), MSX1 (mesoderm) and microtubule-associated protein 2 (ectoderm), and by immunostaining for α-fetoprotein (endoderm), α-smooth muscle actin (mesoderm), and β-III-tubulin (ectoderm). The iPSCs formed teratoma containing the structures developed from all three germ-cell layers in severe combined immune-deficiency mice. Thus, by comparing the aging process of parental TIG-1 cells and the differentiation process of iPSC-derived fibrocytes to fibroblasts, we can reveal the exact differences in processes between senescence and terminal differentiation.

Echinococcus granulosus equinus: an ultrastructural study of murine tissue response to hydatid cysts.

PubMed

Richards, K S; Arme, C; Bridges, J F

1983-06-01

Peritoneal hydatids of Echinococcus granulosus equinus of 9 months standing in BALB/c mice occurred as free cysts or cysts within cyst masses. Both showed wide variation in size and in host tissue response, and all had a well-developed laminated layer separating the host tissue response from the germinal layer. In the smallest cyst-mass cysts the host tissue response was present as remnants of the initial cellular attack involving eosinophils. Slightly larger cyst-mass cysts possessed a primary macrophage invasion which phagocytosed the remnants of the initial attack and also, though to little effect, the laminated layer material. In the largest cyst-mass cysts a second macrophage invasion, of monocyte origin, had commenced and transformation stages of these cells to macrophages were observed. No fibroblasts surrounded individual cyst-mass cysts but they were present around the cyst mass, encapsulating it and possibly preventing further host cell invasion. Thus, the host tissue response around individual cyst-mass cysts remained 'preserved' at an early stage such as existed at the time of encapsulation. Small free cysts showed a primary macrophage invasion and transformation stages of cells of a secondary infiltration of peritoneal origin. Peripheral to the macrophages were fibroblasts demonstrating limited fibrinogenesis, and each cyst was surrounded by a layer of mesothelial cells. Large free cysts, also delimited by a mesothelial layer, possessed peripheral connective tissue, a deep fibrous layer and a monolayer of very compressed macrophages lying adjacent to the laminated layer. It is emphasized that an understanding of the host tissue response in cysts of different sizes and from different locations is an essential pre-requisite for the design of experimental studies.

A metro-access integrated network with all-optical virtual private network function using DPSK/ASK modulation format

NASA Astrophysics Data System (ADS)

Tian, Yue; Leng, Lufeng; Su, Yikai

2008-11-01

All-optical virtual private network (VPN), which offers dedicated optical channels to connect users within a VPN group, is considered a promising approach to efficient internetworking with low latency and enhanced security implemented in the physical layer. On the other hand, time-division multiplexed (TDM) / wavelength-division multiplexed (WDM) network architecture based on a feeder-ring with access-tree topology, is considered a pragmatic migration scenario from current TDM-PONs to future WDM-PONs and a potential convergence scheme for access and metropolitan networks, due to its efficiently shared hardware and bandwidth resources. All-optical VPN internetworking in such a metro-access integrated structure is expected to cover a wider service area and therefore is highly desirable. In this paper, we present a TDM/WDM metro-access integrated network supporting all-optical VPN internetworking among ONUs in different sub- PONs based on orthogonal differential-phase-shift keying (DPSK) / amplitude-shift keying (ASK) modulation format. In each ONU, no laser but a single Mach-Zehnder modulator (MZM) is needed for the upstream and VPN signal generation, which is cost-effective. Experiments and simulations are performed to verify its feasibility as a potential solution to the future access service.

Development of dry coal feeders

NASA Technical Reports Server (NTRS)

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.

Faculty Articulation with Feeder High Schools and Local Employers.

ERIC Educational Resources Information Center

Parrott, Marietta

As a first step in developing an articulation plan with feeder high schools, a College of the Sequoias (COS) task force developed and distributed a survey to all full-time faculty members to determine if individual faculty members were articulating with feeder high schools and local businesses, and if they would be willing to participate in an…

Ornithologists by Design: Kindergarteners Design, Construct, and Evaluate Bird Feeders

ERIC Educational Resources Information Center

Shorter, Angela; Segers, Marcia

2016-01-01

How can an engineer design a bird feeder that attracts many birds? This question resulted from kindergarten students' observations of the bird feeders in their school's bird sanctuary. The challenging question is the heart of project-based learning (PBL), a teaching strategy in which students tackle real-world problems and design projects to solve…

Preliminary evaluation of feeder and lint slide moisture addition on ginning, fiber quality, and textile processing of western cotton

USDA-ARS?s Scientific Manuscript database

The objective of this study was to evaluate the effects of moisture addition at the gin stand feeder conditioning hopper and/or the battery condenser slide on gin performance and Western cotton fiber quality and textile processing. The test treatments included no moisture addition, feeder hopper hum...

19. DETAIL OF STAMP BATTERY AUTOMATIC FEEDER, LOOKING EAST. THIS ...

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

19. DETAIL OF STAMP BATTERY AUTOMATIC FEEDER, LOOKING EAST. THIS IS THE MIDDLE OF THREE FEEDERS, ONE FOR EACH STAMP BATTERY. THE CHUTE (UPPER RIGHT) INTRODUCED THE CRUSHED ORE FROM THE ORE BIN. FLOW WAS CONTROLLED BY A SLIDING DOOR ON THE UPPER LEVEL. - Skidoo Mine, Park Route 38 (Skidoo Road), Death Valley Junction, Inyo County, CA

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

PubMed

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 feeding of hens in the ECH did not increase with increasing feeder space. However, it is worth noting that lower feeder space may lead to aggression or frustration which was not quantified in the current study.

Accuracy of software-assisted detection of tumour feeders in transcatheter hepatic chemoembolization using three target definition protocols.

PubMed

Iwazawa, J; Ohue, S; Hashimoto, N; Mitani, T

2014-02-01

To compare the accuracy of computer software analysis using three different target-definition protocols to detect tumour feeder vessels for transarterial chemoembolization of hepatocellular carcinoma. C-arm computed tomography (CT) data were analysed for 81 tumours from 57 patients who had undergone chemoembolization using software-assisted detection of tumour feeders. Small, medium, and large-sized targets were manually defined for each tumour. The tumour feeder was verified when the target tumour was enhanced on selective C-arm CT of the investigated vessel during chemoembolization. The sensitivity, specificity, and accuracy of the three protocols were evaluated and compared. One hundred and eight feeder vessels supplying 81 lesions were detected. The sensitivity of the small, medium, and large target protocols was 79.8%, 91.7%, and 96.3%, respectively; specificity was 95%, 88%, and 50%, respectively; and accuracy was 87.5%, 89.9%, and 74%, respectively. The sensitivity was significantly higher for the medium (p = 0.003) and large (p < 0.001) target protocols than for the small target protocol. The specificity and accuracy were higher for the small (p < 0.001 and p < 0.001, respectively) and medium (p < 0.001 and p < 0.001, respectively) target protocols than for the large target protocol. The overall accuracy of software-assisted automated feeder analysis in transarterial chemoembolization for hepatocellular carcinoma is affected by the target definition size. A large target definition increases sensitivity and decreases specificity in detecting tumour feeders. A target size equivalent to the tumour size most accurately predicts tumour feeders. Copyright © 2013 The Royal College of Radiologists. Published by Elsevier Ltd. All rights reserved.

Foraging scent marks of bumblebees: footprint cues rather than pheromone signals

NASA Astrophysics Data System (ADS)

Wilms, Jessica; Eltz, Thomas

2008-02-01

In their natural habitat foraging bumblebees refuse to land on and probe flowers that have been recently visited (and depleted) by themselves, conspecifics or other bees, which increases their overall rate of nectar intake. This avoidance is often based on recognition of scent marks deposited by previous visitors. While the term ‘scent mark’ implies active labelling, it is an open question whether the repellent chemicals are pheromones actively and specifically released during flower visits, or mere footprints deposited unspecifically wherever bees walk. To distinguish between the two possibilities, we presented worker bumblebees ( Bombus terrestris) with three types of feeders in a laboratory experiment: unvisited control feeders, passive feeders with a corolla that the bee had walked over on its way from the nest (with unspecific footprints), and active feeders, which the bee had just visited and depleted, but which were immediately refilled with sugar water (potentially with specific scent marks). Bumblebees rejected both active and passive feeders more frequently than unvisited controls. The rate of rejection of passive feeders was only slightly lower than that of active feeders, and this difference vanished completely when passive corollas were walked over repeatedly on the way from the nest. Thus, mere footprints were sufficient to emulate the repellent effect of an actual feeder visit. In confirmation, glass slides on which bumblebees had walked on near the nest entrance accumulated hydrocarbons (alkanes and alkenes, C23 to C31), which had previously been shown to elicit repellency in flower choice experiments. We conclude that repellent scent marks are mere footprints, which foraging bees avoid when they encounter them in a foraging context.

Derivation and characterization of putative embryonic stem cells from cloned rabbit embryos.

PubMed

Intawicha, Payungsuk; Siriboon, Chawalit; Chen, Chien-Hong; Chiu, Yung-Tsung; Lin, Tzu-An; Kere, Michel; Lo, Neng-Wen; Lee, Kun-Hsiung; Chang, Li-Yung; Chiang, Hsing-I; Ju, Jyh-Cherng

2016-10-15

The present study aimed to establish embryonic stem (ES) cell lines, i.e., ntES cells, using rabbit blastocyst stage embryos cloned by somatic cell nuclear transfer. First, we investigated the development of cloned rabbit embryos reconstructed with normal fibroblasts and fibroblasts transfected with enhanced green fluorescence protein (eGFP). Blastocyst rates were 27.4% and 23.9%, respectively, for the embryos reconstructed with normal fibroblasts and fibroblasts transfected with eGFP compared with that from the parthenogenetic group (43.1%). One ntES cell line was established from embryos reconstructed with eGFP-transfected fibroblasts (1 of 17, 5.9%), and three ntES cell lines were derived from those with normal fibroblasts (3 of 17, 17.6%). All the ntES cell lines retained alkaline phosphatase activity and expressed ES cell-specific markers SSEA-4, Oct-4, TRA-1-60, and TRA-1-81. The pluripotency was further confirmed by reverse transcription-polymerase chain reaction analyses of Oct-4, Nanog, and Sox-2 expressions in ntES cell lines. The differentiation capacity of ntES cells was also examined in vitro and in vivo, by which these ntES cell lines were able to differentiate into all three germ layers through embryoid bodies and teratomas. In conclusion, it is apparent that the efficiency of ntES cells derived using eGFP-transfected donor cells is lower than that with nontransfected, normal fibroblasts donor cells. Similar to those from parthenogenetic embryos, all ntES cell lines derived from cloned rabbit embryos are able to express pluripotency markers and retain their capability to differentiate into various cell lineages both in vitro and in vivo. Copyright © 2016 Elsevier Inc. All rights reserved.

Bacterial populations and adaptations in the mucus layers on living corals

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

Ducklow, H.W.; Mitchell, R.

1979-07-01

The external mucus layers of the stony coral Porites astreoides and the soft corals Palythoa sp. and Heteroxenia fuscesens are inhabited by communities of marine heterotrophic bacteria. Population levels of bacteria in coral mucus may be regulated by the self-cleaning behavior of the host. Bacterial populations in coral mucus respond to stresses applied to the host coral by growing to higher population levels in the mucus, indicating that these are populations of viable organisms closely attuned to host metabolism. Members of these microbial populations utilize the mucus compounds and may play a role in processing coral mucus for reef detritusmore » feeders. One such species, Vibrio alginolyticus, grows rapidly on Heteroxenia mucus, is attracted to dissolved mucus, and possesses a mechanism to maintain itself on the coral surface.« less

Voltage-Load Sensitivity Matrix Based Demand Response for Voltage Control in High Solar Penetration Distribution Feeders

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

Zhu, Xiangqi; Wang, Jiyu; Mulcahy, David

This paper presents a voltage-load sensitivity matrix (VLSM) based voltage control method to deploy demand response resources for controlling voltage in high solar penetration distribution feeders. The IEEE 123-bus system in OpenDSS is used for testing the performance of the preliminary VLSM-based voltage control approach. A load disaggregation process is applied to disaggregate the total load profile at the feeder head to each load nodes along the feeder so that loads are modeled at residential house level. Measured solar generation profiles are used in the simulation to model the impact of solar power on distribution feeder voltage profiles. Different casemore » studies involving various PV penetration levels and installation locations have been performed. Simulation results show that the VLSM algorithm performance meets the voltage control requirements and is an effective voltage control strategy.« less

Broadcasting satellite feeder links - Characteristics and planning

NASA Technical Reports Server (NTRS)

Kiebler, J. W.

1982-01-01

The paper presents the results of recent studies by the Feeder Link Sub-Working Group of the FCC Advisory Committee for the 1983 Regional Administrative Radio Conference (RARC). These studies conclude that specification of a few key parameters will make feeder link planning relatively straightforward. Feeder links can be located anywhere within a country if satellite orbit locations are separated by 10 deg for adjacent service areas and key parameter values presented in the paper are adopted. Colocated satellites serving a common service area need special attention to attain sufficient isolation between a desired channel and its adjacent cross-polarized channels and alternate co-polarized channels. In addition to presenting planning conclusions by the Advisory Committee, the paper presents and analyzes actions of the International Radio Consultative Committee's Conference Planning Meeting (CPM) concerning feeder links. The CPM reached conclusions similar to, and compatible with, those of the Advisory Committee.

Consequences of Feeder Delays for the Success of A380 Operations

NASA Technical Reports Server (NTRS)

Ruehle, Jens; Goetsch, Bjoern; Koch, Benjamin

2006-01-01

Due to existing slot and infrastructure constraints at international hub-and-spoke airports, an increase in feeder traffic seems only possible if larger feeder aircraft are used. Using a case study of Lufthansa German Airlines at Frankfurt International Airport, three possible A380 routes (Beijing, Tokyo-Narita, Los Angeles) were examined to assess the extent to which delays of feeder traffic may impact the economic performance of very large aircraft. On the basis of today s delays and anticipated traffic growth in the future, we found that between 9.5% and 13.5% of connecting passengers are unable to transfer to their respective intercontinental flights. In addition, the results demonstrate that a further increase in delays can be detrimental to the profitable operation of very large aircraft, as demonstrated by two out of three simulated routes. We suggest options for airlines operating very large aircraft to counteract the negative impacts of feeder delays.

Response of human corneal fibroblasts on silk film surface patterns.

PubMed

Gil, Eun Seok; Park, Sang-Hyug; Marchant, Jeff; Omenetto, Fiorenzo; Kaplan, David L

2010-06-11

Transparent, biodegradable, mechanically robust, and surface-patterned silk films were evaluated for the effect of surface morphology on human corneal fibroblast (hCF) cell proliferation, orientation, and ECM deposition and alignment. A series of dimensionally different surface groove patterns were prepared from optically graded glass substrates followed by casting poly(dimethylsiloxane) (PDMS) replica molds. The features on the patterned silk films showed an array of asymmetric triangles and displayed 37-342 nm depths and 445-3 582 nm widths. hCF DNA content on all patterned films were not significantly different from that on flat silk films after 4 d in culture. However, the depth and width of the grooves influenced cell alignment, while the depth differences affected cell orientation; overall, deeper and narrower grooves induced more hCF orientation. Over 14 d in culture, cell layers and actin filament organization demonstrated that confluent hCFs and their cytoskeletal filaments were oriented along the direction of the silk film patterned groove axis. Collagen type V and proteoglycans (decorin and biglycan), important markers of corneal stromal tissue, were highly expressed with alignment. Understanding corneal stromal fibroblast responses to surface features on a protein-based biomaterial applicable in vivo for corneal repair potential suggests options to improve corneal tissue mimics. Further, the approaches provide fundamental biomaterial designs useful for bioengineering oriented tissue layers, an endemic feature in most biological tissue structures that lead to critical tissue functions.

Effects of bale feeder type and supplementation of monensin on hay waste, intake, and performance of beef cattle

USDA-ARS?s Scientific Manuscript database

The effects of feeder type and supplemental monensin on hay utilization in beef cows was investigated using 56 crossbred beef cows (BW= 494 ± 50 kg; BCS= 5.2 ± 0.5) in a split-plot treatment arrangement with a completely randomized design. Supplement treatment served as the main plot and feeder desi...

High pressure rotary piston coal feeder

NASA Technical Reports Server (NTRS)

Gardner, J. F.; Gencsoy, H. T.; Strimbeck, D. C.

1977-01-01

This feeder concept uniquely combines the functions of solids feeding, metering, and pressurization into one compact system. Success with the rotary-piston concept would provide a lower-cost alternative to lock-hopper systems. The design of the feeder is presented, with special emphasis on the difficult problem of seal design. Initial tests will be to check seal performance. Subsequent tests will evaluate solids-feeding ability.

Low Spillage Metabolic Feeder

NASA Technical Reports Server (NTRS)

Evans, JuliAnn (Inventor); Gundo, Daniel P. (Inventor); Harper, Jennifer S. (Inventor); Mulenburg, Gerald M. (Inventor); Skundberg, Thomas L. (Inventor)

1996-01-01

An animal feeder for use in a metabolic cage is introduced. The feeder includes a confined passageway and an adjustable notched gate proceeding a food cup. The gate is adjusted so that the entry area to the food cup approximates the cross sectional head area of the animal. Food ejected from the food cup by a caged animal is dropped through a grate into a spill tray.

7 CFR 1230.113 - Collection and remittance of assessments for the sale of feeder pigs and market hogs.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 10 2011-01-01 2011-01-01 false Collection and remittance of assessments for the sale of feeder pigs and market hogs. 1230.113 Section 1230.113 Agriculture Regulations of the Department... pigs and market hogs. Pursuant to the provisions of § 1230.71, purchasers of feeder pigs or market hogs...

7 CFR 1230.113 - Collection and remittance of assessments for the sale of feeder pigs and market hogs.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 10 2014-01-01 2014-01-01 false Collection and remittance of assessments for the sale of feeder pigs and market hogs. 1230.113 Section 1230.113 Agriculture Regulations of the Department... pigs and market hogs. Pursuant to the provisions of § 1230.71, purchasers of feeder pigs or market hogs...

7 CFR 1230.113 - Collection and remittance of assessments for the sale of feeder pigs and market hogs.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 10 2012-01-01 2012-01-01 false Collection and remittance of assessments for the sale of feeder pigs and market hogs. 1230.113 Section 1230.113 Agriculture Regulations of the Department... pigs and market hogs. Pursuant to the provisions of § 1230.71, purchasers of feeder pigs or market hogs...

7 CFR 1230.113 - Collection and remittance of assessments for the sale of feeder pigs and market hogs.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 10 2013-01-01 2013-01-01 false Collection and remittance of assessments for the sale of feeder pigs and market hogs. 1230.113 Section 1230.113 Agriculture Regulations of the Department... pigs and market hogs. Pursuant to the provisions of § 1230.71, purchasers of feeder pigs or market hogs...

7 CFR 1230.113 - Collection and remittance of assessments for the sale of feeder pigs and market hogs.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 10 2010-01-01 2010-01-01 false Collection and remittance of assessments for the sale of feeder pigs and market hogs. 1230.113 Section 1230.113 Agriculture Regulations of the Department... pigs and market hogs. Pursuant to the provisions of § 1230.71, purchasers of feeder pigs or market hogs...

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

NASA Astrophysics Data System (ADS)

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.

High precision during food recruitment of experienced (reactivated) foragers in the stingless bee Scaptotrigona mexicana (Apidae, Meliponini)

NASA Astrophysics Data System (ADS)

Sánchez, Daniel; Nieh, James C.; Hénaut, Yann; Cruz, Leopoldo; Vandame, Rémy

Several studies have examined the existence of recruitment communication mechanisms in stingless bees. However, the spatial accuracy of location-specific recruitment has not been examined. Moreover, the location-specific recruitment of reactivated foragers, i.e., foragers that have previously experienced the same food source at a different location and time, has not been explicitly examined. However, such foragers may also play a significant role in colony foraging, particularly in small colonies. Here we report that reactivated Scaptotrigona mexicana foragers can recruit with high precision to a specific food location. The recruitment precision of reactivated foragers was evaluated by placing control feeders to the left and the right of the training feeder (direction-precision tests) and between the nest and the training feeder and beyond it (distance-precision tests). Reactivated foragers arrived at the correct location with high precision: 98.44% arrived at the training feeder in the direction trials (five-feeder fan-shaped array, accuracy of at least +/-6° of azimuth at 50 m from the nest), and 88.62% arrived at the training feeder in the distance trials (five-feeder linear array, accuracy of at least +/-5 m or +/-10% at 50 m from the nest). Thus, S. mexicana reactivated foragers can find the indicated food source at a specific distance and direction with high precision, higher than that shown by honeybees, Apis mellifera, which do not communicate food location at such close distances to the nest.

Number of hummingbird visits determines flower mite abundance on hummingbird feeders.

PubMed

Márquez-Luna, Ubaldo; Vázquez González, María Magdalena; Castellanos, Ignacio; Ortiz-Pulido, Raúl

2016-08-01

Members of several genera of mites from the family Melicharidae (Mesostigmata) use hummingbirds as transport host to move from flower to flower, where they feed on pollen and nectar. The factors that influence hummingbird flower mite abundance on host plant flowers are not currently known. Here we tested whether hummingbird flower mite abundance on an artificial nectar source is determined by number of hummingbird visits, nectar energy content or species richness of visiting hummingbirds. We conducted experiments employing hummingbird feeders with sucrose solutions of low, medium, and high energy concentrations, placed in a xeric shrubland. In the first experiment, we recorded the number of visiting hummingbirds and the number of visiting hummingbird species, as well as the abundance of hummingbird flower mites on each feeder. Feeders with the highest sucrose concentration had the most hummingbird visits and the highest flower mite abundances; however, there was no significant effect of hummingbird species richness on mite abundance. In the second experiment, we recorded flower mite abundance on feeders after we standardized the number of hummingbird visits to them. Abundance of flower mites did not differ significantly between feeders when we controlled for hummingbird visits. Our results suggest that nectar energy concentration determines hummingbird visits, which in turn determines flower mite abundance in our feeders. Our results do not support the hypothesis that mites descend from hummingbird nostrils more on richer nectar sources; however, it does not preclude the possibility that flower mites select for nectar concentration at other spatial and temporal scales.

47 CFR 25.258 - Sharing between NGSO MSS Feeder links Stations and GSO FSS services in the 29.25-29.5 GHz Bands.

Code of Federal Regulations, 2012 CFR

2012-10-01

.... (a) Operators of NGSO MSS feeder link earth stations and GSO FSS earth stations in the band 29.25 to... MSS feeder link earth station complexes, that will minimize instances of unacceptable interference to the GSO FSS space stations. Earth station licensees operating with GSO FSS systems shall be capable of...

47 CFR 25.257 - Special requirements for operations in the band 29.1-29.25 GHz between NGSO MSS and LMDS.

Code of Federal Regulations, 2012 CFR

2012-10-01

... Earth-to-space transmissions from feeder link earth station complexes. A “feeder link earth station complex” may include up to three (3) earth station groups, with each earth station group having up to four... NGSO MSS licensees or applicants pursuant to § 101.147. (b) A maximum of seven (7) feeder link earth...

47 CFR 25.257 - Special requirements for operations in the band 29.1-29.25 GHz between NGSO MSS and LMDS.

Code of Federal Regulations, 2014 CFR

2014-10-01

... Earth-to-space transmissions from feeder link earth station complexes. A “feeder link earth station complex” may include up to three (3) earth station groups, with each earth station group having up to four... NGSO MSS licensees or applicants pursuant to § 101.147. (b) A maximum of seven (7) feeder link earth...

47 CFR 25.257 - Special requirements for operations in the band 29.1-29.25 GHz between NGSO MSS and LMDS.

Code of Federal Regulations, 2011 CFR

2011-10-01

... Earth-to-space transmissions from feeder link earth station complexes. A “feeder link earth station complex” may include up to three (3) earth station groups, with each earth station group having up to four... NGSO MSS licensees or applicants pursuant to § 101.147. (b) A maximum of seven (7) feeder link earth...

47 CFR 25.257 - Special requirements for operations in the band 29.1-29.25 GHz between NGSO MSS and LMDS.

Code of Federal Regulations, 2010 CFR

2010-10-01

... Earth-to-space transmissions from feeder link earth station complexes. A “feeder link earth station complex” may include up to three (3) earth station groups, with each earth station group having up to four... NGSO MSS licensees or applicants pursuant to § 101.147. (b) A maximum of seven (7) feeder link earth...

47 CFR 25.258 - Sharing between NGSO MSS Feeder links Stations and GSO FSS services in the 29.25-29.5 GHz Bands.

Code of Federal Regulations, 2011 CFR

2011-10-01

.... (a) Operators of NGSO MSS feeder link earth stations and GSO FSS earth stations in the band 29.25 to... MSS feeder link earth station complexes, that will minimize instances of unacceptable interference to the GSO FSS space stations. Earth station licensees operating with GSO FSS systems shall be capable of...

47 CFR 25.258 - Sharing between NGSO MSS Feeder links Stations and GSO FSS services in the 29.25-29.5 GHz Bands.

Code of Federal Regulations, 2013 CFR

2013-10-01

.... (a) Operators of NGSO MSS feeder link earth stations and GSO FSS earth stations in the band 29.25 to... MSS feeder link earth station complexes, that will minimize instances of unacceptable interference to the GSO FSS space stations. Earth station licensees operating with GSO FSS systems shall be capable of...

47 CFR 25.223 - Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS.

Code of Federal Regulations, 2013 CFR

2013-10-01

... feeder link earth stations in the 17/24 GHz BSS. 25.223 Section 25.223 Telecommunication FEDERAL....223 Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS. (a) This section applies to all applications for earth station licenses in the 17/24 GHz BSS frequency...

47 CFR 25.223 - Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS.

Code of Federal Regulations, 2011 CFR

2011-10-01

... feeder link earth stations in the 17/24 GHz BSS. 25.223 Section 25.223 Telecommunication FEDERAL....223 Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS. (a) This section applies to all applications for earth station licenses in the 17/24 GHz BSS frequency...

47 CFR 25.258 - Sharing between NGSO MSS Feeder links Stations and GSO FSS services in the 29.25-29.5 GHz Bands.

Code of Federal Regulations, 2010 CFR

2010-10-01

.... (a) Operators of NGSO MSS feeder link earth stations and GSO FSS earth stations in the band 29.25 to... MSS feeder link earth station complexes, that will minimize instances of unacceptable interference to the GSO FSS space stations. Earth station licensees operating with GSO FSS systems shall be capable of...

47 CFR 25.223 - Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS.

Code of Federal Regulations, 2012 CFR

2012-10-01

... feeder link earth stations in the 17/24 GHz BSS. 25.223 Section 25.223 Telecommunication FEDERAL....223 Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS. (a) This section applies to all applications for earth station licenses in the 17/24 GHz BSS frequency...

47 CFR 25.258 - Sharing between NGSO MSS Feeder links Stations and GSO FSS services in the 29.25-29.5 GHz Bands.

Code of Federal Regulations, 2014 CFR

2014-10-01

.... (a) Operators of NGSO MSS feeder link earth stations and GSO FSS earth stations in the band 29.25 to... MSS feeder link earth station complexes, that will minimize instances of unacceptable interference to the GSO FSS space stations. Earth station licensees operating with GSO FSS systems shall be capable of...

47 CFR 25.257 - Special requirements for operations in the band 29.1-29.25 GHz between NGSO MSS and LMDS.

Code of Federal Regulations, 2013 CFR

2013-10-01

... Earth-to-space transmissions from feeder link earth station complexes. A “feeder link earth station complex” may include up to three (3) earth station groups, with each earth station group having up to four... NGSO MSS licensees or applicants pursuant to § 101.147. (b) A maximum of seven (7) feeder link earth...

47 CFR 25.223 - Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS.

Code of Federal Regulations, 2010 CFR

2010-10-01

... feeder link earth stations in the 17/24 GHz BSS. 25.223 Section 25.223 Telecommunication FEDERAL....223 Off-axis EIRP spectral density limits for feeder link earth stations in the 17/24 GHz BSS. (a) This section applies to all applications for earth station licenses in the 17/24 GHz BSS frequency...

The Effects of a Modified Treatment Package with and without Feeder Modeling on One Child's Acceptance of Novel Foods

ERIC Educational Resources Information Center

Seiverling, Laura; Harclerode, Whitney; Williams, Keith

2014-01-01

The purpose of this study was to examine if sequential presentation with feeder modeling would lead to an increase in bites accepted of new foods compared to sequential presentation without feeder modeling in a typically developing 4-year-old boy with food selectivity. The participant's acceptance of novel foods increased both in the modeling and…

Novel developmental biology-based protocol of embryonic stem cell differentiation to morphologically sound and functional yet immature hepatocytes.

PubMed

Bukong, Terence N; Lo, Tracie; Szabo, Gyongyi; Dolganiuc, Angela

2012-05-01

Liver diseases are common in the United States and often require liver transplantation; however, donated organs are limited and thus alternative sources for liver cells are in high demand. Embryonic stem cells (ESC) can provide a continuous and readily available source of liver cells. ESC differentiation to liver cells is yet to be fully understood and comprehensive differentiation protocols are yet to be defined. Here, we aimed to achieve human (h)ESC differentiation into mature hepatocytes using defined recombinant differentiation factors and metabolites. Embryonic stem cell H1 line was sub-cultured on feeder layer. We induced hESCs into endodermal differentiation succeeded by early/late hepatic specification and finally into hepatocyte maturation using step combinations of Activin A and fibroblast growth factor (FGF)-2 for 7 days; followed by FGF-4 and bone morphogenic protein 2 (BMP2) for 7 days, succeeded by FGF-10 + hepatocyte growth factor 4 + epidermal growth factor for 14 days. Specific inhibitors/stimulators were added sequentially throughout differentiation. Cells were analysed by PCR, flow cytometry, microscopy or functional assays. Our hESC differentiation protocol resulted in viable cells with hepatocyte shape and morphology. We observed gradual changes in cell transcriptome, including up-regulation of differentiation-promoting GATA4, GATA6, POU5F1 and HNF4 transcription factors, steady levels of stemness-promoting SOX-2 and low levels of Nanog, as defined by PCR. The hESC-derived hepatocytes expressed alpha-antitrypsin, CD81, cytokeratin 8 and low density lipoprotein (LDL) receptor. The levels of alpha-fetoprotein and proliferation marker Ki-67 in hESC-derived hepatocytes remained elevated. Unlike stem cells, the hESC-derived hepatocytes performed LDL uptake, produced albumin and alanine aminotransferase and had functional alcohol dehydrogenase. We report a novel protocol for hESC differentiation into morphological and functional yet immature hepatocytes as an alternative method for hepatocyte generation. © 2012 John Wiley & Sons A/S.

Ultrastructural study of the primary olfactory pathway in Macaca fascicularis.

PubMed

Herrera, Loren P; Casas, Carlos E; Bates, Margaret L; Guest, James D

2005-08-08

Olfactory ensheathing glial cells (OEGs) interact with a wide repertoire of cell types and support extension of olfactory axons (OAs) within the olfactory pathway. OEGs are thought to exclude OAs from contact with all other cells between the olfactory epithelium and the glomerulus of the olfactory bulb. These properties have lead to testing to determine whether OEGs support axonal growth following transplantation. The cellular interactions of transplanted OEGs will probably resemble those that occur within the normal pathway where interactions between OEGs and fibroblasts are prominent. No previous primate studies have focused on these interactions, knowledge of which is important if clinical application is envisioned. We describe the detailed intercellular interactions of OAs with supporting cells throughout the olfactory epithelium, the lamina propria, the fila olfactoria, and the olfactory nerve layer by using transmission electron microscopy in adult Macaca fascicularis. Patterns of OEG ensheathment and variations of the endo- and perineurium formed by olfactory nerve fibroblasts are described. OAs mainly interacted with horizontal basal cells, OEGs, and astrocytes. At both transitional ends of the pathway seamless intercellular interactions were observed, and fibroblast processes were absent. Perineurial cells produced surface basal lamina; however, endoneurial, epineurial, and meningeal fibroblasts did not. Perineurial cells contained intermediate filaments and were distinct from other fibroblasts and meningeal cells. OAs had direct contacts with astrocytes near the glia limitans. The properties of OEGs differed depending on whether astrocytic or fibroblastic processes were present. This indicates the importance of the cellular milieu in the structure and function of OEGs in primates.

TRPV2 channel inhibitors attenuate fibroblast differentiation and contraction mediated by keratinocyte-derived TGF-β1 in an in vitro wound healing model of rats.

PubMed

Ishii, Taro; Uchida, Kunitoshi; Hata, Shozaburo; Hatta, Mitsutoki; Kita, Tomo; Miyake, Yuki; Okamura, Kazuhiko; Tamaoki, Sachio; Ishikawa, Hiroyuki; Yamazaki, Jun

2018-06-01

Keratinocytes release several factors that are involved in wound contracture and scar formation. We previously reported that a three-dimensional reconstruction model derived from rat skin represents a good wound healing model. We characterized the role of transient receptor potential (TRP) channels in the release of transforming growth factor (TGF)-β1 from keratinocytes and the differentiation of fibroblasts to identify possible promising pharmacological approaches to prevent scar formation and contractures. The three-dimensional culture model was made from rat keratinocytes seeded on a collagen gel in which dermal fibroblasts had been embedded. Among the TRP channel inhibitors tested, the TRPV2 inhibitors SKF96365 and tranilast attenuated most potently keratinocyte-dependent and - independent collagen gel contraction due to TGF-β signaling as well as TGF-β1 release from keratinocytes and α-smooth muscle actin production in myofibroblasts. Besides the low amounts detected in normal dermis, TRPV2 mRNA and protein levels were increased after fibroblasts were embedded in the gel. TRPV2 was also expressed in the epidermis and keratinocyte layers of the model. Both inhibitors and TRPV2 siRNA attenuated the intracellular increase of Ca 2+ induced by the TRPV agonist 2-aminoethoxydiphenyl borate in TGF-β1-pretreated fibroblasts. This is the first study to show that compounds targeting TRPV2 channels ameliorate wound contraction through the inhibition of TGF-β1 release and the differentiation of dermal fibroblasts in a culture model. Copyright © 2018. Published by Elsevier B.V.

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.

PubMed

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 < 0.05). Pigs fed the D30 treatment had greater (P < 0.05) ATTD for DM and GE for both collection periods compared with those on the D60 treatment. In summary, 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 yield because of increasing intestinal weights when pigs are fed diets containing increasing DDGS inclusion rates. Dry matter and energy digestibility were greater in pigs fed the lower DDGS treatment.

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

PubMed

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 < 0.01) for SF than for CF or CFL. However, feeder design did not influence the other performance and 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 < 0.05) rumen pH compared with CF and CFL bulls. On d 7, the acetate to propionate ratio from SF was greater (P < 0.05) than for CFL or CF. At d 7, NEFA of SF were greater (P < 0.05) compared with CF and CFL. Bulls fed with CF have the greatest (P < 0.01) concentrate disappearance velocity followed by bulls fed with CFL and finally by bulls fed with SF, and this was associated with different feeding behaviors. Bulls on SF spent more time (P < 0.05) eating straw and exhibited fewer (P < 0.05) displacements at concentrate feeder than CF and CFL bulls. The CFL bulls exhibited (P < 0.01) more attempted mounts and tended (P = 0.10) to exhibit more completed mounts than CF bulls. In conclusion, both alternative feeder designs (CFL and SF) are good strategies to reduce total concentrate consumption without impairing performance, rumen health, or 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.

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.

PubMed

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-day egg production in combination with 10.4 cm/pullet rearing but not with 7.0 cm/pullet rearing space. In a similar manner, hen mortality was greater in the 7.0 to 10.3 cm/female feeder space combination that the 10.4 to 10.3 cm/female combination. © 2014 Poultry Science Association Inc.

Effects of protein-coated nanofibers on conformation of gingival fibroblast spheroids: potential utility for connective tissues regeneration.

PubMed

Kaufman, Gili; Whitescarver, Ryan; Nunes, Laiz; Palmer, Xavier-Lewis; Skrtic, Drago; Tutak, Wojtek

2017-10-09

Deep wounds in the gingiva caused by trauma or surgery require a rapid and robust healing of connective tissues. We propose utilizing gas-brushed nanofibers coated with collagen and fibrin for that purpose. Our hypotheses are that protein-coated nanofibers will: (i) attract and mobilize cells in various spatial orientations, and (ii) regulate the expression levels of specific extracellular matrix (ECM)-associated proteins, determining the initial conformational nature of dense and soft connective tissues. Gingival fibroblast monolayers and 3D spheroids were cultured on ECM substrate and covered with gas-blown poly-(DL-lactide-co-glycolide) (PLGA) nanofibers (uncoated/coated with collagen and fibrin). Cell attraction and rearrangement was followed by F-actin staining and confocal microscopy. Thicknesses of the cell layers, developed within the nanofibers, were quantified by imageJ software. The expression of collagen1α1 chain (Col1α1), fibronectin, and metalloproteinase 2 (MMP2) encoding genes was determined by quantitative reverse transcription analysis. Collagen- and fibrin- coated nanofibers induced cell migration toward fibers and supported cellular growth within the scaffolds. Both proteins affected the spatial rearrangement of fibroblasts by favoring packed cell clusters or intermittent cell spreading. These cell arrangements resembled the structural characteristic of dense and soft connective tissues, respectively. Within 3 days of incubation, fibroblast spheroids interacted with the fibers and grew robustly by increasing their thickness compared to monolayers. While the ECM key components, such as fibronectin and MMP2 encoding genes, were expressed in both protein groups, Col1α1 was predominantly expressed in bundled fibroblasts grown on collagen fibers. This enhanced expression of collagen1 is typical for dense connective tissue. Based on results of this study, our gas-blown, collagen- and fibrin-coated PLGA nanofibers are viable candidates for engineering soft and dense connective tissues with the required structural characteristics and functions needed for wound healing applications. Rapid regeneration of these layers should enhance healing of open wounds in a harsh oral environment. © 2017 IOP Publishing Ltd.

Three-dimensional model of an ultramafic feeder system to the Nikolai Greenstone mafic large igneous province, central Alaska Range

USGS Publications Warehouse

Glen, J.M.G.; Schmidt, J.M.; Connard, G.G.

2011-01-01

The Amphitheater Mountains and southern central Alaska Range expose a thick sequence of Triassic Nikolai basalts that is underlain by several mafic-ultramafic complexes, the largest and best exposed being the Fish Lake and Tangle (FL-T) mafic-ultramafic sills that flank the Amphitheater Mountains synform. Three-dimensional (3-D) modeling of gravity and magnetic data reveals details of the structure of the Amphitheater Mountains, such as the orientation and thickness of Nikolai basalts, and the geometry of the FL-T intrusions. The 3-D model (50 ?? 70 km) includes the full geographic extent of the FL-T complexes and consists of 11 layers. Layer surfaces and properties (density and magnetic susceptibility) were modified by forward and inverse methods to reduce differences between the observed and calculated gravity and magnetic grids. The model suggests that the outcropping FL-T sills are apparently connected and traceable at depth and reveals variations in thickness, shape, and orientation of the ultramafic bodies that may identify paths of magma flow. The model shows that a significant volume (2000 km3) of ultramafic material occurs in the subsurface, gradually thickening and plunging westward to depths exceeding 4 km. This deep ultramafic material is interpreted as the top of a keel or root system that supplied magma to the Nikolai lavas and controlled emplacement of related magmatic intrusions. The presence of this deep, keel-like structure, and asymmetry of the synform, supports a sag basin model for development of the Amphitheater Mountains structure and reveals that the feeders to the Nikolai are much more extensive than previously known. Copyright 2011 by the American Geophysical Union.

Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

PubMed Central

Gardell, Alison M.; Qin, Qin; Rice, Robert H.; Li, Johnathan; Kültz, Dietmar

2014-01-01

Fish cell cultures are becoming more widely used models for investigating molecular mechanisms of physiological response to environmental challenge. In this study, we derived two immortalized Mozambique tilapia (Oreochromis mossambicus) cell lines from brain (OmB) and lip epithelium (OmL), and compared them to a previously immortalized bulbus arteriosus (TmB) cell line. The OmB and OmL cell lines were generated without or with Rho-associated kinase (ROCK) inhibitor/3T3 feeder layer supplementation. Although both approaches were successful, ROCK inhibitor/feeder layer supplementation was found to offer the advantages of selecting for epithelial-like cell type and decreasing time to immortalization. After immortalization (≥ passage 5), we characterized the proteomes of the newly derived cell lines (OmB and OmL) using LCMS and identified several unique cell markers for each line. Subsequently, osmotolerance for each of the three cell lines following acute exposure to elevated sodium chloride was evaluated. The acute maximum osmotolerance of these tilapia cell lines (>700 mOsm/kg) was markedly higher than that of any other known vertebrate cell line, but was significantly higher in the epithelial-like OmL cell line. To validate the physiological relevance of these tilapia cell lines, we quantified the effects of acute hyperosmotic challenge (450 mOsm/kg and 700 mOsm/kg) on the transcriptional regulation of two enzymes involved in biosynthesis of the compatible organic osmolyte, myo-inositol. Both enzymes were found to be robustly upregulated in all three tilapia cell lines. Therefore, the newly established tilapia cells lines represent valuable tools for studying molecular mechanisms involved in the osmotic stress response of euryhaline fish. PMID:24797371

Analysis of PV Advanced Inverter Functions and Setpoints under Time Series Simulation.

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

Seuss, John; Reno, Matthew J.; Broderick, Robert Joseph

Utilities are increasingly concerned about the potential negative impacts distributed PV may have on the operational integrity of their distribution feeders. Some have proposed novel methods for controlling a PV system's grid - tie inverter to mitigate poten tial PV - induced problems. This report investigates the effectiveness of several of these PV advanced inverter controls on improving distribution feeder operational metrics. The controls are simulated on a large PV system interconnected at several locations within two realistic distribution feeder models. Due to the time - domain nature of the advanced inverter controls, quasi - static time series simulations aremore » performed under one week of representative variable irradiance and load data for each feeder. A para metric study is performed on each control type to determine how well certain measurable network metrics improve as a function of the control parameters. This methodology is used to determine appropriate advanced inverter settings for each location on the f eeder and overall for any interconnection location on the feeder.« less

Analysis of Feeder Bus Network Design and Scheduling Problems

PubMed Central

Almasi, Mohammad Hadi; Karim, Mohamed Rehan

2014-01-01

A growing concern for public transit is its inability to shift passenger's mode from private to public transport. In order to overcome this problem, a more developed feeder bus network and matched schedules will play important roles. The present paper aims to review some of the studies performed on Feeder Bus Network Design and Scheduling Problem (FNDSP) based on three distinctive parts of the FNDSP setup, namely, problem description, problem characteristics, and solution approaches. The problems consist of different subproblems including data preparation, feeder bus network design, route generation, and feeder bus scheduling. Subsequently, descriptive analysis and classification of previous works are presented to highlight the main characteristics and solution methods. Finally, some of the issues and trends for future research are identified. This paper is targeted at dealing with the FNDSP to exhibit strategic and tactical goals and also contributes to the unification of the field which might be a useful complement to the few existing reviews. PMID:24526890

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

NASA Astrophysics Data System (ADS)

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

2016-02-01

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

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

NASA Astrophysics Data System (ADS)

Funamoto, Daichi; Sugiura, Shinji

2017-02-01

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

Social and nonsocial behaviours of sex- and age-matched enclosed and free-ranging rhesus monkeys (Macaca mulatta).

PubMed

Baulu, J; Redmond, D E

1980-01-01

The behavioural profiles (time budgeting of social and nonsocial activities) and the frequencies of major social interactions of corral-enclosed rhesus monkeys were compared with sex- and age-matched free-ranging rhesus monkeys on La Cueva Island, Puerto Rico. All animals (n = 32) were provisioned ad libitum at specific feeder sites. The occurrence of 14 behaviours around feeders was compared with their occurrence away from the feeders by noting the location of each monkey relative to the feeder at the time of observation. An analysis of variance between free-ranging versus corral-enclosed groups and within groups by location (around or away from the feeder) revealed significant differences in several behavioural categories, including foraging, lookout, inactive, dominant, submissive, allogrooming, social contact, social initiative, active, and passive behaviours. When the effect of limited food distribution sites was analyzed by comparing data recorded away from the feeding sites, there were remarkably few differences between the groups.

A new angle on microscopic suspension feeders near boundaries.

PubMed

Pepper, Rachel E; Roper, Marcus; Ryu, Sangjin; Matsumoto, Nobuyoshi; Nagai, Moeto; Stone, Howard A

2013-10-15

Microscopic sessile suspension feeders are a critical component in aquatic ecosystems, acting as an intermediate trophic stage between bacteria and higher eukaryotic taxa. Because they live attached to boundaries, it has long been thought that recirculation of the feeding currents produced by sessile suspension feeders inhibits their ability to access fresh fluid. However, previous models for the feeding flows of these organisms assume that they feed by pushing fluid perpendicular to surfaces they live upon, whereas we observe that sessile suspension feeders often feed at an angle to these boundaries. Using experiments and calculations, we show that living suspension feeders (Vorticella) likely actively regulate the angle that they feed relative to a substratum. We then use theory and simulations to show that angled feeding increases nutrient and particle uptake by reducing the reprocessing of depleted water. This work resolves an open question of how a key class of suspension-feeding organisms escapes physical limitations associated with their sessile lifestyle. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Nutrient compensatory foraging in a free-living social insect

NASA Astrophysics Data System (ADS)

Christensen, Keri L.; Gallacher, Anthony P.; Martin, Lizzie; Tong, Desmond; Elgar, Mark A.

2010-10-01

The geometric framework model predicts that animal foraging decisions are influenced by their dietary history, with animals targeting a combination of essential nutrients through compensatory foraging. We provide experimental confirmation of nutrient-specific compensatory foraging in a natural, free-living population of social insects by supplementing their diet with sources of protein- or carbohydrate-rich food. Colonies of the ant Iridomyrmex suchieri were provided with feeders containing food rich in either carbohydrate or protein for 6 days, and were then provided with a feeder containing the same or different diet. The patterns of recruitment were consistent with the geometric framework: while feeders with a carbohydrate diet typically attracted more workers than did feeders with protein diet, the difference in recruitment between the two nutrients was smaller if the colonies had had prior access to carbohydrate than protein. Further, fewer ants visited feeders if the colony had had prior access to protein than to carbohydrates, suggesting that the larvae play a role in worker foraging behaviour.

An enhanced SOCP-based method for feeder load balancing using the multi-terminal soft open point in active distribution networks

DOE PAGES

Ji, Haoran; Wang, Chengshan; Li, Peng; ...

2017-09-20

The integration of distributed generators (DGs) exacerbates the feeder power flow fluctuation and load unbalanced condition in active distribution networks (ADNs). The unbalanced feeder load causes inefficient use of network assets and network congestion during system operation. The flexible interconnection based on the multi-terminal soft open point (SOP) significantly benefits the operation of ADNs. The multi-terminal SOP, which is a controllable power electronic device installed to replace the normally open point, provides accurate active and reactive power flow control to enable the flexible connection of feeders. An enhanced SOCP-based method for feeder load balancing using the multi-terminal SOP is proposedmore » in this paper. Furthermore, by regulating the operation of the multi-terminal SOP, the proposed method can mitigate the unbalanced condition of feeder load and simultaneously reduce the power losses of ADNs. Then, the original non-convex model is converted into a second-order cone programming (SOCP) model using convex relaxation. In order to tighten the SOCP relaxation and improve the computation efficiency, an enhanced SOCP-based approach is developed to solve the proposed model. Finally, case studies are performed on the modified IEEE 33-node system to verify the effectiveness and efficiency of the proposed method.« less

Object categorization by wild ranging birds-Winter feeder experiments.

PubMed

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.

An enhanced SOCP-based method for feeder load balancing using the multi-terminal soft open point in active distribution networks

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

Ji, Haoran; Wang, Chengshan; Li, Peng

The integration of distributed generators (DGs) exacerbates the feeder power flow fluctuation and load unbalanced condition in active distribution networks (ADNs). The unbalanced feeder load causes inefficient use of network assets and network congestion during system operation. The flexible interconnection based on the multi-terminal soft open point (SOP) significantly benefits the operation of ADNs. The multi-terminal SOP, which is a controllable power electronic device installed to replace the normally open point, provides accurate active and reactive power flow control to enable the flexible connection of feeders. An enhanced SOCP-based method for feeder load balancing using the multi-terminal SOP is proposedmore » in this paper. Furthermore, by regulating the operation of the multi-terminal SOP, the proposed method can mitigate the unbalanced condition of feeder load and simultaneously reduce the power losses of ADNs. Then, the original non-convex model is converted into a second-order cone programming (SOCP) model using convex relaxation. In order to tighten the SOCP relaxation and improve the computation efficiency, an enhanced SOCP-based approach is developed to solve the proposed model. Finally, case studies are performed on the modified IEEE 33-node system to verify the effectiveness and efficiency of the proposed method.« less

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

PubMed

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 future hiPSC-based clinical trials or therapies and for drug screening.

Characterization and evaluation of whey protein-based biofilms as substrates for in vitro cell cultures.

PubMed

Gilbert, Vanessa; Rouabhia, Mahmoud; Wang, Hongxum; Arnould, Anne-Lise; Remondetto, Gabriel; Subirade, Muriel

2005-12-01

Whey proteins-based biofilms were prepared using different plasticizers in order to obtain a biomaterial for the human keratinocytes and fibroblasts in vitro culture. The film properties were evaluated by Fourier Transform Infrared Spectroscopy (FTIR) technique and mechanical tests. A relationship was found between the decrease of intermolecular hydrogen bond strength and film mechanical behavior changes, expressed by a breaking stress and Young modulus values diminishing. These results allow stating that the film molecular configuration could induce dissimilarities in its mechanical properties. The films toxicity was assessed by evaluating the cutaneous cells adherence, growth, proliferation and structural stratification. Microscopic observation demonstrated that both keratinocytes and fibroblasts adhered to the biofilms. The trypan blue exclusion test showed that keratinocytes grew at a significantly high rate on all the biofilms. Structural analysis demonstrated that keratinocytes stratified when cultured on the whey protein-based biofilms and gave rise to multi-layered epidermal structures. The most organized epidermis was obtained with whey protein isolate/DEG biofilm. This structure had a well-organized basal layer under supra-basal and corneous layers. This study demonstrated that whey proteins, an inexpensive renewable resource which can be obtained readily, were non-toxic to cutaneous cells and thus they could be useful substrates for a variety of biomedical applications, including tissue engineering.

Assessment of cleaning and disinfection in Salmonella-contaminated poultry layer houses using qualitative and semi-quantitative culture techniques.

PubMed

Wales, Andrew; Breslin, Mark; Davies, Robert

2006-09-10

Salmonella infection of laying flocks in the UK is predominantly a problem of the persistent contamination of layer houses and associated wildlife vectors by Salmonella Enteritidis. Methods for its control and elimination include effective cleaning and disinfection of layer houses between flocks, and it is important to be able to measure the success of such decontamination. A method for the environmental detection and semi-quantitative enumeration of salmonellae was used and compared with a standard qualitative method, in 12 Salmonella-contaminated caged layer houses before and after cleaning and disinfection. The quantitative technique proved to have comparable sensitivity to the standard method, and additionally provided insights into the numerical Salmonella challenge that replacement flocks would encounter. Elimination of S. Enteritidis was not achieved in any of the premises examined although substantial reductions in the prevalence and numbers of salmonellae were demonstrated, whilst in others an increase in contamination was observed after cleaning and disinfection. Particular problems with feeders and wildlife vectors were highlighted. The use of a quantitative method assisted the identification of problem areas, such as those with a high initial bacterial load or those experiencing only a modest reduction in bacterial count following decontamination.

Technical note: Method for isolation of the bovine sweat gland and conditions for in vitro culture.

PubMed

Hamzaoui, S; Burger, C A; Collier, J L; Collier, R J

2018-05-01

Apocrine sweat glands in bovine skin are involved in thermoregulation. Human, horse, and sheep sweat gland epithelial cells have been isolated and grown in vitro. The present study was conducted to identify a method to isolate bovine sweat glands and culture apocrine bovine sweat gland epithelial cells in vitro. Mechanical shearing, collagenase digestion, centrifugation, and neutral red staining were used to identify and isolate the apocrine glands from skin. Bovine sweat glands in situ and after isolation comprised 2 major cell types consisting of a single layer of cuboidal epithelial cells resting on a layer of myoepithelial cells. In situ, the glands were embedded in a collagen matrix primarily comprising fibroblasts, and some of these cells were also present in the isolated material. The isolated material was transferred to complete medium (keratinocyte serum-free medium, bovine pituitary extract, and human recombinant epidermal growth factor + 2.5% fetal bovine serum) in a T 25 flask (Falcon, Franklin Lakes, NJ) with media film and then incubated at 37°C for 24 h. After sweat glands adhered to the bottom of the flask, an additional 2 mL of complete medium was added and the medium was changed every 3 d. Isolated apocrine sweat glands and bovine sweat gland epithelial cells were immunostained for cytokeratin and fibroblast specific protein, indicating fibroblast-free cultures. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Cardiac myocyte diversity and a fibroblast network in the junctional region of the zebrafish heart revealed by transmission and serial block-face scanning electron microscopy.

PubMed

Lafontant, Pascal J; Behzad, Ali R; Brown, Evelyn; Landry, Paul; Hu, Norman; Burns, Alan R

2013-01-01

The zebrafish has emerged as an important model of heart development and regeneration. While the structural characteristics of the developing and adult zebrafish ventricle have been previously studied, little attention has been paid to the nature of the interface between the compact and spongy myocardium. Here we describe how these two distinct layers are structurally and functionally integrated. We demonstrate by transmission electron microscopy that this interface is complex and composed primarily of a junctional region occupied by collagen, as well as a population of fibroblasts that form a highly complex network. We also describe a continuum of uniquely flattened transitional cardiac myocytes that form a circumferential plate upon which the radially-oriented luminal trabeculae are anchored. In addition, we have uncovered within the transitional ring a subpopulation of markedly electron dense cardiac myocytes. At discrete intervals the transitional cardiac myocytes form contact bridges across the junctional space that are stabilized through localized desmosomes and fascia adherentes junctions with adjacent compact cardiac myocytes. Finally using serial block-face scanning electron microscopy, segmentation and volume reconstruction, we confirm the three-dimensional nature of the junctional region as well as the presence of the sheet-like fibroblast network. These ultrastructural studies demonstrate the previously unrecognized complexity with which the compact and spongy layers are structurally integrated, and provide a new basis for understanding development and regeneration in the zebrafish heart.

Localization of basic fibroblast growth factor binding sites in the chick embryonic neural retina.

PubMed

Cirillo, A; Arruti, C; Courtois, Y; Jeanny, J C

1990-12-01

We have investigated the localization of basic fibroblast growth factor (bFGF) binding sites during the development of the neural retina in the chick embryo. The specificity of the affinity of bFGF for its receptors was assessed by competition experiments with unlabelled growth factor or with heparin, as well as by heparitinase treatment of the samples. Two different types of binding sites were observed in the neural retina by light-microscopic autoradiography. The first type, localized mainly to basement membranes, was highly sensitive to heparitinase digestion and to competition with heparin. It was not developmentally regulated. The second type of binding site, resistant to heparin competition, appeared to be associated with retinal cells from the earliest stages studied (3-day-old embryo, stages 21-22 of Hamburger and Hamilton). Its distribution was found to vary during embryonic development, paralleling layering of the neural retina. Binding of bFGF to the latter sites was observed throughout the retinal neuroepithelium at early stages but displayed a distinct pattern at the time when the inner and outer plexiform layers were formed. During the development of the inner plexiform layer, a banded pattern of bFGF binding was observed. These bands, lying parallel to the vitreal surface, seemed to codistribute with the synaptic bands existing in the inner plexiform layer. The presence of intra-retinal bFGF binding sites whose distribution varies with embryonic development suggests a regulatory mechanism involving differential actions of bFGF on neural retinal cells.

Structure of a Multilayer Nanofilm To Increase the Encapsulation Efficiency of Basic Fibroblast Growth Factor.

PubMed

Han, Uiyoung; Hong, Jinkee

2018-03-05

In this study, we established the structure of a multilayer nanofilm that more efficiently encapsulates basic fibroblast growth factor (bFGF). First, a positively charged layer material was selected from biocompatible polymers such as collagen (Col), poly(beta-amino ester) (Poly2), and chitosan (Chi), while considering the film thickness. We then investigated the change in bFGF encapsulation efficiency when the multilayer structure was changed from a tetralayer to a trilayer. As a result, we obtained a highly improved bFGF encapsulation efficiency in the nanofilm using a positively charged layer formed by a blend of Col and Poly2 and a negatively charged poly(acrylic acid) (PAA) layer within a trilayered structure. In particular, we found that a significant amount of adsorbed bFGF was desorbed again during the film fabrication process of a tetralayered nanofilm. In the conventional nanofilm, bFGF was regarded as a polycation and formed a multilayer nanofilm that was composed of a tetralayered structure and was represented as (polycation/polyanion/bFGF/polyanion) n where n = number of repeated tetralayers. Here, we suggested that bFGF should not be considered a polycation, rather it should be considered as a small quantity of molecule that exists between the polyanion and polycation layers. In this case, the nanofilm is composed of repeating units of (polycation/polyanion/bFGF/polycation/polyanion), because the amount of adsorbed bFGF is considerably lower than that of other building blocks.

Effects of diet form and feeder adjustment on growth performance of nursery and finishing pigs.

PubMed

Nemechek, J E; Tokach, M D; Dritz, S S; Fruge, E D; Hansen, E L; Goodband, R D; DeRouchey, J M; Woodworth, J C

2015-08-01

Three experiments were conducted to determine the effects of feeder adjustment and diet form on growth performance of nursery (Exp. 1 and 2) and finishing (Exp. 3) pigs. Treatments were arranged as a 2 × 3 factorial with the main effects of feeder adjustment and diet form. The 2 feeder adjustments were a narrow and wide feeder adjustment (minimum gap opening of 1.27 and 2.54 cm, respectively). The 3 diet forms were meal, poor-quality pellets (70% pellets and 30% fines for Exp. 1 and 2 and 50% pellets and 50% fines for Exp. 3), and screened pellets with minimal fines (3 to 10%). In Exp. 1, 210 pigs (initially 11.9 kg BW) were used in a 21-d trial with 7 pigs per pen and 5 pens per treatment. No feeder adjustment × diet form interactions were observed. There were no differences in ADG, ADFI, or G:F due to feeder adjustment. Pigs fed the meal diet had increased ( < 0.05) ADG and ADFI compared with pigs fed the poor-quality or screened pellets. Pigs fed meal or poor-quality pellets had decreased ( < 0.05) G:F compared with pigs fed screened pellets. In Exp. 2, 1,005 nursery pigs (initially 14.1 kg BW) were used in a 28-d trial with 26 to 28 pigs per pen and 6 pens per treatment. Pigs fed from the narrow feeder adjustment had decreased ( < 0.05) ADG and ADFI compared with pigs fed from the wide adjustment with no differences in G:F. Pigs fed the meal diet had decreased ( < 0.05) ADG compared with pigs fed poor-quality or screened pellets. Pigs fed meal or poor-quality pellets had decreased ( < 0.05) G:F compared with pigs fed screened pellets. In Exp. 3, 246 pigs (initially 56.8 kg BW) were used in a 69-d trial with 5 pens per treatment and 6 or 7 pigs per pen. Overall, ADFI decreased ( < 0.05) and G:F increased ( < 0.05) for pigs fed from the narrow adjusted feeders compared with the wide adjustment with no differences in ADG. Overall, pigs fed meal diets tended to have decreased ( < 0.10) ADG and had decreased ( < 0.05) G:F compared with pigs fed screened pellets; ADG and G:F in those fed poor-quality pellets were intermediate. Feeding meal or poor-quality pellets increased ( < 0.05) ADFI compared with pigs fed screened pellets. In conclusion, feeding nursery pigs from a wide feeder gap may increase ADG and ADFI with no negative effects on G:F. For finishing pigs, reducing feeder gap reduced feed disappearance and improved G:F. In all experiments, the greatest G:F improvements from pelleting were observed when the percentage of fines was minimized.

Modern Grid Initiative Distribution Taxonomy Final Report

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

Schneider, Kevin P.; Chen, Yousu; Chassin, David P.

2008-11-01

This is the final report for the development of a toxonomy of prototypical electrical distribution feeders. Two of the primary goals of the Department of Energy's (DOE) Modern Grid Initiative (MGI) are 'to accelerate the modernization of our nation's electricity grid' and to 'support demonstrations of systems of key technologies that can serve as the foundation for an integrated, modern power grid'. A key component to the realization of these goals is the effective implementation of new, as well as existing, 'smart grid technologies'. Possibly the largest barrier that has been identified in the deployment of smart grid technologies ismore » the inability to evaluate how their deployment will affect the electricity infrastructure, both locally and on a regional scale. The inability to evaluate the impacts of these technologies is primarily due to the lack of detailed electrical distribution feeder information. While detailed distribution feeder information does reside with the various distribution utilities, there is no central repository of information that can be openly accessed. The role of Pacific Northwest National Laboratory (PNNL) in the MGI for FY08 was to collect distribution feeder models, in the SynerGEE{reg_sign} format, from electric utilities around the nation so that they could be analyzed to identify regional differences in feeder design and operation. Based on this analysis PNNL developed a taxonomy of 24 prototypical feeder models in the GridLAB-D simulations environment that contain the fundamental characteristics of non-urban core, radial distribution feeders from the various regions of the U.S. Weighting factors for these feeders are also presented so that they can be used to generate a representative sample for various regions within the United States. The final product presented in this report is a toolset that enables the evaluation of new smart grid technologies, with the ability to aggregate their effects to regional and national levels. The distribution feeder models presented in this report are based on actual utility models but do not contain any proprietary or system specific information. As a result, the models discussed in this report can be openly distributed to industry, academia, or any interested entity, in order to facilitate the ability to evaluate smart grid technologies.« less

Characterization of cultivated murine lacrimal gland epithelial cells

PubMed Central

Kobayashi, Shinya; Kawashima, Motoko; Okada, Naoko; Mishima, Kenji; Saito, Ichiro; Ito, Masataka; Shimmura, Shigeto; Tsubota, Kazuo

2012-01-01

Purpose To date, mouse lacrimal gland epithelial cells have been cultured successfully but only in cases involving newborn mouse lacrimal glands. In this work, we attempted to cultivate and characterize adult mouse lacrimal gland epithelial cells. Methods Lacrimal glands were removed from newborn mice (C57B/6) and isolated lacrimal gland epithelial cells were seeded onto tissue culture treated or low adherent culture dishes in Cnt-07 culture medium with or without cholera toxin. Cultivated cells were characterized by immunostaining with pan-cytokeratin, α-smooth muscle actin, and lactoferrin antibodies. Lacrimal gland cells from 7-week-old green fluorescent protein (GFP) and non-GFP (C57B/6) mice were mixed and seeded onto uncoated dishes to assess sphere-forming efficiency. Cells were also seeded onto 3T3 cell feeder layers to assess colony forming efficiency. Results Lacrimal gland epithelial cells were selectively cultured with cholera toxin, and cell type was verified by pan-cytokeratin and α-smooth muscle actin immunostaining. Sphere formation from single cells of adult mice was observed using specific medium and low adherent culture dishes. These cells could also undergo colony formation on 3T3 feeder cells. Conclusions Adult mouse lacrimal gland epithelial cells were successfully cultivated in cholera toxin-containing medium, and were observed to form spheres from single cells. PMID:22665974

Process-Based Expansion and Neural Differentiation of Human Pluripotent Stem Cells for Transplantation and Disease Modeling

PubMed Central

Stover, Alexander E.; Brick, David J.; Nethercott, Hubert E.; Banuelos, Maria G.; Sun, Lei; O’Dowd, Diane K.; Schwartz, Philip H.

2014-01-01

Robust strategies for developing patient-specific, human, induced pluripotent stem cell (iPSC)-based therapies of the brain require an ability to derive large numbers of highly defined neural cells. Recent progress in iPSC culture techniques includes partial-to-complete elimination of feeder layers, use of defined media, and single-cell passaging. However, these techniques still require embryoid body formation or coculture for differentiation into neural stem cells (NSCs). In addition, none of the published methodologies has employed all of the advances in a single culture system. Here we describe a reliable method for long-term, single-cell passaging of PSCs using a feeder-free, defined culture system that produces confluent, adherent PSCs that can be differentiated into NSCs. To provide a basis for robust quality control, we have devised a system of cellular nomenclature that describes an accurate genotype and phenotype of the cells at specific stages in the process. We demonstrate that this protocol allows for the efficient, large-scale, cGMP-compliant production of transplantable NSCs from all lines tested. We also show that NSCs generated from iPSCs produced with the process described are capable of forming both glia defined by their expression of S100β and neurons that fire repetitive action potentials. PMID:23893392

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

PubMed

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.

Can rats solve a simple version of the traveling salesman problem?

PubMed

Bures, J; Buresová, O; Nerad, L

1992-12-31

Whereas correct tours through the radial arm maze are almost equally long, free choice mazes with multiple goals scattered in an open field allow the animal to select the shortest one from a multitude of correct tours. Thirteen rats were trained (at 10 trials per day) to visit an array of cylindrical feeders in an open field (40 x 100 cm) with reward available only when visiting the last feeder of the set. In Expt. 1 with eight feeders arranged in five different configurations the rats made after 10 days of training 1 error in the first 8 choices and incidence of errorless trials was about 20%. In Expt. 2. the use of six feeders in a rectangular (A) or double triangle (B) configuration increased the incidence of errorless trials to 60%. Expt. 3 showed that performance in the 6-feeder maze was significantly impaired by 6 mg/kg ketamine or 0.25 mg/kg scopolamine but not by lower dosages of these drugs. Tours generated on errorless trials (each feeder visited only once) during 10 days of Expt. 2 were analyzed. Six places can be visited in 6! = 720 different closed tours the lengths of which (in arbitrary units) range from 6.00 to 10.12 for A and from 6.83 to 10.47 for B. Whereas random generation of correct routes yielded only 5% of the shortest tours, they were clearly preferred by rats (41% in A and 45% in B). The apparent proficiency of rats in this optimization problem is probably not due to cognitive comparison of the possible correct routes but rather to following a simple rule 'Always go to the nearest not yet visited feeder'.

Niemann-Pick disease, Type C: evidence for the deficiency of an activating factor stimulating sphingomyelin and glucocerebroside degradation.

PubMed

Christomanou, H

1980-10-01

1) Qualitative lipid analyses by thin-layer chromatography of 4 Niemann-Pick type C spleens confirmed sphingomyelin accumulation together with increase in the amount of glucocerebroside. 2) In the presence of crude sodium taurocholate as detergent, sphingomyelin degradation rates of normal and Niemann-Pick type C-cultured fibroblasts were fairly close under standard conditions at pH 5.0. In the absence of sodium taurocholate, sphingomyelinase activity was optimal at pH 4.0. Sphingomyelinase activities of fibroblasts from two patients with Niemann-Pick disease type C measured without detergent, were about 30% of that of controls. 3) Extracts from Gaucher spleen heated to 90 degrees C and devoid of sphingomyelinase activity stimulated at the optimal pH of 4.0 sphingomyelin degradation by cultured normal fibroblasts (2--4-fold, Niemann-Pick type C fibroblasts (5--9-fold), whereas similarly treated extracts from Niemann-Pick type C spleen showed no stimulation of sphingomyelin catabolism. Heated extracts from normal human spleen exhibited a smaller stimulation than that shown by Gaucher spleen. This stimulating effect could not be observed in fibroblasts from patients suffering from Niemann-Pick type B (sphingomyelinase defect). 4) Heat-treated extracts of Gaucher spleen were fractionated by ion exchange chromatography, isoelectric focusing and gel filtration. The active fractions obtained by these procedures stimulated sphingomyelin as well as glucocerebroside degradation and were absent from the corresponding Niemann-Pick type C preparations. Enriched activator preparations of Gaucher spleen stimulated sphingomyelinase activity of Niemann-Pick type C fibroblasts 25--38-fold and that of normal cells 3-fold. 5) The activating factor had an isoelectric point of 4.0 and an apparent molecular weight, as estimated by gel filtration, of 25000. Treatment with pronase E abolished its activity.

Reprogramming of fibroblasts from older women with pelvic floor disorders alters cellular behavior associated with donor age.

PubMed

Wen, Yan; Wani, Prachi; Zhou, Lu; Baer, Tom; Phadnis, Smruti Madan; Reijo Pera, Renee A; Chen, Bertha

2013-02-01

We aimed to derive induced pluripotent stem cell (iPSC) lines from vaginal fibroblasts from older women with pelvic organ prolapse. We examined the effect of donor age on iPSCs and on the cells redifferentiated from these iPSCs. Vaginal fibroblasts were isolated from younger and older subjects for reprogramming. iPSCs were generated simultaneously using an excisable polycistronic lentiviral vector expressing Oct4, Klf4, Sox2, and cMyc. The pluripotent markers of iPSCs were confirmed by immunocytochemistry and quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Spectral karyotyping was performed. The ability of the iPSCs to differentiate into three germ layers was confirmed by embryoid body and teratoma formation. Senescence marker (p21, p53, and Bax) expressions were determined by qRT-PCR and Western blot. The iPSCs were redifferentiated to fibroblasts and were evaluated with senescence-associated β-galactosidase (SA) activity and mitotic index using time-lapse dark-field microscopy. iPSCs derived from both the younger and older subjects expressed pluripotency markers and showed normal karyotype and positive teratoma assays. There was no significant difference in expression of senescence and apoptosis markers (p21, p53, and Bax) in iPSCs derived from the younger subject compared with the older subject. Furthermore, fibroblasts redifferentiated from these iPSCs did not differ in SA activity or mitotic index. We report successful derivation of iPSCs from women with pelvic organ prolapse. Older age did not interfere with successful reprogramming. Donor age differences were not observed in these iPSCs using standard senescence markers, and donor age did not appear to affect cell mitotic activity in fibroblasts redifferentiated from iPSCs.

47 CFR 25.250 - Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 47 Telecommunication 2 2014-10-01 2014-10-01 false Sharing between NGSO MSS Feeder links Earth....250 Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands. (a) NGSO MSS applicants shall be licensed to operate in the 29.1-29.5 GHz band for Earth-to-space...

47 CFR 25.250 - Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 47 Telecommunication 2 2013-10-01 2013-10-01 false Sharing between NGSO MSS Feeder links Earth....250 Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands. (a) NGSO MSS applicants shall be licensed to operate in the 29.1-29.5 GHz band for Earth-to-space...

47 CFR 25.250 - Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 47 Telecommunication 2 2012-10-01 2012-10-01 false Sharing between NGSO MSS Feeder links Earth....250 Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands. (a) NGSO MSS applicants shall be licensed to operate in the 29.1-29.5 GHz band for Earth-to-space...

47 CFR 25.250 - Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 47 Telecommunication 2 2010-10-01 2010-10-01 false Sharing between NGSO MSS Feeder links Earth....250 Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands. (a) NGSO MSS applicants shall be licensed to operate in the 29.1-29.5 GHz band for Earth-to-space...

47 CFR 25.250 - Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 47 Telecommunication 2 2011-10-01 2011-10-01 false Sharing between NGSO MSS Feeder links Earth....250 Sharing between NGSO MSS Feeder links Earth Stations in the 19.3-19.7 GHz and 29.1-29.5 GHz Bands. (a) NGSO MSS applicants shall be licensed to operate in the 29.1-29.5 GHz band for Earth-to-space...

Conceptual Design and Analysis of Cold Mass Support of the CS3U Feeder for the ITER

NASA Astrophysics Data System (ADS)

Zhu, Yinfeng; Song, Yuntao; Zhang, Yuanbin; Wang, Zhongwei

2013-06-01

In the International Thermonuclear Experimental Reactor (ITER) project, the feeders are one of the most important and critical systems. To convey the power supply and the coolant for the central solenoid (CS) magnet, 6 sets of CS feeders are employed, which consist mainly of an in-cryostat feeder (ICF), a cryostat feed-through (CFT), an S-bend box (SBB), and a coil terminal box (CTB). To compensate the displacements of the internal components of the CS feeders during operation, sliding cold mass supports consisting of a sled plate, a cylindrical support, a thermal shield, and an external ring are developed. To check the strength of the developed cold mass supports of the CS3U feeder, electromagnetic analysis of the two superconducting busbars is performed by using the CATIA V5 and ANSYS codes based on parametric technology. Furthermore, the thermal-structural coupling analysis is performed based on the obtained results, except for the stress concentration, and the max. stress intensity is lower than the allowable stress of the selected material. It is found that the conceptual design of the cold mass support can satisfy the required functions under the worst case of normal working conditions. All these performed activities will provide a firm technical basis for the engineering design and development of cold mass supports.

Characterization of a Liver Organoid Tissue Composed of Hepatocytes and Fibroblasts in Dense Collagen Fibrils

PubMed Central

Tamai, Miho; Adachi, Eijiro

2013-01-01

The adult liver is wrapped in a connective tissue sheet called the liver capsule, which consists of collagen fibrils and fibroblasts. In this study, we set out to construct a liver organoid tissue that would be comparable to the endogenous liver, using a bioreactor. In vitro liver organoid tissue was generated by combining collagen fibrils, fibroblasts, and primary murine hepatocytes or Hep G2 on a mesh of poly-lactic acid fabric using a bioreactor. Then, the suitability of this liver organoid tissue for transplantation was tested by implanting the constructs into partially hepatectomized BALB/cA-nu/nu mice. As determined by using scanning and transmission electron microscopes, the liver organoid tissues were composed of densely packed collagen fibrils with fibroblasts and aggregates of oval or spherical hepatocytes. Angiogenesis was induced after the transplantation, and blood vessels connected the liver organoid tissue with the surrounding tissue. Thus, a novel approach was applied to generate transplantable liver organoid tissue within a condensed collagen fibril matrix. These results suggested that a dense collagen network populated with fibroblasts can hold a layer of concentrated hepatocytes, providing a three-dimensional microenvrionment suitable for the reestablishment of cell–cell and cell–extracellular matrix (ECM) interactions, and resulting in the maintenance of their liver-specific functions. This liver organoid tissue may be useful for the study of intrahepatic functions of various cells, cytokines, and ECMs, and may fulfill the fundamental requirements of a donor tissue. PMID:23815236

30 CFR 75.1001 - Overcurrent protection.

Code of Federal Regulations, 2011 CFR

2011-07-01

... MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Trolley Wires and Trolley Feeder Wires § 75.1001 Overcurrent protection. [Statutory Provisions] Trolley wires and trolley feeder wires shall be provided with...

30 CFR 75.1001 - Overcurrent protection.

Code of Federal Regulations, 2010 CFR

2010-07-01

... MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Trolley Wires and Trolley Feeder Wires § 75.1001 Overcurrent protection. [Statutory Provisions] Trolley wires and trolley feeder wires shall be provided with...

The influence of laser alloying on the structure and mechanical properties of AlMg5Si2Mn surface layers

NASA Astrophysics Data System (ADS)

Pakieła, W.; Tański, T.; Brytan, Z.; Labisz, K.

2016-04-01

The goal of this paper was focused on investigation of microstructure and properties of surface layer produced during laser surface treatment of aluminium alloy by high-power fibre laser. The performed laser treatment involves remelting and feeding of Inconel 625 powder into the aluminium surface. As a base metal was used aluminium alloy AlMg5Si2Mn. The Inconel powder was injected into the melt pool and delivered by a vacuum feeder at a constant rate of 4.5 g/min. The size of Inconel alloying powder was in the range 60-130 µm. In order to remelt the aluminium alloy surface, the fibre laser of 3 kW laser beam power has been used. The linear laser scan rate of the beam was set 0.5 m/min. Based on performed investigations, it was possible to obtain the layer consisting of heat-affected zone, transition zone and remelted zone, without cracks and defects having much higher hardness value compared to the non-alloyed material.

30 CFR 75.1000 - Cutout switches.

Code of Federal Regulations, 2011 CFR

2011-07-01

... MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Trolley Wires and Trolley Feeder Wires § 75.1000 Cutout switches. [Statutory Provisions] Trolley wires and trolley feeder wires, shall be provided with cutout...

30 CFR 75.1000 - Cutout switches.

Code of Federal Regulations, 2010 CFR

2010-07-01

... MANDATORY SAFETY STANDARDS-UNDERGROUND COAL MINES Trolley Wires and Trolley Feeder Wires § 75.1000 Cutout switches. [Statutory Provisions] Trolley wires and trolley feeder wires, shall be provided with cutout...

Migration and Differentiation of GFP-transplanted Bone Marrow-derived Cells into Experimentally Induced Periodontal Polyp in Mice.

PubMed

Matsuda, Saeka; Shoumura, Masahito; Osuga, Naoto; Tsujigiwa, Hidetsugu; Nakano, Keisuke; Okafuji, Norimasa; Ochiai, Takanaga; Hasegawa, Hiromasa; Kawakami, Toshiyuki

2016-01-01

Perforation of floor of the dental pulp is often encountered during root canal treatment in routine clinical practice of dental caries. If perforation were large, granulation tissue would grow to form periodontal polyp. Granulation tissue consists of proliferating cells however their origin is not clear. It was shown that the cells in granulation tissue are mainly from migration of undifferentiated mesenchymal cells of the bone marrow. Hence, this study utilized GFP bone marrow transplantation mouse model. The floor of the pulp chamber in maxillary first molar was perforated using ½ dental round bur. Morphological assessment was carried out by micro CT and microscopy and GFP cell mechanism was further assessed by immunohistochemistry using double fluorescent staining with GFP-S100A4; GFP-Runx2 and GFP-CD31. Results of micro CT revealed alveolar bone resorption and widening of periodontal ligament. Histopathological examination showed proliferation of fibroblasts with some round cells and blood vessels in the granulation tissue. At 2 weeks, the outermost layer of the granulation tissue was lined by squamous cells with distinct intercellular bridges. At 4 weeks, the granulation tissue became larger than the perforation and the outermost layer was lined by relatively typical stratified squamous epithelium. Double immunofluorescent staining of GFP and Runx2 revealed that both proteins were expressed in spindle-shaped cells. Double immunofluorescent staining of GFP and CD31 revealed that both proteins were expressed in vascular endothelial cells in morphologically distinct vessels. The results suggest that fibroblasts, periodontal ligament fibroblasts and blood vessels in granulation tissue were derived from transplanted-bone marrow cells. Thus, essential growth of granulation tissue in periodontal polyp was caused by the migration of undifferentiated mesenchymal cells derived from bone marrow, which differentiated into fibroblasts and later on differentiated into other cells in response to injury.

Identification of Novel Targets of the Human Cell Cycle Regulatory Protein Cdc34

DTIC Science & Technology

1999-07-01

centrifugal elutriation, with a purity of -80% as shown by micro - ethyl acetate and separated on thin-layer chromatography plates (Whatman, scopic...Spain. (2) Servicio Bioquimica, Hosp. La Paz. Madrid. ICER protein is elevated in mHR6b-/- (murine Rad6B) fibroblasts Spain. and in human cells

Engineering bioartificial tracheal tissue using hybrid fibroblast-mesenchymal stem cell cultures in collagen hydrogels.

PubMed

Naito, Hiroshi; Tojo, Takashi; Kimura, Michitaka; Dohi, Yoshiko; Zimmermann, Wolfram-Hubertus; Eschenhagen, Thomas; Taniguchi, Shigeki

2011-02-01

We aimed at providing the first in vitro and in vivo proof-of-concept for a novel tracheal tissue engineering technology. We hypothesized that bioartificial trachea (BT) could be generated from fibroblast and collagen hydrogels, mechanically supported by osteogenically-induced mesenchymal stem cells (MSC) in ring-shaped 3D-hydrogel cultures, and applied in an experimental model of rat trachea injury. Tube-shaped tissue was constructed from mixtures of rat fibroblasts and collagen in custom-made casting molds. The tissue was characterized histologically and mechanically. Ring-shaped tissue was constructed from mixtures of rat MSCs and collagen and fused to the tissue-engineered tubes to function as reinforcement. Stiffness of the biological reinforcement was enhanced by induction of osteogeneic differentiation in MSCs. Osteogenic differentiation was evaluated by assessment of osteocalcin (OC) secretion, quantification of calcium (Ca) deposit, and mechanical testing. Finally, BT was implanted to bridge a surgically-induced tracheal defect. A three-layer tubular tissue structure composed of an interconnected network of fibroblasts was constructed. Tissue collapse was prevented by the placement of MSC-containing ring-shaped tissue reinforcement around the tubular constructs. Osteogenic induction resulted in high OC secretion, high Ca deposit, and enhanced construct stiffness. Ultimately, when BT was implanted, recipient rats were able to breathe spontaneously.

Differential Response of Human Adipose Tissue-Derived Mesenchymal Stem Cells, Dermal Fibroblasts, and Keratinocytes to Burn Wound Exudates: Potential Role of Skin-Specific Chemokine CCL27

PubMed Central

van den Broek, Lenie J.; Kroeze, Kim L.; Waaijman, Taco; Breetveld, Melanie; Sampat-Sardjoepersad, Shakun C.; Niessen, Frank B.; Middelkoop, Esther; Scheper, Rik J.

2014-01-01

Many cell-based regenerative medicine strategies toward tissue-engineered constructs are currently being explored. Cell–cell interactions and interactions with different biomaterials are extensively investigated, whereas very few studies address how cultured cells will interact with soluble wound-healing mediators that are present within the wound bed after transplantation. The aim of this study was to determine how adipose tissue-derived mesenchymal stem cells (ASC), dermal fibroblasts, and keratinocytes will react when they come in contact with the deep cutaneous burn wound bed. Burn wound exudates isolated from deep burn wounds were found to contain many cytokines, including chemokines and growth factors related to inflammation and wound healing. Seventeen mediators were identified by ELISA (concentration range 0.0006–9 ng/mg total protein), including the skin-specific chemokine CCL27. Burn wound exudates activated both ASC and dermal fibroblasts, but not keratinocytes, to increase secretion of CXCL1, CXCL8, CCL2, and CCL20. Notably, ASC but not fibroblasts or keratinocytes showed significant increased secretion of vascular endothelial growth factor (5-fold) and interleukin-6 (253-fold), although when the cells were incorporated in bi-layered skin substitute (SS) these differences were less pronounced. A similar discrepancy between ASC and dermal fibroblast mono-cultures was observed when recombinant human-CCL27 was used instead of burn wound exudates. Although CCL27 did not stimulate the secretion of any of the wound-healing mediators by keratinocytes, these cells, in contrast to ASC or dermal fibroblasts, showed increased proliferation and migration. Taken together, these results indicate that on transplantation, keratinocytes are primarily activated to promote wound closure. In contrast, dermal fibroblasts and, in particular, ASC respond vigorously to factors present in the wound bed, leading to increased secretion of angiogenesis/granulation tissue formation factors. Our findings have implications for the choice of cell type (ASC or dermal fibroblast) to be used in regenerative medicine strategies and indicate the importance of taking into account interactions with the wound bed when developing advanced therapies for difficult-to-close cutaneous wounds. PMID:23980822

Distribution Feeder Modeling for Time-Series Simulation of Voltage Management Strategies: Preprint

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

Giraldez Miner, Julieta I; Gotseff, Peter; Nagarajan, Adarsh

This paper presents techniques to create baseline distribution models using a utility feeder from Hawai'ian Electric Company. It describes the software-to-software conversion, steady-state, and time-series validations of a utility feeder model. It also presents a methodology to add secondary low-voltage circuit models to accurately capture the voltage at the customer meter level. This enables preparing models to perform studies that simulate how customer-sited resources integrate into legacy utility distribution system operations.

Ex vivo preservation and expansion of human limbal epithelial stem cells on amniotic membrane cultures.

PubMed

Meller, D; Pires, R T F; Tseng, S C G

2002-04-01

Amniotic membrane (AM) transplantation effectively expands the remaining limbal epithelial stem cells in patients with partial limbal stem cell deficiency. The authors investigated whether this action could be produced ex vivo. The outgrowth rate on AM was compared among explants derived from human limbus, peripheral cornea, and central cornea. For outgrowth of human limbal epithelial cells (HLEC), cell cycle kinetics were measured by BrdU labelling for 1 or 7 days, of which the latter was also chased in primary cultures, secondary 3T3 fibroblast cultures, and in athymic Balb/c mice following a brief treatment with a phorbol ester. Epithelial morphology was studied by histology and transmission electron microscopy, and phenotype was defined by immunostaining with monoclonal antibodies to keratins and mucins. Outgrowth rate was 0/22 (0%) and 2/24 (8.3%) for central and peripheral corneal explants, respectively, but was 77/80 (96.2%) for limbal explants (p <0.0001). 24 hour BrdU labelling showed a uniformly low (that is, less than 5%) labelling index in 65% of the limbal explants, but a mixed pattern with areas showing a high (that is, more than 40%) labelling index in 35% of limbal explants, and in all (100%) peripheral corneal explants. Continuous BrdU labelling for 7 days detected a high labelling index in 61.5% of the limbal explants with the remainder still retaining a low labelling index. A number of label retaining cells were noted after 7 day labelling followed by 14 days of chase in primary culture or by 21 days of chase after transplantation to 3T3 fibroblast feeder layers. After exposure to phorbol 12-myristate 13-acetate for 24 hours and 7 day labelling, HLEC transplanted in athymic mice still showed a number of label retaining basal cells after 9 days of chase. HLEC cultured on AM were strongly positive for K14 keratin and MUC4 and slightly positive in suprabasal cells for K3 keratin but negative for K12 keratin, AMEM2, and MUC5AC. After subcutaneous implantation in athymic mice, the resultant epithelium was markedly stratified and the basal epithelial cells were strongly positive for K14 keratin, while the suprabasal epithelial cells were strongly positive for K3 keratin and MUC4, and the entire epithelium was negative for K12 keratin and MUC5A/C. These data support the notion that AM cultures preferentially preserve and expand limbal epithelial stem cells that retain their in vivo properties of slow cycling, label retaining, and undifferentiation. This finding supports the feasibility of ex vivo expansion of limbal epithelial stem cells for treating patients with total limbal stem cell deficiency using a small amount of donor limbal tissue.

21 CFR 520.1448a - Monensin blocks.

Code of Federal Regulations, 2011 CFR

2011-04-01

...) Limitations. Block to be fed free choice to pasture cattle (slaughter, stocker, feeder, and dairy and beef.... Blocks to be fed free choice to pasture cattle (slaughter, stocker, feeder, and dairy and beef...

11. MOVABLE BED SEDIMENTATION MODELS. AUTOMATIC SEDIMENT FEEDER DESIGNED AND ...

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

11. MOVABLE BED SEDIMENTATION MODELS. AUTOMATIC SEDIMENT FEEDER DESIGNED AND BUILT BY WES. - Waterways Experiment Station, Hydraulics Laboratory, Halls Ferry Road, 2 miles south of I-20, Vicksburg, Warren County, MS

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

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

Allehyani, Ahmed; Beshir, Mohammed

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 eachmore » node. After that, the impact of electric vehicles on distribution feeders that do not have voltage regulators will appear.« less

IEEE 342 Node Low Voltage Networked Test System

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

Schneider, Kevin P.; Phanivong, Phillippe K.; Lacroix, Jean-Sebastian

The IEEE Distribution Test Feeders provide a benchmark for new algorithms to the distribution analyses community. The low voltage network test feeder represents a moderate size urban system that is unbalanced and highly networked. This is the first distribution test feeder developed by the IEEE that contains unbalanced networked components. The 342 node Low Voltage Networked Test System includes many elements that may be found in a networked system: multiple 13.2kV primary feeders, network protectors, a 120/208V grid network, and multiple 277/480V spot networks. This paper presents a brief review of the history of low voltage networks and how theymore » evolved into the modern systems. This paper will then present a description of the 342 Node IEEE Low Voltage Network Test System and power flow results.« less

Comparing the engineering program feeders from SiF and convention models

NASA Astrophysics Data System (ADS)

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.

Complex memories in honeybees: can there be more than two?

PubMed

Reinhard, Judith; Srinivasan, Mandyam V; Zhang, Shaowu

2006-04-01

Foraging honeybees are likely to learn visual and chemical cues associated with many different food sources. Here, we explore how many such sources can be memorized and recalled. Marked bees were trained to visit two (or three) sugar feeders, each placed at a different outdoor location and carrying a different scent. We then tested the ability of the bees to recall these locations and fly to them, when the training scents were blown into the hive, and the scents and food at the feeders were removed. When trained on two feeder locations, each associated with a different scent, the bees could correctly recall the location associated with each scent. However, this ability broke down when the number of scents and feeder locations was increased to three. Performance was partially restored when each of the three training feeders was endowed with an additional cue, namely, a distinct colour. Our results suggest that bees can recall a maximum of two locations when each is associated with a different scent. However, this number can be increased if the scent cues are augmented by visual cues. These findings have implications for the ways in which associations are established and laid down in honeybee memory.

Optic flow informs distance but not profitability for honeybees.

PubMed

Shafir, Sharoni; Barron, Andrew B

2010-04-22

How do flying insects monitor foraging efficiency? Honeybees (Apis mellifera) use optic flow information as an odometer to estimate distance travelled, but here we tested whether optic flow informs estimation of foraging costs also. Bees were trained to feeders in flight tunnels such that bees experienced the greatest optic flow en route to the feeder closest to the hive. Analyses of dance communication showed that, as expected, bees indicated the close feeder as being further, but they also indicated this feeder as the more profitable, and preferentially visited this feeder when given a choice. We show that honeybee estimates of foraging cost are not reliant on optic flow information. Rather, bees can assess distance and profitability independently and signal these aspects as separate elements of their dances. The optic flow signal is sensitive to the nature of the environment travelled by the bee, and is therefore not a good index of flight energetic costs, but it provides a good indication of distance travelled for purpose of navigation and communication, as long as the dancer and recruit travel similar routes. This study suggests an adaptive dual processing system in honeybees for communicating and navigating distance flown and for evaluating its energetic costs.

Optic flow informs distance but not profitability for honeybees

PubMed Central

Shafir, Sharoni; Barron, Andrew B.

2010-01-01

How do flying insects monitor foraging efficiency? Honeybees (Apis mellifera) use optic flow information as an odometer to estimate distance travelled, but here we tested whether optic flow informs estimation of foraging costs also. Bees were trained to feeders in flight tunnels such that bees experienced the greatest optic flow en route to the feeder closest to the hive. Analyses of dance communication showed that, as expected, bees indicated the close feeder as being further, but they also indicated this feeder as the more profitable, and preferentially visited this feeder when given a choice. We show that honeybee estimates of foraging cost are not reliant on optic flow information. Rather, bees can assess distance and profitability independently and signal these aspects as separate elements of their dances. The optic flow signal is sensitive to the nature of the environment travelled by the bee, and is therefore not a good index of flight energetic costs, but it provides a good indication of distance travelled for purpose of navigation and communication, as long as the dancer and recruit travel similar routes. This study suggests an adaptive dual processing system in honeybees for communicating and navigating distance flown and for evaluating its energetic costs. PMID:20018787

Energy efficiency analysis of two-sided feed scheme of DC traction network with high asymmetry of feeders parameters

NASA Astrophysics Data System (ADS)

Abramov, E. Y.; Sopov, V. I.

2017-10-01

In a given research using the example of traction network area with high asymmetry of power supply parameters, the sequence of comparative assessment of power losses in DC traction network with parallel and traditional separated operating modes of traction substation feeders was shown. Experimental measurements were carried out under these modes of operation. The calculation data results based on statistic processing showed the power losses decrease in contact network and the increase in feeders. The changes proved to be critical ones and this demonstrates the significance of potential effects when converting traction network areas into parallel feeder operation. An analytical method of calculation the average power losses for different feed schemes of the traction network was developed. On its basis, the dependences of the relative losses were obtained by varying the difference in feeder voltages. The calculation results showed unreasonableness transition to a two-sided feed scheme for the considered traction network area. A larger reduction in the total power loss can be obtained with a smaller difference of the feeders’ resistance and / or a more symmetrical sectioning scheme of contact network.

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

PubMed

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.

View northwest, discharge basin, floor, showing cement cross beams built ...

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

View northwest, discharge basin, floor, showing cement cross beams built on stone bases - Glens Falls Feeder, Sluice, Along south side of Glens Falls Feeder between locks 10 & 20, Hudson Falls, Washington County, NY

Reconstruction of living bilayer human skin equivalent utilizing human fibrin as a scaffold.

PubMed

Mazlyzam, A L; Aminuddin, B S; Fuzina, N H; Norhayati, M M; Fauziah, O; Isa, M R; Saim, L; Ruszymah, B H I

2007-05-01

Our aim of this study was to develop a new methodology for constructing a bilayer human skin equivalent to create a more clinical compliance skin graft composite for the treatment of various skin defects. We utilized human plasma derived fibrin as the scaffold for the development of a living bilayer human skin equivalent: fibrin-fibroblast and fibrin-keratinocyte (B-FF/FK SE). Skin cells from six consented patients were culture-expanded to passage 1. For B-FF/FK SE formation, human fibroblasts were embedded in human fibrin matrix and subsequently another layer of human keratinocytes in human fibrin matrix was stacked on top. The B-FF/FK SE was then transplanted to athymic mice model for 4 weeks to evaluate its regeneration and clinical performance. The in vivo B-FF/FK SE has similar properties as native human skin by histological analysis and expression of basal Keratin 14 gene in the epidermal layer and Collagen type I gene in the dermal layer. Electron microscopy analysis of in vivo B-FF/FK SE showed well-formed and continuous epidermal-dermal junction. We have successfully developed a technique to engineer living bilayer human skin equivalent using human fibrin matrix. The utilization of culture-expanded human skin cells and fibrin matrix from human blood will allow a fully autologous human skin equivalent construction.

Spacelab 4: Primate experiment support hardware

NASA Astrophysics Data System (ADS)

Fusco, P. R.; Peyran, R. J.

1984-05-01

A squirrel monkey feeder and automatic urine collection system were designed to fly on the Spacelab 4 Shuttle Mission presently scheduled for January 1986. Prototypes of the feeder and urine collection systems were fabricated and extensively tested on squirrel monkeys at the National Aeronautics and Space Administration's (NASA) Ames Research Center (ARC). The feeder design minimizes impact on the monkey's limited space in the cage and features improved reliability and biocompatibility over previous systems. The urine collection system is the first flight qualified, automatic urine collection device for squirrel monkeys. Flight systems are currently being fabricated.

Spacelab 4: Primate experiment support hardware

NASA Technical Reports Server (NTRS)

Fusco, P. R.; Peyran, R. J.

1984-01-01

A squirrel monkey feeder and automatic urine collection system were designed to fly on the Spacelab 4 Shuttle Mission presently scheduled for January 1986. Prototypes of the feeder and urine collection systems were fabricated and extensively tested on squirrel monkeys at the National Aeronautics and Space Administration's (NASA) Ames Research Center (ARC). The feeder design minimizes impact on the monkey's limited space in the cage and features improved reliability and biocompatibility over previous systems. The urine collection system is the first flight qualified, automatic urine collection device for squirrel monkeys. Flight systems are currently being fabricated.

In vitro reconstruction of human junctional and sulcular epithelium

PubMed Central

Dabija-Wolter, G; Bakken, V; Cimpan, M R; Johannessen, A C; Costea, D E

2013-01-01

BACKGROUND The aim of this study was to develop and characterize standardized in vitro three-dimensional organotypic models of human junctional epithelium (JE) and sulcular epithelium (SE). METHODS Organotypic models were constructed by growing human normal gingival keratinocytes on top of collagen matrices populated with gingival fibroblasts (GF) or periodontal ligament fibroblasts (PLF). Tissues obtained were harvested at different time points and assessed for epithelial morphology, proliferation (Ki67), expression of JE-specific markers (ODAM and FDC-SP), cytokeratins (CK), transglutaminase, filaggrin, and basement membrane proteins (collagen IV and laminin1). RESULTS The epithelial component in 3- and 5-day organotypics showed limited differentiation and expressed Ki-67, ODAM, FDC-SP, CK 8, 13, 16, 19, and transglutaminase in a similar fashion to control JE samples. PLF supported better than GF expression of CK19 and suprabasal proliferation, although statistically significant only at day 5. Basement membrane proteins started to be deposited only from day 5. The rate of proliferating cells as well as the percentage of CK19-expressing cells decreased significantly in 7- and 9-day cultures. Day 7 organotypics presented higher number of epithelial cell layers, proliferating cells in suprabasal layers, and CK expression pattern similar to SE. CONCLUSION Both time in culture and fibroblast type had impact on epithelial phenotype. Five-day cultures with PLF are suggested as JE models, 7-day cultures with PLF or GF as SE models, while 9-day cultures with GF as gingival epithelium (GE) models. Such standard, reproducible models represent useful tools to study periodontal bacteria–host interactions in vitro. PMID:22947066

Existence of c-Kit negative cells with ultrastructural features of interstitial cells of Cajal in the subserosal layer of the W/Wv mutant mouse colon

PubMed Central

Tamada, Hiromi; Kiyama, Hiroshi

2015-01-01

Interstitial cells of Cajal (ICC) are mesenchymal cells that are distributed along the gastrointestinal tract and function as pacemaker cells or intermediary cells between nerves and smooth muscle cells. ICC express a receptor tyrosine kinase c-Kit, which is an established marker for ICC. The c-kit gene is allelic with the murine white-spotting locus (W), and some ICC subsets were reported to be missing in heterozygous mutant W/Wv mice carrying W and Wv mutated alleles. In this study, the characterization of interstitial cells in the subserosal layer of W/Wv mice was analyzed by immunohistochemistry and electron microscopy. In the proximal and distal colon of W/Wv mutant mice, no c-Kit-positive cells were detected in the subserosal layer by immunohistochemistry. By electron microscopy, the interstitial cells, which were characterized by the existence of caveolae, abundant mitochondria and gap junctions, were observed in the W/Wv mutant colon. The morphological characteristics were comparable to those of the multipolar c-Kit positive ICC seen in the subserosa of proximal and distal colon of wild-type mice. Fibroblasts were also located in the same layers, but the morphology of the fibroblasts was distinguishable from that of ICC in wild type mice or of ICC-like cells in W/Wv mutant mice. Collectively, it is concluded that c-Kit-negative interstitial cells showing a typical ICC ultrastructure exist in the proximal and distal colon of W/Wv mutant mice. PMID:26727725

Existence of c-Kit negative cells with ultrastructural features of interstitial cells of Cajal in the subserosal layer of the W/W(v) mutant mouse colon.

PubMed

Tamada, Hiromi; Kiyama, Hiroshi

2015-01-01

Interstitial cells of Cajal (ICC) are mesenchymal cells that are distributed along the gastrointestinal tract and function as pacemaker cells or intermediary cells between nerves and smooth muscle cells. ICC express a receptor tyrosine kinase c-Kit, which is an established marker for ICC. The c-kit gene is allelic with the murine white-spotting locus (W), and some ICC subsets were reported to be missing in heterozygous mutant W/W(v) mice carrying W and W(v) mutated alleles. In this study, the characterization of interstitial cells in the subserosal layer of W/W(v) mice was analyzed by immunohistochemistry and electron microscopy. In the proximal and distal colon of W/W(v) mutant mice, no c-Kit-positive cells were detected in the subserosal layer by immunohistochemistry. By electron microscopy, the interstitial cells, which were characterized by the existence of caveolae, abundant mitochondria and gap junctions, were observed in the W/W(v) mutant colon. The morphological characteristics were comparable to those of the multipolar c-Kit positive ICC seen in the subserosa of proximal and distal colon of wild-type mice. Fibroblasts were also located in the same layers, but the morphology of the fibroblasts was distinguishable from that of ICC in wild type mice or of ICC-like cells in W/W(v) mutant mice. Collectively, it is concluded that c-Kit-negative interstitial cells showing a typical ICC ultrastructure exist in the proximal and distal colon of W/W(v) mutant mice.

Existence of c-Kit negative cells with ultrastructural features of interstitial cells of Cajal in the subserosal layer of the W/Wv mutant mouse colon.

PubMed

Tamada, Hiromi; Kiyama, Hiroshi

2015-01-01

Interstitial cells of Cajal (ICC) are mesenchymal cells that are distributed along the gastrointestinal tract and function as pacemaker cells or intermediary cells between nerves and smooth muscle cells. ICC express a receptor tyrosine kinase c-Kit, which is an established marker for ICC. The c-kit gene is allelic with the murine white-spotting locus (W), and some ICC subsets were reported to be missing in heterozygous mutant W/Wv mice carrying W and Wv mutated alleles. In this study, the characterization of interstitial cells in the subserosal layer of W/Wv mice was analyzed by immunohistochemistry and electron microscopy. In the proximal and distal colon of W/Wv mutant mice, no c-Kit-positive cells were detected in the subserosal layer by immunohistochemistry. By electron microscopy, the interstitial cells, which were characterized by the existence of caveolae, abundant mitochondria and gap junctions, were observed in the W/Wv mutant colon.The morphological characteristics were comparable to those of the multipolar c-Kit positive ICC seen in the subserosa of proximal and distal colon of wild-type mice. Fibroblasts were also located in the same layers,but the morphology of the fibroblasts was distinguishable from that of ICC in wild type mice or of ICC-like cells in W/Wv mutant mice. Collectively, it is concluded that c-Kit-negative interstitial cells showing a typical ICC ultrastructure exist in the proximal and distal colon of W/Wv mutant mice.

View southeast, interior of second drop, foundation and footwall and ...

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

View southeast, interior of second drop, foundation and footwall and sidewalls, showing failed lower arch - Glens Falls Feeder, Sluice, Along south side of Glens Falls Feeder between locks 10 & 20, Hudson Falls, Washington County, NY

Extremum Seeking Control of Smart Inverters for VAR Compensation

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

Arnold, Daniel; Negrete-Pincetic, Matias; Stewart, Emma

2015-09-04

Reactive power compensation is used by utilities to ensure customer voltages are within pre-defined tolerances and reduce system resistive losses. While much attention has been paid to model-based control algorithms for reactive power support and Volt Var Optimization (VVO), these strategies typically require relatively large communications capabilities and accurate models. In this work, a non-model-based control strategy for smart inverters is considered for VAR compensation. An Extremum Seeking control algorithm is applied to modulate the reactive power output of inverters based on real power information from the feeder substation, without an explicit feeder model. Simulation results using utility demand informationmore » confirm the ability of the control algorithm to inject VARs to minimize feeder head real power consumption. In addition, we show that the algorithm is capable of improving feeder voltage profiles and reducing reactive power supplied by the distribution substation.« less

Mass extinctions: Ecological selectivity and primary production

NASA Astrophysics Data System (ADS)

Rhodes, Melissa Clark; Thayer, Charles W.

1991-09-01

If mass extinctions were caused by reduced primary productivity, then extinctions should be concentrated among animals with starvation-susceptible feeding modes, active lifestyles, and high-energy budgets. The stratigraphic ranges (by stage) of 424 genera of bivalves and 309 genera of articulate brachiopods suggest that there was an unusual reduction of primary productivity at the Cretaceous/Tertiary (K/T) boundary extinction. For bivalves at the K/T, there were (1) selective extinction of suspension feeders and other susceptible trophic categories relative to deposit feeders and other resistant categories, and (2) among suspension feed-ers, selective extinction of bivalves with active locomotion. During the Permian-Triassic (P/Tr) extinction and Jurassic background time, extinction rates among suspension feeders were greater for articulate brachiopods than for bivalves. But during the K/T event, extinction rates of articulates and suspension-feeding bivalves equalized, possibly because the low-energy budgets of articulates gave them an advantage when food was scarce.

Subjective value of risky foods for individual domestic chicks: a hierarchical Bayesian model.

PubMed

Kawamori, Ai; Matsushima, Toshiya

2010-05-01

For animals to decide which prey to attack, the gain and delay of the food item must be integrated in a value function. However, the subjective value is not obtained by expected profitability when it is accompanied by risk. To estimate the subjective value, we examined choices in a cross-shaped maze with two colored feeders in domestic chicks. When tested by a reversal in food amount or delay, chicks changed choices similarly in both conditions (experiment 1). We therefore examined risk sensitivity for amount and delay (experiment 2) by supplying one feeder with food of fixed profitability and the alternative feeder with high- or low-profitability food at equal probability. Profitability varied in amount (groups 1 and 2 at high and low variance) or in delay (group 3). To find the equilibrium, the amount (groups 1 and 2) or delay (group 3) of the food in the fixed feeder was adjusted in a total of 18 blocks. The Markov chain Monte Carlo method was applied to a hierarchical Bayesian model to estimate the subjective value. Chicks undervalued the variable feeder in group 1 and were indifferent in group 2 but overvalued the variable feeder in group 3 at a population level. Re-examination without the titration procedure (experiment 3) suggested that the subjective value was not absolute for each option. When the delay was varied, the variable option was often given a paradoxically high value depending on fixed alternative. Therefore, the basic assumption of the uniquely determined value function might be questioned.

Feeder systems of acidic lava flows from the Paraná-Etendeka Igneous Province in southern Brazil and their implications for eruption style

NASA Astrophysics Data System (ADS)

de Lima, Evandro Fernandes; Waichel, Breno Leitão; Rossetti, Lucas De Magalhães May; Sommer, Carlos Augusto; Simões, Matheus Silva

2018-01-01

In the Rio Grande do Sul State, southern Brazil, the volcanic sequence of the Paraná-Etendeka Igneous Province consists of pahoehoe and rubbly pahoehoe lava flows with basaltic and basaltic andesitic composition respectively, overlaid by acidic volcanic rocks. The acidic volcanic rocks of the Paraná-Etendeka Igneous Province exhibit textures and structures that can be related to effusive and/or explosive eruptions generating predominantly rheoignimbrites. The huge lava volume related to the emplacement of large igneous provinces implicates on efficient feeder systems that are more commonly observed in continental environments. In the Paraná-Etendeka Igneous Province, feeders of basaltic rocks are exposed in several dyke swarms (Ponta Grossa NW trending, Florianópolis/Skeleton Coast (NW Namibia) N-S trending, Serra do Mar NE trending and Henties Bay/Outjo NE trending). In contrast, the only feeder system proposed to the acidic rocks of the Paraná-Etendeka Igneous Province is the Messum complex in Namibia (Milner et al. 1995). In the study area, the opening of three quarries for the extraction of dimension stones has exposed impressive structures/textures that show the effusive emplacement and the ductile to fragile-ductile magma transition along the acidic feeder dykes. Besides that, magma mixing/mingling processes between two acidic magmas are observed along the dykes. Here we describe new occurrences of acidic feeder dykes, correlate the dykes with acidic flows and discuss their importance to understand the emplacement of the Palmas type acid units in southern Brazil.

2. Photocopy of a drawing (original in the Collection of ...

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

2. Photocopy of a drawing (original in the Collection of the PL&C, Shelf 117, Drawing 155) SECTION OF THE MOODY STREET FEEDER, DECEMBER 30, 1847 - Moody Street Feeder, Moody Street vicinity, Lowell, Middlesex County, MA

30 CFR 77.1801 - Overcurrent protection.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Overcurrent protection. 77.1801 Section 77.1801... Wires and Trolley Feeder Wires § 77.1801 Overcurrent protection. Trolley wires and trolley feeder wires shall be provided with overcurrent protection. ...

Earthicle: The Design of a Conceptually New Type of Particle.

PubMed

Uskoković, Vuk; Pernal, Sebastian; Wu, Victoria M

2017-01-18

The conception and the steps made in the design of a conceptually new type of composite particle, so-called "earthicle", are being described. This particle is meant to roughly mimic the layered structure of the Earth, having zerovalent iron core, silicate mantle, and a thin carbonaceous crust resembling the biosphere and its geological remnants. Particles are made in a stable colloidal form in an aqueous medium, involving chemical precipitation and pyrolysis of citric acid in the solution. The effects of various synthesis parameters were studied, including borohydride and oleate concentrations, APTES/TEOS molar ratio, chemical nature of the carbon precursors, and others. XRD analysis confirmed the predominantly zerovalent iron composition of the core, amorphous silica and crystalline iron silicate/silicide composition of the mesolayer, and the carbonaceous, amorphous graphitic composition of the surface coating. The atomically thin carbon shell was also detected as a distinct shoulder on the broad n-π* absorption resonance and the peak at ∼300 nm, a signature of sp 2 hybridized electronic orbitals and the result of the interband π-π* transition characteristic of graphitic structures. The irregularity of the shape of generally round Fe 0 particles has caused the uniformity of the silica shell to be directly proportional to the particle size. The size of the earthicles ranged from 60 to 500 nm depending on the ionic concentration of the precursors and additives. Silica layer effectively prevented the aggregation of the iron core and increased the biocompatibility of the particles. The point of zero charge first increased from the acidic to the neutral range after coating Fe 0 core with the APTES-functionalized, aminated silica shell and then restored its low value after depositing the carboxylated carbonic crust in a charge-reversal process designed to facilitate the formation of core-multishell structures. Tested on K7M2 osteosarcoma cell line and primary kidney and lung fibroblasts, cytotoxicity was cell-line dependent; however, the trend assessed in both planar and 3D cell culture with respect to the three types of particles, Fe 0 , Fe/SiO 2 , and Fe/SiO 2 /C, was general and independent of the cell line. Thus, the pronounced toxicity of Fe 0 alone became neutralized after the silica layer was coated around Fe 0 . The further addition of the carbonic layer reduced the viability as compared to Fe/SiO 2 , albeit in a statistically significant manner only for K7M2 cell line when compared against the untreated control. Cell response also varied depending on the formulation: while some formulations exhibited lethal effects on kidney fibroblasts, were harmless to lung fibroblasts, and boosted the proliferation of K7M2 osteosarcoma cells, other formulations exhibited the opposite behavior despite being similar in terms of their core/double-shell structure. Compared across three different cancerous cell lines, K7M2 osteosarcoma and U87 and E297 glioblastoma, a similar cell-line dependency in response was observed, yet the viability reduction was consistent for all Fe/SiO 2 /C particles, ranging from 80% to 85% of the untreated control. Carbon surface layer, albeit of graphitic structural nature, was of a markedly more viable character than that of nanosized graphene oxide. The viability of lung fibroblasts incubated with Fe/SiO 2 /C particles was reduced in the presence of the alternating magnetic field of 312.75 A/m and 1 MHz, while the viability reduction caused by Fe/SiO 2 /C particles in kidney fibroblasts and K7M2 cells was converted from statistically insignificant to significant, suggesting that the composite particles could be used for hyperthermia treatments, although their properties should be optimized for a more intense effect. A single-cell immunofluorescent analysis of the interaction of primary kidney fibroblasts and K7M2 osteosarcoma cells with Fe/SiO 2 /C particles demonstrated that the cell uptake and perinuclear localization may be responsible for the necrotic effects. This analysis also showed that composite Fe/SiO 2 /C particles may have the ability to cause the rupture of the cancer cell nucleus while having a harmless effect on the primary cells. Such a promising and selective anticancer activity will be investigated in more detail in future studies.

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

PubMed

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.

Microstructure and properties of laser-clad high-temperature wear-resistant alloys

NASA Astrophysics Data System (ADS)

Yang, Yongqiang

1999-02-01

A 2-kW CO 2 laser with a powder feeder was used to produce alloy coatings with high temperature-wear resistance on the surface of steel substrates. To analyze the microstructure and microchemical composition of the laser-clad layers, a scanning electron microscope (SEM) equipped with an energy dispersive X-ray microanalysis system was employed. X-ray diffraction techniques were applied to characterize the phases formed during the cladding process. The results show that the microstructure of the cladding alloy consists mainly of many dispersed particles (W 2C, (W,Ti)C 1- x, WC), a lamellar eutectic carbide M 12C, and an (f.c.c) matrix. Hardness tested at room and high temperature showed that the laser-clad zone has a moderate room temperature hardness and relatively higher elevated temperature hardness. The application of the laser-clad layer to a hot tool was very successful, and its operational life span was prolonged 1 to 4 times.

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

Code of Federal Regulations, 2014 CFR

2014-10-01

... COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Applications and Licenses Space Stations § 25.147 Licensing provision for NGSO MSS feeder downlinks in the band 6700-6875 MHz. If an NGSO...

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

Code of Federal Regulations, 2012 CFR

2012-10-01

... COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Applications and Licenses Space Stations § 25.147 Licensing provision for NGSO MSS feeder downlinks in the band 6700-6875 MHz. If an NGSO...

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

Code of Federal Regulations, 2010 CFR

2010-10-01

... COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Applications and Licenses Space Stations § 25.147 Licensing provision for NGSO MSS feeder downlinks in the band 6700-6875 MHz. If an NGSO...

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

Code of Federal Regulations, 2011 CFR

2011-10-01

... COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Applications and Licenses Space Stations § 25.147 Licensing provision for NGSO MSS feeder downlinks in the band 6700-6875 MHz. If an NGSO...

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

Code of Federal Regulations, 2013 CFR

2013-10-01

... COMMISSION (CONTINUED) COMMON CARRIER SERVICES SATELLITE COMMUNICATIONS Applications and Licenses Space Stations § 25.147 Licensing provision for NGSO MSS feeder downlinks in the band 6700-6875 MHz. If an NGSO...

Evaluation of Advanced Microwave Landing System Procedures in the New York Terminal Area

DTIC Science & Technology

1991-03-01

sector controller called the CAMRN sector who must then sequence that traffic with multiple feeders from the south before handing off to the final...Right (13R) were all being used by landing traffic, the final controller handled the runway 22 arrivals and the CAMRN controller handled the runway 13R...Feeder Fix AAL678 DC10 H 00:09:00 AAL68 B767 H 00:23:00 AAL588 A300 H 00:27:00 PAA224 A300 H 01:20:00 4/ TWAll L101 H 01:34:00 CAMRN Feeder Fix DAL144

Vertical-Screw-Auger Conveyer Feeder

NASA Technical Reports Server (NTRS)

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.

Cost Benefit and Alternatives Analysis of Distribution Systems with Energy Storage Systems: Preprint

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

Harris, Tom; Nagarajan, Adarsh; Baggu, Murali

This paper explores monetized and non-monetized benefits from storage interconnected to distribution system through use cases illustrating potential applications for energy storage in California's electric utility system. This work supports SDG&E in its efforts to quantify, summarize, and compare the cost and benefit streams related to implementation and operation of energy storage on its distribution feeders. This effort develops the cost benefit and alternatives analysis platform, integrated with QSTS feeder simulation capability, and analyzed use cases to explore the cost-benefit of implementation and operation of energy storage for feeder support and market participation.

Microstrip Patch Antenna And Method

NASA Technical Reports Server (NTRS)

Fink, Patrick W. (Inventor)

2001-01-01

Method and apparatus are provided for a microstrip feeder structure for supplying properly phased signals to each radiator element in a microstrip antenna array that may be utilized for radiating circularly polarized electromagnetic waves. In one disclosed embodiment. the microstrip feeder structure includes a plurality of microstrip sections many or all of which preferably have an electrical length substantially equal to one-quarter wavelength at the antenna operating frequency. The feeder structure provides a low loss feed structure that may be duplicated multiple times through a set of rotations and translations to provide a radiating array of the desired size.

Movement of feeder-using songbirds: the influence of urban features.

PubMed

Cox, Daniel T C; Inger, Richard; Hancock, Steven; Anderson, Karen; Gaston, Kevin J

2016-11-23

Private gardens provide vital opportunities for people to interact with nature. The most popular form of interaction is through garden bird feeding. Understanding how landscape features and seasons determine patterns of movement of feeder-using songbirds is key to maximising the well-being benefits they provide. To determine these patterns we established three networks of automated data loggers along a gradient of greenspace fragmentation. Over a 12-month period we tracked 452 tagged blue tits Cyantistes caeruleus and great tits Parus major moving between feeder pairs 9,848 times, to address two questions: (i) Do urban features within different forms, and season, influence structural (presence-absence of connections between feeders by birds) and functional (frequency of these connections) connectivity? (ii) Are there general patterns of structural and functional connectivity across forms? Vegetation cover increased connectivity in all three networks, whereas the presence of road gaps negatively affected functional but not structural connectivity. Across networks structural connectivity was lowest in the summer when birds maintain breeding territories, however patterns of functional connectivity appeared to vary with habitat fragmentation. Using empirical data this study shows how key urban features and season influence movement of feeder-using songbirds, and we provide evidence that this is related to greenspace fragmentation.

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

NASA Astrophysics Data System (ADS)

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.

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

PubMed

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 <2 L was consumed every 12 h), and exploratory behavior, such as sniffing and licking of the feeder. Data were analyzed using mixed effects 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 stall may influence how quickly calves learn to use an AMF, but that the influence of stall wall design was affected by how easy calves were to train on the feeder upon initial introduction, which may depend in part on certain aspects of calf temperament. For many calves, solid side stalls at an AMF resulted faster in adaption than the steel bar stalls. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Induction of pluripotent stem cells from fibroblast cultures.

PubMed

Takahashi, Kazutoshi; Okita, Keisuke; Nakagawa, Masato; Yamanaka, Shinya

2007-01-01

Clinical application of embryonic stem (ES) cells faces difficulties regarding use of embryos, as well as tissue rejection after implantation. One way to circumvent these issues is to generate pluripotent stem cells directly from somatic cells. Somatic cells can be reprogrammed to an embryonic-like state by the injection of a nucleus into an enucleated oocyte or by fusion with ES cells. However, little is known about the mechanisms underlying these processes. We have recently shown that the combination of four transcription factors can generate ES-like pluripotent stem cells directly from mouse fibroblast cultures. The cells, named induced pluripotent stem (iPS) cells, can be differentiated into three germ layers and committed to chimeric mice. Here we describe detailed methods and tips for the generation of iPS cells.

A density gradient of VAPG peptides on a cell-resisting surface achieves selective adhesion and directional migration of smooth muscle cells over fibroblasts.

PubMed

Yu, Shan; Zuo, Xingang; Shen, Tao; Duan, Yiyuan; Mao, Zhengwei; Gao, Changyou

2018-05-01

Selective adhesion and migration of smooth muscle cells (SMCs) over fibroblasts (FIBs) is required to prevent adventitia fibrosis in vascular regeneration. In this study, a uniform cell-resisting layer of poly(ethylene glycol) (PEG) with a density gradient of azide groups was generated on a substrate by immobilizing two kinds of PEG molecules in a gradient manner. A density gradient of alkynyl-functionalized Val-Ala-Pro-Gly (VAPG) peptides was then prepared on the PEG layer via click chemistry. The VAPG density gradient was characterized by fluorescence imaging, revealing the gradual enhancement of the fluorescent intensity along the substrate direction. The adhesion and mobility of SMCs were selectively enhanced on the VAPG density gradient, leading to directional migration toward the higher peptide density (up to 84%). In contrast, the adhesion and mobility of FIBs were significantly weakened. The net displacement of SMCs also significantly increased compared with that on tissue culture polystyrene (TCPS) and that of FIBs on the gradient. The mitogen-activated protein kinase (MAPK) signaling pathways related to cell migration were studied, showing higher expressions of functional proteins from SMCs on the VAPG-modified surface in a density-dependent manner. For the first time the selective adhesion and directional migration of SMCs over FIBs was achieved by an elaborative design of a gradient surface, leading to a new insight in design of novel vascular regenerative materials. Selective cell adhesion and migration guided by regenerative biomaterials are extremely important for the regeneration of targeted tissues, which can avoid the drawbacks of incorrect and uncontrolled responses of tissue cells to implants. For example, selectivity of smooth muscle cells (SMCs) over fibroblasts (FIBs) is required to prevent adventitia fibrosis in vascular regeneration. Herein we prepare a uniform cell-repelling layer, on which SMCs-selective Val-Ala-Pro-Gly (VAPG) peptides are immobilized in a continuous manner. Selective adhesion and enhanced and directional migration of SMCs over FIBs are achieved by the interplay of cell-repelling layer and gradient SMCs-selective VAPG peptides, paving a new way for the design of novel vascular grafts with enhanced biological performance. Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Freeform inkjet printing of cellular structures with bifurcations.

PubMed

Christensen, Kyle; Xu, Changxue; Chai, Wenxuan; Zhang, Zhengyi; Fu, Jianzhong; Huang, Yong

2015-05-01

Organ printing offers a great potential for the freeform layer-by-layer fabrication of three-dimensional (3D) living organs using cellular spheroids or bioinks as building blocks. Vascularization is often identified as a main technological barrier for building 3D organs. As such, the fabrication of 3D biological vascular trees is of great importance for the overall feasibility of the envisioned organ printing approach. In this study, vascular-like cellular structures are fabricated using a liquid support-based inkjet printing approach, which utilizes a calcium chloride solution as both a cross-linking agent and support material. This solution enables the freeform printing of spanning and overhang features by providing a buoyant force. A heuristic approach is implemented to compensate for the axially-varying deformation of horizontal tubular structures to achieve a uniform diameter along their axial directions. Vascular-like structures with both horizontal and vertical bifurcations have been successfully printed from sodium alginate only as well as mouse fibroblast-based alginate bioinks. The post-printing fibroblast cell viability of printed cellular tubes was found to be above 90% even after a 24 h incubation, considering the control effect. © 2014 Wiley Periodicals, Inc.

Surgical option for the correction of Peyronie's disease: an autologous tissue-engineered endothelialized graft.

PubMed

Imbeault, Annie; Bernard, Geneviève; Ouellet, Gabrielle; Bouhout, Sara; Carrier, Serge; Bolduc, Stéphane

2011-11-01

Surgical treatment is indicated in severe cases of Peyronie's disease. Incision of the plaque with subsequent graft material implantation is the option of choice. Ideal graft tissue is not yet available. To evaluate the use of an autologous tissue-engineered endothelialized graft by the self-assembly method, for tunica albuginea (TA) reconstruction in Peyronie's disease. Two TA models were created. Human fibroblasts were isolated from a skin biopsy and cultured in vitro until formation of fibroblast sheets. After 4 weeks of maturation, human umbilical vein endothelial cells (HUVEC) were seeded on fibroblasts sheets and wrapped around a tubular support to form a cylinder of about 10 layers. After 21 days of tube maturation, HUVEC were seeded into the lumen of the fibroblast tubes for the endothelialized tunica albuginea (ETA). No HUVEC were seeded into the lumen for the TA model. Both constructs were placed under perfusion in a bioreactor for 1 week. Histology, immunohistochemistry, and burst pressure were performed to characterize mature tubular graft. Animal manipulations were also performed to demonstrate the impact of endothelial cells in vivo. Histology showed uniform multilayered fibroblasts. Extracellular matrix, produced entirely by fibroblasts, presented a good staining for collagen 1. Some elastin fibers were also present. For the TA model, anti-human von Willebrand antibody revealed the endothelial cells forming capillary-like structures. TA model reached a burst pressure of 584 mm Hg and ETA model obtained a burst pressure of 719 mm Hg. This tissue-engineered endothelialized tubular graft is structurally similar to normal TA and presents an adequate mechanical resistance. The self-assembly method used and the autologous property of this model could represent an advantage comparatively to other available grafts. Further evaluation including functional testing will be necessary to characterize in vivo implantation and behavior of the graft. © 2011 International Society for Sexual Medicine.

View east, stone sluice, beginning of lower standing section, showing ...

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

View east, stone sluice, beginning of lower standing section, showing third drop, stone pier in center, cement piers to right - Glens Falls Feeder, Sluice, Along south side of Glens Falls Feeder between locks 10 & 20, Hudson Falls, Washington County, NY

System Operations Studies : Feeder System Model. User's Manual.

DOT National Transportation Integrated Search

1982-11-01

The Feeder System Model (FSM) is one of the analytic models included in the System Operations Studies (SOS) software package developed for urban transit systems analysis. The objective of the model is to assign a proportion of the zone-to-zone travel...

Development of Experimental Setup of Metal Rapid Prototyping Machine using Selective Laser Sintering Technique

NASA Astrophysics Data System (ADS)

Patil, S. N.; Mulay, A. V.; Ahuja, B. B.

2018-04-01

Unlike in the traditional manufacturing processes, additive manufacturing as rapid prototyping, allows designers to produce parts that were previously considered too complex to make economically. The shift is taking place from plastic prototype to fully functional metallic parts by direct deposition of metallic powders as produced parts can be directly used for desired purpose. This work is directed towards the development of experimental setup of metal rapid prototyping machine using selective laser sintering and studies the various parameters, which plays important role in the metal rapid prototyping using SLS technique. The machine structure in mainly divided into three main categories namely, (1) Z-movement of bed and table, (2) X-Y movement arrangement for LASER movements and (3) feeder mechanism. Z-movement of bed is controlled by using lead screw, bevel gear pair and stepper motor, which will maintain the accuracy of layer thickness. X-Y movements are controlled using timing belt and stepper motors for precise movements of LASER source. Feeder mechanism is then developed to control uniformity of layer thickness metal powder. Simultaneously, the study is carried out for selection of material. Various types of metal powders can be used for metal RP as Single metal powder, mixture of two metals powder, and combination of metal and polymer powder. Conclusion leads to use of mixture of two metals powder to minimize the problems such as, balling effect and porosity. Developed System can be validated by conducting various experiments on manufactured part to check mechanical and metallurgical properties. After studying the results of these experiments, various process parameters as LASER properties (as power, speed etc.), and material properties (as grain size and structure etc.) will be optimized. This work is mainly focused on the design and development of cost effective experimental setup of metal rapid prototyping using SLS technique which will gives the feel of metal rapid prototyping process and its important parameters.

Proliferation in culture of primordial germ cells derived from embryonic stem cell: induction by retinoic acid.

PubMed

Makoolati, Zohreh; Movahedin, Mansoureh; Forouzandeh-Moghadam, Mehdi

2016-12-01

An in vitro system that supports primordial germ cells (PGCs) survival and proliferation is useful for enhancement of these cells and efficient transplantation in infertility disorders. One approach is cultivation of PGCs under proper conditions that allow self-renewal and proliferation of PGCs. For this purpose, we compared the effects of different concentrations of retinoic acid (RA), and the effect of PGCs co-culture (Co-C) with SIM mouse embryo-derived thioguanine- and ouabain-resistant (STO) cells on the proliferation of embryonic stem cells (ESCs)-derived PGCs. One-day-old embryoid body (EB) was cultured for 4 days in simple culture system in the presence of 5 ng/ml bone morphogenetic protein-4 (BMP4) (SCB group) for PGC induction. For PGC enrichment, ESCs-derived germ cells were cultured for 7 days in the presence of different doses (0-5  μM) of RA, both in the simple and STO Co-C systems. At the end of the culture period, viability and proliferation rates were assessed and expression of mouse vasa homologue (Mvh),  α6 integrin,  β1 integrin, stimulated by retinoic acid 8 (Stra8) and piwi (Drosophila)-like 2 (Piwil2) was evaluated using quantitative PCR. Also, the inductive effects were investigated immunocytochemically with Mvh and cadherin1 (CDH1) on the selected groups. Immunocytochemistry/PCR results showed higher expression of Mvh, the PGC-specific marker, in 3  μM RA concentrations on the top of the STO feeder layer. Meanwhile, assessment of the Stra8 mRNA and CDH1 protein, the specific makers for spermatogonia, showed no significant differences between groups. Based on the results, it seems that in the presence of 3 μM RA on top of the STO feeder layer cells, the majority of the cells transdifferentiated into germ cells were PGCs. © 2016 The Author(s).

In Vitro Differentiation of Human Mesenchymal Stem Cells into Functional Cardiomyocyte-like Cells.

PubMed

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 differentiation strategy can effectively harness the cardiomyogenic potential of young MSCs, such as FTM HUCPVCs, and suggests that in vitro pre-differentiation could be a potential strategy to increase their regenerative efficacy in vivo.

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

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

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

2006-06-30

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

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

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

Lee, In Joon; Chung, Jin Wook, E-mail: chungjw@snu.ac.kr; Yin, Yong Hu

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.more » 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.« less

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

PubMed

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.

Feeding group responses of a Neotropical termite assemblage to rain forest fragmentation.

PubMed

Davies, Richard G

2002-10-01

Biomass collapse and its associated microclimatic stresses within recently isolated rain forest fragments may negatively affect species diversity of most resident taxa. However, for some decomposer organisms, increased resource availability via accompanying tree die-off may effect positive responses, at least for a time, with implications for rates of nutrient cycling and greenhouse gas release. This study investigates the early effects of forest fragmentation on a Neotropical termite assemblage. Numbers of encounters (surrogate for relative abundance) and species richness of wood and leaf-litter feeders, soil feeders, and the whole assemblage, were studied across true forest islands and mainland sites at a hydroelectric reservoir in French Guiana. Results showed no overall effect of fragmentation on either total termite encounters or species richness. However, numbers of encounters and species richness of wood and leaf-litter feeders showed positive responses to forest fragmentation. By contrast, soil feeders showed a negative response for numbers of encounters and no significant effect for species richness. Environmental data suggest that increased tree die-off, and other edge effects associated with biomass collapse, were underway at the time of sampling. Resulting increase in resource availability may therefore explain the positive influence on wood and leaf-litter feeders. A possible decrease in predation pressure from ants with decrease in island size was not tested for, but was a likely effect of the flooded matrix habitat. Fragmentation effects on soil feeder encounters may be due to the energetic and microclimatic constraints of feeding lower down the humification gradient of termite food substrates, but were not sufficient to affect species richness. The patterns revealed suggest that rates of wood decomposition following tree die-off, and of soil nutrient cycling, under different rain forest fragmentation scenarios, merit further study.

A comparison of two systems for chlorinating water in rural Honduras.

PubMed

Henderson, Amy K; Sack, R Bradley; Toledo, Erick

2005-09-01

This study investigated a small subset of the two community water-disinfection systems--hypochlorinators and tablet feeders-in rural Honduras. Levels of residual chlorine were assessed at three locations within the distribution system: the tank, the proximal house, and the distal house. The levels of residual chlorine were compared with the standard guidelines set by the Pan American Health Organization and the International Rural Water Association for potable water that require a minimum of 1.0 (tank), 0.5 (proximal house), and 0.2 (distal house) ppm for each location. The levels of residual chlorine were also compared across systems, e.g. hypochlorinators to tablet feeders. At the tank and proximal house, tablet feeders had significantly higher mean values for levels of residual chlorine (measured in ppm) than hypochlorinators (tank: 1.20 vs 0.67; proximal house: 0.44 vs 0.32, p < 0.001 for both) with no significant difference at the distal house (0.16 vs 0.16). At the tank and proximal house, tablet feeders were more likely to meet recommended standards than hypochlorinators (90.3% vs 13.3%, p < 0.0001 and 41.3% vs 23.7%, p < 0.0001) with a smaller difference seen at the distal house (30.6% vs 27.1%, p = 0.24). The apparent dichotomy in chlorine levels of tablet feeders (e.g. between tank/proximal house and distal house) is discussed. The results suggest that tablet feeders may be more effective than hypochlorinators in supplying clean water in rural, resource-poor settings and possibly serve as an alternative technology for water disinfection. Further research on techniques for empowering and building capacity within community water boards will help organize and introduce sustainable water systems in developing countries.

Curcumin inhibits TGF-β1-induced connective tissue growth factor expression through the interruption of Smad2 signaling in human gingival fibroblasts.

PubMed

Chen, Jung-Tsu; Wang, Chen-Ying; Chen, Min-Huey

2018-01-13

Many fibrotic processes are associated with an increased level of transforming growth factor-β1 (TGF-β1). TGF-β1 can increase synthesis of matrix proteins and enhance secretion of protease inhibitors, resulting in matrix accumulation. Connective tissue growth factor (CTGF) is a downstream profibrotic effector of TGF-β1 and is associated with the fibrosis in several human organs. Curcumin has been applied to reduce matrix accumulation in fibrotic diseases. This study was aimed to evaluate whether curcumin could suppress TGF-β1-induced CTGF expression and its related signaling pathway involving in this inhibitory action in primary human gingival fibroblasts. The differences in CTGF expression among three types of gingival overgrowth and normal gingival tissues were assessed by immunohistochemistry. Gingival fibroblast viability in cultured media with different concentrations of curcumin was studied by MTT assay. The effect of curcumin on TGF-β1-induced CTGF expression in primary human gingival fibroblasts was examined by immunoblotting. Moreover, the proteins involved in TGF-β1 signaling pathways including TGF-β1 receptors and Smad2 were also analyzed by immunoblotting. CTGF was highly expressed in fibroblasts, epithelial cells and some of endothelial cells, smooth muscle cells, and inflammatory cells in phenytoin-induced gingival overgrowth tissues rather than in those of hereditary and inflammatory gingival overgrowth tissues. Moreover, CTGF expression in the epithelial and connective tissue layers was higher in phenytoin-induced gingival overgrowth tissues than in normal gingival tissues. Curcumin was nontoxic and could reduce TGF-β1-induced CTGF expression by attenuating the phosphorylation and nuclear translocation of Smad2. Curcumin can suppress TGF-β1-induced CTGF expression through the interruption of Smad2 signaling. Copyright © 2018. Published by Elsevier B.V.

Container barge feeder service study : Bridgeport, New Haven, New London, Norwich

DOT National Transportation Integrated Search

2001-03-01

The Connecticut Department of Transportation (ConnDOT) has conducted this study to determine the need and opportunity for establishing a Container Barge Feeder Service along Long Island Sound between the Port of New York and New Jersey (NY&NJ) and th...

An in vitro monocyte culture method and establishment of a human monocytic cell line (K63).

PubMed

Kadoi, Katsuyuki

2011-01-01

A novel method of monocyte culture in vitro was developed. The fraction of monocytes was obtained by density centrifugation of heparinised human venous blood samples. Monocytes were suspended in a modified Rosewell Park Memorial Institute medium (RPMI)-1640 (mRPMI) supplemented with 10% non-inactivated autologous serum added to the feeder cells. An avian cell line was used for feeder cells. Only those monocytes that settled on feeder cells grew rapidly at 37°C-38°C into a formation of clumped masses within two to three days. The cell mass was harvested and subcultures were made without feeder cells. A stable cell line (K63) was established from subcultures using a limited dilution method and cell cloning in microplates. K63 cells were adapted for later growth in the mRPMI medium supplemented with 10% foetal calf serum. The cells were well maintained at over 50th passage levels. This method proved to be applicable for monocyte cultures of animals as well.

Evaluating the Magnitude and Duration of Cold Load Pick-up on Residential Distribution Feeders Using Multi-State Load Models

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

Schneider, Kevin P.; Sortomme, Eric; Venkata, S. S.

The increased level of demand that is associated with the restoration of service after an outage, Cold Load Pick-Up (CLPU), can be significantly higher than pre-outage levels, even exceeding the normal distribution feeder peak demand. These high levels of demand can delay restoration efforts and in extreme cases damage equipment. The negative impacts of CLPU can be mitigated with strategies that restore the feeder in sections, minimizing the load current. The challenge for utilities is to manage the current level on critical equipment while minimizing the time to restore service to all customers. Accurately modeling CLPU events is the firstmore » step in developing improved restoration strategies that minimize restoration times. This paper presents a new method for evaluating the magnitude of the CLPU peak, and its duration, using multi-state load models. The use of multi-state load models allows for a more accurate representation of the end-use loads that are present on residential distribution feeders.« less

Controlled cell morphology and liver-specific function of engineered primary hepatocytes by fibroblast layer cell densities.

PubMed

Sakai, Yusuke; Koike, Makiko; Kawahara, Daisuke; Hasegawa, Hideko; Murai, Tomomi; Yamanouchi, Kosho; Soyama, Akihiko; Hidaka, Masaaki; Takatsuki, Mitsuhisa; Fujita, Fumihiko; Kuroki, Tamotsu; Eguchi, Susumu

2018-03-05

Engineered primary hepatocytes, including co-cultured hepatocyte sheets, are an attractive to basic scientific and clinical researchers because they maintain liver-specific functions, have reconstructed cell polarity, and have high transplantation efficiency. However, co-culture conditions regarding engineered primary hepatocytes were suboptimal in promoting these advantages. Here we report that the hepatocyte morphology and liver-specific function levels are controlled by the normal human diploid fibroblast (TIG-118 cell) layer cell density. Primary rat hepatocytes were plated onto TIG-118 cells, previously plated 3 days before at 1.04, 5.21, and 26.1×10 3  cells/cm 2 . Hepatocytes plated onto lower TIG-118 cell densities expanded better during the early culture period. The hepatocytes gathered as colonies and only exhibited small adhesion areas because of the pushing force from proliferating TIG-118 cells. The smaller areas of each hepatocyte result in the development of bile canaliculi. The highest density of TIG-118 cells downregulated albumin synthesis activity of hepatocytes. The hepatocytes may have undergone apoptosis associated with high TGF-β1 concentration and necrosis due to a lack of oxygen. These occurrences were supported by apoptotic chromatin condensation and high expression of both proteins HIF-1a and HIF-1b. Three types of engineered hepatocyte/fibroblast sheets comprising different TIG-118 cell densities were harvested after 4 days of hepatocyte culture and showed a complete cell sheet format without any holes. Hepatocyte morphology and liver-specific function levels are controlled by TIG-118 cell density, which helps to design better engineered hepatocytes for future applications such as in vitro cell-based assays and transplantable hepatocyte tissues. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Anti-Inflammatory Activities of Pentaherbs Formula, Berberine, Gallic Acid and Chlorogenic Acid in Atopic Dermatitis-Like Skin Inflammation.

PubMed

Tsang, Miranda S M; Jiao, Delong; Chan, Ben C L; Hon, Kam-Lun; Leung, Ping C; Lau, Clara B S; Wong, Eric C W; Cheng, Ling; Chan, Carmen K M; Lam, Christopher W K; Wong, Chun K

2016-04-20

Atopic dermatitis (AD) is a common allergic skin disease, characterized by dryness, itchiness, thickening and inflammation of the skin. Infiltration of eosinophils into the dermal layer and presence of edema are typical characteristics in the skin biopsy of AD patients. Previous in vitro and clinical studies showed that the Pentaherbs formula (PHF) consisting of five traditional Chinese herbal medicines, Flos Lonicerae, Herba Menthae, Cortex Phellodendri, Cortex Moutan and Rhizoma Atractylodis at w/w ratio of 2:1:2:2:2 exhibited therapeutic potential in treating AD. In this study, an in vivo murine model with oxazolone (OXA)-mediated dermatitis was used to elucidate the efficacy of PHF. Active ingredients of PHF water extract were also identified and quantified, and their in vitro anti-inflammatory activities on pruritogenic cytokine IL-31- and alarmin IL-33-activated human eosinophils and dermal fibroblasts were evaluated. Ear swelling, epidermis thickening and eosinophils infiltration in epidermal and dermal layers, and the release of serum IL-12 of the murine OXA-mediated dermatitis were significantly reduced upon oral or topical treatment with PHF (all p < 0.05). Gallic acid, chlorogenic acid and berberine contents (w/w) in PHF were found to be 0.479%, 1.201% and 0.022%, respectively. Gallic acid and chlorogenic acid could suppress the release of pro-inflammatory cytokine IL-6 and chemokine CCL7 and CXCL8, respectively, in IL-31- and IL-33-treated eosinophils-dermal fibroblasts co-culture; while berberine could suppress the release of IL-6, CXCL8, CCL2 and CCL7 in the eosinophil culture and eosinophils-dermal fibroblasts co-culture (all p < 0.05). These findings suggest that PHF can ameliorate allergic inflammation and attenuate the activation of eosinophils.

3D tissue formation by stacking detachable cell sheets formed on nanofiber mesh.

PubMed

Kim, Min Sung; Lee, Byungjun; Kim, Hong Nam; Bang, Seokyoung; Yang, Hee Seok; Kang, Seong Min; Suh, Kahp-Yang; Park, Suk-Hee; Jeon, Noo Li

2017-03-23

We present a novel approach for assembling 3D tissue by layer-by-layer stacking of cell sheets formed on aligned nanofiber mesh. A rigid frame was used to repeatedly collect aligned electrospun PCL (polycaprolactone) nanofiber to form a mesh structure with average distance between fibers 6.4 µm. When human umbilical vein endothelial cells (HUVECs), human foreskin dermal fibroblasts, and skeletal muscle cells (C2C12) were cultured on the nanofiber mesh, they formed confluent monolayers and could be handled as continuous cell sheets with areas 3 × 3 cm 2 or larger. Thicker 3D tissues have been formed by stacking multiple cell sheets collected on frames that can be nested (i.e. Matryoshka dolls) without any special tools. When cultured on the nanofiber mesh, skeletal muscle, C2C12 cells oriented along the direction of the nanofibers and differentiated into uniaxially aligned multinucleated myotube. Myotube cell sheets were stacked (upto 3 layers) in alternating or aligned directions to form thicker tissue with ∼50 µm thickness. Sandwiching HUVEC cell sheets with two dermal fibroblast cell sheets resulted in vascularized 3D tissue. HUVECs formed extensive networks and expressed CD31, a marker of endothelial cells. Cell sheets formed on nanofiber mesh have a number of advantages, including manipulation and stacking of multiple cell sheets for constructing 3D tissue and may find applications in a variety of tissue engineering applications.

Analysis of the 60-Hz power system at KSC: The Orsino substation

NASA Technical Reports Server (NTRS)

Kalu, Alex O.

1989-01-01

An analysis of the Orsino Substation, a component (50 percent) of the 60-Hertz electric power system at the Kennedy Space Center, is presented. Presented here are separate single-line diagrams of the sixteen feeder circuits to permit easy access to information on the individual feeders for future planning. The load condition of each feeder and load break switch are presented and a heuristic reliability analysis of the system is performed. Information is given about the system fashion useful for decision making purposes. The beauty of it is in the simplified manner by which information about the system can be obtained.

Methods to Determine Recommended Feeder-Wide Advanced Inverter Settings for Improving Distribution System Performance

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

Rylander, Matthew; Reno, Matthew J.; Quiroz, Jimmy E.

This paper describes methods that a distribution engineer could use to determine advanced inverter settings to improve distribution system performance. These settings are for fixed power factor, volt-var, and volt-watt functionality. Depending on the level of detail that is desired, different methods are proposed to determine single settings applicable for all advanced inverters on a feeder or unique settings for each individual inverter. Seven distinctly different utility distribution feeders are analyzed to simulate the potential benefit in terms of hosting capacity, system losses, and reactive power attained with each method to determine the advanced inverter settings.

An automatic 14-day paste diet feeder for animals

NASA Technical Reports Server (NTRS)

Vasques, Marilyn; Mulenburg, Jerry; Gundo, Dan; Griffith, Jon

1994-01-01

During a centrifuge experiment, any interruption that requires stopping the centrifuge may influence the results. Centrifuges often must be stopped for animal maintenance (food, water and waste removal), especially in cases of timed feedings. To eliminate the need for stopping the centrifuge while still providing timed feeding, an automatic paste diet feeder was developed. The feeder is based on a constant volume concept and can deliver a predetermined amount of paste diet at specified time intervals. This unit was supported by water delivery and waste collection systems. The entire system performed reliably and maintained the animals well for a continuous centrifugation experiment of 14 days.

Systems Operation Studies for Automated Guideway Transit Systems: Feeder Systems Model Functional Specification

DOT National Transportation Integrated Search

1981-01-01

This document specifies the functional requirements for the AGT-SOS Feeder Systems Model (FSM), the type of hardware required, and the modeling techniques employed by the FSM. The objective of the FSM is to map the zone-to-zone transit patronage dema...

View southeast, stone sluice, top of lower standing section, showing ...

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

View southeast, stone sluice, top of lower standing section, showing row of cement piers in center, retaining wall to left, barge canal sluice to right - Glens Falls Feeder, Sluice, Along south side of Glens Falls Feeder between locks 10 & 20, Hudson Falls, Washington County, NY

47. FEEDER CANAL AT WILL'S BASIN. BOATS IN THE FOREGROUND ...

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

47. FEEDER CANAL AT WILL'S BASIN. BOATS IN THE FOREGROUND SHOW THE CONTRAST BETWEEN A FULLY LOADED CANAL BOAT (LEFT) AND AN EMPTY ONE (RIGHT). D, L & W RAILROAD'S DOUBLE INTERSECTION PRATT TRUSS BRIDGE IS VISIBLE IN BACKGROUND. - Morris Canal, Phillipsburg, Warren County, NJ

34. INTERIOR VIEW, SAME AS ABOVE WITH THE FEEDER ROD ...

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

34. INTERIOR VIEW, SAME AS ABOVE WITH THE FEEDER ROD BEING OFFERED INTO THE MACHINE; UNLIKE THE OTHER NAIL CUTTING MACHINES, THE NAIL PLATE MUST BE HAND FLIPPED AND HAND FED WITH THIS MACHINE - LaBelle Iron Works, Thirtieth & Wood Streets, Wheeling, Ohio County, WV

Termites as bioindicators of habitat quality in the caatinga, Brazil: is there agreement between structural habitat variables and the sampled assemblages?

PubMed

Alves, W de F; Mota, A S; Lima, R A A de; Bellezoni, R; Vasconcellos, A

2011-01-01

The composition of termite assemblages was analyzed in three caatinga sites of the Estação Ecológica do Seridó, located in the municipality of Serra Negra do Norte, in the state of Rio Grande do Norte, Brazil. These sites have been subjected to selective logging, and cleared for pasture and farming. A standardized sampling protocol for termite assemblages (30h/person/site) was conducted between September 2007 and February 2009. At each site we measured environmental variables, such as soil pH and organic matter, necromass stock, vegetation height, stem diameter at ankle height (DAH) and the largest and the smallest crown width. Ten species of termites, belonging to eight genera and three families, were found at the three experimental sites. Four feeding groups were sampled: wood-feeders, soil-feeders, wood-soil interface feeders and leaf-feeders. The wood-feeders were dominant in number of species and number of encounters at all sites. In general, the sites were not significantly different in relation to the environmental variables measured. The same pattern was observed for termite assemblages, where no significant differences in species richness, relative abundance and taxonomic and functional composition were observed between the three sites. The agreement between composition of assemblages and environmental variables reinforces the potential of termites as biological indicators of habitat quality.

Four eyes match better than two: Sharing of precise patch-use time among socially foraging domestic chicks.

PubMed

Xin, Qiuhong; Ogura, Yukiko; Matsushima, Toshiya

2017-07-01

To examine how resource competition contributes to patch-use behaviour, we examined domestic chicks foraging in an I-shaped maze equipped with two terminal feeders. In a variable interval schedule, one feeder supplied grains three times more frequently than the other, and the sides were reversed midway through the experiment. The maze was partitioned into two lanes by a transparent wall, so that chicks fictitiously competed without actual interference. Stay time at feeders was compared among three groups. The "single" group contained control chicks; the "pair" group comprised the pairs of chicks tested in the fictitious competition; "mirror" included single chicks accompanied by their respective mirror images. Both "pair" and "mirror" chicks showed facilitated running. In terms of the patch-use ratio, "pair" chicks showed precise matching at approximately 3:1 with significant mutual dependence, whereas "single" and "mirror" chicks showed a comparable under-matching. The facilitated running increased visits to feeders, but failed to predict the patch-use ratio of the subject. At the reversal, quick switching occurred similarly in all groups, but the "pair" chicks revealed a stronger memory-based matching. Perceived competition therefore contributes to precise matching and lasting memory of the better feeder, in a manner dissociated from socially facilitated food search. Copyright © 2017 Elsevier B.V. All rights reserved.

Response of a benthic suspension feeder ( Crassostrea virginica Gmelin) to three centuries of anthropogenic eutrophication in Chesapeake Bay

NASA Astrophysics Data System (ADS)

Kirby, Michael X.; Miller, Henry M.

2005-03-01

Biogenic reefs built by oysters and other suspension feeders are vital components of estuarine ecosystems. By consuming phytoplankton, suspension feeders act to suppress accumulation of organic matter in the water column. Nutrient loading increases the rate of primary production, thereby causing eutrophication. As suspension feeders consume more organic matter from increasing abundance of phytoplankton, their rate of growth should also increase if they are food limited. We show here that the eastern oyster, Crassostrea virginica (Gmelin), from St. Mary's and Patuxent rivers, Chesapeake Bay, grew faster during anthropogenic eutrophication relative to C. virginica before eutrophication. Growth of shell height, shell thickness and adductor muscle increased after eutrophication began in the late 18th century. After 1860, growth decreased, perhaps reflecting the negative effects of hypoxia, harmful algal blooms, disease and fishing on oyster growth. These results are consistent with the view that an increasing supply of phytoplankton resulting from eutrophication enhanced growth of C. virginica between 1760 and 1860, before oyster reefs were degraded by destructive fishing practices between 1870 and 1930. Alternative factors, such as changes in water temperature, salinity, and fishing are less likely to be responsible for this pattern. These results have implications for restoration of oyster reefs in order to mitigate the effects of eutrophication in estuaries, as well as the paleoecological relationship between suspension feeders and paleoproductivity.

View north, stone sluice, head of 40foot break, showing failed ...

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

View north, stone sluice, head of 40-foot break, showing failed arch in center, stone pier in center right, cement piers to upper left, retaining wall in background - Glens Falls Feeder, Sluice, Along south side of Glens Falls Feeder between locks 10 & 20, Hudson Falls, Washington County, NY

Dry piston coal feeder

DOEpatents

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.

76 FR 3655 - Bunker Hill Groundwater Basin, Riverside-Corona Feeder Project, San Bernardino and Riverside...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-20

... DEPARTMENT OF THE INTERIOR Bureau of Reclamation Bunker Hill Groundwater Basin, Riverside-Corona... Draft Environmental Impact Statement (SDEIR/DEIS) for the proposed Riverside-Corona Feeder (RCF) Project... Bernardino, California 92410 Corona Public Library, 650 South Main Street, Corona, California 92882 Riverside...

30 CFR 57.12086 - Location of trolley wire.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Location of trolley wire. 57.12086 Section 57... Underground Only § 57.12086 Location of trolley wire. Trolley and trolley feeder wire shall be installed... limitations would prevent the safe installation or use of such trolley and trolley feeder wire. ...

30 CFR 57.12086 - Location of trolley wire.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Location of trolley wire. 57.12086 Section 57... Underground Only § 57.12086 Location of trolley wire. Trolley and trolley feeder wire shall be installed... limitations would prevent the safe installation or use of such trolley and trolley feeder wire. ...

Optimal foraging by birds: feeder-based experiments for secondary and post-secondary students

USDA-ARS?s Scientific Manuscript database

Optimal foraging theory attempts to explain the foraging patterns observed in animals, including their choice of particular food items and foraging locations. Here, we describe three exercises designed to test hypotheses about food choice and foraging habitat preference using bird feeders. These e...

Theoretical Comparison of Fixed Route Bus and Flexible Route Subscription Bus Feeder Service in Low Density Areas

DOT National Transportation Integrated Search

1975-03-01

parametric variation of demand density was used to compare service level and cost of two alternative systems for providing low density feeder service. Supply models for fixed route and flexible route service were developed and applied to determine ra...

77 FR 13191 - Airworthiness Directives; Bombardier, Inc. Airplanes

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-06

... corrosion and inadequate silver-plating. This AD requires replacing ADG power feeder cables. We are issuing this AD to prevent galvanic corrosion on ADG power feeder cables, which could result in damage to the... damaged due to galvanic corrosion. It was subsequently determined that the silver- plating is inadequate...

Expression of cholesteryl glucoside by heat shock in human fibroblasts

PubMed Central

Kunimoto, Shohko; Kobayashi, Tetsuyuki; Kobayashi, Susumu; Murakami-Murofushi, Kimiko

2000-01-01

ABSTRACT We investigated the heat-induced alteration of glycolipids in human cultured cells, TIG-3 fibroblasts, to show the expression of steryl glucoside by heat shock. A glycolipid band was detected on a thin-layer chromatography plate in lipid extracts from TIG-3 cells exposed to high temperature (42°C) for 15 and 30 minutes, while it was hardly detectable without heat shock. Both cholesterol and glucose were almost exclusively detected by gas liquid chromatography as degradation products of the lipid. The structure of the lipid molecule was elucidated by electrospray mass spectrometry to be a cholesteryl glucoside. This is the first report to show the occurrence of a steryl glucoside in mammalian cells, and this substance is considered to have a significant role in heat shock responses in mammalian cells. PMID:10701833

Constructing a blood vessel on the porous scaffold modified with vascular endothelial growth factor and basic fibroblast growth factor

NASA Astrophysics Data System (ADS)

Sevostyanova, V. V.; Matveeva, V. G.; Antonova, L. V.; Velikanova, E. A.; Shabaev, A. R.; Senokosova, E. A.; Krivkina, E. O.; Vasyukov, G. Yu.; Glushkova, T. V.; Kudryavtseva, Yu. A.; Barbarash, O. L.; Barbarash, L. S.

2016-11-01

Incorporation of the growth factors into biodegradable polymers is a promising approach for the fabrication of tissue-engineered vascular grafts. Here we blended poly(ɛ-caprolactone) (PCL) with poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) following incorporation of either vascular endothelial growth factor (VEGF) or basic fibroblast growth factor (bFGF) and then fabricated electrospun 2 mm diameter vascular grafts. Grafts without the growth factors were used as a control group. Structure of the grafts was assessed utilizing scanning electron microscopy. We further implanted our grafts into rat abdominal aorta for 1 and 3 months with the aim to test endothelialization, cell infiltration, and patency in vivo. Histological and immunofluorescence examination demonstrated enhanced endothelialization and cell infiltration of the grafts with either VEGF or bFGF compared to those without the growth factors. Grafts with VEGF showed higher patency compared to those with bFGF; however, bFGF promoted migration of smooth muscle cells and fibroblasts into the graft. Therefore, we conclude that incorporation of VEGF and bFGF into the inner and medial/outer layer, respectively, can be a promising option for the fabrication of tissue-engineered vascular grafts.

UV radiation induces CXCL5 expression in human skin.

PubMed

Reichert, Olga; Kolbe, Ludger; Terstegen, Lara; Staeb, Franz; Wenck, Horst; Schmelz, Martin; Genth, Harald; Kaever, Volkhard; Roggenkamp, Dennis; Neufang, Gitta

2015-04-01

CXCL5 has recently been identified as a mediator of UVB-induced pain in rodents. To compare and to extend previous knowledge of cutaneous CXCL5 regulation, we performed a comprehensive study on the effects of UV radiation on CXCL5 regulation in human skin. Our results show a dose-dependent increase in CXCL5 protein in human skin after UV radiation. CXCL5 can be released by different cell types in the skin. We presumed that, in addition to immune cells, non-immune skin cells also contribute to UV-induced increase in CXCL5 protein. Analysis of monocultured dermal fibroblasts and keratinocytes revealed that only fibroblasts but not keratinocytes displayed up regulated CXCL5 levels after UV stimulation. Whereas UV treatment of human skin equivalents, induced epidermal CXCL5 mRNA and protein expression. Up regulation of epidermal CXCL5 was independent of keratinocyte differentiation and keratinocyte-keratinocyte interactions in epidermal layers. Our findings provide first evidence on the release of CXCL5 in UV-radiated human skin and the essential role of fibroblast-keratinocyte interaction in the regulation of epidermal CXCL5. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Pentagalloyl glucose increases elastin deposition, decreases reactive oxygen species and matrix metalloproteinase activity in pulmonary fibroblasts under inflammatory conditions.

PubMed

Parasaram, Vaideesh; Nosoudi, Nasim; Chowdhury, Aniqa; Vyavahare, Naren

2018-04-30

Emphysema is characterized by degradation of lung alveoli that leads to poor airflow in lungs. Irreversible elastic fiber degradation by matrix metalloproteinases (MMPs) and reactive oxygen species (ROS) activity leads to loss of elasticity and drives the progression of this disease. We investigated if a polyphenol, pentagalloyl glucose (PGG) can increase elastin production in pulmonary fibroblasts. We also studied the effect of PGG treatment in reducing MMP activity and ROS levels in cells. We exposed rat pulmonary fibroblasts to two different types of inflammatory environments i.e., tumor necrosis factor-α (TNF-α) and cigarette smoke extract (CSE) to mimic the disease. Parameters like lysyl oxidase (LOX) and elastin gene expression, MMP-9 activity in the medium, lysyl oxidase (LOX) activity and ROS levels were studied to assess the effect of PGG on pulmonary fibroblasts. CSE inhibited lysyl oxidase (LOX) enzyme activity that resulted in a decreased elastin formation. Similarly, TNF-α treated cells showed less elastin in the cell layers. Both these agents caused increase in MMP activity and ROS levels in cells. However, when supplemented with PGG treatment along with these two inflammatory agents, we saw a significant increase in elastin deposition, reduction in both MMP activity and ROS levels. Thus PGG, which has anti-inflammatory, anti-oxidant properties coupled with its ability to aid in elastic fiber formation, can be a multifunctional drug to potentially arrest the progression of emphysema. Copyright © 2018 Elsevier Inc. All rights reserved.

Polycistronic lentiviral vector for "hit and run" reprogramming of adult skin fibroblasts to induced pluripotent stem cells.

PubMed

Chang, Chia-Wei; Lai, Yi-Shin; Pawlik, Kevin M; Liu, Kaimao; Sun, Chiao-Wang; Li, Chao; Schoeb, Trenton R; Townes, Tim M

2009-05-01

We report the derivation of induced pluripotent stem (iPS) cells from adult skin fibroblasts using a single, polycistronic lentiviral vector encoding the reprogramming factors Oct4, Sox2, and Klf4. Porcine teschovirus-1 2A sequences that trigger ribosome skipping were inserted between human cDNAs for these factors, and the polycistron was subcloned downstream of the elongation factor 1 alpha promoter in a self-inactivating (SIN) lentiviral vector containing a loxP site in the truncated 3' long terminal repeat (LTR). Adult skin fibroblasts from a humanized mouse model of sickle cell disease were transduced with this single lentiviral vector, and iPS cell colonies were picked within 30 days. These cells expressed endogenous Oct4, Sox2, Nanog, alkaline phosphatase, stage-specific embryonic antigen-1, and other markers of pluripotency. The iPS cells produced teratomas containing tissue derived from all three germ layers after injection into immunocompromised mice and formed high-level chimeras after injection into murine blastocysts. iPS cell lines with as few as three lentiviral insertions were obtained. Expression of Cre recombinase in these iPS cells resulted in deletion of the lentiviral vector, and sequencing of insertion sites demonstrated that remnant 291-bp SIN LTRs containing a single loxP site did not interrupt coding sequences, promoters, or known regulatory elements. These results suggest that a single, polycistronic "hit and run" vector can safely and effectively reprogram adult dermal fibroblasts into iPS cells.

Effect of the Heat-Treated Ti6Al4V Alloy on the Fibroblastic Cell Response

PubMed Central

Chávez-Díaz, Mercedes Paulina; Escudero-Rincón, María Lorenza; Arce-Estrada, Elsa Miriam; Cabrera-Sierra, Román

2017-01-01

Two heat treatments were carried out below (Ti6Al4V800) and above (Ti6Al4V1050) Ti6Al4V beta-phase transformation temperature (980 °C), with the purpose of studying the effect of microstructure on the adhesion and proliferation of fibroblast cells, as well as their electrochemical behavior. These alloys were seeded with 10,000 L929 fibroblast cells and immersed for 7 days in the cell culture at 37 °C, pH 7.40, 5% CO2 and 100% relative humidity. Cell adhesion was characterized by Scanning Electron Microscopy (SEM) and Electrochemical Impedance Spectroscopy (EIS) techniques. Polygonal and elongated cell morphology was observed independent of Ti6Al4V microstructure. Besides, C, O, P, S, Na and Cl signals were detected by Energy Dispersive X-Ray Spectroscopy (EDX), associated with the synthesis of organic compounds excreted by the cells, including protein adsorption from the medium. In certain areas on Ti6Al4V and Ti6Al4V800 alloys, cells were agglomerated (island type), likely related to the globular microstructure; meanwhile, larger cellular coverage is shown for Ti6Al4V1050 alloy, forming more than one layer on the surface, where only Ca was recorded. Impedance diagrams showed a similar passive behavior for the different Ti6Al4V alloys, mainly due to TiO2 overlaying the contribution of the organic compounds excreted by fibroblast cells. PMID:29301205

78 FR 79338 - Airworthiness Directives; Bombardier, Inc. Airplanes

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-30

...We propose to adopt a new airworthiness directive (AD) for certain Bombardier, Inc. Model DHC-8-400 series airplanes. This proposed AD was prompted by reports of missing clamps that are required to provide positive separation between the alternating current (AC) feeder cables and the hydraulic line of the landing gear alternate extension. This proposed AD would require inspecting for missing clamps, and related investigative and corrective actions if necessary. We are proposing this AD to detect and correct chafing of the AC feeder cable. A chafed and arcing AC feeder cable could puncture the adjacent hydraulic line, which, in combination with the use of the alternate extension system, could result in an in-flight fire.

Technologies to Increase PV Hosting Capacity in Distribution Feeders: Preprint

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

Ding, Fei; Mather, Barry; Gotseff, Peter

This paper studies the distributed photovoltaic (PV) hosting capacity in distribution feeders by using the stochastic analysis approach. Multiple scenario simulations are conducted to analyze several factors that affect PV hosting capacity, including the existence of voltage regulator, PV location, the power factor of PV inverter and Volt/VAR control. Based on the conclusions obtained from simulation results, three approaches are then proposed to increase distributed PV hosting capacity, which can be formulated as the optimization problem to obtain the optimal solution. All technologies investigated in this paper utilize only existing assets in the feeder and therefore are implementable for amore » low cost. Additionally, the tool developed for these studies is described.« less

Technologies to Increase PV Hosting Capacity in Distribution Feeders

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

Ding, Fei; Mather, Barry; Gotseff, Peter

This paper studies the distributed photovoltaic (PV) hosting capacity in distribution feeders by using the stochastic analysis approach. Multiple scenario simulations are conducted to analyze several factors that affect PV hosting capacity, including the existence of voltage regulator, PV location, the power factor of PV inverter and Volt/VAR control. Based on the conclusions obtained from simulation results, three approaches are then proposed to increase distributed PV hosting capacity, which can be formulated as the optimization problem to obtain the optimal solution. All technologies investigated in this paper utilize only existing assets in the feeder and therefore are implementable for amore » low cost. Additionally, the tool developed for these studies is described.« less

Petrological constraints on the recycling of mafic crystal mushes, magma ascent and intrusion of braided sills in the Torres del Paine mafic complex (Patagonia)

NASA Astrophysics Data System (ADS)

Leuthold, Julien; Müntener, Othmar; Baumgartner, Lukas; Putlitz, Benita

2014-05-01

Cumulate and crystal mush disruption and reactivation are difficult to recognise in coarse grained shallow plutonic rocks. Mafic minerals included in hornblende and zoned plagioclase provide snapshots of early crystallization and cumulate formation, but are difficult to interpret in terms of the dynamics of magma ascent and possible links between silicic and mafic rock emplacement. We will present the field relations, the microtextures and the mineral chemistry of the Miocene mafic sill complex of the Torres del Paine intrusive complex (Patagonia, Chile) and its sub-vertical feeder-zone. The mafic sill complex was built up by a succession of braided sills of shoshonitic and high-K calc-alkaline porphyritic hornblende-gabbro and fine grained monzodioritic sills. The mafic units were over-accreted over 41±11 ka, underplating the overlying granite. Local diapiric structures and felsic magma accumulation between sills indicate limited separation of intercumulus liquid from the mafic sills. Anhedral hornblende cores, with olivine + clinopyroxene ± plagioclase ± apatite inclusions, crystallized at temperatures >900°C and pressures of ~300 to ~500 MPa. The corresponding rims and monzodiorite matrix crystallized at <830°C, ~70 MPa. This abrupt compositional variation suggests stability and instability of hornblende during mafic roots recycling and subsequent decompression. The near lack of intercumulus crystals in the sub-vertical feeder zone layered gabbronorite and pyroxene-hornblende gabbronorite stocks testifies that melt is more efficiently extracted than in sills, resulting in a cumulate signature in the feeding system. The emplacement age of the sill complex topmost granitic unit is identical, within uncertainties, to the feeder zone mafic cumulates. Granitic liquids formed by AFC processes and were extracted at high temperature (T>950°C) from the middle crust reservoir to the emplacement level. We show that hornblende-plagioclase thermobarometry is a useful monitor for the determination of segregation conditions of granitic magmas from gabbroic crystal mushes, and for monitoring the evolution of shallow crustal magmatic crystallization, decompression and cooling.

Distribution of free gas and 3D mirror image structures beneath Sevastopol mud volcano, Black sea, from 3D high resolution wide-angle seismic data

NASA Astrophysics Data System (ADS)

Krabbenhoeft, A.; Papenberg, C. A.; Klaeschen, D.; Bialas, J.

2016-12-01

The goal of this study is to image the sub-seafloor structure beneath the Sevastopol mud volcano (SMV), Sorokin Trough, SE of the Crimean peninsula, Black Sea. The focus lies on structures of/within the feeder channel, the distribution of gas and gas hydrates, and their relation to fluid migration zones in sediments. This study concentrates on a 3D high resolution seismic grid (7 km x 2.5 km) recorded with 13 ocean bottom stations (OBS). The 3D nature of the experiment results from the geometry of 68 densely spaced (25/50 m) profiles, as well as the cubical configuration of the densely spaced receivers on the seafloor ( 300 m station spacing). The seismic profiles are typically longer than 6 km which results in large offsets for the reflections of the OBS. This enables the study of the seismic velocities of the sub-seafloor sediments and additionally large offset incident analysis.The 3D Kirchhoff mirror image time migration, applied to all OBS sections including all shots from all profiles, leads to a spatial image of the sub-seafloor. Here, the migration was applied with the velocity distribution of 1.49 km/s in the water column, 1.5 km/s below the seafloor (bsf) increasing to 2 km/s for the deeper sediments at 2 s bsf. Acoustic blanking occurs beneath the south-easterly located OBS and is associated with the feeder channel of the mud volcano. There, gas from depth can vertically migrate to the seafloor and on its way to the surface horizontally distribute patchily within sediment layers. High amplitude reflections are not observed as continuous reflections, but in a patchy distribution. They are associated with accumulations of gas. Also structures exist within the feeder channel of the SMV.3D mirror imaging proves to be a good tool to seismically image structures compared with 2D streamer seismics, especially steep dipping reflectors and structures which are otherwise obscured by signal scattering, i.e structures associated with fluid migration paths.

Splashing, feeding, contracting: Drop impact and fluid dynamics of Vorticella

NASA Astrophysics Data System (ADS)

Pepper, Rachel E.

This thesis comprises two main topics: understanding drop impact and splashing, and studying the feeding and contracting of the microorganism Vorticella. In Chapter 1, we study the effect of substrate compliance on the splash threshold of a liquid drop using an elastic membrane under variable tension. We find that splashing can be suppressed by reducing this tension. Measurements of the velocity and acceleration of the spreading drop after impact indicate that the splashing behavior is set at very early times after, or possibly just before, impact, far before the actual splash occurs. We also provide a model for the tension dependence of the splashing threshold. In Chapter 2, we study the evolution of the ejected liquid sheet, or lamella, created after impact of a liquid drop onto a solid surface using high-speed video. We find that the lamella rim thickness is always much larger than the boundary layer thickness, and that this thickness decreases with increasing impact speed. We also observe an unusual plateau behavior in thickness versus time at higher impact speeds as we approach the splash threshold. In Chapter 3, we show through calculations, simulations, and experiments that the eddies often observed near sessile filter feeders are due to the presence of nearby boundaries. We model the common filter feeder Vorticella, and also track particles around live feeding Vorticella to determine the experimental flow field. Our models are in good agreement both with each other and with the experiments. We also provide simple approximate equations to predict experimental eddy sizes due to boundaries. In Chapter 4, we show through calculations that filter feeders such as Vorticella can greatly enhance their nutrient uptake by feeding at an angle rather than perpendicular to a substrate. We also show experimental evidence that living Vorticella use this strategy. Finally, in Chapter 5, we discuss possible future directions for these projects, including potential insights from a close examination of lamella behavior at the splash threshold, and calculations to determine if Vorticella contract rapidly towards the substrate to which they are attached in order to mix the surrounding fluid.

Long-term culture of chicken primordial germ cells isolated from embryonic blood and production of germline chimaeric chickens.

PubMed

Naito, Mitsuru; Harumi, Takashi; Kuwana, Takashi

2015-02-01

Production of germline chimaeric chickens by the transfer of cultured primordial germ cells (PGC) is a useful system for germline manipulation. A novel culture system was developed for chicken PGC isolated from embryonic blood. The isolated PGC were cultured on feeder cells derived from chicken embryonic fibroblast. The cultured PGC formed colonies and they proliferated about 300-times during the first 30 days. The cultured PGC retained the ability to migrate to recipient gonads and were also chicken VASA homologue (CVH)-positive. Female PGC were present in the mixed-sex PGC populations cultured for more than 90 days and gave rise to viable offspring efficiently via germline chimaeric chickens. Male cultured PGC were transferred to recipient embryos and produced putative chimaeric chickens. The DNA derived from the cultured PGC was detected in the sperm samples of male putative chimaeric chickens, but no donor derived offspring were obtained. Donor-derived offspring were also obtained from germline chimaeric chickens by the transfer of frozen-thawed cultured PGC. The culture method for PGC developed in the present study is useful for manipulation of the germline in chickens, such as preservation of genetic resources and gene transfer. Copyright © 2014 Elsevier B.V. All rights reserved.

Conserved Role of bFGF and a Divergent Role of LIF for Pluripotency Maintenance and Survival in Canine Pluripotent Stem Cells.

PubMed

Luo, Jiesi; Cibelli, Jose B

2016-09-19

Dogs have been widely used as a preclinical model for human disease. With the successful generation of canine induced pluripotent stem cells (ciPSCs), the biomedical community has a unique opportunity to study therapeutic interventions using autologous stem cells that can benefit dogs and humans. Unlike mice and human pluripotent cells, which are leukemia inhibitory factor (LIF)- and basic fibroblast growth factor (bFGF)-dependent, respectively, dog iPSCs require both growth factors simultaneously. In an effort to elucidate the role of each factor in the control of ciPSC self-renewal, we performed a series of experiments aiming at understanding the signaling pathways activated by them. We found that bFGF regulates pluripotency by indirectly activating the SMAD2/3 pathway in the presence of feeder cells, exclusively targeting NANOG expression, and inhibiting spontaneous differentiation toward ectoderm and mesoderm. LIF activates the JAK-STAT3 pathway but does not function in the typical manner described in mouse naïve embryonic stem cells. These results show that a unique mechanism for maintenance of pluripotency is present in ciPSC. These findings should be taken into account when establishing stem cell differentiation protocols and may provide more insight into pluripotency regulation in species other than mice and humans.

46 CFR 58.25-55 - Overcurrent protection for steering-gear systems.

Code of Federal Regulations, 2010 CFR

2010-10-01

... overcurrent protection except short-circuit protection that is instantaneous and rated at 400% to 500% of— (1... for steering-gear systems. (a) Each feeder circuit for steering must be protected by a circuit breaker... motor for an alternating-current motor. (b) No feeder circuit for steering may have any overcurrent...

26 CFR 1.1402(a)-14 - Options available to farmers in computing net earnings from self-employment for taxable years...

Code of Federal Regulations, 2013 CFR

2013-04-01

... held for breeding or dairy purposes, and $600 from the sale of a tractor. The income from the sale of... the sale of feeder cattle, which C bought for $500. The income from the sale of the feeder cattle is...

26 CFR 1.1402(a)-14 - Options available to farmers in computing net earnings from self-employment for taxable years...

Code of Federal Regulations, 2011 CFR

2011-04-01

... held for breeding or dairy purposes, and $600 from the sale of a tractor. The income from the sale of... the sale of feeder cattle, which C bought for $500. The income from the sale of the feeder cattle is...

26 CFR 1.1402(a)-14 - Options available to farmers in computing net earnings from self-employment for taxable years...

Code of Federal Regulations, 2014 CFR

2014-04-01

... held for breeding or dairy purposes, and $600 from the sale of a tractor. The income from the sale of... the sale of feeder cattle, which C bought for $500. The income from the sale of the feeder cattle is...

26 CFR 1.1402(a)-14 - Options available to farmers in computing net earnings from self-employment for taxable years...

Code of Federal Regulations, 2012 CFR

2012-04-01

... held for breeding or dairy purposes, and $600 from the sale of a tractor. The income from the sale of... the sale of feeder cattle, which C bought for $500. The income from the sale of the feeder cattle is...

View southeast, stone sluice, top of upper standing section, showing ...

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

View southeast, stone sluice, top of upper standing section, showing cement piers over arch in foreground, foot of break at third drop in center, retaining wall to left, barge canal sluice to right - Glens Falls Feeder, Sluice, Along south side of Glens Falls Feeder between locks 10 & 20, Hudson Falls, Washington County, NY

77 FR 70355 - Airworthiness Directives; The Boeing Company Airplanes

Federal Register 2010, 2011, 2012, 2013, 2014

2012-11-26

... leakage zone) or heat damage to the APU power feeder cable, insulation blankets, or pressure bulkhead...) of the NPRM requires repair of the APU power feeder, insulation blankets, and clamps, if no primer... bulletin, which states, ``If visual indications of heat damage are found, do steps 6.c through 6.f...

30 CFR 77.1800 - Cutout switches.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 30 Mineral Resources 1 2013-07-01 2013-07-01 false Cutout switches. 77.1800 Section 77.1800... Wires and Trolley Feeder Wires § 77.1800 Cutout switches. Trolley wires and trolley feeder wires shall be provided with cutout switches at intervals of not more than 2,000 feet and near the beginning of...

30 CFR 77.1800 - Cutout switches.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 30 Mineral Resources 1 2012-07-01 2012-07-01 false Cutout switches. 77.1800 Section 77.1800... Wires and Trolley Feeder Wires § 77.1800 Cutout switches. Trolley wires and trolley feeder wires shall be provided with cutout switches at intervals of not more than 2,000 feet and near the beginning of...

30 CFR 77.1800 - Cutout switches.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Cutout switches. 77.1800 Section 77.1800... Wires and Trolley Feeder Wires § 77.1800 Cutout switches. Trolley wires and trolley feeder wires shall be provided with cutout switches at intervals of not more than 2,000 feet and near the beginning of...

30 CFR 77.1800 - Cutout switches.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 30 Mineral Resources 1 2014-07-01 2014-07-01 false Cutout switches. 77.1800 Section 77.1800... Wires and Trolley Feeder Wires § 77.1800 Cutout switches. Trolley wires and trolley feeder wires shall be provided with cutout switches at intervals of not more than 2,000 feet and near the beginning of...

46 CFR 58.25-65 - Feeder circuits.

Code of Federal Regulations, 2013 CFR

2013-10-01

... seconds of loss of power from the vessel's service switchboard; (2) Comes from an independent source of power in the steering-gear compartment; (3) Is used for no other purpose; and (4) Has a capacity for one... feeder circuit must have a current-carrying capacity of— (1) 125% of the rated full-load current rating...

46 CFR 58.25-65 - Feeder circuits.

Code of Federal Regulations, 2011 CFR

2011-10-01

... seconds of loss of power from the vessel's service switchboard; (2) Comes from an independent source of power in the steering-gear compartment; (3) Is used for no other purpose; and (4) Has a capacity for one... feeder circuit must have a current-carrying capacity of— (1) 125% of the rated full-load current rating...

46 CFR 58.25-65 - Feeder circuits.

Code of Federal Regulations, 2014 CFR

2014-10-01

... seconds of loss of power from the vessel's service switchboard; (2) Comes from an independent source of power in the steering-gear compartment; (3) Is used for no other purpose; and (4) Has a capacity for one... feeder circuit must have a current-carrying capacity of— (1) 125% of the rated full-load current rating...

46 CFR 58.25-65 - Feeder circuits.

Code of Federal Regulations, 2012 CFR

2012-10-01

... seconds of loss of power from the vessel's service switchboard; (2) Comes from an independent source of power in the steering-gear compartment; (3) Is used for no other purpose; and (4) Has a capacity for one... feeder circuit must have a current-carrying capacity of— (1) 125% of the rated full-load current rating...

46 CFR 58.25-65 - Feeder circuits.

Code of Federal Regulations, 2010 CFR

2010-10-01

... seconds of loss of power from the vessel's service switchboard; (2) Comes from an independent source of power in the steering-gear compartment; (3) Is used for no other purpose; and (4) Has a capacity for one... feeder circuit must have a current-carrying capacity of— (1) 125% of the rated full-load current rating...

78 FR 35314 - Availability of Final Environmental Impact Statement; Bunker Hill Groundwater Basin, Riverside...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-12

... DEPARTMENT OF THE INTERIOR Bureau of Reclamation [A10-1999-6000-100-00-0-0-3, 3501000] Availability of Final Environmental Impact Statement; Bunker Hill Groundwater Basin, Riverside-Corona Feeder... proposed Riverside-Corona Feeder Project. DATES: The Bureau of Reclamation will not make a decision on the...

Counseling in the Elementary Feeder Schools.

ERIC Educational Resources Information Center

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.…

40 CFR Table 4 to Subpart Kkkkk of... - Requirements for Performance Tests

Code of Federal Regulations, 2010 CFR

2010-07-01

... block average pressure drop values for the three test runs, and determine and record the 3-hour block... limit for the limestone feeder setting Data from the limestone feeder during the performance test You must ensure that you maintain an adequate amount of limestone in the limestone hopper, storage bin...

Energy Systems Integration News | Energy Systems Integration Facility |

Science.gov Websites

distribution feeder models for use in hardware-in-the-loop (HIL) experiments. Using this method, a full feeder ; proposes an additional control loop to improve frequency support while ensuring stable operation. The and Frequency Deviation," also proposes an additional control loop, this time to smooth the wind

Exploring associations between international trade and environmental factors with establishment patterns of exotic Scolytinae

Treesearch

Lorenzo Marini; Robert A. Haack; Robert J. Rabaglia; Edoardo Petrucco Toffolo; Andrea Battisti; Massimo Faccoli

2011-01-01

Although invasion of exotic ambrosia beetles (fungus feeders) and bark beetles (phloem feeders) (Coleoptera: Curculionidae: Scolytinae) is considered a major threat to forest health worldwide, no studies have quantitatively investigated the anthropogenic and environmental factors shaping the biogeographical patterns of invasion by these insects across large spatial...

Blood feeding behavior of the stable fly

USDA-ARS?s Scientific Manuscript database

Stable fly is a fly that looks similar to a house fly but both sexes are blood feeders. Blood is required for successful fertilization and development of eggs. Bites are painful but there is usually no pain after the fly stops feeding. The stable fly is a persistent feeder and will continue trying t...

75 FR 78758 - Prohibited Transaction Exemptions From Certain Prohibited Transaction Restrictions

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-16

..., between the DB Torus Japan Master Portfolio (the Master Fund), in which the assets of a client employee...) Prior to investing in DB Torus Japan Fund Ltd. (hereinafter the Feeder Fund, the vehicle through which... DB Torus Japan Fund Ltd. (hereinafter the Feeder Fund, the vehicle through which investments in the...

Coal gasification system with a modulated on/off control system

DOEpatents

Fasching, George E.

1984-01-01

A modulated control system is provided for improving regulation of the bed level in a fixed-bed coal gasifier into which coal is fed from a rotary coal feeder. A nuclear bed level gauge using a cobalt source and an ion chamber detector is used to detect the coal bed level in the gasifier. The detector signal is compared to a bed level set point signal in a primary controller which operates in proportional/integral modes to produce an error signal. The error signal is modulated by the injection of a triangular wave signal of a frequency of about 0.0004 Hz and an amplitude of about 80% of the primary deadband. The modulated error signal is fed to a triple-deadband secondary controller which jogs the coal feeder speed up or down by on/off control of a feeder speed change driver such that the gasifier bed level is driven toward the set point while preventing excessive cycling (oscillation) common in on/off mode automatic controllers of this type. Regulation of the bed level is achieved without excessive feeder speed control jogging.

Improvement of automatic fish feeder machine design

NASA Astrophysics Data System (ADS)

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.

Mathematical analysis of the honeybee waggle dance.

PubMed

Okada, R; Ikeno, H; Kimura, T; Ohashi, Mizue; Aonuma, H; Ito, E

2012-01-01

A honeybee informs her nestmates of the location of a flower by doing a waggle dance. The waggle dance encodes both the direction of and distance to the flower from the hive. To reveal how the waggle dance benefits the colony, we created a Markov model of bee foraging behavior and performed simulation experiments by incorporating the biological parameters that we obtained from our own observations of real bees as well as from the literature. When two feeders were each placed 400 m away from the hive in different directions, a virtual colony in which honeybees danced and correctly transferred information (a normal, real bee colony) made significantly greater numbers of successful visits to the feeders compared to a colony with inaccurate information transfer. Howerer, when five feeders were each located 400 m from the hive, the inaccurate information transfer colony performed better than the normal colony. These results suggest that dancing's ability to communicate accurate information depends on the number of feeders. Furthermore, because non-dancing colonies always made significantly fewer visits than those two colonies, we concluded that dancing behavior is beneficial for hives' ability to visit food sources.

Preliminary analysis of hub and spoke air freight distribution system

NASA Technical Reports Server (NTRS)

Whitehead, A. H., Jr.

1978-01-01

A brief analysis is made of the hub and spoke air freight distribution system which would employ less than 15 hub centers world wide with very large advanced distributed-load freighters providing the line-haul delivery between hubs. This system is compared to a more conventional network using conventionally-designed long-haul freighters which travel between numerous major airports. The analysis calculates all of the transportation costs, including handling charges and pickup and delivery costs. The results show that the economics of the hub/spoke system are severely compromised by the extensive use of feeder aircraft to deliver cargo into and from the large freighter terminals. Not only are the higher costs for the smaller feeder airplanes disadvantageous, but their use implies an additional exchange of cargo between modes compared to truck delivery. The conventional system uses far fewer feeder airplanes, and in many cases, none at all. When feeder aircraft are eliminated from the hub/spoke system, however, that system is universally more economical than any conventional system employing smaller line-haul aircraft.

Alternatives to the 15% Rule

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

Broderick, Robert Joseph; Quiroz, Jimmy Edward; Reno, Matthew J.

2015-11-01

The third solicitation of the California Solar Initiative (CSI) Research, Development, Demonstration and Deployment (RD&D) Program established by the California Public Utility Commission (CPUC) is supporting the Electric Power Research Institute (EPRI), National Renewable Energy Laboratory (NREL), and Sandia National Laboratories (SNL) with collaboration from Pacific Gas and Electric (PG&E), Southern California Edison (SCE), and San Diego Gas and Electric (SDG&E), in research to improve the Utility Application Review and Approval process for interconnecting distributed energy resources to the distribution system. Currently this process is the most time - consuming of any step on the path to generating power onmore » the distribution system. This CSI RD&D solicitation three project has completed the tasks of collecting data from the three utilities, clustering feeder characteristic data to attain representative feeders, detailed modeling of 16 representative feeders, analysis of PV impacts to those feeders, refinement of current screening processes, and validation of those suggested refinements. In this report each task is summarized to produce a final summary of all components of the overall project.« less

Role of Nectin-1 and Herpesvirus Entry Mediator as Cellular Receptors for Herpes Simplex Virus 1 on Primary Murine Dermal Fibroblasts.

PubMed

Petermann, Philipp; Rahn, Elena; Thier, Katharina; Hsu, Mei-Ju; Rixon, Frazer J; Kopp, Sarah J; Knebel-Mörsdorf, Dagmar

2015-09-01

The cellular proteins nectin-1 and herpesvirus entry mediator (HVEM) can both mediate the entry of herpes simplex virus 1 (HSV-1). We have recently shown how these receptors contribute to infection of skin by investigating HSV-1 entry into murine epidermis. Ex vivo infection studies reveal nectin-1 as the primary receptor in epidermis, whereas HVEM has a more limited role. Although the epidermis represents the outermost layer of skin, the contribution of nectin-1 and HVEM in the underlying dermis is still open. Here, we analyzed the role of each receptor during HSV-1 entry in murine dermal fibroblasts that were deficient in expression of either nectin-1 or HVEM or both receptors. Because infection was not prevented by the absence of either nectin-1 or HVEM, we conclude that they can act as alternative receptors. Although HVEM was found to be highly expressed on fibroblasts, entry was delayed in nectin-1-deficient cells, suggesting that nectin-1 acts as the more efficient receptor. In the absence of both receptors, entry was strongly delayed leading to a much reduced viral spread and virus production. These results suggest an unidentified cellular component that acts as alternate but inefficient receptor for HSV-1 on dermal fibroblasts. Characterization of the cellular entry mechanism suggests that HSV-1 can enter dermal fibroblasts both by direct fusion with the plasma membrane and via endocytic vesicles and that this is not dependent on the presence or absence of nectin-1. Entry was also shown to require dynamin and cholesterol, suggesting comparable entry pathways in keratinocytes and dermal fibroblasts. Herpes simplex virus (HSV) is a human pathogen which infects its host via mucosal surfaces or abraded skin. To understand how HSV-1 overcomes the protective barrier of mucosa or skin and reaches its receptors in tissue, it is essential to know which receptors contribute to the entry into individual skin cells. Previously, we have explored the contribution of nectin-1 and herpesvirus entry mediator (HVEM) as receptors for HSV-1 entry into murine epidermis, where keratinocytes form the major cell type. Since the underlying dermis consists primarily of fibroblasts, we have now extended our study of HSV-1 entry to dermal fibroblasts isolated from nectin-1- or HVEM-deficient mice or from mice deficient in both receptors. Our results demonstrate a role for both nectin-1 and HVEM as receptors and suggest a further receptor which appears much less efficient. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Likeability of Garden Birds: Importance of Species Knowledge & Richness in Connecting People to Nature.

PubMed

Cox, Daniel T C; Gaston, Kevin J

2015-01-01

Interacting with nature is widely recognised as providing many health and well-being benefits. As people live increasingly urbanised lifestyles, the provision of food for garden birds may create a vital link for connecting people to nature and enabling them to access these benefits. However, it is not clear which factors determine the pleasure that people receive from watching birds at their feeders. These may be dependent on the species that are present, the abundance of individuals and the species richness of birds around the feeders. We quantitatively surveyed urban households from towns in southern England to determine the factors that influence the likeability of 14 common garden bird species, and to assess whether people prefer to see a greater abundance of individuals or increased species richness at their feeders. There was substantial variation in likeability across species, with songbirds being preferred over non-songbirds. Species likeability increased for people who fed birds regularly and who could name the species. We found a strong correlation between the number of species that a person could correctly identify and how connected to nature they felt when they watched garden birds. Species richness was preferred over a greater number of individuals of the same species. Although we do not show causation this study suggests that it is possible to increase the well-being benefits that people gain from watching birds at their feeders. This could be done first through a human to bird approach by encouraging regular interactions between people and their garden birds, such as through learning the species names and providing food. Second, it could be achieved through a bird to human approach by increasing garden songbird diversity because the pleasure that a person receives from watching an individual bird at a feeder is dependent not only on its species but also on the diversity of birds at the feeder.

Likeability of Garden Birds: Importance of Species Knowledge & Richness in Connecting People to Nature

PubMed Central

Cox, Daniel T. C.; Gaston, Kevin J.

2015-01-01

Interacting with nature is widely recognised as providing many health and well-being benefits. As people live increasingly urbanised lifestyles, the provision of food for garden birds may create a vital link for connecting people to nature and enabling them to access these benefits. However, it is not clear which factors determine the pleasure that people receive from watching birds at their feeders. These may be dependent on the species that are present, the abundance of individuals and the species richness of birds around the feeders. We quantitatively surveyed urban households from towns in southern England to determine the factors that influence the likeability of 14 common garden bird species, and to assess whether people prefer to see a greater abundance of individuals or increased species richness at their feeders. There was substantial variation in likeability across species, with songbirds being preferred over non-songbirds. Species likeability increased for people who fed birds regularly and who could name the species. We found a strong correlation between the number of species that a person could correctly identify and how connected to nature they felt when they watched garden birds. Species richness was preferred over a greater number of individuals of the same species. Although we do not show causation this study suggests that it is possible to increase the well-being benefits that people gain from watching birds at their feeders. This could be done first through a human to bird approach by encouraging regular interactions between people and their garden birds, such as through learning the species names and providing food. Second, it could be achieved through a bird to human approach by increasing garden songbird diversity because the pleasure that a person receives from watching an individual bird at a feeder is dependent not only on its species but also on the diversity of birds at the feeder. PMID:26560968

Antituberculosis nanodelivery system with controlled-release properties based on para-amino salicylate–zinc aluminum-layered double-hydroxide nanocomposites

PubMed Central

Saifullah, Bullo; Hussein, Mohd Zobir; Hussein-Al-Ali, Samer Hasan; Arulselvan, Palanisamy; Fakurazi, Sharida

2013-01-01

We report the intercalation and characterization of para-amino salicylic acid (PASA) into zinc/aluminum-layered double hydroxides (ZLDHs) by two methods, direct and indirect, to form nanocomposites: PASA nanocomposite prepared by a direct method (PASA-D) and PASA nanocomposite prepared by an indirect method (PASA-I). Powder X-ray diffraction, Fourier-transform infrared spectroscopy, and thermogravimetric analysis revealed that the PASA drugs were accommodated within the ZLDH interlayers. The anions of the drug were accommodated as an alternate monolayer (along the long-axis orientation) between ZLDH interlayers. Drug loading was estimated to be 22.8% and 16.6% for PASA-D and PASA-I, respectively. The in vitro release properties of the drug were investigated in physiological simulated phosphate-buffered saline solution of pH 7.4 and 4.8. The release followed the pseudo-second-order model for both nanocomposites. Cell viability (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide [MTT] assays) was assessed against normal human lung fibroblast MRC-5 and 3T3 mouse fibroblast cells at 24, 48, and 72 hours. The results showed that the nanocomposite formulations did not possess any cytotoxicity, at least up to 72 hours. PMID:24255593

PCL-PGLA composite tubular scaffold preparation and biocompatibility investigation.

PubMed

Mo, X; Weber, H-J; Ramakrishna, S

2006-08-01

The objective of this paper was to fabricate a biodegradable tubular scaffold for small diameter (d<6 mm) blood vessel tissue engineering. The tube scaffold needed a porous wall for cell attachment, proliferation and tissue regeneration with its degradation. A novel method given in this paper was to coat a porous layer of poly (epsilon-caprolactone) (PCL) on the outside of a poly (glycolic-co-lactic acid) (PGLA with GA:LA=90:10) fiber braided tube to give a PCL-PGLA composite. The PGLA tube was fabricated using a braiding machine by inserting a Teflon tube with the desired diameter in center of the 20 spindles, which are the carriers of PGLA fibers. Changing the diameter of the Teflon tube can vary the inner diameter of a braided PGLA tube. Thermally induced phase separation method was used for PCL solution coating on the surface of the PGLA braided tube. Controlling the polymer concentration, non-solvent addition and quenching temperature generated the pore structures, with pore sizes ranging from 10-30 microm. The fibroblast cells were seeded on the tubular scaffold and cultured in vitro for the biocompatibility investigation. Histology results showed that the fibroblast cells proliferated on the interconnected pore of the PCL porous layer in 1 week.

LOADING REEL AND PERFECTO STRIP STOCK FEEDER FOR #84 WATERBURYFARREL ...

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

LOADING REEL AND PERFECTO STRIP STOCK FEEDER FOR #84 WATERBURY-FARREL (U.S. GOVERNMENT) PRESS. THIS CONTINUOUS-FEED, 2-DRAW, 100 TON PRESS IS ONE OF TWO IN THE U.S. UNDER CONTRACT WITH THE DEPARTMENT OF DEFENSE FOR PRODUCTION OF BULLET JACKETS AND CARTRIDGE CASINGS. - American Brass Foundry, 70 Sayre Street, Buffalo, Erie County, NY

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

USDA-ARS?s Scientific Manuscript database

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

Feeding the Elite: The Evolution of Elite Pathways from Star High Schools to Elite Universities

ERIC Educational Resources Information Center

LeTendre, Gerald K.; Gonzalez, Roger Geertz; Nomi, Takako

2006-01-01

During the last 50 years, private "feeder" schools in Japan came to dominate entry into elite colleges. Intense organizational competition shaped the organizational environment and changed the pathways available to social elites. Compared to Japan, elite private feeders in the US have failed to dominate pathways into elite colleges. In…

ADJUSTABLE OUTPUT RATE CHEMICAL FEEDING EQUIPMENT FOR SWIMMING POOLS. NATIONAL SANITATION FOUNDATION STANDARD NUMBER 19.

ERIC Educational Resources Information Center

National Sanitation Foundation, Ann Arbor, MI.

THE SCOPE OF THIS STANDARD COVERS ADJUSTABLE OUTPUT RATE CHEMICAL FEEDERS, WHETHER USED FOR SOLUTIONS, SLURRIES OR SOLIDS. IT ALSO INCLUDES AUXILIARY EQUIPMENT SUCH AS PUMPS, STRAINERS, TUBING CONNECTIONS, TANKS, INJECTION FITTINGS AND OTHER REQUIRED COMPONENTS. THE FEEDERS DESCRIBED ARE INTENDED TO BE DESIGNED AND USED SPECIFICALLY FOR CHEMICAL…

Norm-Optimal ILC Applied to a High-Speed Rack Feeder

NASA Astrophysics Data System (ADS)

Schindele, Dominik; Aschemann, Harald; Ritzke, Jöran

2010-09-01

Rack feeders as automated conveying systems for high bay rackings are of high practical importance. To shorten the transport times by using trajectories with increased kinematic values accompanying control measures for a reduction of the excited structural vibrations are necessary. In this contribution, the model-based design of a norm-optimal iterative learning control structure is presented. The rack feeder is modelled as an elastic multibody system. For the mathematical description of the bending deflections a Ritz ansatz is introduced. The tracking control design is performed separately for both axes using decentralised state space representations. Both the achievable performance and the resulting tracking accuracy of the proposed control concept are shown by measurement results from the experimental set-up.

Voltage Impacts of Utility-Scale Distributed Wind

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

Allen, A.

2014-09-01

Although most utility-scale wind turbines in the United States are added at the transmission level in large wind power plants, distributed wind power offers an alternative that could increase the overall wind power penetration without the need for additional transmission. This report examines the distribution feeder-level voltage issues that can arise when adding utility-scale wind turbines to the distribution system. Four of the Pacific Northwest National Laboratory taxonomy feeders were examined in detail to study the voltage issues associated with adding wind turbines at different distances from the sub-station. General rules relating feeder resistance up to the point of turbinemore » interconnection to the expected maximum voltage change levels were developed. Additional analysis examined line and transformer overvoltage conditions.« less

Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM).

PubMed

Xiong, Kai; Zhou, Yan; Hyttel, Poul; Bolund, Lars; Freude, Kristine Karla; Luo, Yonglun

2016-11-01

Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4 and C-MYC. We have validated that the reprogramming cassette is silenced in the SAM iPSC clones. Expression of pluripotency genes (OCT4, SOX2, LIN28A, NANOG, GDF3, SSEA4, and TRA-1-60), differentiation potential to all three germ layers, and normal karyotypes are validated. These SAM-iPSCs provide a novel, useful tool to investigate genetic regulation of stem cell proliferation and differentiation through CRISPR-mediated activation of endogenous genes. Copyright © 2016 Michael Boutros, German Cancer Research Center, Heidelberg, Germany. Published by Elsevier B.V. All rights reserved.

Considerations for an Earth Relay Satellite with RF and Optical Trunklines

NASA Technical Reports Server (NTRS)

Israel, David J.

2016-01-01

Support for user platforms through the use of optical links to geosynchronous relay spacecraft are expected to be part of the future space communications architecture. The European Data Relay Satellite System (EDRS) has its first node, EDRS-A, in orbit. The EDRS architecture includes space-to-space optical links with a Ka-Band feeder link or trunkline. NASA's Laser Communications Relay Demonstration (LCRD) mission, originally baselined to support a space-to-space optical link relayed with an optical trunkline, has added an Radio Frequency (RF) trunkline. The use of an RF trunkline avoids the outages suffered by an optical trunkline due to clouds, but an RF trunkline will be bandwidth limited. A space relay architecture with both RF and optical trunklines could relay critical realtime data, while also providing a high data volume capacity. This paper considers the relay user scenarios that could be supported, and the implications to the space relay system and operations. System trades such as the amount of onboard processing and storage required, the use of link layer switching vs. network layer routing, and the use of Delay/Disruption Tolerant Networking (DTN) are discussed.

Porous PLGA microspheres tailored for dual delivery of biomolecules via layer-by-layer assembly.

PubMed

Go, Dewi P; Palmer, Jason A; Mitchell, Geraldine M; Gras, Sally L; O'Connor, Andrea J

2015-05-01

Tissue engineering is a complex and dynamic process that requires varied biomolecular cues to promote optimal tissue growth. Consequently, the development of delivery systems capable of sequestering more than one biomolecule with controllable release profiles is a key step in the advancement of this field. This study develops multilayered polyelectrolyte films incorporating alpha-melanocyte stimulating hormone (α-MSH), an anti-inflammatory molecule, and basic fibroblast growth factor (bFGF). The layers were successfully formed on macroporous poly lactic-co-glycolic acid microspheres produced using a combined inkjet and thermally induced phase separation technique. Release profiles could be varied by altering layer properties including the number of layers and concentrations of layering molecules. α-MSH and bFGF were released in a sustained manner and the bioactivity of α-MSH was shown to be preserved using an activated macrophage cell assay in vitro. The system performance was also tested in vivo subcutaneously in rats. The multilayered microspheres reduced the inflammatory response induced by a carrageenan stimulus 6 weeks after implantation compared to the non-layered microspheres without the anti-inflammatory and growth factors, demonstrating the potential of such multilayered constructs for the controlled delivery of bioactive molecules. © 2014 Wiley Periodicals, Inc.

Morphologic differentiation of colon carcinoma cell lines HT-29 and HT-29KM in rotating-wall vessels

NASA Technical Reports Server (NTRS)

Goodwin, T. J.; Jessup, J. M.; Wolf, D. A.

1992-01-01

A new low shear stress microcarrier culture system has been developed at NASA's Johnson Space Center that permits three-dimensional tissue culture. Two established human colon adenocarcinoma cell lines, HT-29, an undifferentiated, and HT-29KM, a stable, moderately differentiated subline of HT-29, were grown in new tissue culture bioreactors called Rotating-Wall Vessels (RWVs). RWVs are used in conjunction with multicellular cocultivation to develop a unique in vitro tissue modeling system. Cells were cultivated on Cytodex-3 microcarrier beads, with and without mixed normal human colonic fibroblasts, which served as the mesenchymal layer. Culture of the tumor lines in the absence of fibroblasts produced spheroidlike growth and minimal differentiation. In contrast, when tumor lines were co-cultivated with normal colonic fibroblasts, initial growth was confined to the fibroblast population until the microcarriers were covered. The tumor cells then commenced proliferation at an accelerated rate, organizing themselves into three-dimensional tissue masses that achieved 1.0- to 1.5-cm diameters. The masses displayed glandular structures, apical and internal glandular microvilli, tight intercellular junctions, desmosomes, cellular polarity, sinusoid development, internalized mucin, and structural organization akin to normal colon crypt development. Differentiated samples were subjected to transmission and scanning electron microscopy and histologic analysis, revealing embryoniclike mesenchymal cells lining the areas around the growth matrices. Necrosis was minimal throughout the tissue masses. These data suggest that the RWV affords a new model for investigation and isolation of growth, regulatory, and structural processes within neoplastic and normal tissue.

Derivation and Characterization of Induced Pluripotent Stem Cells from Equine Fibroblasts

PubMed Central

Breton, Amandine; Sharma, Ruchi; Diaz, Andrea Catalina; Parham, Alea Gillian; Graham, Audrey; Neil, Claire; Whitelaw, Christopher Bruce; Milne, Elspeth

2013-01-01

Pluripotent stem cells offer unprecedented potential not only for human medicine but also for veterinary medicine, particularly in relation to the horse. Induced pluripotent stem cells (iPSCs) are particularly promising, as they are functionally similar to embryonic stem cells and can be generated in vitro in a patient-specific manner. In this study, we report the generation of equine iPSCs from skin fibroblasts obtained from a foal and reprogrammed using viral vectors coding for murine Oct4, Sox2, c-Myc, and Klf4 sequences. The reprogrammed cell lines were morphologically similar to iPSCs reported from other species and could be stably maintained over more than 30 passages. Immunostaining and polymerase chain reaction analyses revealed that these cell lines expressed an array of endogenous markers associated with pluripotency, including OCT4, SOX2, NANOG, REX1, LIN28, SSEA1, SSEA4, and TRA1-60. Furthermore, under the appropriate conditions, the equine iPSCs readily formed embryoid bodies and differentiated in vitro into cells expressing markers of ectoderm, mesoderm, and endoderm, and when injected into immunodeficient mice, gave raise to tumors containing differentiated derivatives of the 3 germ layers. Finally, we also reprogrammed fibroblasts from a 2-year-old horse. The reprogrammed cells were similar to iPSCs derived from neonatal fibroblasts in terms of morphology, expression of pluripotency markers, and differentiation ability. The generation of these novel cell lines constitutes an important step toward the understanding of pluripotency in the horse, and paves the way for iPSC technology to potentially become a powerful research and clinical tool in veterinary biomedicine. PMID:22897112

Pirfenidone Nanoparticles Improve Corneal Wound Healing and Prevent Scarring Following Alkali Burn

PubMed Central

Chowdhury, Sushovan; Guha, Rajdeep; Trivedi, Ruchit; Kompella, Uday B.; Konar, Aditya; Hazra, Sarbani

2013-01-01

Purpose To evaluate the effects of pirfenidone nanoparticles on corneal re-epithelialization and scarring, major clinical challenges after alkali burn. Methods Effect of pirfenidone on collagen I and α-smooth muscle actin (α-SMA) synthesis by TGFβ induced primary corneal fibroblast cells was evaluated by immunoblotting and immunocytochemistry. Pirfenidone loaded poly (lactide-co-glycolide) (PLGA) nanoparticles were prepared, characterized and their cellular entry was examined in primary corneal fibroblast cells by fluorescence microscopy. Alkali burn was induced in one eye of Sprague Dawley rats followed by daily topical treatment with free pirfenidone, pirfenidone nanoparticles or vehicle. Corneal re-epithelialization was assessed daily by flourescein dye test; absence of stained area indicated complete re-epithelialization and the time for complete re-epithelialization was determined. Corneal haze was assessed daily for 7 days under slit lamp microscope and graded using a standard method. After 7 days, collagen I deposition in the superficial layer of cornea was examined by immunohistochemistry. Results Pirfenidone prevented (P<0.05) increase in TGF β induced collagen I and α-SMA synthesis by corneal fibroblasts in a dose dependent manner. Pirfenidone could be loaded successfully within PLGA nanoparticles, which entered the corneal fibroblasts within 5 minutes. Pirfenidone nanoparticles but not free pirfenidone significantly (P<0.05) reduced collagen I level, corneal haze and the time for corneal re-epithelialization following alkali burn. Conclusion Pirfenidone decreases collagen synthesis and prevents myofibroblast formation. Pirfenidone nanoparticles improve corneal wound healing and prevent fibrosis. Pirfenidone nanoparticles are of potential value in treating corneal chemical burns and other corneal fibrotic diseases. PMID:23940587