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Sample records for fiber cells

  1. Coated metal fiber coalescing cell

    SciTech Connect

    Rutz, W.D.; Swain, R.J.

    1980-12-23

    A cell is described for coalescing oil droplets dispersed in a water emulsion including an elongated perforated tube core into which the emulsion is injected, layers of oleophilic plastic covered metal mat wound about the core through which the emulsion is forced to pass, the fibers of the metal mat being covered by oleophilic plastic such as vinyl, acrylic, polypropylene, polyethylene, polyvinyl chloride, the metal being in the form of layers of expanded metal or metal fibers, either aluminum or stainless steel. In manufacturing the cell a helix wound wire is formed around the cylindrical plastic coated metal to retain it in place and resist pressure drop of fluid flowing through the metal fibers. In addition, the preferred arrangement includes the use of an outer sleeve formed of a mat of fibrous material such as polyester fibers, acrylic fibers, modacrylic fibers and mixtures thereof.

  2. Cells on foam and fiber

    SciTech Connect

    Clyde, R.

    1995-11-01

    Cells growing on high area foam and when a screen is put around the foam, it is made heavier so it can be fluidized. When foam is rotated in a half full RBC, drops are formed and mass transfer of oxygen to drops in much faster. Most fungi and some mammalian cells need oxygen. Corrugated fibers with holes in the valleys also produce drops. White rot fungus needs oxygen and it degrades many chlorine compounds, azo dyes, and TNT. Old cardboard boxes are readily available and when buried in soil, oxygen is entrapped. In a lake, the boxes expose high area. Fibers have high surface area for immobilizing cells and when the fibers are rotated, fast reactions occur, converting one chemical to another. Sugar has been fermented to alcohol in 10-15 minutes. Ethanol has high octane and does not need lead. Old cars and trucks still use lead and high levels have been found in the drinking water of several large cities. Bacteria on fibers can remove lead in a few seconds. When an RBC of plain fiber discs is rotated and a light shone in the top the light hits a thin moving film to degrade chlorine compounds. Microbes and light remove sulfur from oil. Calcium magnesium acetate is a non corrosive road deicer. Salt on roads causes millions of dollars damage to bridges and cars. An inexpensive reactor has been made for organization studies of mammalian and plant cells. A magnet is near the bottom but not touching and oxygen is put on the top where there is no seal that can leak.

  3. Cell death regulates muscle fiber number.

    PubMed

    Sarkissian, Tatevik; Arya, Richa; Gyonjyan, Seda; Taylor, Barbara; White, Kristin

    2016-07-01

    Cell death can have both cell autonomous and non-autonomous roles in normal development. Previous studies have shown that the central cell death regulators grim and reaper are required for the developmentally important elimination of stem cells and neurons in the developing central nervous system (CNS). Here we show that cell death in the nervous system is also required for normal muscle development. In the absence of grim and reaper, there is an increase in the number of fibers in the ventral abdominal muscles in the Drosophila adult. This phenotype can be partially recapitulated by inhibition of cell death specifically in the CNS, indicating a non-autonomous role for neuronal death in limiting muscle fiber number. We also show that FGFs produced in the cell death defective nervous system are required for the increase in muscle fiber number. Cell death in the muscle lineage during pupal stages also plays a role in specifying fiber number. Our work suggests that FGFs from the CNS act as a survival signal for muscle founder cells. Thus, proper muscle fiber specification requires cell death in both the nervous system and in the developing muscle itself. PMID:27131625

  4. Glycoproteome of Elongating Cotton Fiber Cells*

    PubMed Central

    Kumar, Saravanan; Kumar, Krishan; Pandey, Pankaj; Rajamani, Vijayalakshmi; Padmalatha, Kethireddy Venkata; Dhandapani, Gurusamy; Kanakachari, Mogilicherla; Leelavathi, Sadhu; Kumar, Polumetla Ananda; Reddy, Vanga Siva

    2013-01-01

    Cotton ovule epidermal cell differentiation into long fibers primarily depends on wall-oriented processes such as loosening, elongation, remodeling, and maturation. Such processes are governed by cell wall bound structural proteins and interacting carbohydrate active enzymes. Glycosylation plays a major role in the structural, functional, and localization aspects of the cell wall and extracellular destined proteins. Elucidating the glycoproteome of fiber cells would reflect its wall composition as well as compartmental requirement, which must be system specific. Following complementary proteomic approaches, we have identified 334 unique proteins comprising structural and regulatory families. Glycopeptide-based enrichment followed by deglycosylation with PNGase F and A revealed 92 unique peptides containing 106 formerly N-linked glycosylated sites from 67 unique proteins. Our results showed that structural proteins like arabinogalactans and carbohydrate active enzymes were relatively more abundant and showed stage- and isoform-specific expression patterns in the differentiating fiber cell. Furthermore, our data also revealed the presence of heterogeneous and novel forms of structural and regulatory glycoproteins. Comparative analysis with other plant glycoproteomes highlighted the unique composition of the fiber glycoproteome. The present study provides the first insight into the identity, abundance, diversity, and composition of the glycoproteome within single celled cotton fibers. The elucidated composition also indirectly provides clues about unicellular compartmental requirements underlying single cell differentiation. PMID:24019148

  5. Hybrid solar cell on a carbon fiber.

    PubMed

    Grynko, Dmytro A; Fedoryak, Alexander N; Smertenko, Petro S; Dimitriev, Oleg P; Ogurtsov, Nikolay A; Pud, Alexander A

    2016-12-01

    In this work, a method to assemble nanoscale hybrid solar cells in the form of a brush of radially oriented CdS nanowire crystals around a single carbon fiber is demonstrated for the first time. A solar cell was assembled on a carbon fiber with a diameter of ~5-10 μm which served as a core electrode; inorganic CdS nanowire crystals and organic dye or polymer layers were successively deposited on the carbon fiber as active components resulting in a core-shell photovoltaic structure. Polymer, dye-sensitized, and inverted solar cells have been prepared and compared with their analogues made on the flat indium-tin oxide electrode. PMID:27216603

  6. Hybrid solar cell on a carbon fiber

    NASA Astrophysics Data System (ADS)

    Grynko, Dmytro A.; Fedoryak, Alexander N.; Smertenko, Petro S.; Dimitriev, Oleg P.; Ogurtsov, Nikolay A.; Pud, Alexander A.

    2016-05-01

    In this work, a method to assemble nanoscale hybrid solar cells in the form of a brush of radially oriented CdS nanowire crystals around a single carbon fiber is demonstrated for the first time. A solar cell was assembled on a carbon fiber with a diameter of ~5-10 μm which served as a core electrode; inorganic CdS nanowire crystals and organic dye or polymer layers were successively deposited on the carbon fiber as active components resulting in a core-shell photovoltaic structure. Polymer, dye-sensitized, and inverted solar cells have been prepared and compared with their analogues made on the flat indium-tin oxide electrode.

  7. Hollow-fiber H2/O2 fuel cell

    NASA Technical Reports Server (NTRS)

    Ingham, J. D.; Lawson, D. D.

    1977-01-01

    Dual-membrane hollow-fiber electrode increases reliability and lowers costs. Leakage of fuel or oxidizer through fiber does not result in failure; excess product water migrates into electrolyte where it is removed by evaporation or distillation; constant exposure of fiber to electrolyte eliminates problems of drying and consequent failure; reference electrode monitors current collectors and overall cell performance.

  8. Rotational streaming in fiber cells and its role in translocation.

    PubMed

    Worley, J F

    1968-10-01

    All visible protoplasmic streaming in sections of various plant stems was reversibly stopped by 2,4-dinitrophenol (DNP). Sections contained epidermal, cortical, and fiber cell types. Cells treated with DNP retained their semipermeability as evidenced by their plasmolysis in sucrose solutions. Washing out the DNP resulted in the rapid resumption of protoplasmic streaming in all 3 cell types. Both the rate of movement of sodium fluorescein and the shape of the advancing dye front were greatly altered by DNP treatment. Dye transport was decreased in the fibers and little affected in cortical cells. The results suggest that rotational streaming accelerates the translocation of soluble substances in fiber cells. PMID:16656950

  9. Hollow fiber clinostat for simulating microgravity in cell culture

    NASA Technical Reports Server (NTRS)

    Rhodes, Percy H. (Inventor); Miller, Teresa Y. (Inventor); Snyder, Robert S. (Inventor)

    1992-01-01

    A clinostat for simulating microgravity on cell systems carried in a fiber fixedly mounted in a rotatable culture vessel is disclosed. The clinostat is rotated horizontally along its longitudinal axis to simulate microgravity or vertically as a control response. Cells are injected into the fiber and the ends of the fiber are sealed and secured to spaced end pieces of a fiber holder assembly which consists of the end pieces, a hollow fiber, a culture vessel, and a tension spring with three alignment pins. The tension spring is positioned around the culture vessel with its ends abutting the end pieces for alignment of the spring. After the fiber is secured, the spring is decompressed to maintain tension on the fiber while it is being rotated. This assures that the fiber remains aligned along the axis of rotation. The fiber assembly is placed in the culture vessel and culture medium is added. The culture vessel is then inserted into the rotatable portion of the clinostat and subjected to rotate at selected rpms. The internal diameter of the hollow fiber determines the distance the cells are from the axis of rotation.

  10. Integrating perovskite solar cells into a flexible fiber.

    PubMed

    Qiu, Longbin; Deng, Jue; Lu, Xin; Yang, Zhibin; Peng, Huisheng

    2014-09-22

    Perovskite solar cells have triggered a rapid development of new photovoltaic devices because of high energy conversion efficiencies and their all-solid-state structures. To this end, they are particularly useful for various wearable and portable electronic devices. Perovskite solar cells with a flexible fiber structure were now prepared for the first time by continuously winding an aligned multiwalled carbon nanotube sheet electrode onto a fiber electrode; photoactive perovskite materials were incorporated in between them through a solution process. The fiber-shaped perovskite solar cell exhibits an energy conversion efficiency of 3.3%, which remained stable on bending. The perovskite solar cell fibers may be woven into electronic textiles for large-scale application by well-developed textile technologies. PMID:25047870

  11. Fibronectin Fiber Extension Decreases Cell Spreading and Migration.

    PubMed

    Hubbard, Brant; Buczek-Thomas, Jo Ann; Nugent, Matthew A; Smith, Michael L

    2016-08-01

    The extracellular matrix (ECM) is present in a range of molecular conformations and intermolecular arrangements. Fibronectin (Fn) molecules that constitute fibers within the ECM can exist in a variety of conformations that result from both mechanical stress and chemical factors such as allosteric binding partners. The long-standing hypothesis that conformational changes regulate the binding of cells to Fn fibers has only been tested for mutated molecules of Fn and has yet to be fully evaluated with Fn fibers. Using time-lapse microscopy we examined how mechanical extension of single fibers of Fn affects the adhesion and migration of endothelial cells. Using this single fiber adhesion technique, we show that high levels of mechanical strain applied to Fn fibers decreases the rates of both cell spreading and cell migration. These data indicate a fundamental cellular response to mechanical strain in the ECM that might have important implications for understanding how cells are recruited during tissue development and repair. J. Cell. Physiol. 231: 1728-1736, 2016. © 2015 Wiley Periodicals, Inc. PMID:26621030

  12. De-esterified Pectins in the Cell Walls of Cotton Fiber: A Study of Fiber Mutants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the wild-type cotton (DP 5690), the cell walls of elongating cotton fibers are bilayered, with the outer layer enriched in de-esterified homogalacturonan (HGA), and an inner layer enriched in xyloglucans and cellulose. This bilayer is conspicuously absent in the cell walls of the ovule epidermal...

  13. Dynamics of Cancer Cell near Collagen Fiber Chain

    NASA Astrophysics Data System (ADS)

    Kim, Jihan; Sun, Bo

    Cell migration is an integrated process that is important in life. Migration is essential for embryonic development as well as homeostatic processes such as wound healing and immune responses. When cell migrates through connective extracellular matrix (ECM), it applies cellular traction force to ECM and senses the rigidity of their local environment. We used human breast cancer cell (MDA-MB-231) which is highly invasive and applies strong traction force to ECM. As cancer cell applies traction force to type I collage-based ECM, it deforms collagen fibers near the surface. Patterns of deforming collagen fibers are significantly different with pairs of cancer cells compared to a single cancer cell. While a pair of cancer cells within 60 um creates aligned collagen fiber chains between them permanently, a single cancer cell does not form any fiber chains. In this experiment we measured a cellular response and an interaction between a pair of cells through the chain. Finally, we analyzed correlation of directions between cancer cell migration and the collagen chain alignment.

  14. Regulation of auxin on secondary cell wall cellulose biosynthesis in developing cotton fibers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cotton (Gossypium hirsutum L.) fibers are unicellular trichomes that differentiate from epidermal cells of developing cotton ovules. Mature fibers exhibit thickened secondary walls composed of nearly pure cellulose. Cotton fiber development is divided into four overlapping phases, 1) initiation sta...

  15. Biomimetic spinning of silk fibers and in situ cell encapsulation.

    PubMed

    Cheng, Jie; Park, DoYeun; Jun, Yesl; Lee, JaeSeo; Hyun, Jinho; Lee, Sang-Hoon

    2016-07-01

    In situ embedding of sensitive materials (e.g., cells and proteins) in silk fibers without damage presents a significant challenge due to the lack of mild and efficient methods. Here, we report the development of a microfluidic chip-based method for preparation of meter-long silk fibroin (SF) hydrogel fibers by mimicking the silkworm-spinning process. For the spinning of SF fibers, alginate was used as a sericin-like material to induce SF phase separation and entrap liquid SFs, making it possible to shape the outline of SF-based fibers under mild physicochemical conditions. L929 fibroblasts were encapsulated in the fibric hydrogel and displayed excellent viability. Cell-laden SF fibric hydrogels prepared using our method offer a new type of SF-based biomedical device with potential utility in biomedicine. PMID:27296229

  16. Titanium diboride ceramic fiber composites for Hall-Heroult cells

    DOEpatents

    Besmann, T.M.; Lowden, R.A.

    1990-05-29

    An improved cathode structure is described for Hall-Heroult cells for the electrolytic production of aluminum metal. This cathode structure is a preform fiber base material that is infiltrated with electrically conductive titanium diboride using chemical vapor infiltration techniques. The structure exhibits good fracture toughness, and is sufficiently resistant to attack by molten aluminum. Typically, the base can be made from a mat of high purity silicon carbide fibers. Other ceramic or carbon fibers that do not degrade at temperatures below about 1000 C can be used.

  17. Titanium diboride ceramic fiber composites for Hall-Heroult cells

    DOEpatents

    Besmann, Theodore M.; Lowden, Richard A.

    1990-01-01

    An improved cathode structure for Hall-Heroult cells for the electrolytic production of aluminum metal. This cathode structure is a preform fiber base material that is infiltrated with electrically conductive titanium diboride using chemical vapor infiltration techniques. The structure exhibits good fracture toughness, and is sufficiently resistant to attack by molten aluminum. Typically, the base can be made from a mat of high purity silicon carbide fibers. Other ceramic or carbon fibers that do not degrade at temperatures below about 1000 deg. C can be used.

  18. CHARACTERIZATION OF ALVEOLAR EPITHELIAL CELLS CULTURED IN SEMIPERMEABLE HOLLOW FIBERS

    PubMed Central

    Grek, Christina L.; Newton, Danforth A.; Qiu, Yonhzhi; Wen, Xuejun; Spyropoulos, Demetri D.; Baatz, John E.

    2012-01-01

    Cell culture methods commonly used to represent alveolar epithelial cells in vivo have lacked airflow, a 3-dimensional air-liquid interface, and dynamic stretching characteristics of native lung tissue—physiological parameters critical for normal phenotypic gene expression and cellular function. Here the authors report the development of a selectively semipermeable hollow fiber culture system that more accurately mimics the in vivo microenvironment experienced by mammalian distal airway cells than in conventional or standard air-liquid interface culture. Murine lung epithelial cells (MLE-15) were cultured within semipermeable polyurethane hollow fibers and introduced to controlled airflow through the microfiber interior. Under these conditions, MLE-15 cells formed confluent monolayers, demonstrated a cuboidal morphology, formed tight junctions, and produced and secreted surfactant proteins. Numerous lamellar bodies and microvilli were present in MLE-15 cells grown in hollow fiber culture. Conversely, these alveolar type II cell characteristics were reduced in MLE-15 cells cultured in conventional 2D static culture systems. These data support the hypothesis that MLE-15 cells grown within our microfiber culture system in the presence of airflow maintain the phenotypic characteristics of type II cells to a higher degree than those grown in standard in vitro cell culture models. Application of our novel model system may prove advantageous for future studies of specific gene and protein expression involving alveolar epithelial or bronchiolar epithelial cells. PMID:19263283

  19. Dual fiber microprobe for mapping elemental distributions in biological cells

    DOEpatents

    Martin, Rodger C [Powell, TN; Martin, Madhavi Z [Powell, TN

    2007-07-31

    Laser-induced breakdown spectroscopy (LIBS) is applied on a microscale for in situ elemental analysis and spatial mapping in biological cells. A high power laser beam is focused onto a cell surface using a dual branching optical fiber probe for optical excitation of the cell constituents. Dual spectrometers and ICCD detectors capture the emission spectra from the excited cell(s). Repeated probing or repositioning of the laser beam with respect to the cell can provide 2-D or 3-D mapping of the cell.

  20. Cotton fiber: a powerful single-cell model for cell wall and cellulose research

    PubMed Central

    Haigler, Candace H.; Betancur, Lissete; Stiff, Michael R.; Tuttle, John R.

    2012-01-01

    Cotton fibers are single-celled extensions of the seed epidermis. They can be isolated in pure form as they undergo staged differentiation including primary cell wall synthesis during elongation and nearly pure cellulose synthesis during secondary wall thickening. This combination of features supports clear interpretation of data about cell walls and cellulose synthesis in the context of high throughput modern experimental technologies. Prior contributions of cotton fiber to building fundamental knowledge about cell walls will be summarized and the dynamic changes in cell wall polymers throughout cotton fiber differentiation will be described. Recent successes in using stable cotton transformation to alter cotton fiber cell wall properties as well as cotton fiber quality will be discussed. Futurec prospects to perform experiments more rapidly through altering cotton fiberwall properties via virus-induced gene silencing will be evaluated. PMID:22661979

  1. Fiber

    MedlinePlus

    ... it can help with weight control. Fiber aids digestion and helps prevent constipation . It is sometimes used ... fiber attracts water and turns to gel during digestion. This slows digestion. Soluble fiber is found in ...

  2. Polymer Solar Cells: Solubility Controls Fiber Network Formation.

    PubMed

    van Franeker, Jacobus J; Heintges, Gaël H L; Schaefer, Charley; Portale, Giuseppe; Li, Weiwei; Wienk, Martijn M; van der Schoot, Paul; Janssen, René A J

    2015-09-16

    The photoactive layer of polymer solar cells is commonly processed from a four-component solution, containing a semiconducting polymer and a fullerene derivative dissolved in a solvent-cosolvent mixture. The nanoscale dimensions of the polymer-fullerene morphology that is formed upon drying determines the solar cell performance, but the fundamental processes that govern the size of the phase-separated polymer and fullerene domains are poorly understood. Here, we investigate morphology formation of an alternating copolymer of diketopyrrolopyrrole and a thiophene-phenyl-thiophene oligomer (PDPPTPT) with relatively long 2-decyltetradecyl (DT) side chains blended with [6,6]-phenyl-C71-butyric acid methyl ester. During solvent evaporation the polymer crystallizes into a fibrous network. The typical width of these fibers is analyzed by quantification of transmission electron microscopic images, and is mainly determined by the solubility of the polymer in the cosolvent and the molecular weight of the polymer. A higher molecular weight corresponds to a lower solubility and film processing results in a smaller fiber width. Surprisingly, the fiber width is not related to the drying rate or the amount of cosolvent. We have made solar cells with fiber widths ranging from 28 to 68 nm and found an inverse relation between fiber width and photocurrent. Finally, by mixing two cosolvents, we develop a ternary solvent system to tune the fiber width. We propose a model based on nucleation-and-growth which can explain these measurements. Our results show that the width of the semicrystalline polymer fibers is not the result of a frozen dynamical state, but determined by the nucleation induced by the polymer solubility. PMID:26306585

  3. Satellite cell depletion prevents fiber hypertrophy in skeletal muscle.

    PubMed

    Egner, Ingrid M; Bruusgaard, Jo C; Gundersen, Kristian

    2016-08-15

    The largest mammalian cells are the muscle fibers, and they have multiple nuclei to support their large cytoplasmic volumes. During hypertrophic growth, new myonuclei are recruited from satellite stem cells into the fiber syncytia, but it was recently suggested that such recruitment is not obligatory: overload hypertrophy after synergist ablation of the plantaris muscle appeared normal in transgenic mice in which most of the satellite cells were abolished. When we essentially repeated these experiments analyzing the muscles by immunohistochemistry and in vivo and ex vivo imaging, we found that overload hypertrophy was prevented in the satellite cell-deficient mice, in both the plantaris and the extensor digitorum longus muscles. We attribute the previous findings to a reliance on muscle mass as a proxy for fiber hypertrophy, and to the inclusion of a significant number of regenerating fibers in the analysis. We discuss that there is currently no model in which functional, sustainable hypertrophy has been unequivocally demonstrated in the absence of satellite cells; an exception is re-growth, which can occur using previously recruited myonuclei without addition of new myonuclei. PMID:27531949

  4. Oxide Fiber Cathode Materials for Rechargeable Lithium Cells

    NASA Technical Reports Server (NTRS)

    Rice, Catherine E.; Welker, Mark F.

    2008-01-01

    LiCoO2 and LiNiO2 fibers have been investigated as alternatives to LiCoO2 and LiNiO2 powders used as lithium-intercalation compounds in cathodes of rechargeable lithium-ion electrochemical cells. In making such a cathode, LiCoO2 or LiNiO2 powder is mixed with a binder [e.g., poly(vinylidene fluoride)] and an electrically conductive additive (usually carbon) and the mixture is pressed to form a disk. The binder and conductive additive contribute weight and volume, reducing the specific energy and energy density, respectively. In contrast, LiCoO2 or LiNiO2 fibers can be pressed and sintered to form a cathode, without need for a binder or a conductive additive. The inter-grain contacts of the fibers are stronger and have fewer defects than do those of powder particles. These characteristics translate to increased flexibility and greater resilience on cycling and, consequently, to reduced loss of capacity from cycle to cycle. Moreover, in comparison with a powder-based cathode, a fiber-based cathode is expected to exhibit significantly greater ionic and electronic conduction along the axes of the fibers. Results of preliminary charge/discharge-cycling tests suggest that energy densities of LiCoO2- and LiNiO2-fiber cathodes are approximately double those of the corresponding powder-based cathodes.

  5. Carbon fiber enhanced bioelectricity generation in soil microbial fuel cells.

    PubMed

    Li, Xiaojing; Wang, Xin; Zhao, Qian; Wan, Lili; Li, Yongtao; Zhou, Qixing

    2016-11-15

    The soil microbial fuel cell (MFC) is a promising biotechnology for the bioelectricity recovery as well as the remediation of organics contaminated soil. However, the electricity production and the remediation efficiency of soil MFC are seriously limited by the tremendous internal resistance of soil. Conductive carbon fiber was mixed with petroleum hydrocarbons contaminated soil and significantly enhanced the performance of soil MFC. The maximum current density, the maximum power density and the accumulated charge output of MFC mixed carbon fiber (MC) were 10, 22 and 16 times as high as those of closed circuit control due to the carbon fiber productively assisted the anode to collect the electron. The internal resistance of MC reduced by 58%, 83% of which owed to the charge transfer resistance, resulting in a high efficiency of electron transfer from soil to anode. The degradation rates of total petroleum hydrocarbons enhanced by 100% and 329% compared to closed and opened circuit controls without the carbon fiber respectively. The effective range of remediation and the bioelectricity recovery was extended from 6 to 20cm with the same area of air-cathode. The mixed carbon fiber apparently enhanced the bioelectricity generation and the remediation efficiency of soil MFC by means of promoting the electron transfer rate from soil to anode. The use of conductively functional materials (e.g. carbon fiber) is very meaningful for the remediation and bioelectricity recovery in the bioelectrochemical remediation. PMID:27162144

  6. Electrospun fiber membranes enable proliferation of genetically modified cells

    PubMed Central

    Borjigin, Mandula; Eskridge, Chris; Niamat, Rohina; Strouse, Bryan; Bialk, Pawel; Kmiec, Eric B

    2013-01-01

    Polycaprolactone (PCL) and its blended composites (chitosan, gelatin, and lecithin) are well-established biomaterials that can enrich cell growth and enable tissue engineering. However, their application in the recovery and proliferation of genetically modified cells has not been studied. In the study reported here, we fabricated PCL-biomaterial blended fiber membranes, characterized them using physicochemical techniques, and used them as templates for the growth of genetically modified HCT116-19 colon cancer cells. Our data show that the blended polymers are highly miscible and form homogenous electrospun fiber membranes of uniform texture. The aligned PCL nanofibers support robust cell growth, yielding a 2.5-fold higher proliferation rate than cells plated on standard plastic plate surfaces. PCL-lecithin fiber membranes yielded a 2.7-fold higher rate of proliferation, while PCL-chitosan supported a more modest growth rate (1.5-fold higher). Surprisingly, PCL-gelatin did not enhance cell proliferation when compared to the rate of cell growth on plastic surfaces. PMID:23467983

  7. Tissue engineering the retinal ganglion cell nerve fiber layer.

    PubMed

    Kador, Karl E; Montero, Ramon B; Venugopalan, Praseeda; Hertz, Jonathan; Zindell, Allison N; Valenzuela, Daniel A; Uddin, Mohammed S; Lavik, Erin B; Muller, Kenneth J; Andreopoulos, Fotios M; Goldberg, Jeffrey L

    2013-06-01

    Retinal degenerative diseases, such as glaucoma and macular degeneration, affect millions of people worldwide and ultimately lead to retinal cell death and blindness. Cell transplantation therapies for photoreceptors demonstrate integration and restoration of function, but transplantation into the ganglion cell layer is more complex, requiring guidance of axons from transplanted cells to the optic nerve head in order to reach targets in the brain. Here we create a biodegradable electrospun (ES) scaffold designed to direct the growth of retinal ganglion cell (RGC) axons radially, mimicking axon orientation in the retina. Using this scaffold we observed an increase in RGC survival and no significant change in their electrophysiological properties. When analyzed for alignment, 81% of RGCs were observed to project axons radially along the scaffold fibers, with no difference in alignment compared to the nerve fiber layer of retinal explants. When transplanted onto retinal explants, RGCs on ES scaffolds followed the radial pattern of the host retinal nerve fibers, whereas RGCs transplanted directly grew axons in a random pattern. Thus, the use of this scaffold as a cell delivery device represents a significant step towards the use of cell transplant therapies for the treatment of glaucoma and other retinal degenerative diseases. PMID:23489919

  8. Tissue Engineering the Retinal Ganglion Cell Nerve Fiber Layer

    PubMed Central

    Kador, Karl E.; Montero, Ramon B.; Venugopalan, Praseeda; Hertz, Jonathan; Zindell, Allison N.; Valenzuela, Daniel A.; Uddin, Mohammed S.; Lavik, Erin B.; Muller, Kenneth J.; Andreopoulos, Fotios M.; Goldberg, Jeffrey L.

    2013-01-01

    Retinal degenerative diseases, such as glaucoma and macular degeneration, affect millions of people worldwide and ultimately lead to retinal cell death and blindness. Cell transplantation therapies for photoreceptors demonstrate integration and restoration of function, but transplantation into the ganglion cell layer is more complex, requiring guidance of axons from transplanted cells to the optic nerve head in order to reach targets in the brain. Here we create a biodegradable electrospun (ES) scaffold designed to direct the growth of retinal ganglion cell (RGC) axons radially, mimicking axon orientation in the retina. Using this scaffold we observed an increase in RGC survival and no significant change in their electrophysiological properties. When analyzed for alignment, 81% of RGCs were observed to project axons radially along the scaffold fibers, with no difference in alignment compared to the nerve fiber layer of retinal explants. When transplanted onto retinal explants, RGCs on ES scaffolds followed the radial pattern of the host retinal nerve fibers, whereas RGCs transplanted directly grew axons in a random pattern. Thus, the use of this scaffold as a cell delivery device represents a significant step towards the use of cell transplant therapies for the treatment of glaucoma and other retinal degenerative diseases. PMID:23489919

  9. Caveolae internalization repairs wounded cells and muscle fibers

    PubMed Central

    Corrotte, Matthias; Almeida, Patricia E; Tam, Christina; Castro-Gomes, Thiago; Fernandes, Maria Cecilia; Millis, Bryan A; Cortez, Mauro; Miller, Heather; Song, Wenxia; Maugel, Timothy K; Andrews, Norma W

    2013-01-01

    Rapid repair of plasma membrane wounds is critical for cellular survival. Muscle fibers are particularly susceptible to injury, and defective sarcolemma resealing causes muscular dystrophy. Caveolae accumulate in dystrophic muscle fibers and caveolin and cavin mutations cause muscle pathology, but the underlying mechanism is unknown. Here we show that muscle fibers and other cell types repair membrane wounds by a mechanism involving Ca2+-triggered exocytosis of lysosomes, release of acid sphingomyelinase, and rapid lesion removal by caveolar endocytosis. Wounding or exposure to sphingomyelinase triggered endocytosis and intracellular accumulation of caveolar vesicles, which gradually merged into larger compartments. The pore-forming toxin SLO was directly visualized entering cells within caveolar vesicles, and depletion of caveolin inhibited plasma membrane resealing. Our findings directly link lesion removal by caveolar endocytosis to the maintenance of plasma membrane and muscle fiber integrity, providing a mechanistic explanation for the muscle pathology associated with mutations in caveolae proteins. DOI: http://dx.doi.org/10.7554/eLife.00926.001 PMID:24052812

  10. Fiber-laser-based photoacoustic microscopy and melanoma cell detection

    PubMed Central

    Wang, Yu; Maslov, Konstantin; Zhang, Yu; Hu, Song; Yang, Lihmei; Xia, Younan; Liu, Jian; Wang, Lihong V.

    2011-01-01

    For broad applications in biomedical research involving functional dynamics and clinical studies, a photoacoustic microscopy system should be compact, stable, and fast. In this work, we use a fiber laser as the photoacoustic irradiation source to meet these goals. The laser system measures 45×56×13 cm3. The stability of the laser is attributed to the intrinsic optical fiber-based light amplification and output coupling. Its 50-kHz pulse repetition rate enables fast scanning or extensive signal averaging. At the laser wavelength of 1064 nm, the photoacoustic microscope still has enough sensitivity to image small blood vessels while providing high optical absorption contrast between melanin and hemoglobin. Label-free melanoma cells in flowing bovine blood are imaged in vitro, yielding measurements of both cell size and flow speed. PMID:21280901

  11. Fiber-laser-based photoacoustic microscopy and melanoma cell detection.

    PubMed

    Wang, Yu; Maslov, Konstantin; Zhang, Yu; Hu, Song; Yang, Lihmei; Xia, Younan; Liu, Jian; Wang, Lihong V

    2011-01-01

    For broad applications in biomedical research involving functional dynamics and clinical studies, a photoacoustic microscopy system should be compact, stable, and fast. In this work, we use a fiber laser as the photoacoustic irradiation source to meet these goals. The laser system measures 45×56×13 cm3. The stability of the laser is attributed to the intrinsic optical fiber-based light amplification and output coupling. Its 50-kHz pulse repetition rate enables fast scanning or extensive signal averaging. At the laser wavelength of 1064 nm, the photoacoustic microscope still has enough sensitivity to image small blood vessels while providing high optical absorption contrast between melanin and hemoglobin. Label-free melanoma cells in flowing bovine blood are imaged in vitro, yielding measurements of both cell size and flow speed. PMID:21280901

  12. Hollow fibers - Their applications to the study of mammalian cell function

    NASA Technical Reports Server (NTRS)

    Hymer, W. C.; Angeline, M.; Harkness, J.; Chu, M.; Grindleland, R.

    1984-01-01

    The use of hollow fiber technology in cell culture and transplantation is examined. The morphologies of encapsulated pituitary cells before and after implantation into the rat are defined. Implantation experiments using hollow fibers to study mammalian cell functions are described. Consideration is given to examining somatotroph, prolactin, prostrate, fibroblast, and retinal cell functions. These experiments demonstrate that hollow fiber technology is applicable for studying mammalian cell functions.

  13. Electrospinning of unidirectionally and orthogonally aligned thermoplastic polyurethane nanofibers: fiber orientation and cell migration.

    PubMed

    Mi, Hao-Yang; Salick, Max R; Jing, Xin; Crone, Wendy C; Peng, Xiang-Fang; Turng, Lih-Sheng

    2015-02-01

    Unidirectionally and orthogonally aligned thermoplastic polyurethane (TPU) nanofibers were electrospun using a custom-built electrospinning device. The unidirectionally aligned fibers were collected using two parallel copper plates, and the orthogonally aligned fibers were collected using two orthogonal sets of parallel copper plates with alternate negative connections. Carbon nanotubes (CNT) and polyacrylic acid (PAA) were added to modify the polymer solution. It was found that both CNT and PAA were capable of increasing solution conductivity. The TPU/PAA fiber showed the highest degree of fiber orientation with more than 90% of the fibers having an orientation angle between -10° and 10° for unidirectionally aligned fibers, and for orthogonally aligned fibers, the orientation angle of 50% fibers located between -10° and 10° and 48% fibers located between 80° and 100°. Viability assessment of 3T3 fibroblasts cultured on TPU/PAA fibers suggested that the material was cytocompatible. The cells' orientation and migration direction closely matched the fibers' orientation. The cell migration velocity and distance were both enhanced with the guidance of fibers compared with cells cultured on random fibers and common tissue culture plastic. Controlling cell migration velocity and directionality may provide ways to influence differentiation and gene expression and systems that would allow further exploration of wound repair and metastatic cell behavior. PMID:24771704

  14. E-spun composite fibers of collagen and dragline silk protein: fiber mechanics, biocompatibility, and application in stem cell differentiation.

    PubMed

    Zhu, Bofan; Li, Wen; Lewis, Randolph V; Segre, Carlo U; Wang, Rong

    2015-01-12

    Biocomposite matrices with high mechanical strength, high stability, and the ability to direct matrix-specific stem cell differentiation are essential for the reconstruction of lesioned tissues in tissue engineering and cell therapeutics. Toward this end, we used the electrospinning technique to fabricate well-aligned composite fibers from collagen and spider dragline silk protein, obtained from the milk of transgenic goats, mimicking the native extracellular matrix (ECM) on a similar scale. Collagen and the dragline silk proteins were found to mix homogeneously at all ratios in the electrospun (E-spun) fibers. As a result, the ultimate tensile strength and elasticity of the fibers increased monotonically with silk percentage, whereas the stretchability was slightly reduced. Strikingly, we found that the incorporation of silk proteins to collagen dramatically increased the matrix stability against excessive fiber swelling and shape deformation in cell culture medium. When human decidua parietalis placental stem cells (hdpPSCs) were seeded on the collagen-silk matrices, the matrices were found to support cell proliferation at a similar rate as that of the pure collagen matrix, but they provided cell adhesion with reduced strengths and induced cell polarization at varied levels. Matrices containing 15 and 30 wt % silk in collagen (CS15, CS30) were found to induce a level of neural differentiation comparable to that of pure collagen. In particular, CS15 matrix induced the highest extent of cell polarization and promoted the development of extended 1D neural filaments strictly in-line with the aligned fibers. Taking the increased mechanical strength and fiber stability into consideration, CS15 and CS30 E-spun fibers offer better alternatives to pure collagen fibers as scaffolds that can be potentially utilized in neural tissue repair and the development of future nanobiodevices. PMID:25405355

  15. E-Spun Composite Fibers of Collagen and Dragline Silk Protein: Fiber Mechanics, Biocompatibility, and Application in Stem Cell Differentiation

    PubMed Central

    2015-01-01

    Biocomposite matrices with high mechanical strength, high stability, and the ability to direct matrix-specific stem cell differentiation are essential for the reconstruction of lesioned tissues in tissue engineering and cell therapeutics. Toward this end, we used the electrospinning technique to fabricate well-aligned composite fibers from collagen and spider dragline silk protein, obtained from the milk of transgenic goats, mimicking the native extracellular matrix (ECM) on a similar scale. Collagen and the dragline silk proteins were found to mix homogeneously at all ratios in the electrospun (E-spun) fibers. As a result, the ultimate tensile strength and elasticity of the fibers increased monotonically with silk percentage, whereas the stretchability was slightly reduced. Strikingly, we found that the incorporation of silk proteins to collagen dramatically increased the matrix stability against excessive fiber swelling and shape deformation in cell culture medium. When human decidua parietalis placental stem cells (hdpPSCs) were seeded on the collagen–silk matrices, the matrices were found to support cell proliferation at a similar rate as that of the pure collagen matrix, but they provided cell adhesion with reduced strengths and induced cell polarization at varied levels. Matrices containing 15 and 30 wt % silk in collagen (CS15, CS30) were found to induce a level of neural differentiation comparable to that of pure collagen. In particular, CS15 matrix induced the highest extent of cell polarization and promoted the development of extended 1D neural filaments strictly in-line with the aligned fibers. Taking the increased mechanical strength and fiber stability into consideration, CS15 and CS30 E-spun fibers offer better alternatives to pure collagen fibers as scaffolds that can be potentially utilized in neural tissue repair and the development of future nanobiodevices. PMID:25405355

  16. Carbon nanotube fibers are compatible with Mammalian cells and neurons.

    PubMed

    Dubin, R A; Callegari, G; Kohn, J; Neimark, A

    2008-03-01

    We demonstrate the biocompatibility of carbon nanotube fibers (CNFs) fabricated from single-wall carbon nanotubes. Produced by a particle-coagulation spinning process, CNFs are "hair-like" conductive microwires, which uniquely combine properties of porous nanostructured scaffolds, high-area electrodes, and permeable microfluidic conduits. We report that CNFs are nontoxic and support the attachment, spreading, and growth of mammalian cells and the extension of processes from neurons in vitro. Our findings suggest that CNF may be employed for an electrical interfacing of nerve cells and external devices. PMID:18334451

  17. Survivin expression is associated with lens epithelial cell proliferation and fiber cell differentiation

    PubMed Central

    Mansergh, Fiona C.; Boulton, Michael E.; Gunhaga, Lena

    2012-01-01

    Purpose Survivin (Birc5) is the smallest member of the inhibitor of apoptosis (IAP) protein family, which regulates the cell cycle/apoptosis balance. The purpose of this study was to examine Survivin expression in the embryonic chick lens, in chick lens epithelial cell cultures, and in the postnatal mouse lens. Methods Survivin expression was examined using a combination of quantitative real-time polymerase chain reaction, western blotting, and immunocytochemistry. To correlate Survivin expression with the timing of proliferation, we determined the profile of cell proliferation in the developing lens using the cell cycle marker proliferating cell nuclear antigen (PCNA) in quantitative western blotting and immunocytochemistry studies. We also examined the expression of PCNA and the extent of denucleation using terminal deoxynucleotidyl transferase (TdT)-mediated biotin-dUTP nick-end labeling (TUNEL) of lentoids (lens fiber-like cells) during chick lens epithelial cell differentiation in vitro. Results At embryonic day (ED) 4, Survivin immunostaining was present in two pools in lens epithelial cells and fiber cells: cytoplasmic and nuclear. The nuclear staining became more pronounced as the lens epithelial cells differentiated into lens fiber cells. At ED12, Survivin staining was observed in lens fiber cell nuclei containing marginalized chromatin, indicative of early denucleation events. Using western blotting, Survivin expression peaked at ED6, diminishing thereafter. This profile of expression correlated with the events in chick lens epithelial cell cultures: i) increased Survivin expression was associated with an increase in PCNA staining up to day 6 of culture and ii) downregulation of Survivin expression at day 8 of culture was coincident with a dramatic decrease in PCNA staining and an increase in TdT-mediated biotin-dUTP nick-end labeling in lentoids. In early postnatal mouse lenses, Survivin and PCNA were highly expressed and decreased thereafter during

  18. Cell attachment to hydrogel-electrospun fiber mat composite materials.

    PubMed

    Han, Ning; Johnson, Jed K; Bradley, Patrick A; Parikh, Kunal S; Lannutti, John J; Winter, Jessica O

    2012-01-01

    Hydrogels, electrospun fiber mats (EFMs), and their composites have been extensively studied for tissue engineering because of their physical and chemical similarity to native biological systems. However, while chemically similar, hydrogels and electrospun fiber mats display very different topographical features. Here, we examine the influence of surface topography and composition of hydrogels, EFMs, and hydrogel-EFM composites on cell behavior. Materials studied were composed of synthetic poly(ethylene glycol) (PEG) and poly(ethylene glycol)-poly(ε-caprolactone) (PEGPCL) hydrogels and electrospun poly(caprolactone) (PCL) and core/shell PCL/PEGPCL constituent materials. The number of adherent cells and cell circularity were most strongly influenced by the fibrous nature of materials (e.g., topography), whereas cell spreading was more strongly influenced by material composition (e.g., chemistry). These results suggest that cell attachment and proliferation to hydrogel-EFM composites can be tuned by varying these properties to provide important insights for the future design of such composite materials. PMID:24955629

  19. Effect of fiber diameter and alignment of electrospun polyurethane meshes on mesenchymal progenitor cells.

    PubMed

    Bashur, Chris A; Shaffer, Robyn D; Dahlgren, Linda A; Guelcher, Scott A; Goldstein, Aaron S

    2009-09-01

    Effective strategies to guide cell alignment and the deposition of an oriented extracellular matrix are critical for the development of anisotropic engineered tissues suitable for the repair of ligament defects. Electrospinning is a promising means to create meshes that can align adherent cells, but the effect of fiber mesh architecture on differentiation has not been examined closely. Therefore, the goal of this study was to determine the effect of fiber diameter and the degree of fiber alignment on mesenchymal progenitor cell morphology, proliferation, and ligament gene expression. Specifically, a poly(ester urethane)urea elastomer was electrospun onto rigid supports under conditions designed to independently vary the mean fiber diameter (from 0.28 to 2.3 microm) and the degree of fiber alignment. Bone marrow stromal cells--seeded onto supported meshes--adhered to and proliferated on all surfaces. Cells assumed a more spindle-shaped morphology with increasing fiber diameter and degree of fiber alignment, and oriented parallel to fibers on aligned meshes. Expression of the ligament markers collagen 1alpha1, decorin, and tenomodulin appeared to be sensitive to fiber diameter and greatest on the smallest fibers. Concurrently, expression of the transcription factor scleraxis appeared to decrease with increasing fiber alignment. These results suggest that the formation of a ligament-like tissue on electrospun scaffolds is enhanced when the scaffolds consist of aligned submicron fibers. PMID:19292650

  20. Relationship between cell stiffness and stress fiber amount, assessed by simultaneous atomic force microscopy and live-cell fluorescence imaging.

    PubMed

    Gavara, Núria; Chadwick, Richard S

    2016-06-01

    Actomyosin stress fibers, one of the main components of the cell's cytoskeleton, provide mechanical stability to adherent cells by applying and transmitting tensile forces onto the extracellular matrix (ECM) at the sites of cell-ECM adhesion. While it is widely accepted that changes in spatial and temporal distribution of stress fibers affect the cell's mechanical properties, there is no quantitative knowledge on how stress fiber amount and organization directly modulate cell stiffness. We address this key open question by combining atomic force microscopy with simultaneous fluorescence imaging of living cells, and combine for the first time reliable quantitative parameters obtained from both techniques. We show that the amount of myosin and (to a lesser extent) actin assembled in stress fibers directly modulates cell stiffness in adherent mouse fibroblasts (NIH3T3). In addition, the spatial distribution of stress fibers has a second-order modulatory effect. In particular, the presence of either fibers located in the cell periphery, aligned fibers or thicker fibers gives rise to reinforced cell stiffness. Our results provide basic and significant information that will help design optimal protocols to regulate the mechanical properties of adherent cells via pharmacological interventions that alter stress fiber assembly or via micropatterning techniques that restrict stress fiber spatial organization. PMID:26206449

  1. Are dietary fiber-induced alterations in colonic epithelial cell proliferation predictive of fiber's effect on colon cancer?

    PubMed

    Whiteley, L O; Klurfeld, D M

    2000-01-01

    Alterations in cell proliferation of the colon have been observed as a result of changes in amount and type of dietary fiber and in relation to risk of developing colon cancer. Although some human observational and intervention studies contribute to the database, most information results from experiments on rodents. Because of numerous contradictory reports linking dietary fiber, cell proliferation, and colon cancer, we undertook a critical review of existing methods in an attempt to explain the inconsistencies. Although there may be some individual types of dietary fiber that protect against chemically induced colon cancer, dietary fiber as a single entity does not appear to afford any consistent protection. Because of significant differences in experimental protocols among laboratories, it is not yet possible to state with certainty that increases in cell proliferation, induced by fiber consumption, are predictive of increased tumorigenesis. Much of what has been observed and interpreted as elevation of risk may simply be normal homeostatic changes in cell proliferation. Even though fermentation to short-chain fatty acids is a mechanistically attractive hypothesis to explain why fiber modulates cytokinetics, data do not consistently support short-chain fatty acids as biological intermediates in risk of colon cancer. The state of the art in this field has not yet progressed to the point where a clear effect of dietary fiber on cytokinetics and colon carcinogenesis can be assessed with any degree of certainty. Additional markers of apoptosis, differentiation, and cell-cell communication may be required for a more accurate analysis of the relation among fiber, cytokinetics, and colon cancer. PMID:10890023

  2. Electrospinning of unidirectionally and orthogonally aligned thermoplastic polyurethane nanofibers: Fiber orientation and cell migration

    PubMed Central

    Mi, Hao-Yang; Salick, Max R.; Jing, Xin; Crone, Wendy C.; Peng, Xiang-Fang; Turng, Lih-Sheng

    2015-01-01

    Unidirectionally and orthogonally aligned thermoplastic polyurethane (TPU) nanofibers were electrospun using a custom-built electrospinning device. The unidirectionally aligned fibers were collected using two parallel copper plates, and the orthogonally aligned fibers were collected using two orthogonal sets of parallel copper plates with alternate negative connections. Carbon nanotubes (CNT) and polyacrylic acid (PAA) were added to modify the polymer solution. It was found that both CNT and PAA were capable of increasing solution conductivity. The TPU/PAA fiber showed the highest degree of fiber orientation with more than 90% of the fibers having an orientation angle between −10° and 10° for unidirectionally aligned fibers, and for orthogonally aligned fibers, the orientation angle of 50% fibers located between −10° and 10° and 48% fibers located between 80° and 100°. Viability assessment of 3T3 fibroblasts cultured on TPU/PAA fibers suggested that the material was cytocompatible. The cells’ orientation and migration direction closely matched the fibers’ orientation. The cell migration velocity and distance were both enhanced with the guidance of fibers compared with cells cultured on random fibers and common tissue culture plastic. Controlling cell migration velocity and directionality may provide ways to influence differentiation and gene expression and systems that would allow further exploration of wound repair and metastatic cell behavior. PMID:24771704

  3. Functional analyses of cotton (Gossypium hirsutum L.) immature fiber (im) mutant reveal that fiber cell wall development is associated with sensitivity to stress.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Cotton fiber maturity refers the degree of fiber cell wall development and is an important factor for determining commercial value of cotton. The molecular mechanism regulating the fiber cell wall development has not been well characterized. Microscopic image analysis of the cross-sect...

  4. Fiber

    MedlinePlus

    ... broccoli, spinach, and artichokes legumes (split peas, soy, lentils, etc.) almonds Look for the fiber content of ... salsa, taco sauce, and cheese for dinner. Add lentils or whole-grain barley to your favorite soups. ...

  5. Fiber

    MedlinePlus

    ... short period of time can cause intestinal gas ( flatulence ), bloating , and abdominal cramps . This problem often goes ... 213. National Research Council. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and ...

  6. Continual Cell Deformation Induced via Attachment to Oriented Fibers Enhances Fibroblast Cell Migration

    PubMed Central

    Qin, Sisi; Ricotta, Vincent; Simon, Marcia; Clark, Richard A. F.; Rafailovich, Miriam H.

    2015-01-01

    Fibroblast migration is critical to the wound healing process. In vivo, migration occurs on fibrillar substrates, and previous observations have shown that a significant time lag exists before the onset of granulation tissue. We therefore conducted a series of experiments to understand the impact of both fibrillar morphology and migration time. Substrate topography was first shown to have a profound influence. Fibroblasts preferentially attach to fibrillar surfaces, and orient their cytoplasm for maximal contact with the fiber edge. In the case of en-mass cell migration out of an agarose droplet, fibroblasts on flat surfaces emerged with an enhanced velocity, v = 52μm/h, that decreases to the single cell value, v = 28μm/h within 24 hours and remained constant for at least four days. Fibroblasts emerging on fibrillar surfaces emerged with the single cell velocity, which remained constant for the first 24 hours and then increased reaching a plateau with more than twice the initial velocity within the next three days. The focal adhesions were distributed uniformly in cells on flat surfaces, while on the fibrillar surface they were clustered along the cell periphery. Furthermore, the number of focal adhesions for the cells on the flat surfaces remained constant, while it decreased on the fibrillar surface during the next three days. The deformation of the cell nuclei was found to be 50% larger on the fiber surfaces for the first 24 hours. While the mean deformation remained constant on the flat surface, it increased for the next three days by 24% in cells on fibers. On the fourth day, large actin/myosin fibers formed in cells on fibrillar surfaces only and coincided with a change from the standard migration mechanism involving extension of lamellipodia, and retraction of the rear, to one involving strong contractions oriented along the fibers and centered about the nucleus. PMID:25774792

  7. On the mechanism of cell internalization of chrysotile fibers: An immunocytochemical and ultrastructural study

    SciTech Connect

    Malorni, W.; Iosi, F.; Falchi, M.; Donelli, G. )

    1990-08-01

    Human breast carcinoma cells (CG5) and human laryngeal carcinoma cells (HEp-2) were exposed to 10 and 50 {mu}ml (about 5 {mu}m) chrysotile asbestos fibers. Morphological and ultrastructural changes were evaluated by means of immunocytochemistry and by scanning and transmission electron microscopy. The authors attention was focused on the mechanisms of cell internalization and on transport of chrysotile fibers. The fibers appeared to penetrate the cell cytoplasm and to be translocated in proximity of the nucleus. Small chrysotile fibers could also be found inside the nucleus of interphase cells. Involvement of the main cytoskeletal components, i.e., microfilaments, intermediate filaments, and microtubules, in the cytotoxicity of chrysotile fibers was also evaluated. Their findings suggest that after fiber penetration, a rearrangement of the cytoskeletal apparatus occurs. It has also been observed that small fibers remain associated with the cytoskeletal framework, which can thus play a role in asbestos intracytoplasmic translocation in epithelial cells. Furthermore, after the cell has completely recovered its morphology, fiber internalization ultimately seems to lead to the formation of giant multinucleated cells. These data could be indicative of an interaction occurring between asbestos fibers and the normal mitotic process.

  8. Cell-mediated fiber recruitment drives extracellular matrix mechanosensing in engineered fibrillar microenvironments

    PubMed Central

    Baker, Brendon M.; Trappmann, Britta; Wang, William Y.; Sakar, Mahmut S.; Kim, Iris L.; Shenoy, Vivek B.; Burdick, Jason A.; Chen, Christopher S.

    2015-01-01

    To investigate how cells sense stiffness in settings structurally similar to native extracellular matrices (ECM), we designed a synthetic fibrous material with tunable mechanics and user-defined architecture. In contrast to flat hydrogel surfaces, these fibrous materials recapitulated cell-matrix interactions observed with collagen matrices including stellate cell morphologies, cell-mediated realignment of fibers, and bulk contraction of the material. While increasing the stiffness of flat hydrogel surfaces induced mesenchymal stem cell spreading and proliferation, increasing fiber stiffness instead suppressed spreading and proliferation depending on network architecture. Lower fiber stiffness permitted active cellular forces to recruit nearby fibers, dynamically increasing ligand density at the cell surface and promoting the formation of focal adhesions and related signaling. These studies demonstrate a departure from the well-described relationship between material stiffness and spreading established with hydrogel surfaces, and introduce fiber recruitment as a novel mechanism by which cells probe and respond to mechanics in fibrillar matrices. PMID:26461445

  9. Cell proliferation on PVA/sodium alginate and PVA/poly(γ-glutamic acid) electrospun fiber.

    PubMed

    Yang, Jen Ming; Yang, Jhe Hao; Tsou, Shu Chun; Ding, Chian Hua; Hsu, Chih Chin; Yang, Kai Chiang; Yang, Chun Chen; Chen, Ko Shao; Chen, Szi Wen; Wang, Jong Shyan

    2016-09-01

    To overcome the obstacles of easy dissolution of PVA nanofibers without crosslinking treatment and the poor electrospinnability of the PVA cross-linked nanofibers via electrospinning process, the PVA based electrospun hydrogel nanofibers are prepared with post-crosslinking method. To expect the electrospun hydrogel fibers might be a promising scaffold for cell culture and tissue engineering applications, the evaluation of cell proliferation on the post-crosslinking electrospun fibers is conducted in this study. At beginning, poly(vinyl alcohol) (PVA), PVA/sodium alginate (PVASA) and PVA/poly(γ-glutamic acid) (PVAPGA) electrospun fibers were prepared by electrospinning method. The electrospun PVA, PVASA and PVAPGA nanofibers were treated with post-cross-linking method with glutaraldehyde (Glu) as crosslinking agent. These electrospun fibers were characterized with thermogravimetry analysis (TGA) and their morphologies were observed with a scanning electron microscope (SEM). To support the evaluation and explanation of cell growth on the fiber, the study of 3T3 mouse fibroblast cell growth on the surface of pure PVA, SA, and PGA thin films is conducted. The proliferation of 3T3 on the electrospun fiber surface of PVA, PVASA, and PVAPGA was evaluated by seeding 3T3 fibroblast cells on these crosslinked electrospun fibers. The cell viability on electrospun fibers was conducted with water-soluble tetrazolium salt-1 assay (Cell Proliferation Reagent WST-1). The morphology of the cells on the fibers was also observed with SEM. The results of WST-1 assay revealed that 3T3 cells cultured on different electrospun fibers had similar viability, and the cell viability increased with time for all electrospun fibers. From the morphology of the cells on electrospun fibers, it is found that 3T3 cells attached on all electrospun fiber after 1day seeded. Cell-cell communication was noticed on day 3 for all electrospun fibers. Extracellular matrix (ECM) productions were found and

  10. Smooth Muscle-Like Tissue Constructs with Circumferentially Oriented Cells Formed by the Cell Fiber Technology

    PubMed Central

    Hsiao, Amy Y.; Okitsu, Teru; Onoe, Hiroaki; Kiyosawa, Mahiro; Teramae, Hiroki; Iwanaga, Shintaroh; Kazama, Tomohiko; Matsumoto, Taro; Takeuchi, Shoji

    2015-01-01

    The proper functioning of many organs and tissues containing smooth muscles greatly depends on the intricate organization of the smooth muscle cells oriented in appropriate directions. Consequently controlling the cellular orientation in three-dimensional (3D) cellular constructs is an important issue in engineering tissues of smooth muscles. However, the ability to precisely control the cellular orientation at the microscale cannot be achieved by various commonly used 3D tissue engineering building blocks such as spheroids. This paper presents the formation of coiled spring-shaped 3D cellular constructs containing circumferentially oriented smooth muscle-like cells differentiated from dedifferentiated fat (DFAT) cells. By using the cell fiber technology, DFAT cells suspended in a mixture of extracellular proteins possessing an optimized stiffness were encapsulated in the core region of alginate shell microfibers and uniformly aligned to the longitudinal direction. Upon differentiation induction to the smooth muscle lineage, DFAT cell fibers self-assembled to coiled spring structures where the cells became circumferentially oriented. By changing the initial core-shell microfiber diameter, we demonstrated that the spring pitch and diameter could be controlled. 21 days after differentiation induction, the cell fibers contained high percentages of ASMA-positive and calponin-positive cells. Our technology to create these smooth muscle-like spring constructs enabled precise control of cellular alignment and orientation in 3D. These constructs can further serve as tissue engineering building blocks for larger organs and cellular implants used in clinical treatments. PMID:25734774

  11. Smooth muscle-like tissue constructs with circumferentially oriented cells formed by the cell fiber technology.

    PubMed

    Hsiao, Amy Y; Okitsu, Teru; Onoe, Hiroaki; Kiyosawa, Mahiro; Teramae, Hiroki; Iwanaga, Shintaroh; Kazama, Tomohiko; Matsumoto, Taro; Takeuchi, Shoji

    2015-01-01

    The proper functioning of many organs and tissues containing smooth muscles greatly depends on the intricate organization of the smooth muscle cells oriented in appropriate directions. Consequently controlling the cellular orientation in three-dimensional (3D) cellular constructs is an important issue in engineering tissues of smooth muscles. However, the ability to precisely control the cellular orientation at the microscale cannot be achieved by various commonly used 3D tissue engineering building blocks such as spheroids. This paper presents the formation of coiled spring-shaped 3D cellular constructs containing circumferentially oriented smooth muscle-like cells differentiated from dedifferentiated fat (DFAT) cells. By using the cell fiber technology, DFAT cells suspended in a mixture of extracellular proteins possessing an optimized stiffness were encapsulated in the core region of alginate shell microfibers and uniformly aligned to the longitudinal direction. Upon differentiation induction to the smooth muscle lineage, DFAT cell fibers self-assembled to coiled spring structures where the cells became circumferentially oriented. By changing the initial core-shell microfiber diameter, we demonstrated that the spring pitch and diameter could be controlled. 21 days after differentiation induction, the cell fibers contained high percentages of ASMA-positive and calponin-positive cells. Our technology to create these smooth muscle-like spring constructs enabled precise control of cellular alignment and orientation in 3D. These constructs can further serve as tissue engineering building blocks for larger organs and cellular implants used in clinical treatments. PMID:25734774

  12. Compact and Robust Refilling and Connectorization of Hollow Core Photonic Crystal Fiber Gas Reference Cells

    NASA Technical Reports Server (NTRS)

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

    2007-01-01

    This slide presentation reviews a method for refilling and connectorization of hollow core photonic crystal fiber gas reference cells. Thees hollow-core photonic crystal fiber allow optical propagation in air or vacuum and are for use as gas reference cell is proposed and demonstrated. It relies on torch-sealing a quartz filling tube connected to a mechanical splice between regular and hollow-core fibers.

  13. PTEN deletion from adult-generated dentate granule cells disrupts granule cell mossy fiber axon structure

    PubMed Central

    LaSarge, Candi L.; Santos, Victor R; Danzer, Steve C.

    2015-01-01

    Dysregulation of the mTOR-signaling pathway is implicated in the development of temporal lobe epilepsy. In mice, deletion of PTEN from hippocampal dentate granule cells leads to mTOR hyperactivation and promotes the rapid onset of spontaneous seizures. The mechanism by which these abnormal cells initiate epileptogenesis, however, is unclear. PTEN-knockout granule cells develop abnormally, exhibiting morphological features indicative of increased excitatory input. If these cells are directly responsible for seizure genesis, it follows that they should also possess increased output. To test this prediction, dentate granule cell axon morphology was quantified in control and PTEN-knockout mice. Unexpectedly, PTEN deletion increased giant mossy fiber bouton spacing along the axon length, suggesting reduced innervation of CA3. Increased width of the mossy fiber axon pathway in stratum lucidum, however, which likely reflects an unusual increase in mossy fiber axon collateralization in this region, offset the reduction in boutons per axon length. These morphological changes predicts a net increase in granule cell >> CA3 innervation. Increased diameter of axons from PTEN-knockout cells would further enhance granule cell >> CA3 communication. Altogether, these findings suggest that amplified information flow through the hippocampal circuit contributes to seizure occurrence in the PTEN-knockout mouse model of temporal lobe epilepsy. PMID:25600212

  14. The Potential to Improve Cell Infiltration in Composite Fiber-Aligned Electrospun Scaffolds by the Selective Removal of Sacrificial Fibers

    PubMed Central

    Baker, Brendon M.; Gee, Albert O.; Metter, Robert B.; Nathan, Ashwin S.; Marklein, Ross L.; Burdick, Jason A.; Mauck, Robert L.

    2008-01-01

    Aligned electrospun scaffolds are a promising tool for engineering fibrous musculoskeletal tissues as they reproduce the mechanical anisotropy of these tissues and can direct ordered neo-tissue formation. However, these scaffolds suffer from a slow cellular infiltration rate, likely due in part to their dense fiber packing. We hypothesized that cell ingress could be expedited in scaffolds by increasing porosity, while at the same time preserving overall scaffold anisotropy. To test this hypothesis, poly(ε-caprolactone) (a slow-degrading polyester) and poly(ethylene oxide) (a water-soluble polymer) were co-electrospun from two separate spinnerets to form dual-polymer composite fiber-aligned scaffolds. Adjusting fabrication parameters produced aligned scaffolds with a full range of sacrificial (PEO) fiber contents. Tensile properties of scaffolds were a function of the ratio of PCL to PEO in the composite scaffolds, and were altered in a predictable fashion with removal of the PEO component. When seeded with mesenchymal stem cells (MSCs), increases in the starting sacrificial fraction (and porosity) improved cell infiltration and distribution after three weeks in culture. In pure PCL scaffolds, cells lined the scaffold periphery, while scaffolds containing >50% sacrificial PEO content had cells present throughout the scaffold. These findings indicate that cell infiltration can be expedited in dense fibrous assemblies with the removal of sacrificial fibers. This strategy may enhance in vitro and in vivo formation and maturation of a functional constructs for fibrous tissue engineering. PMID:18313138

  15. Adenovirus type 2 expresses fiber in monkey-human hybrids and reconstructed cells

    SciTech Connect

    Zorn, G.A.; Anderson, C.W.

    1981-02-01

    Adenovirus type 2 protein expression was measured by indirect immunofluorescence in monkey-human hybrids and in cells reconstructed from monkey and human cell karyoplasts and cytoplasts. Monkey-human hybrid clones infected with adenovirus type 2 expressed fiber protein, whereas infected monkey cells alone did not. Hybrids constructed after the parental monkey cells were infected with adenovirus type 2 demonstrated that fiber synthesis in these cells could be rescued by fusion to uninfected human cells. Thus, human cells contain a dominant factor that acts in trans and overcomes the inability of monkey cells to synthesize fiber. These results are consistent with the hypothesis that the block to adenovirus replication in monkey cells involves a nuclear event that prevents the formation of functional mRNA for some late viral proteins including fiber polypeptide.

  16. Observation of Chinese Hamster Ovary Cells retained inside the non-woven fiber matrix of the CellTank bioreactor

    PubMed Central

    Zhang, Ye; Chotteau, Véronique

    2015-01-01

    This data article shows how the recombinant Chinese Hamster Ovary (CHO) cells are located in the interstices of the matrix fibers of a CellTank bioreactor after completion of a perfusion culture, supporting the article entitled “Very high cell density perfusion of CHO cells anchored in a non-woven matrix-based bioreactor” by Zhang et al. [1]. It provides a visualization of the cell distribution in the non-woven fiber matrix in a deeper view. PMID:26958613

  17. Dissecting Regional Variations in Stress Fiber Mechanics in Living Cells with Laser Nanosurgery

    SciTech Connect

    Tanner, Kandice; Boudreau, Aaron; Bissell, Mina J; Kumar, Sanjay

    2010-03-02

    The ability of a cell to distribute contractile stresses across the extracellular matrix in a spatially heterogeneous fashion underlies many cellular behaviors, including motility and tissue assembly. Here we investigate the biophysical basis of this phenomenon by using femtosecond laser nanosurgery to measure the viscoelastic recoil and cell-shape contributions of contractile stress fibers (SFs) located in specific compartments of living cells. Upon photodisruption and recoil, myosin light chain kinase-dependent SFs located along the cell periphery display much lower effective elasticities and higher plateau retraction distances than Rho-associated kinase-dependent SFs located in the cell center, with severing of peripheral fibers uniquely triggering a dramatic contraction of the entire cell within minutes of fiber irradiation. Image correlation spectroscopy reveals that when one population of SFs is pharmacologically dissipated, actin density flows toward the other population. Furthermore, dissipation of peripheral fibers reduces the elasticity and increases the plateau retraction distance of central fibers, and severing central fibers under these conditions triggers cellular contraction. Together, these findings show that SFs regulated by different myosin activators exhibit different mechanical properties and cell shape contributions. They also suggest that some fibers can absorb components and assume mechanical roles of other fibers to stabilize cell shape.

  18. Aberrant phenotype and transcriptome expression during fiber cell wall thickening caused by the mutation of the Im gene in immature fiber (im) mutant in Gossypium hirsutum L

    PubMed Central

    2014-01-01

    Background The immature fiber (im) mutant of Gossypium hirsutum L. is a special cotton fiber mutant with non-fluffy fibers. It has low dry weight and fineness of fibers due to developmental defects in fiber secondary cell wall (SCW). Results We compared the cellulose content in fibers, thickness of fiber cell wall and fiber transcriptional profiling during SCW development in im mutant and its near-isogenic wild-type line (NIL) TM-1. The im mutant had lower cellulose content and thinner cell walls than TM-1 at same fiber developmental stage. During 25 ~ 35 day post-anthesis (DPA), sucrose content, an important carbon source for cellulose synthesis, was also significantly lower in im mutant than in TM-1. Comparative analysis of fiber transcriptional profiling from 13 ~ 25 DPA indicated that the largest transcriptional variations between the two lines occurred at the onset of SCW development. TM-1 began SCW biosynthesis approximately at 16 DPA, whereas the same fiber developmental program in im mutant was delayed until 19 DPA, suggesting an asynchronous fiber developmental program between TM-1 and im mutant. Functional classification and enrichment analysis of differentially expressed genes (DEGs) between the two NILs indicated that genes associated with biological processes related to cellulose synthesis, secondary cell wall biogenesis, cell wall thickening and sucrose metabolism, respectively, were significantly up-regulated in TM-1. Twelve genes related to carbohydrate metabolism were validated by quantitative reverse transcription PCR (qRT-PCR) and confirmed a temporal difference at the earlier transition and SCW biosynthesis stages of fiber development between TM-1 and im mutant. Conclusions We propose that Im is an important regulatory gene influencing temporal differences in expression of genes related to fiber SCW biosynthesis. This study lays a foundation for cloning the Im gene, elucidating molecular mechanism of fiber SCW development and further

  19. Sliding Fibers: Slidable, Injectable, and Gel-like Electrospun Nanofibers as Versatile Cell Carriers.

    PubMed

    Lee, Slgirim; Yun, Seokhwan; Park, Kook In; Jang, Jae-Hyung

    2016-03-22

    Designing biomaterial systems that can mimic fibrous, natural extracellular matrix is crucial for enhancing the efficacy of various therapeutic tools. Herein, a smart technology of three-dimensional electrospun fibers that can be injected in a minimally invasive manner was developed. Open surgery is currently the only route of administration of conventional electrospun fibers into the body. Coordinating electrospun fibers with a lubricating hydrogel produced fibrous constructs referred to as slidable, injectable, and gel-like (SLIDING) fibers. These SLIDING fibers could pass smoothly through a catheter and fill any cavity while maintaining their fibrous morphology. Their injectable features were derived from their distinctive rheological characteristics, which were presumably caused by the combinatorial effects of mobile electrospun fibers and lubricating hydrogels. The resulting injectable fibers fostered a highly favorable environment for human neural stem cell (hNSC) proliferation and neurosphere formation within the fibrous structures without compromising hNSC viability. SLIDING fibers demonstrated superior performance as cell carriers in animal stroke models subjected to the middle cerebral artery occlusion (MCAO) stroke model. In this model, SLIDING fiber application extended the survival rate of administered hNSCs by blocking microglial infiltration at the early, acute inflammatory stage. The development of SLIDING fibers will increase the clinical significance of fiber-based scaffolds in many biomedical fields and will broaden their applicability. PMID:26885937

  20. Fiber/collagen composites for ligament tissue engineering: influence of elastic moduli of sparse aligned fibers on mesenchymal stem cells.

    PubMed

    Thayer, Patrick S; Verbridge, Scott S; Dahlgren, Linda A; Kakar, Sanjeev; Guelcher, Scott A; Goldstein, Aaron S

    2016-08-01

    Electrospun microfibers are attractive for the engineering of oriented tissues because they present instructive topographic and mechanical cues to cells. However, high-density microfiber networks are too cell-impermeable for most tissue applications. Alternatively, the distribution of sparse microfibers within a three-dimensional hydrogel could present instructive cues to guide cell organization while not inhibiting cell behavior. In this study, thin (∼5 fibers thick) layers of aligned microfibers (0.7 μm) were embedded within collagen hydrogels containing mesenchymal stem cells (MSCs), cultured for up to 14 days, and assayed for expression of ligament markers and imaged for cell organization. These microfibers were generated through the electrospinning of polycaprolactone (PCL), poly(ester-urethane) (PEUR), or a 75/25 PEUR/PCL blend to produce microfiber networks with elastic moduli of 31, 15, and 5.6 MPa, respectively. MSCs in composites containing 5.6 MPa fibers exhibited increased expression of the ligament marker scleraxis and the contractile phenotype marker α-smooth muscle actin versus the stiffer fiber composites. Additionally, cells within the 5.6 MPa microfiber composites were more oriented compared to cells within the 15 and 31 MPa microfiber composites. Together, these data indicate that the mechanical properties of microfiber/collagen composites can be tuned for the engineering of ligament and other target tissues. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1894-1901, 2016. PMID:27037972

  1. Engineering cholesterol-based fibers for antibody immobilization and cell capture

    NASA Astrophysics Data System (ADS)

    Cohn, Celine

    In 2015, the United States is expected to have nearly 600,000 deaths attributed to cancer. Of these 600,000 deaths, 90% will be a direct result of cancer metastasis, the spread of cancer throughout the body. During cancer metastasis, circulating tumor cells (CTCs) are shed from primary tumors and migrate through bodily fluids, establishing secondary cancer sites. As cancer metastasis is incredibly lethal, there is a growing emphasis on developing "liquid biopsies" that can screen peripheral blood, search for and identify CTCs. One popular method for capturing CTCs is the use of a detection platform with antibodies specifically suited to recognize and capture cancer cells. These antibodies are immobilized onto the platform and can then bind and capture cells of interest. However, current means to immobilize antibodies often leave them with drastically reduced function. The antibodies are left poorly suited for cell capture, resulting in low cell capture efficiencies. This body of work investigates the use of lipid-based fibers to immobilize proteins in a way that retains protein function, ultimately leading to increased cell capture efficiencies. The resulting increased efficiencies are thought to arise from the retained three-dimensional structure of the protein as well as having a complete coating of the material surface with antibodies that are capable of interacting with their antigens. It is possible to electrospin cholesterol-based fibers that are similar in design to the natural cell membrane, providing proteins a more natural setting during immobilization. Such fibers have been produced from cholesterol-based cholesteryl succinyl silane (CSS). These fibers have previously illustrated a keen aptitude for retaining protein function and increasing cell capture. Herein the work focuses on three key concepts. First, a model is developed to understand the immobilization mechanism used by electrospun CSS fibers. The antibody immobilization and cell capturing

  2. Engineering cholesterol-based fibers for antibody immobilization and cell capture

    NASA Astrophysics Data System (ADS)

    Cohn, Celine

    In 2015, the United States is expected to have nearly 600,000 deaths attributed to cancer. Of these 600,000 deaths, 90% will be a direct result of cancer metastasis, the spread of cancer throughout the body. During cancer metastasis, circulating tumor cells (CTCs) are shed from primary tumors and migrate through bodily fluids, establishing secondary cancer sites. As cancer metastasis is incredibly lethal, there is a growing emphasis on developing "liquid biopsies" that can screen peripheral blood, search for and identify CTCs. One popular method for capturing CTCs is the use of a detection platform with antibodies specifically suited to recognize and capture cancer cells. These antibodies are immobilized onto the platform and can then bind and capture cells of interest. However, current means to immobilize antibodies often leave them with drastically reduced function. The antibodies are left poorly suited for cell capture, resulting in low cell capture efficiencies. This body of work investigates the use of lipid-based fibers to immobilize proteins in a way that retains protein function, ultimately leading to increased cell capture efficiencies. The resulting increased efficiencies are thought to arise from the retained three-dimensional structure of the protein as well as having a complete coating of the material surface with antibodies that are capable of interacting with their antigens. It is possible to electrospin cholesterol-based fibers that are similar in design to the natural cell membrane, providing proteins a more natural setting during immobilization. Such fibers have been produced from cholesterol-based cholesteryl succinyl silane (CSS). These fibers have previously illustrated a keen aptitude for retaining protein function and increasing cell capture. Herein the work focuses on three key concepts. First, a model is developed to understand the immobilization mechanism used by electrospun CSS fibers. The antibody immobilization and cell capturing

  3. Climbing fibers mediate vestibular modulation of both "complex" and "simple spikes" in Purkinje cells.

    PubMed

    Barmack, N H; Yakhnitsa, V

    2015-10-01

    Climbing and mossy fibers comprise two distinct afferent paths to the cerebellum. Climbing fibers directly evoke a large multispiked action potential in Purkinje cells termed a "complex spike" (CS). By logical exclusion, the other class of Purkinje cell action potential, termed "simple spike" (SS), has often been attributed to activity conveyed by mossy fibers and relayed to Purkinje cells through granule cells. Here, we investigate the relative importance of climbing and mossy fiber pathways in modulating neuronal activity by recording extracellularly from Purkinje cells, as well as from mossy fiber terminals and interneurons in folia 8-10. Sinusoidal roll-tilt vestibular stimulation vigorously modulates the discharge of climbing and mossy fiber afferents, Purkinje cells, and interneurons in folia 9-10 in anesthetized mice. Roll-tilt onto the side ipsilateral to the recording site increases the discharge of both climbing fibers (CSs) and mossy fibers. However, the discharges of SSs decrease during ipsilateral roll-tilt. Unilateral microlesions of the beta nucleus (β-nucleus) of the inferior olive blocks vestibular modulation of both CSs and SSs in contralateral Purkinje cells. The blockage of SSs occurs even though primary and secondary vestibular mossy fibers remain intact. When mossy fiber afferents are damaged by a unilateral labyrinthectomy (UL), vestibular modulation of SSs in Purkinje cells ipsilateral to the UL remains intact. Two inhibitory interneurons, Golgi and stellate cells, could potentially contribute to climbing fiber-induced modulation of SSs. However, during sinusoidal roll-tilt, only stellate cells discharge appropriately out of phase with the discharge of SSs. Golgi cells discharge in phase with SSs. When the vestibularly modulated discharge is blocked by a microlesion of the inferior olive, the modulated discharge of CSs and SSs is also blocked. When the vestibular mossy fiber pathway is destroyed, vestibular modulation of ipsilateral CSs and

  4. Fiber optic SERS-based plasmonics nanobiosensing in single living cells

    NASA Astrophysics Data System (ADS)

    Scaffidi, Jonathan P.; Gregas, Molly K.; Seewaldt, Victoria; Vo-Dinh, Tuan

    2009-05-01

    We describe the development of small molecule-sensitive plasmonics-active fiber-optic nanoprobes suitable for intracellular bioanalysis in single living human cells using surface-enhanced Raman scattering (SERS) detection. The practical utility of SERS-based fiber-optic nanoprobes is illustrated by measurements of intracellular pH in HMEC- 15/hTERT immortalized "normal" human mammary epithelial cells and PC-3 human prostate cancer cells. The results indicate that fiber-optic nanoprobe insertion and interrogation provide a sensitive and selective means to monitor biologically-relevant small molecules at the single cell level.

  5. High performance methanol-oxygen fuel cell with hollow fiber electrode

    NASA Technical Reports Server (NTRS)

    Lawson, Daniel D. (Inventor); Ingham, John D. (Inventor)

    1983-01-01

    A methanol/air-oxygen fuel cell including an electrode formed by open-ended ion-exchange hollow fibers having a layer of catalyst deposited on the inner surface thereof and a first current collector in contact with the catalyst layer. A second current collector external of said fibers is provided which is immersed along with the hollow fiber electrode in an aqueous electrolyte body. Upon passage of air or oxygen through the hollow fiber electrode and introduction of methanol into the aqueous electrolyte, a steady current output is obtained. Two embodiments of the fuel cell are disclosed. In the first embodiment the second metal electrode is displaced away from the hollow fiber in the electrolyte body while in the second embodiment a spiral-wrap electrode is provided about the outer surface of the hollow fiber electrode.

  6. Cotton Fiber Cell Walls of Gossypium hirsutum and Gossypium barbadense Have Differences Related to Loosely-Bound Xyloglucan

    PubMed Central

    Avci, Utku; Pattathil, Sivakumar; Singh, Bir; Brown, Virginia L.; Hahn, Michael G.; Haigler, Candace H.

    2013-01-01

    Cotton fiber is an important natural textile fiber due to its exceptional length and thickness. These properties arise largely through primary and secondary cell wall synthesis. The cotton fiber of commerce is a cellulosic secondary wall surrounded by a thin cuticulated primary wall, but there were only sparse details available about the polysaccharides in the fiber cell wall of any cotton species. In addition, Gossypium hirsutum (Gh) fiber was known to have an adhesive cotton fiber middle lamella (CFML) that joins adjacent fibers into tissue-like bundles, but it was unknown whether a CFML existed in other commercially important cotton fibers. We compared the cell wall chemistry over the time course of fiber development in Gh and Gossypium barbadense (Gb), the two most important commercial cotton species, when plants were grown in parallel in a highly controlled greenhouse. Under these growing conditions, the rate of early fiber elongation and the time of onset of secondary wall deposition were similar in fibers of the two species, but as expected the Gb fiber had a prolonged elongation period and developed higher quality compared to Gh fiber. The Gb fibers had a CFML, but it was not directly required for fiber elongation because Gb fiber continued to elongate rapidly after CFML hydrolysis. For both species, fiber at seven ages was extracted with four increasingly strong solvents, followed by analysis of cell wall matrix polysaccharide epitopes using antibody-based Glycome Profiling. Together with immunohistochemistry of fiber cross-sections, the data show that the CFML of Gb fiber contained lower levels of xyloglucan compared to Gh fiber. Xyloglucan endo-hydrolase activity was also higher in Gb fiber. In general, the data provide a rich picture of the similarities and differences in the cell wall structure of the two most important commercial cotton species. PMID:23457548

  7. Effects of asbestos fibers on cell division, cell survival, and formation of thioguanine-resistant mutants in Chinese hamster ovary cells

    SciTech Connect

    Kenne, K.; Ljungquist, S.; Ringertz, N.R.

    1986-04-01

    The ability of crocidolite fibers to induce point mutations and mitotic abnormalities in Chinese hamster ovary (CHO) cells was examined in cell cultures. The purpose has been to study the possibilities for establishing in vitro test methods to quantify genetic damage induced by asbestos and other mineral fibers. Results obtained with the CHO/hypoxanthine guanine phosphoribosyl transferase system indicated that crocidolite fibers per se do not significantly increase the number of thioguanine-resistant mutants. Crocidolite fibers also failed to potentiate the mutagenicity of benzo(a)pyrene. Time-lapse cinematography and microscopy showed that asbestos (crocidolite) fibers were markedly cytotoxic. Among surviving cells some underwent abnormal cell divisions which resulted in multi- and micronucleate cells. Many cells that contained a few asbestos fibers, however, underwent mitosis and successfully formed two mononucleate daughter cells capable of further divisions. Individual, fiber-containing cells were examined by time-lapse television recordings for 4-5 days. During this time period some cells underwent six divisions and generated an almost normal number of daughter cells. Cells which contained fibers that were longer or equivalent to the diameter of the mitotic cell (20 ..mu..m), showed different forms of mitotic abnormalities. The frequency of multinucleate cells was drastically increased following exposure to asbestos fibers. Only rarely, however, did these cells divide to produce viable daughter cells capable of continued cell multiplication. The frequency of multinucleate cells was dependent on the dose of exposure to asbestos fibers and could possible be used as an index of the degree of mitotic disturbances induced by mineral fibers.

  8. Understanding the Relationship between Cotton Fiber Properties and Non-Cellulosic Cell Wall Polysaccharides

    PubMed Central

    Rajasundaram, Dhivyaa; Runavot, Jean-Luc; Guo, Xiaoyuan; Willats, William G. T.; Meulewaeter, Frank; Selbig, Joachim

    2014-01-01

    A detailed knowledge of cell wall heterogeneity and complexity is crucial for understanding plant growth and development. One key challenge is to establish links between polysaccharide-rich cell walls and their phenotypic characteristics. It is of particular interest for some plant material, like cotton fibers, which are of both biological and industrial importance. To this end, we attempted to study cotton fiber characteristics together with glycan arrays using regression based approaches. Taking advantage of the comprehensive microarray polymer profiling technique (CoMPP), 32 cotton lines from different cotton species were studied. The glycan array was generated by sequential extraction of cell wall polysaccharides from mature cotton fibers and screening samples against eleven extensively characterized cell wall probes. Also, phenotypic characteristics of cotton fibers such as length, strength, elongation and micronaire were measured. The relationship between the two datasets was established in an integrative manner using linear regression methods. In the conducted analysis, we demonstrated the usefulness of regression based approaches in establishing a relationship between glycan measurements and phenotypic traits. In addition, the analysis also identified specific polysaccharides which may play a major role during fiber development for the final fiber characteristics. Three different regression methods identified a negative correlation between micronaire and the xyloglucan and homogalacturonan probes. Moreover, homogalacturonan and callose were shown to be significant predictors for fiber length. The role of these polysaccharides was already pointed out in previous cell wall elongation studies. Additional relationships were predicted for fiber strength and elongation which will need further experimental validation. PMID:25383868

  9. Effectiveness of two synthetic fiber filters for removing white cells from AS-1 red cells.

    PubMed

    Pikul, F J; Farrar, R P; Boris, M B; Estok, L; Marlo, D; Wildgen, M; Chaplin, H

    1989-09-01

    Two commercially available synthetic fiber filters were studied for their effectiveness at removing white cells (WBCs) from AS-1-preserved red cells (RBCs) stored less than or equal to 14 days. In all, 65 filtrations were performed. An automated microprocessor-controlled hydraulic system designed for use with cellulose acetate fiber filters was employed to prepare filtered RBCs before release for transfusion. Studies were also carried out on polyester fiber filters, which are designed to be used in-line during transfusion. Residual WBCs were below the accurate counting range of Coulter counters and of conventional manual chamber counts. An isosmotic ammonium chloride RBC lysis method, plus a modified chamber counting technique, permitted a 270-fold increase over the number of WBCs counted by the conventional manual method. For the polyester fiber-filtered products, residual WBCs per unit were not affected by speed of filtration, prior length of storage, or mechanical tapping during filtration. The effectiveness of WBC removal (mean 99.7%), total residual WBCs (means, 4.8 and 5.5 x 10(6], and RBC recovery (mean, 93%) was the same for both filters. The majority of residual WBCs were lymphocytes. WBC removal and RBC recovery were strikingly superior to results reported with nonfiltration methods. PMID:2505411

  10. Climbing Fiber Regulation of Spontaneous Purkinje Cell Activity and Cerebellum-Dependent Blink Responses123

    PubMed Central

    Bengtsson, Fredrik

    2016-01-01

    Abstract It has been known for a long time that GABAergic Purkinje cells in the cerebellar cortex, as well as their target neurons in the cerebellar nuclei, are spontaneously active. The cerebellar output will, therefore, depend on how input is integrated into this spontaneous activity. It has been shown that input from climbing fibers originating in the inferior olive controls the spontaneous activity in Purkinje cells. While blocking climbing fiber input to the Purkinje cells causes a dramatic increase in the firing rate, increased climbing fiber activity results in reduced Purkinje cell activity. However, the exact calibration of this regulation has not been examined systematically. Here we examine the relation between climbing fiber stimulation frequency and Purkinje cell activity in unanesthetized decerebrated ferrets. The results revealed a gradual suppression of Purkinje cell activity, starting at climbing fiber stimulation frequencies as low as 0.5 Hz. At 4 Hz, Purkinje cells were completely silenced. This effect lasted an average of 2 min after the stimulation rate was reduced to a lower level. We also examined the effect of sustained climbing fiber stimulation on overt behavior. Specifically, we analyzed conditioned blink responses, which are known to be dependent on the cerebellum, while stimulating the climbing fibers at different frequencies. In accordance with the neurophysiological data, the conditioned blink responses were suppressed at stimulation frequencies of ≥4 Hz. PMID:26839917

  11. Light Weight Design Nickel-Alkaline Cells Using Fiber Electrodes

    NASA Technical Reports Server (NTRS)

    Pickett, David F.; Willis, Bob; Britton, Doris; Saelens, Johan

    2005-01-01

    Using fiber electrode technology, currently produced by Bekaert Corporation (Bekaert), Electro Energy, Inc., (EEI) Mobile Energy Products Group (formerly, Eagle-Picher Technologies, LLC., Power Systems Department) in Colorado Springs, CO has demonstrated that it is feasible to manufacture flight weight nickel-hydrogen cells having about twice the specific energy (80 vs. 40 watt-hr/kg) as state-of-the-art nickel-hydrogen cells that are flown on geosynchronous communications satellites. Although lithium-ion battery technology has made large in-roads to replace the nickel-alkaline technology (nickel-cadmium, nickel-metal hydride), the technology offered here competes with lithium-ion weight and offers alternatives not present in the lithium-ion chemistry such as ability to undergo continuous overcharge, reversal on discharge and sustain rate capability sufficient to start automotive and aircraft engines at subzero temperatures. In development to date seven 50 ampere-hour nickel-hydrogen have been constructed, acceptance tested and briefly tested in a low earth orbit (LEO) cycle regime. The effort was jointly funded by Electro Energy, Inc. and NASA Glenn Research Center, Cleveland, OH. Five of the seven cells have been shipped to NASA GRC for further cycle testing. Two of the cells experienced failure due to internal short circuits during initial cycle testing at EEL Destructive Physical Analysis (DPA) of one of the cells has shown the failure mode to be due to inadequate hydrogen catalyst electrodes that were not capacity balanced with the higher energy density nickel oxide electrodes. In the investigators opinion, rebuild of the cells using proper electrode balance would result in cells that could sustain over 30,000 cycles at moderate depths-of-discharge in a LEO regime or endure over 20 years of geosynchronous orbit (GEO) cycling while realizing a two-fold increase in specific energy for the battery or a 1.1 kg weight savings per 50 ampere-hour cell. Additional

  12. A model for cell density effect on stress fiber alignment and collective directional migration

    NASA Astrophysics Data System (ADS)

    Abeddoust, Mohammad; Shamloo, Amir

    2015-12-01

    In this study, numerical simulation of collective cell migration is presented in order to mimic the group migration of endothelial cells subjected to the concentration gradients of a biochemical factor. The developed 2D model incorporates basic elements of the cell, including both the cell membrane and the cell cytoskeleton, based on a viscoelastic cell mechanic model. Various cell processes—including cell random walk, cell-cell interactions, cell chemotaxis, and cellular cytoskeleton rearrangements—are considered and analyzed in our developed model. After validating the model by using available experimental data, the model is used to investigate various important parameters during collective cell chemotaxis, such as cell density, cytoskeleton organization, stress fiber reorientations, and intracellular forces. The results suggest that increasing the cell density causes the cell-cell interactions to affect the orientation of stress fibers throughout the cytoskeleton and makes the stress fibers more aligned in the direction of the imposed concentration gradient. This improved alignment of the stress fibers correlates with the intensification of the intracellular forces transferred in the gradient direction; this improves the cell group migration. Comparison of the obtained results with available experimental observations of collective chemotaxis of endothelial cells shows an interesting agreement.

  13. Comparative transcriptome analysis of epithelial and fiber cells in newborn mouse lenses with RNA sequencing

    PubMed Central

    Hoang, Thanh V.; Kumar, Praveen Kumar Raj; Sutharzan, Sreeskandarajan; Tsonis, Panagiotis A.; Liang, Chun

    2014-01-01

    Purpose The ocular lens contains only two cell types: epithelial cells and fiber cells. The epithelial cells lining the anterior hemisphere have the capacity to continuously proliferate and differentiate into lens fiber cells that make up the large proportion of the lens mass. To understand the transcriptional changes that take place during the differentiation process, high-throughput RNA-Seq of newborn mouse lens epithelial cells and lens fiber cells was conducted to comprehensively compare the transcriptomes of these two cell types. Methods RNA from three biologic replicate samples of epithelial and fiber cells from newborn FVB/N mouse lenses was isolated and sequenced to yield more than 24 million reads per sample. Sequence reads that passed quality filtering were mapped to the reference genome using Genomic Short-read Nucleotide Alignment Program (GSNAP). Transcript abundance and differential gene expression were estimated using the Cufflinks and DESeq packages, respectively. Gene Ontology enrichment was analyzed using GOseq. RNA-Seq results were compared with previously published microarray data. The differential expression of several biologically important genes was confirmed using reverse transcription (RT)-quantitative PCR (qPCR). Results Here, we present the first application of RNA-Seq to understand the transcriptional changes underlying the differentiation of epithelial cells into fiber cells in the newborn mouse lens. In total, 6,022 protein-coding genes exhibited differential expression between lens epithelial cells and lens fiber cells. To our knowledge, this is the first study identifying the expression of 254 long intergenic non-coding RNAs (lincRNAs) in the lens, of which 86 lincRNAs displayed differential expression between the two cell types. We found that RNA-Seq identified more differentially expressed genes and correlated with RT-qPCR quantification better than previously published microarray data. Gene Ontology analysis showed that genes

  14. Functional analyses of cotton (Gossypium hirsutum L.) immature fiber (im) mutant infer that fiber cell wall development is associated with stress responses

    PubMed Central

    2013-01-01

    Background Cotton fiber maturity is an important factor for determining the commercial value of cotton. How fiber cell wall development affects fiber maturity is not well understood. A comparison of fiber cross-sections showed that an immature fiber (im) mutant had lower fiber maturity than its near isogenic wild type, Texas marker-1 (TM-1). The availability of the im mutant and TM-1 provides a unique way to determine molecular mechanisms regulating cotton fiber maturity. Results Transcriptome analysis showed that the differentially expressed genes (DEGs) in the im mutant fibers grown under normal stress conditions were similar to those in wild type cotton fibers grown under severe stress conditions. The majority of these DEGs in the im mutant were related to stress responses and cellular respiration. Stress is known to reduce the activity of a classical respiration pathway responsible for energy production and reactive oxygen species (ROS) accumulation. Both energy productions and ROS levels in the im mutant fibers are expected to be reduced if the im mutant is associated with stress responses. In accord with the prediction, the transcriptome profiles of the im mutant showed the same alteration of transcriptional regulation that happened in energy deprived plants in which expressions of genes associated with cell growth processes were reduced whereas expressions of genes associated with recycling and transporting processes were elevated. We confirmed that ROS production in developing fibers from the im mutant was lower than that from the wild type. The lower production of ROS in the im mutant fibers might result from the elevated levels of alternative respiration induced by stress. Conclusion The low degree of fiber cell wall thickness of the im mutant fibers is associated with deregulation of the genes involved in stress responses and cellular respiration. The reduction of ROS levels and up-regulation of the genes involved in alternative respirations suggest that

  15. The Hagfish Gland Thread Cell: A Fiber-Producing Cell Involved in Predator Defense.

    PubMed

    Fudge, Douglas S; Schorno, Sarah

    2016-01-01

    Fibers are ubiquitous in biology, and include tensile materials produced by specialized glands (such as silks), extracellular fibrils that reinforce exoskeletons and connective tissues (such as chitin and collagen), as well as intracellular filaments that make up the metazoan cytoskeleton (such as F-actin, microtubules, and intermediate filaments). Hagfish gland thread cells are unique in that they produce a high aspect ratio fiber from cytoskeletal building blocks within the confines of their cytoplasm. These threads are elaborately coiled into structures that readily unravel when they are ejected into seawater from the slime glands. In this review we summarize what is currently known about the structure and function of gland thread cells and we speculate about the mechanism that these cells use to produce a mechanically robust fiber that is almost one hundred thousand times longer than it is wide. We propose that a key feature of this mechanism involves the unidirectional rotation of the cell's nucleus, which would serve to twist disorganized filaments into a coherent thread and impart a torsional stress on the thread that would both facilitate coiling and drive energetic unravelling in seawater. PMID:27258313

  16. Development of secondary cell wall in cotton fibers as examined with Fourier transform-infrared spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Our presentation will focus on continuing efforts to examine secondary cell wall development in cotton fibers using infrared Spectroscopy. Cotton fibers harvested at 18, 20, 24, 28, 32, 36 and 40 days after flowering were examined using attenuated total reflection Fourier transform-infrared (ATR FT-...

  17. Compact and Robust Refilling and Connectorization of Hollow Core Photonic Crystal Fiber Gas Reference Cells

    NASA Technical Reports Server (NTRS)

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

    2007-01-01

    A simple method for evacuating, refilling and connectorizing hollow-core photonic crystal fiber for use asgas reference cell is proposed and demonstrated. It relies on torch-sealing a quartz filling tube connected to amechanical splice between regular and hollow-core fibers.

  18. Topographical effects on fiber-mediated microRNA delivery to control oligodendroglial precursor cells development.

    PubMed

    Diao, Hua Jia; Low, Wei Ching; Lu, Q Richard; Chew, Sing Yian

    2015-11-01

    Effective remyelination in the central nervous system (CNS) facilitates the reversal of disability in patients with demyelinating diseases such as multiple sclerosis. Unfortunately until now, effective strategies of controlling oligodendrocyte (OL) differentiation and maturation remain limited. It is well known that topographical and biochemical signals play crucial roles in modulating cell fate commitment. Therefore, in this study, we explored the combined effects of scaffold topography and sustained gene silencing on oligodendroglial precursor cell (OPC) development. Specifically, microRNAs (miRs) were incorporated onto electrospun polycaprolactone (PCL) fiber scaffolds with different fiber diameters and orientations. Regardless of fiber diameter and orientation, efficient knockdown of differentiation inhibitory factors were achieved by either topography alone (up to 70%) or fibers integrated with miR-219 and miR-338 (up to 80%, p < 0.05). Small fiber promoted OPC differentiation by inducing more RIP(+) cells (p < 0.05) while large fiber promoted OL maturation by inducing more MBP(+) cells (p < 0.05). Random fiber enhanced more RIP(+) cells than aligned fibers (p < 0.05), regardless of fiber diameter. Upon miR-219/miR-338 incorporation, 2 μm aligned fibers supported the most MBP(+) cells (∼17%). These findings indicated that the coupling of substrate topographic cues with efficient gene silencing by sustained microRNA delivery is a promising way for directing OPC maturation in neural tissue engineering and controlling remyelination in the CNS. PMID:26310106

  19. Cell Attachment and Viability Study of PCL Nano-fiber Modified by Cold Atmospheric Plasma.

    PubMed

    Atyabi, Seyed Mohammad; Sharifi, Fereshteh; Irani, Shiva; Zandi, Mojgan; Mivehchi, Houri; Nagheh, Zahra

    2016-06-01

    The field of tissue engineering is an emerging discipline which applies the basic principles of life sciences and engineering to repair and restore living tissues and organs. The purpose of this study was to investigate the effect of cold and non-thermal plasma surface modification of poly (ϵ-caprolactone) (PCL) scaffolds on fibroblast cell behavior. Nano-fiber PCL was fabricated through electrospinning technique, and some fibers were then treated by cold and non-thermal plasma. The cell-biomaterial interactions were studied by culturing the fibroblast cells on nano-fiber PCL. Scaffold biocompatibility test was assessed using an inverted microscope. The growth and proliferation of fibroblast cells on nano-fiber PCL were analyzed by MTT viability assay. Cellular attachment on the nano-fiber and their morphology were evaluated using scanning electron microscope. The result of cell culture showed that nano-fiber could support the cellular growth and proliferation by developing three-dimensional topography. The present study demonstrated that the nano-fiber surface modification with cold plasma sharply enhanced the fibroblast cell attachment. Thus, cold plasma surface modification greatly raised the bioactivity of scaffolds. PMID:27286857

  20. Excimer laser channel creation in polyethersulfone hollow fibers for compartmentalized in vitro neuronal cell culture scaffolds.

    PubMed

    Brayfield, Candace A; Marra, Kacey G; Leonard, John P; Tracy Cui, X; Gerlach, Jörg C

    2008-03-01

    Hollow fiber scaffolds that compartmentalize axonal processes from their cell bodies can enable neuronal cultures with directed neurite outgrowth within a three-dimensional (3-D) space for controlling neuronal cell networking in vitro. Controllable 3-D neuronal networks in vitro could provide tools for studying neurobiological events. In order to create such a scaffold, polyethersulfone (PES) microporous hollow fibers were ablated with a KrF excimer laser to generate specifically designed channels for directing neurite outgrowth into the luminal compartments of the fibers. Excimer laser modification is demonstrated as a reproducible method to generate 5microm diameter channels within PES hollow fiber walls that allow compartmentalization of neuronal cell bodies from their axons. Laser modification of counterpart flat sheet PES membranes with peak surface fluences of 1.2Jcm(-2) results in increased hydrophobicity and laminin adsorption on the surface compared with the unmodified PES surface. This is correlated to enhanced PC12 cell adhesion with increasing fluence onto laser-modified PES membrane surfaces coated with laminin when compared with unmodified surfaces. Adult rat neural progenitor cells differentiated on PES fibers with laser-created channels resulted in spontaneous cell process growth into the channels of the scaffold wall while preventing entrance of cell bodies. Therefore, laser-modified PES fibers serve as scaffolds with channels conducive to directing neuronal cell process growth. These hollow fiber scaffolds can potentially be used in combination with perfusion and oxygenation hollow fiber membrane sets to construct a hollow fiber-based 3-D bioreactor for controlling and studying in vitro neuronal networking in three dimensions between compartmentalized cultures. PMID:18060849

  1. Human induced pluripotent stem cell-derived fiber-shaped cardiac tissue on a chip.

    PubMed

    Morimoto, Y; Mori, S; Sakai, F; Takeuchi, S

    2016-06-21

    We propose a method for the production of a fiber-shaped three-dimensional (3D) cellular construct of human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) for the quantification of the contractile force. By culturing the cardiomyocytes in a patterned hydrogel structure with fixed edges, we succeeded in fabricating hiPS-CM fibers with aligned cardiomyocytes. The fiber generated contractile force along the fiber direction due to the hiPS-CM alignment, and we were able to measure its contractile force accurately. Furthermore, to demonstrate the drug reactivity of hiPS-CM fibers, the changes in the contractile frequency and force following treatment with isoproterenol and propranolol were observed. We believe that hiPS-CM fibers will be a useful tool for pharmacokinetic analyses during drug development. PMID:27217209

  2. Prismatic sealed nickel-cadmium batteries utilizing fiber structured electrodes. I - New advances in cell design

    NASA Astrophysics Data System (ADS)

    Haschka, Friedrich; Benczur-Urmossy, Gabor; Anderman, Menahem

    Prismatic sealed Ni/Cd cells of fiber structured electrodes offer the potential to fully recharge a battery in a uniquely short time. It was demonstrated that the cells show excellent cycle life. The design is not restricted to 20 Ah rated capacity. Cells of 50 Ah have been built and tested in an electric hybrid vehicle. A specially designed ultra high-power cell of 45 Ah rated capacity for APU cranking in commerical aircraft supplies 50 percent more peak power than vented Ni/Cd sintered plate aircraft cells. The fiber structured sealed FNC-RECOM cell will not require any maintenance.

  3. The Hagfish Gland Thread Cell: A Fiber-Producing Cell Involved in Predator Defense

    PubMed Central

    Fudge, Douglas S.; Schorno, Sarah

    2016-01-01

    Fibers are ubiquitous in biology, and include tensile materials produced by specialized glands (such as silks), extracellular fibrils that reinforce exoskeletons and connective tissues (such as chitin and collagen), as well as intracellular filaments that make up the metazoan cytoskeleton (such as F-actin, microtubules, and intermediate filaments). Hagfish gland thread cells are unique in that they produce a high aspect ratio fiber from cytoskeletal building blocks within the confines of their cytoplasm. These threads are elaborately coiled into structures that readily unravel when they are ejected into seawater from the slime glands. In this review we summarize what is currently known about the structure and function of gland thread cells and we speculate about the mechanism that these cells use to produce a mechanically robust fiber that is almost one hundred thousand times longer than it is wide. We propose that a key feature of this mechanism involves the unidirectional rotation of the cell’s nucleus, which would serve to twist disorganized filaments into a coherent thread and impart a torsional stress on the thread that would both facilitate coiling and drive energetic unravelling in seawater. PMID:27258313

  4. Delineating the glycoproteome of elongating cotton fiber cells

    PubMed Central

    Kumar, Saravanan; Pandey, Pankaj; Kumar, Krishan; Rajamani, Vijayalakshmi; Padmalatha, Kethireddy Venkata; Dhandapani, Gurusamy; Kanakachari, Mogilicherla; Leelavathi, Sadhu; Kumar, Polumetla Ananda; Reddy, Vanga Siva

    2015-01-01

    The data presented here delineates the glycoproteome component in the elongating cotton fiber cells attained using complementary proteomic approaches followed by protein and N-linked glycosylation site identification (Kumar et al., 2013) [1]. Utilizing species specific protein sequence databases in proteomic approaches often leads to additional information that may not be obtained using cross-species databases. In this context we have reanalyzed our glycoproteome dataset with the Gossypium arboreum, Gossypium raimondii (version 2.0) and Gossypium hirsutum protein databases that has led to the identification of 21 N-linked glycosylation sites and 18 unique glycoproteins that were not reported in our previous study. The 1D PAGE and solution based glycoprotein identification data is publicly available at the ProteomeXchange Consortium via the PRIDE partner repository (Vizcaíno et al., 2013) [2] using the dataset identifier PXD000178 and the 2D PAGE based protein identification and glycopeptide approach based N-linked glycosylation site identification data is available at the ProteomeXchange Consortium via the PRIDE partner repository (Vizcaíno et al., 2013) [2] using the dataset identifier PXD002849. PMID:26693171

  5. Fiber biology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cotton fiber cells arising from seed epidermis is the most important agricultural textile commodity in the world. To produce fully mature fibers, approximately two months of fiber developmental process are required. The timing of four distinctive fiber development stages consisting of initiation, ...

  6. Fiber-optic Raman sensing of cell proliferation probes and molecular vibrations: Brain-imaging perspective

    NASA Astrophysics Data System (ADS)

    Doronina-Amitonova, Lyubov V.; Fedotov, Il'ya V.; Ivashkina, Olga I.; Zots, Marina A.; Fedotov, Andrei B.; Anokhin, Konstantin V.; Zheltikov, Aleksei M.

    2012-09-01

    Optical fibers are employed to sense fingerprint molecular vibrations in ex vivo experiments on the whole brain and detect cell proliferation probes in a model study on a quantitatively controlled solution. A specifically adapted spectral filtering procedure is shown to allow the Raman signal from molecular vibrations of interest to be discriminated against the background from the fiber, allowing a highly sensitive Raman detection of the recently demonstrated EdU (5-ethynyl-2'-deoxyuridine) labels of DNA synthesis in cells.

  7. Cell shape, spreading symmetry, and the polarization of stress-fibers in cells

    NASA Astrophysics Data System (ADS)

    Zemel, A.; Rehfeldt, F.; Brown, A. E. X.; Discher, D. E.; Safran, S. A.

    2010-05-01

    The active regulation of cellular forces during cell adhesion plays an important role in the determination of cell size, shape, and internal structure. While on flat, homogeneous and isotropic substrates some cells spread isotropically, others spread anisotropically and assume elongated structures. In addition, in their native environment as well as in vitro experiments, the cell shape and spreading asymmetry can be modulated by the local distribution of adhesive molecules and topography of the environment. We present a simple elastic model and experiments on stem cells to explain the variation of cell size with the matrix rigidity. In addition, we predict the experimental consequences of two mechanisms of acto-myosin polarization and focus here on the effect of the cell spreading asymmetry on the regulation of the stress-fiber alignment in the cytoskeleton. We show that when cell spreading is sufficiently asymmetric the alignment of acto-myosin forces in the cell increases monotonically with the matrix rigidity; however, in general this alignment is non-monotonic, as shown previously. These results highlight the importance of the symmetry characteristics of cell spreading in the regulation of cytoskeleton structure and suggest a mechanism by which different cell types may acquire different morphologies and internal structures in different mechanical environments.

  8. Removing PAH`s with cells on fibers

    SciTech Connect

    Clyde, R.

    1996-12-31

    There are over 1,500 sites contaminated with polycyclic aromatic hydrocarbons from coal gas plants. White rot fungi degrade PAH`s in soil, but the problem is to supply oxygen needed for growth of the fungus. When old cardboard boxes are buried with the fungus, oxygen is entrapped in the corrugations. A method for growing the fungus quickly is also described. Pseudomonade also degrade PAH and several strains of this bacterium have been grown on fibers. The fibers have high area, and when Celite is entrapped in the fibers, more area is provided.

  9. Porous, single crystalline titanium nitride nanoplates grown on carbon fibers: excellent counter electrodes for low-cost, high performance, fiber-shaped dye-sensitized solar cells.

    PubMed

    Chen, Liang; Dai, Hui; Zhou, Yong; Hu, Yingjie; Yu, Tao; Liu, Jianguo; Zou, Zhigang

    2014-11-28

    An excellent, platinum free fiber counter electrode (CE) was successfully fabricated, consisting of porous, single crystalline titanium nitride (TiN) nanoplates grown on carbon fibers (CF). The fiber-shaped dye-sensitized solar cells (FDSSCs) based on the TiN-CF CE show a high conversion efficiency of 7.20%, comparable or even superior to that of the Pt wire (6.23%). PMID:25068835

  10. Heteromannan and Heteroxylan Cell Wall Polysaccharides Display Different Dynamics During the Elongation and Secondary Cell Wall Deposition Phases of Cotton Fiber Cell Development.

    PubMed

    Hernandez-Gomez, Mercedes C; Runavot, Jean-Luc; Guo, Xiaoyuan; Bourot, Stéphane; Benians, Thomas A S; Willats, William G T; Meulewaeter, Frank; Knox, J Paul

    2015-09-01

    The roles of non-cellulosic polysaccharides in cotton fiber development are poorly understood. Combining glycan microarrays and in situ analyses with monoclonal antibodies, polysaccharide linkage analyses and transcript profiling, the occurrence of heteromannan and heteroxylan polysaccharides and related genes in developing and mature cotton (Gossypium spp.) fibers has been determined. Comparative analyses on cotton fibers at selected days post-anthesis indicate different temporal and spatial regulation of heteromannan and heteroxylan during fiber development. The LM21 heteromannan epitope was more abundant during the fiber elongation phase and localized mainly in the primary cell wall. In contrast, the AX1 heteroxylan epitope occurred at the transition phase and during secondary cell wall deposition, and localized in both the primary and the secondary cell walls of the cotton fiber. These developmental dynamics were supported by transcript profiling of biosynthetic genes. Whereas our data suggest a role for heteromannan in fiber elongation, heteroxylan is likely to be involved in the regulation of cellulose deposition of secondary cell walls. In addition, the relative abundance of these epitopes during fiber development varied between cotton lines with contrasting fiber characteristics from four species (G. hirsutum, G. barbadense, G. arboreum and G. herbaceum), suggesting that these non-cellulosic polysaccharides may be involved in determining final fiber quality and suitability for industrial processing. PMID:26187898

  11. Heteromannan and Heteroxylan Cell Wall Polysaccharides Display Different Dynamics During the Elongation and Secondary Cell Wall Deposition Phases of Cotton Fiber Cell Development

    PubMed Central

    Hernandez-Gomez, Mercedes C.; Runavot, Jean-Luc; Guo, Xiaoyuan; Bourot, Stéphane; Benians, Thomas A.S.; Willats, William G.T.; Meulewaeter, Frank; Knox, J. Paul

    2015-01-01

    The roles of non-cellulosic polysaccharides in cotton fiber development are poorly understood. Combining glycan microarrays and in situ analyses with monoclonal antibodies, polysaccharide linkage analyses and transcript profiling, the occurrence of heteromannan and heteroxylan polysaccharides and related genes in developing and mature cotton (Gossypium spp.) fibers has been determined. Comparative analyses on cotton fibers at selected days post-anthesis indicate different temporal and spatial regulation of heteromannan and heteroxylan during fiber development. The LM21 heteromannan epitope was more abundant during the fiber elongation phase and localized mainly in the primary cell wall. In contrast, the AX1 heteroxylan epitope occurred at the transition phase and during secondary cell wall deposition, and localized in both the primary and the secondary cell walls of the cotton fiber. These developmental dynamics were supported by transcript profiling of biosynthetic genes. Whereas our data suggest a role for heteromannan in fiber elongation, heteroxylan is likely to be involved in the regulation of cellulose deposition of secondary cell walls. In addition, the relative abundance of these epitopes during fiber development varied between cotton lines with contrasting fiber characteristics from four species (G. hirsutum, G. barbadense, G. arboreum and G. herbaceum), suggesting that these non-cellulosic polysaccharides may be involved in determining final fiber quality and suitability for industrial processing. PMID:26187898

  12. Dynamic changes in stress fiber expression in rat uterine vein endothelial cells associated with pregnancy.

    PubMed

    Sago, H; Sugimoto, K; Fujii, S; Iinuma, K; Yamashita, K; Kitagawa, M; Terashima, Y

    1993-09-01

    En face endothelial preparations of rat uterine vein were stained with rhodamine-phalloidin to investigate the dynamics of stress fiber expression during pregnancy. In prepregnant animals, somewhat plump, spindle-like endothelial cells of the uterine vein had only a few short stress fibers. With the progress of pregnancy, however, many long stress fibers appeared within the elongated endothelial cells. Within 2 hr after delivery, these stress fibers became dramatically decreased in number as the cells reverted from an elongated to a plump shape and returned to the prepregnancy level by 14 days postpartum. The uterine vein showed a significant increase in length during pregnancy and quickly shortened after delivery. Thus, expression of stress fibers in endothelial cells of the uterine vein seems to be related to the tension loaded on this vessel during its elongation in parallel with the marked growth of the uterine body during pregnancy. This study shows that stress fibers are dynamic structures that may serve to maintain endothelial cell integrity during the exertion of tensile stress on the vessel wall. PMID:8246817

  13. Unique and Analogous Functions of Aquaporin 0 for Fiber Cell Architecture and Ocular Lens Transparency

    PubMed Central

    Kumari, S. Sindhu; Eswaramoorthy, Subramaniam; Mathias, Richard T.; Varadaraj, Kulandaiappan

    2011-01-01

    Aquaporin (AQP) 1 and AQP0 water channels are expressed in lens epithelial and fiber cells, respectively, facilitating fluid circulation for nourishing the avascular lens to maintain transparency. Even though AQP0 water permeability is 40-fold less than AQP1, AQP0 is selectively expressed in the fibers. Delimited AQP0 fiber expression is attributed to a unique structural role as an adhesion protein. To validate this notion, we determined if wild type (WT) lens ultrastructure and fiber cell adhesion are different in AQP0−/−, and TgAQP1+/+/AQP0−/− mice that transgenically express AQP1 (TgAQP1) in fiber cells without AQP0 (AQP0−/−). In WT, lenses were transparent with ‘Y’ sutures. Fibers contained opposite end curvature, lateral interdigitations and hexagonal shape, and were arranged as concentric growth shells. AQP0−/− lenses were cataractous, lacked ‘Y’ sutures, ordered packing and well-defined lateral interdigitations. TgAQP1+/+/AQP0−/− lenses showed improvement in transparency and lateral interdigitations in the outer cortex while inner cortex and nuclear fibers were severely disintegrated. Transmission electron micrographs exhibited tightly packed fiber cells in WT whereas AQP0−/− and TgAQP1+/+/AQP0−/− lenses had wide extracellular spaces. Fibers were easily separable by teasing in AQP0−/− and TgAQP1+/+/AQP0−/− lenses compared to WT. Our data suggest that the increased water permeability through AQP1 does not compensate for loss of AQP0 expression in TgAQP1+/+/AQP0−/− mice. Fiber cell AQP0 expression is required to maintain their organization, which is a requisite for lens transparency. AQP0 appears necessary for cell-to-cell adhesion and thereby to minimize light scattering since in the AQP0−/− and TgAQP1+/+/AQP0−/− lenses, fiber cell disorganization was evident. PMID:21511033

  14. Progress in the development of the hollow fiber sodium-sulfur secondary cell

    NASA Technical Reports Server (NTRS)

    Levine, C. A.

    1975-01-01

    This report describes the development and status of the sodium-sulfur secondary cell which uses fine hollow glass fibers as the electrolyte. Laboratory size cells containing up to 7000 fibers and having capacities up to 5 ampere-hours have been built and operated. These cells have been run at various cycle depths up to 95% of capacity. Lifetime does not depend on depth of discharge up to at least 50% depth and possibly deeper. Rates of charge and discharge of the nominally one hour cells have been varied from three times the design rate to 0.05 times the design rate. Smaller cells operate with essentially no internal resistance increase during their lifetimes of over four months on continuous charge/discharge at the one hour rate. Larger cells assembled with somewhat different mechanical assembly techniques have shorter lives. Two types of failure modes are observed: progressive weakening and breaking of the fibers inside the cell assembly, and fiber breakage at the fiber/tube sheet interface.

  15. Doxazosin treatment alters stromal cell behavior and increases elastic system fibers deposition in rat prostate.

    PubMed

    Delella, Flávia Karina; Felisbino, Sérgio Luis

    2010-10-01

    Doxazosin (DOX), an α-adrenoceptor antagonist, induces the relaxation of smooth muscle cell tonus and reduces the clinical symptoms of benign prostatic hyperplasia (BPH). However, the effects of DOX in the prostate stromal microenvironment are not fully known. In a previous study, we showed that DOX treatment for 30 days increased deposition of collagen fibers in the three rat prostatic lobes. Herein, we investigated the effects of DOX on stromal cell ultrastructure and elastic fiber deposition. Adult Wistar rats were treated with DOX (25 mg/kg/day); and the ventral, dorsal, and anterior prostates were excised at 30 days of treatment. The prostatic lobes were submitted to histochemical and stereological-morphometric analyze and transmission electron microscopy (TEM). Histochemical staining plus stereological analysis of the elastic fiber system showed that DOX-treated prostatic lobes presented more elaunin and elastic fibers than controls, mainly in the ventral lobe. Ultrastructural analysis showed that fibroblasts and smooth muscle cells from DOX-treated prostates presented active synthetic phenotypes, evidenced by enlarged rough endoplasmic reticulum and Golgi apparatus cisterns, and confirmed the observation of thickened elaunin fibers. Our findings suggest that, under α-adrenergic blockade by DOX, the fibroblasts become more active and smooth muscle cells shift from a predominantly contractile to a more synthetic phenotype. The deposition of collagen and elastic system fibers in the prostatic stroma may counterbalance the absence of smooth muscle tone during α-blockers treatment. PMID:20155861

  16. Effects of Aramid, a high strength synthetic fiber, on respiratory cells in vitro.

    PubMed

    Marsh, J P; Mossman, B T; Driscoll, K E; Schins, R F; Borm, P J

    1994-01-01

    Industry continues to develop synthetic fibers for new technologies and as replacements for asbestos, a toxic and carcinogenic fiber. To determine whether the in vitro effects of the aromatic polyamide fiber, Aramid (Kevlar, Twaron), resembled those induced by asbestos, fibers were surveyed for (1) cytotoxicity as measured by total cell protein, and (2) proliferative capacity as measured by [3H]thymidine incorporation, colony forming efficiency (CFE), and ornithine decarboxylase (ODC) activity in two target cells of mineral dust-induced lung damage, hamster tracheal epithelial (HTE) cells and rat lung (RL90) fibroblasts. Results of cytotoxicity tests indicated that Aramid was as toxic to HTE and RL90 cells as were crocidolite and chrysotile asbestos when expressed on both an equal mass and equal fiber number basis. In HTE cells, Aramid caused a statistically significant increase in [3H]thymidine incorporation and CFE and produced a dose-dependent induction of ODC enzyme activity. Proliferative effects by asbestos or Aramid were not observed in RL90 fibroblasts. Thus, when tested over a respirable size range, Aramid exhibited many of the same effects on epithelial cells in vitro as did asbestos, including increased radiolabeled nucleotide incorporation into DNA and induction of ODC enzyme activity. PMID:8062644

  17. Noradrenergic modulation of the parallel fiber-Purkinje cell synapse in mouse cerebellum.

    PubMed

    Lippiello, Pellegrino; Hoxha, Eriola; Volpicelli, Floriana; Lo Duca, Giuseppina; Tempia, Filippo; Miniaci, Maria Concetta

    2015-02-01

    The signals arriving to Purkinje cells via parallel fibers are essential for all tasks in which the cerebellum is involved, including motor control, learning new motor skills and calibration of reflexes. Since learning also requires the activation of adrenergic receptors, we investigated the effects of adrenergic receptor agonists on the main plastic site of the cerebellar cortex, the parallel fiber-Purkinje cell synapse. Here we show that noradrenaline serves as an endogenous ligand for both α1-and α2-adrenergic receptors to produce synaptic depression between parallel fibers and Purkinje cells. On the contrary, PF-EPSCs were potentiated by the β-adrenergic receptor agonist isoproterenol. This short-term potentiation was postsynaptically expressed, required protein kinase A, and was mimicked by the β2-adrenoceptor agonist clenbuterol, suggesting that the β2-adrenoceptors mediate the noradrenergic facilitation of synaptic transmission between parallel fibers and Purkinje cells. Moreover, β-adrenoceptor activation lowered the threshold for cerebellar long-term potentiation induced by 1 Hz parallel fiber stimulation. The presence of both α and β adrenergic receptors on Purkinje cells suggests the existence of bidirectional mechanisms of regulation allowing the noradrenergic afferents to refine the signals arriving to Purkinje cells at particular arousal states or during learning. PMID:25218865

  18. Effect of dietary fibers on losartan uptake and transport in Caco-2 cells.

    PubMed

    Iwazaki, Ayano; Takahashi, Naho; Miyake, Reiko; Hiroshima, Yuka; Abe, Mariko; Yasui, Airi; Imai, Kimie

    2016-05-01

    The objective of this study was to assess the effect of dietary fibers on the transport of losartan, an angiotensin II type 1 receptor blocker, in small intestinal cells. Using Caco-2 cells in vitro, losartan uptake and transport were evaluated in the presence of various fibers (cellulose, chitosan, sodium alginate and glucomannan). Dietary fibers caused a decrease in the uptake of losartan, with chitosan causing a significant reduction. Chitosan and glucomannan significantly reduced the transport of losartan, while cellulose or sodium alginate did not. Dietary fibers also reduced the level of free losartan; however, this did not correlate with the observed reduction in losartan uptake and transport. In summary, chitosan had the greatest inhibitory effect on losartan uptake and transport, and this potential interaction should be considered in patients taking losartan. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26748460

  19. Three-dimensional hierarchical cultivation of human skin cells on bio-adaptive hybrid fibers.

    PubMed

    Planz, Viktoria; Seif, Salem; Atchison, Jennifer S; Vukosavljevic, Branko; Sparenberg, Lisa; Kroner, Elmar; Windbergs, Maike

    2016-07-11

    The human skin comprises a complex multi-scale layered structure with hierarchical organization of different cells within the extracellular matrix (ECM). This supportive fiber-reinforced structure provides a dynamically changing microenvironment with specific topographical, mechanical and biochemical cell recognition sites to facilitate cell attachment and proliferation. Current advances in developing artificial matrices for cultivation of human cells concentrate on surface functionalizing of biocompatible materials with different biomolecules like growth factors to enhance cell attachment. However, an often neglected aspect for efficient modulation of cell-matrix interactions is posed by the mechanical characteristics of such artificial matrices. To address this issue, we fabricated biocompatible hybrid fibers simulating the complex biomechanical characteristics of native ECM in human skin. Subsequently, we analyzed interactions of such fibers with human skin cells focusing on the identification of key fiber characteristics for optimized cell-matrix interactions. We successfully identified the mediating effect of bio-adaptive elasto-plastic stiffness paired with hydrophilic surface properties as key factors for cell attachment and proliferation, thus elucidating the synergistic role of these parameters to induce cellular responses. Co-cultivation of fibroblasts and keratinocytes on such fiber mats representing the specific cells in dermis and epidermis resulted in a hierarchical organization of dermal and epidermal tissue layers. In addition, terminal differentiation of keratinocytes at the air interface was observed. These findings provide valuable new insights into cell behaviour in three-dimensional structures and cell-material interactions which can be used for rational development of bio-inspired functional materials for advanced biomedical applications. PMID:27241237

  20. Polymeric optical fiber tweezers as a tool for single cell micro manipulation and sensing

    NASA Astrophysics Data System (ADS)

    Rodrigues Ribeiro, R. S.; Soppera, O.; Guerreiro, A.; Jorge, P. A...

    2015-09-01

    In this paper a new type of polymeric fiber optic tweezers for single cell manipulation is reported. The optical trapping of a yeast cell using a polymeric micro lens fabricated by guided photo polymerization at the fiber tip is demonstrated. The 2D trapping of the yeast cells is analyzed and maximum optical forces on the pN range are calculated. The experimental results are supported by computational simulations using a FDTD method. Moreover, new insights on the potential for simultaneous sensing and optical trapping, are presented.

  1. Processing and properties of multiscale cellular thermoplastic fiber reinforced composite (CellFRC)

    NASA Astrophysics Data System (ADS)

    Sorrentino, L.; Cafiero, L.; D'Auria, M.; Iannace, S.

    2015-12-01

    High performance fiber reinforced polymer composites are made by embedding high strength/modulus fibers in a polymeric matrix. They are a class of materials that owe its success to the impressive specific mechanical properties with respect to metals. In many weight-sensitive applications, where high mechanical properties and low mass are required, properties per unit of mass are more important than absolute properties and further weight reduction is desirable. A route to reach this goal could be the controlled induction of porosity into the polymeric matrix, while still ensuring load transfer to the reinforcing fibers and fiber protection from the environment. Cellular lightweight fiber reinforced composites (CellFRC) were prepared embedding gas bubbles of controlled size within a high performance thermoplastic matrix reinforced with continuous fibers. Pores were induced after the composite was first saturated with CO2 and then foamed by using an in situ foaming/shaping technology based on compression moulding with adjustable mould cavities. The presence of micro- or submicro-sized cells in the new CellFRC reduced the apparent density of the structure and led to significant improvements of its impact properties. Both structural and functional performances were further improved through the use of a platelet-like nanofiller (Expanded Graphite) dispersed into the matrix.

  2. Cotton fiber tips have diverse morphologies and show evidence of apical cell wall synthesis.

    PubMed

    Stiff, Michael R; Haigler, Candace H

    2016-01-01

    Cotton fibers arise through highly anisotropic expansion of a single seed epidermal cell. We obtained evidence that apical cell wall synthesis occurs through examining the tips of young elongating Gossypium hirsutum (Gh) and G. barbadense (Gb) fibers. We characterized two tip types in Gh fiber (hemisphere and tapered), each with distinct apical diameter, central vacuole location, and distribution of cell wall components. The apex of Gh hemisphere tips was enriched in homogalacturonan epitopes, including a relatively high methyl-esterified form associated with cell wall pliability. Other wall components increased behind the apex including cellulose and the α-Fuc-(1,2)-β-Gal epitope predominantly found in xyloglucan. Gb fibers had only one narrow tip type featuring characters found in each Gh tip type. Pulse-labeling of cell wall glucans indicated wall synthesis at the apex of both Gh tip types and in distal zones. Living Gh hemisphere and Gb tips ruptured preferentially at the apex upon treatment with wall degrading enzymes, consistent with newly synthesized wall at the apex. Gh tapered tips ruptured either at the apex or distantly. Overall, the results reveal diverse cotton fiber tip morphologies and support primary wall synthesis occurring at the apex and discrete distal regions of the tip. PMID:27301434

  3. Cotton fiber tips have diverse morphologies and show evidence of apical cell wall synthesis

    PubMed Central

    Stiff , Michael R.; Haigler, Candace H.

    2016-01-01

    Cotton fibers arise through highly anisotropic expansion of a single seed epidermal cell. We obtained evidence that apical cell wall synthesis occurs through examining the tips of young elongating Gossypium hirsutum (Gh) and G. barbadense (Gb) fibers. We characterized two tip types in Gh fiber (hemisphere and tapered), each with distinct apical diameter, central vacuole location, and distribution of cell wall components. The apex of Gh hemisphere tips was enriched in homogalacturonan epitopes, including a relatively high methyl-esterified form associated with cell wall pliability. Other wall components increased behind the apex including cellulose and the α-Fuc-(1,2)-β-Gal epitope predominantly found in xyloglucan. Gb fibers had only one narrow tip type featuring characters found in each Gh tip type. Pulse-labeling of cell wall glucans indicated wall synthesis at the apex of both Gh tip types and in distal zones. Living Gh hemisphere and Gb tips ruptured preferentially at the apex upon treatment with wall degrading enzymes, consistent with newly synthesized wall at the apex. Gh tapered tips ruptured either at the apex or distantly. Overall, the results reveal diverse cotton fiber tip morphologies and support primary wall synthesis occurring at the apex and discrete distal regions of the tip. PMID:27301434

  4. Neighboring Parenchyma Cells Contribute to Arabidopsis Xylem Lignification, while Lignification of Interfascicular Fibers Is Cell Autonomous[W

    PubMed Central

    Smith, Rebecca A.; Schuetz, Mathias; Roach, Melissa; Mansfield, Shawn D.; Ellis, Brian; Samuels, Lacey

    2013-01-01

    Lignin is a critical structural component of plants, providing vascular integrity and mechanical strength. Lignin precursors (monolignols) must be exported to the extracellular matrix where random oxidative coupling produces a complex lignin polymer. The objectives of this study were twofold: to determine the timing of lignification with respect to programmed cell death and to test if nonlignifying xylary parenchyma cells can contribute to the lignification of tracheary elements and fibers. This study demonstrates that lignin deposition is not exclusively a postmortem event, but also occurs prior to programmed cell death. Radiolabeled monolignols were not detected in the cytoplasm or vacuoles of tracheary elements or neighbors. To experimentally define which cells in lignifying tissues contribute to lignification in intact plants, a microRNA against CINNAMOYL CoA-REDUCTASE1 driven by the promoter from CELLULOSE SYNTHASE7 (ProCESA7:miRNA CCR1) was used to silence monolignol biosynthesis specifically in cells developing lignified secondary cell walls. When monolignol biosynthesis in ProCESA7:miRNA CCR1 lines was silenced in the lignifying cells themselves, but not in the neighboring cells, lignin was still deposited in the xylem secondary cell walls. Surprisingly, a dramatic reduction in cell wall lignification of extraxylary fiber cells demonstrates that extraxylary fibers undergo cell autonomous lignification. PMID:24096341

  5. High efficiency, broadband solar cell architectures based on arrays of volumetrically distributed narrowband photovoltaic fibers.

    PubMed

    O'Connor, Brendan; Nothern, Denis; Pipe, Kevin P; Shtein, Max

    2010-09-13

    We propose a novel solar cell architecture consisting of multiple fiber-based photovoltaic (PV) cells. Each PV fiber element is designed to maximize the power conversion efficiency within a narrow band of the incident solar spectrum, while reflecting other spectral components through the use of optical microcavity effects and distributed Bragg reflector (DBR) coatings. Combining PV fibers with complementary absorption and reflection characteristics into volume-filling arrays enables spectrally tuned modules having an effective dispersion element intrinsic to the architecture, resulting in high external quantum efficiency over the incident spectrum. While this new reflective tandem architecture is not limited to one particular material system, here we apply the concept to organic PV (OPV) cells that use a metal-organic-metal-dielectric layer structure, and calculate the expected performance of such arrays. Using realistic material properties for organic absorbers, transport layers, metallic electrodes, and DBR coatings, 17% power conversion efficiency can be reached. PMID:21165073

  6. Modulation of Dendritic-Epithelial Cell Responses against Sphingomonas Paucimobilis by Dietary Fibers.

    PubMed

    Bermudez-Brito, Miriam; Faas, Marijke M; de Vos, Paul

    2016-01-01

    Non-fermenting Gram-negative bacilli, such as Sphingomonas paucimobilis (S.paucimobilis), are among the most widespread causes of nosocomial infections. Up to now, no definitive guidelines exist for antimicrobial therapy for S. paucimobilis infections. As we have shown that some dietary fibers exhibit pronounced immune-regulatory properties, we hypothesized that specific immune active dietary fibers might modulate the responses against S. paucimobilis. We studied the immunomodulatory effects of dietary fibers against S. paucimobilis on cytokine release and maturation of human dendritic cells (DCs) in co-cultures of DCs and intestinal epithelial cells (IECs). S. paucimobilis infection resulted in increased release of pro-inflammatory cytokines and chemokines by DCs/IECs; these effects were strongly attenuated by specific dietary fibers. Chicory inulin, sugar beet pectin, and both starches had the strongest regulatory effects. IL-12 and TNF-α were drastically diminished upon exposure to chicory inulin and sugar beet pectin, or both starches. High-maize 260, was more effective in the reduction of chemokine release than the others fibers tested. In summary, chicory inulin, sugar beet pectin, High-maize 260, and Novelose 330 attenuate S. paucimobilis-induced cytokines. These results demonstrate that dietary fibers with a specific chemical composition can be used to manage immune responses against pathogens such as S. paucimobilis. PMID:27452116

  7. Spider silk fibers spun from soluble recombinant silk produced in mammalian cells.

    PubMed

    Lazaris, Anthoula; Arcidiacono, Steven; Huang, Yue; Zhou, Jiang-Feng; Duguay, Francois; Chretien, Nathalie; Welsh, Elizabeth A; Soares, Jason W; Karatzas, Costas N

    2002-01-18

    Spider silks are protein-based "biopolymer" filaments or threads secreted by specialized epithelial cells as concentrated soluble precursors of highly repetitive primary sequences. Spider dragline silk is a flexible, lightweight fiber of extraordinary strength and toughness comparable to that of synthetic high-performance fibers. We sought to "biomimic" the process of spider silk production by expressing in mammalian cells the dragline silk genes (ADF-3/MaSpII and MaSpI) of two spider species. We produced soluble recombinant (rc)-dragline silk proteins with molecular masses of 60 to 140 kilodaltons. We demonstrated the wet spinning of silk monofilaments spun from a concentrated aqueous solution of soluble rc-spider silk protein (ADF-3; 60 kilodaltons) under modest shear and coagulation conditions. The spun fibers were water insoluble with a fine diameter (10 to 40 micrometers) and exhibited toughness and modulus values comparable to those of native dragline silks but with lower tenacity. Dope solutions with rc-silk protein concentrations >20% and postspinning draw were necessary to achieve improved mechanical properties of the spun fibers. Fiber properties correlated with finer fiber diameter and increased birefringence. PMID:11799236

  8. Modulation of Dendritic-Epithelial Cell Responses against Sphingomonas Paucimobilis by Dietary Fibers

    PubMed Central

    Bermudez-Brito, Miriam; Faas, Marijke M; de Vos, Paul

    2016-01-01

    Non-fermenting Gram-negative bacilli, such as Sphingomonas paucimobilis (S.paucimobilis), are among the most widespread causes of nosocomial infections. Up to now, no definitive guidelines exist for antimicrobial therapy for S. paucimobilis infections. As we have shown that some dietary fibers exhibit pronounced immune-regulatory properties, we hypothesized that specific immune active dietary fibers might modulate the responses against S. paucimobilis. We studied the immunomodulatory effects of dietary fibers against S. paucimobilis on cytokine release and maturation of human dendritic cells (DCs) in co-cultures of DCs and intestinal epithelial cells (IECs). S. paucimobilis infection resulted in increased release of pro-inflammatory cytokines and chemokines by DCs/IECs; these effects were strongly attenuated by specific dietary fibers. Chicory inulin, sugar beet pectin, and both starches had the strongest regulatory effects. IL-12 and TNF-α were drastically diminished upon exposure to chicory inulin and sugar beet pectin, or both starches. High-maize 260, was more effective in the reduction of chemokine release than the others fibers tested. In summary, chicory inulin, sugar beet pectin, High-maize 260, and Novelose 330 attenuate S. paucimobilis-induced cytokines. These results demonstrate that dietary fibers with a specific chemical composition can be used to manage immune responses against pathogens such as S. paucimobilis. PMID:27452116

  9. Screening and quantification of anticancer compounds in traditional Chinese medicine by hollow fiber cell fishing and hollow fiber liquid/solid-phase microextraction.

    PubMed

    Wang, Caiyun; Hu, Shuang; Chen, Xuan; Bai, Xiaohong

    2016-05-01

    Hollow fiber cell fishing, based on HepG-2, SKOV-3, and ACHN cancer cells, and hollow fiber liquid/solid microextraction with HPLC were developed and introduced for researching the anticancer activity of Rhizoma Curcumae Longae, Radix Curcumae, and Rhizoma Curcumae. The structures of curcumin, demethoxycurcumin, and bisdemethoxycurcumin screened were identified and their contents were determined. The compound target fishing factors and cell apoptosis rates under the effect of the three medicines were determined. The binding sites (cell membrane and cell organelle) and binding target (phospholipase C) on the cell were researched. Hollow fiber liquid/solid-phase microextraction mechanism was analyzed and expounded. Before the application, cell seeding time, growth state and survival rate, compound nonspecific binding, positive and negative controls, repeatability in hollow fiber cell fishing with high-performance liquid chromatography; extraction solvent, sample pH, salt concentration, agitation speed, extraction time, temperature and sample volume in hollow fiber liquid/solid-phase microextraction with high-performance liquid chromatography were investigated. The results demonstrated that the proposed strategy is a simple and quick method to identify bioactive compounds at the cellular level as well as determine their contents (particularly trace levels of the bioactive compounds), analyze multicompound and multitarget entirety effects, and elucidate the efficacious material base in traditional medicine. PMID:26987300

  10. Unique and analogous functions of aquaporin O for fiber cell architecture and ocular lens transparency

    SciTech Connect

    Kumari, S.S.; Eswaramoorthy, S.; Mathias, R. T.; Varadaraj, K.

    2011-09-01

    Aquaporin (AQP) 1 and AQP0 water channels are expressed in lens epithelial and fiber cells, respectively, facilitating fluid circulation for nourishing the avascular lens to maintain transparency. Even though AQP0 water permeability is 40-fold less than AQP1, AQP0 is selectively expressed in the fibers. Delimited AQP0 fiber expression is attributed to a unique structural role as an adhesion protein. To validate this notion, we determined if wild type (WT) lens ultrastructure and fibercell adhesion are different in AQP0{sup -/-}, and TgAQP1{sup +/+}/AQP0{sup -/-} mice that transgenically express AQP1 (TgAQP1) in fibercells without AQP0 (AQP0{sup -/-}). In WT, lenses were transparent with 'Y' sutures. Fibers contained opposite end curvature, lateral interdigitations, hexagonal shape, and were arranged as concentric growth shells. AQP0{sup -/-}lenses were cataractous, lacked 'Y' sutures, ordered packing and well-defined lateral interdigitations. TgAQP1{sup +/+}/AQP0{sup -/-} lenses showed improvement in transparency and lateral interdigitations in the outer cortex while inner cortex and nuclear fibers were severely disintegrated. Transmission electron micrographs exhibited tightly packed fibercells in WT whereas AQP0{sup -/-} and TgAQP1{sup +/+}/AQP0{sup -/-}lenses had wide extracellular spaces. Fibers were easily separable by teasing in AQP0{sup -/-} and TgAQP1{sup +/+}/AQP0{sup -/-}lenses compared to WT. Our data suggest that the increased water permeability through AQP1 does not compensate for loss of AQP0 expression in TgAQP1{sup +/+}/AQP0{sup -/-} mice. Fibercell AQP0 expression is required to maintain their organization, which is a requisite for lenstransparency. AQP0 appears necessary for cell-to-cell adhesion and thereby to minimize light scattering since in the AQP0{sup -/-} and TgAQP1{sup +/+}/AQP0{sup -/-} lenses, fiber cell disorganization was evident.

  11. Enhanced Transduction and Replication of RGD-Fiber Modified Adenovirus in Primary T Cells

    PubMed Central

    Sengupta, Sadhak; Ulasov, Ilya V.; Thaci, Bart; Ahmed, Atique U.; Lesniak, Maciej S.

    2011-01-01

    Background Adenoviruses are often used as vehicles to mediate gene delivery for therapeutic purposes, but their research scope in hematological cells remains limited due to a narrow choice of host cells that express the adenoviral receptor (CAR). T cells, which are attractive targets for gene therapy of numerous diseases, remain resistant to adenoviral infection because of the absence of CAR expression. Here, we demonstrate that this resistance can be overcome when murine or human T cells are transduced with an adenovirus incorporating the RGD-fiber modification (Ad-RGD). Methodology/Principal Finding A luciferase-expressing replication-deficient Ad-RGD infected 3-fold higher number of activated primary T cells than an adenovirus lacking the RGD-fiber modification in vitro. Infection with replication-competent Ad-RGD virus also caused increased cell cycling, higher E1A copy number and enriched hexon antigen expression in both human and murine T cells. Transduction with oncolytic Ad-RGD also resulted in higher titers of progeny virus and enhanced the killing of T cells. In vivo, 35–45% of splenic T cells were transduced by Ad-RGD. Conclusions Collectively, our results prove that a fiber modified Ad-RGD successfully transduces and replicates in primary T cells of both murine and human origin. PMID:21464908

  12. In Situ Liquid Cell Observations of Asbestos Fiber Diffusion in Water.

    PubMed

    Wu, Lei; Ortiz, Carlos; Xu, Ye; Willenbring, Jane; Jerolmack, Douglas

    2015-11-17

    We present real-time observations of the diffusion of individual asbestos fibers in water. We first scaled up a technique for fluorescent tagging and imaging of chrysotile asbestos fibers and prepared samples with a distribution of fiber lengths ranging from 1 to 20 μm. Experiments were then conducted by placing a 20, 100, or 150 ppm solution of these fibers in a liquid cell mounted on a spinning-disk confocal microscope. Using automated elliptical-particle detection methods, we determined the translation and rotation and two-dimensional (2D) trajectories of thousands of diffusing chrysotile fibers. We find that fiber diffusion is size-dependent and in reasonable agreement with theoretical predictions for the Brownian motion of rods. This agreement is remarkable given that experiments involved non-idealized particles at environmentally relevant concentrations in a confined cell, in which particle-particle and particle-wall interactions might be expected to cause deviations from theory. Experiments also confirmed that highly elongated chrysotile fibers exhibit anisotropic diffusion at short time scales, a predicted effect that may have consequences for aggregate formation and transport of asbestos in confined spaces. The examined fibers vary greatly in their lengths and were prepared from natural chrysotile. Our findings thus indicate that the diffusion rates of a wide range of natural colloidal particles can be predicted from theory, so long as the particle aspect ratio is properly taken into account. This is an important first step for understanding aggregate formation and transport of non-spherical contaminant particles, in the environment and in vivo. PMID:26461183

  13. KCC2 expression supersedes NKCC1 in mature fiber cells in mouse and rabbit lenses

    PubMed Central

    Kasinathan, Chinnaswamy

    2015-01-01

    Purpose Na-K-Cl cotransporter 1 (NKCC1) and K-Cl cotransporter 2 (KCC2) have fundamental roles in neuron differentiation that are integrated with gamma-aminobutyric acid (GABA) and glutamate receptors, GABA synthesized by GAD25/65/67 encoded by GAD1/GAD2 genes, and GABA transporters (GATs). Cells in the eye lens express at least 13 GABA receptor subunits, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl D-aspartate (NMDA) glutamate receptors, GAD1/GAD2, GAT1–4 and vGAT, and NKCC1. NKCC1:KCC2 ratios determine the switch in GABA actions from trophic/growth promoting early in development to their classic inhibitory roles in adult neurons. Lens epithelial cells cover the anterior surface and differentiate to elongated fiber cells in the lens interior with comparable morphology and sub-cellular structures as neurons. NKCC1 is expressed before KCC2 in neuron development and increases cell chloride, which stimulates differentiation and process formation. Subsequently, KCC2 increases and extrudes cell chloride linked with maturation. KCC2 has an additional structural moonlighting role interacting with F-actin scaffolding in dendritic spine morphogenesis. We examined KCC2 versus NKCC1 spatial expression in relation to fiber cell developmental status within the lens. Methods Immunofluorescence and immunoblots were used to detect expression in mouse and rabbit lenses. Results NKCC1 was restricted to peripheral elongating lens fiber cells in young adult mouse and rabbit lenses. Lens KCC2 expression included the major KCC2b neuronal isoform and was detected in interior fiber cells with decreased NKCC1 expression and localized at the membranes. Lens expression of RE-1 silencing transcription factor (REST) regulated KCC2 is consistent with GAD1 and GAD2, several GABA and glutamate receptor subunits, miR-124, and other REST-regulated genes expressed in lenses. Conclusions NKCC1 in peripheral elongating fiber cells is superseded by KCC2 expression in

  14. [INVITED] Cell sensing with near-infrared plasmonic optical fiber sensors

    NASA Astrophysics Data System (ADS)

    Caucheteur, Christophe; Malachovska, Viera; Ribaut, Clotilde; Wattiez, Ruddy

    2016-04-01

    Surface Plasmon resonance (SPR) optical fiber biosensors are a miniaturized counterpart to the bulky prism configuration that offer remote operation in very small volumes of analyte. They have the potential to yield in situ (or even possibly in vivo) molecular detection. They usually result from a gold-coated fiber segment for which the core-guided light is brought into contact with the surrounding medium. Recently, SPR excitation was achieved with tilted fiber Bragg gratings (TFBGs) photo-imprinted in the fiber core and surrounded by a thin gold layer. These gratings probe the surrounding medium with near-infrared narrowband (~100 pm linewidth) resonances, which enhances both the penetration depth of the evanescent field in the external medium and the wavelength resolution of the interrogation. They also constitute the unique configuration able to probe all the fiber cladding modes individually, with high Q-factors. We use these unique spectral features in this work to target and detect extracellular membrane receptors in native membranes of different human epithelial cell lines. A differential diagnosis has been demonstrated between two systems, a cell line with overexpressed membrane receptors (the positive control) and another cell line with a low number of these receptors (a negative control). Such results bring an important step towards the demonstration of in situ diagnosis.

  15. Fibers in the Extracellular Matrix Enable Long-Range Stress Transmission between Cells

    PubMed Central

    Ma, Xiaoyue; Schickel, Maureen E.; Stevenson, Mark D.; Sarang-Sieminski, Alisha L.; Gooch, Keith J.; Ghadiali, Samir N.; Hart, Richard T.

    2013-01-01

    Cells can sense, signal, and organize via mechanical forces. The ability of cells to mechanically sense and respond to the presence of other cells over relatively long distances (e.g., ∼100 μm, or ∼10 cell-diameters) across extracellular matrix (ECM) has been attributed to the strain-hardening behavior of the ECM. In this study, we explore an alternative hypothesis: the fibrous nature of the ECM makes long-range stress transmission possible and provides an important mechanism for long-range cell-cell mechanical signaling. To test this hypothesis, confocal reflectance microscopy was used to develop image-based finite-element models of stress transmission within fibroblast-seeded collagen gels. Models that account for the gel’s fibrous nature were compared with homogenous linear-elastic and strain-hardening models to investigate the mechanisms of stress propagation. Experimentally, cells were observed to compact the collagen gel and align collagen fibers between neighboring cells within 24 h. Finite-element analysis revealed that stresses generated by a centripetally contracting cell boundary are concentrated in the relatively stiff ECM fibers and are propagated farther in a fibrous matrix as compared to homogeneous linear elastic or strain-hardening materials. These results support the hypothesis that ECM fibers, especially aligned ones, play an important role in long-range stress transmission. PMID:23561517

  16. Fibers in the extracellular matrix enable long-range stress transmission between cells.

    PubMed

    Ma, Xiaoyue; Schickel, Maureen E; Stevenson, Mark D; Sarang-Sieminski, Alisha L; Gooch, Keith J; Ghadiali, Samir N; Hart, Richard T

    2013-04-01

    Cells can sense, signal, and organize via mechanical forces. The ability of cells to mechanically sense and respond to the presence of other cells over relatively long distances (e.g., ∼100 μm, or ∼10 cell-diameters) across extracellular matrix (ECM) has been attributed to the strain-hardening behavior of the ECM. In this study, we explore an alternative hypothesis: the fibrous nature of the ECM makes long-range stress transmission possible and provides an important mechanism for long-range cell-cell mechanical signaling. To test this hypothesis, confocal reflectance microscopy was used to develop image-based finite-element models of stress transmission within fibroblast-seeded collagen gels. Models that account for the gel's fibrous nature were compared with homogenous linear-elastic and strain-hardening models to investigate the mechanisms of stress propagation. Experimentally, cells were observed to compact the collagen gel and align collagen fibers between neighboring cells within 24 h. Finite-element analysis revealed that stresses generated by a centripetally contracting cell boundary are concentrated in the relatively stiff ECM fibers and are propagated farther in a fibrous matrix as compared to homogeneous linear elastic or strain-hardening materials. These results support the hypothesis that ECM fibers, especially aligned ones, play an important role in long-range stress transmission. PMID:23561517

  17. Fiber-optic control and thermometry of single-cell thermosensation logic.

    PubMed

    Fedotov, I V; Safronov, N A; Ermakova, Yu G; Matlashov, M E; Sidorov-Biryukov, D A; Fedotov, A B; Belousov, V V; Zheltikov, A M

    2015-01-01

    Thermal activation of transient receptor potential (TRP) cation channels is one of the most striking examples of temperature-controlled processes in cell biology. As the evidence indicating the fundamental role of such processes in thermosensation builds at a fast pace, adequately accurate tools that would allow heat receptor logic behind thermosensation to be examined on a single-cell level are in great demand. Here, we demonstrate a specifically designed fiber-optic probe that enables thermal activation with simultaneous online thermometry of individual cells expressing genetically encoded TRP channels. This probe integrates a fiber-optic tract for the delivery of laser light with a two-wire microwave transmission line. A diamond microcrystal fixed on the fiber tip is heated by laser radiation transmitted through the fiber, providing a local heating of a cell culture, enabling a well-controlled TRP-assisted thermal activation of cells. Online local temperature measurements are performed by using the temperature-dependent frequency shift of optically detected magnetic resonance, induced by coupling the microwave field, delivered by the microwave transmission line, to nitrogen--vacancy centers in the diamond microcrystal. Activation of TRP channels is verified by using genetically encoded fluorescence indicators, visualizing an increase in the calcium flow through activated TRP channels. PMID:26563494

  18. Fiber-optic control and thermometry of single-cell thermosensation logic

    PubMed Central

    Fedotov, I.V.; Safronov, N.A.; Ermakova, Yu.G.; Matlashov, M.E.; Sidorov-Biryukov, D.A.; Fedotov, A.B.; Belousov, V.V.; Zheltikov, A.M.

    2015-01-01

    Thermal activation of transient receptor potential (TRP) cation channels is one of the most striking examples of temperature-controlled processes in cell biology. As the evidence indicating the fundamental role of such processes in thermosensation builds at a fast pace, adequately accurate tools that would allow heat receptor logic behind thermosensation to be examined on a single-cell level are in great demand. Here, we demonstrate a specifically designed fiber-optic probe that enables thermal activation with simultaneous online thermometry of individual cells expressing genetically encoded TRP channels. This probe integrates a fiber-optic tract for the delivery of laser light with a two-wire microwave transmission line. A diamond microcrystal fixed on the fiber tip is heated by laser radiation transmitted through the fiber, providing a local heating of a cell culture, enabling a well-controlled TRP-assisted thermal activation of cells. Online local temperature measurements are performed by using the temperature-dependent frequency shift of optically detected magnetic resonance, induced by coupling the microwave field, delivered by the microwave transmission line, to nitrogen—vacancy centers in the diamond microcrystal. Activation of TRP channels is verified by using genetically encoded fluorescence indicators, visualizing an increase in the calcium flow through activated TRP channels. PMID:26563494

  19. Fiber-optic control and thermometry of single-cell thermosensation logic

    NASA Astrophysics Data System (ADS)

    Fedotov, I. V.; Safronov, N. A.; Ermakova, Yu. G.; Matlashov, M. E.; Sidorov-Biryukov, D. A.; Fedotov, A. B.; Belousov, V. V.; Zheltikov, A. M.

    2015-11-01

    Thermal activation of transient receptor potential (TRP) cation channels is one of the most striking examples of temperature-controlled processes in cell biology. As the evidence indicating the fundamental role of such processes in thermosensation builds at a fast pace, adequately accurate tools that would allow heat receptor logic behind thermosensation to be examined on a single-cell level are in great demand. Here, we demonstrate a specifically designed fiber-optic probe that enables thermal activation with simultaneous online thermometry of individual cells expressing genetically encoded TRP channels. This probe integrates a fiber-optic tract for the delivery of laser light with a two-wire microwave transmission line. A diamond microcrystal fixed on the fiber tip is heated by laser radiation transmitted through the fiber, providing a local heating of a cell culture, enabling a well-controlled TRP-assisted thermal activation of cells. Online local temperature measurements are performed by using the temperature-dependent frequency shift of optically detected magnetic resonance, induced by coupling the microwave field, delivered by the microwave transmission line, to nitrogen—vacancy centers in the diamond microcrystal. Activation of TRP channels is verified by using genetically encoded fluorescence indicators, visualizing an increase in the calcium flow through activated TRP channels.

  20. Dual membrane hollow fiber fuel cell and method of operating same

    NASA Technical Reports Server (NTRS)

    Ingham, J. D.; Lawson, D. D. (Inventor)

    1978-01-01

    A gaseous fuel cell is described which includes a pair of electrodes formed by open-ended, ion-exchange hollow fibers, each having a layer of metal catalyst deposited on the inner surface and large surface area current collectors such as braided metal mesh in contact with the metal catalyst layer. A fuel cell results when the electrodes are immersed in electrolytes and electrically connected. As hydrogen and oxygen flow through the bore of the fibers, oxidation and reduction reactions develop an electrical potential. Since the hollow fiber configuration provides large electrode area per unit volume and intimate contact between fuel and oxidizer at the interface, and due to the low internal resistance of the electrolyte, high power densities can be obtained.

  1. Extracellular Recordings of Patterned Human Pluripotent Stem Cell-Derived Cardiomyocytes on Aligned Fibers

    PubMed Central

    Minami, Itsunari; Yu, Leqian; Nakajima, Minako; Qiao, Jing; Shimono, Ken; Nakatsuji, Norio; Kotera, Hitetoshi; Chen, Yong

    2016-01-01

    Human induced pluripotent stem cell (hiPSC) derived cardiomyocytes (CMs) hold high potential for use in drug assessment and myocardial regeneration. To create tissue-like constructs of CMs for extracellular monitoring, we placed aligned fibers (AFs) on the surface of a microelectrode array and then seeded hiPSC-CMs for subsequent monitoring for 14 days. As expected, the CMs organized into anisotropic and matured tissue and the extracellular recordings showed reduced premature beating higher signal amplitude and a higher probability of T-wave detection as compared to the culture without fibers. The CMs on the aligned fibers samples also exhibited anisotropic propagation of the field potential. These results therefore suggest that the hiPSC-CMs cultured on AFs can be used more reliably for cell based assays. PMID:27446217

  2. Liquid core optical fiber total reflection cell as a colorimetric detector for flow injection analysis

    SciTech Connect

    Fujiwara, K.; Fuwa, K.

    1985-05-01

    A hollow fiber (250 ..mu..m i.d.) was used as a colorimetric cell for detecting iodine absorption. To attain total reflection of source light inside the capillary, carbon disulfide was used as a solvent which constitutes the fiber core. A funnel-shaped glass was used for efficiency condensing the light source emission into an aperture of hollow fiber; a low-power tungsten lamp was usable as the light source. With a 5-m cell, 0.1 ..mu..g of I/mL (10 ng of I) can be detected based on the iodine absorption at 540 nm when the solution was injected into the carbon disulfide flow. An automated detection system of iodide ion was also constructed. 11 references, 8 figures.

  3. Extracellular Recordings of Patterned Human Pluripotent Stem Cell-Derived Cardiomyocytes on Aligned Fibers.

    PubMed

    Li, Junjun; Minami, Itsunari; Yu, Leqian; Tsuji, Kiyotaka; Nakajima, Minako; Qiao, Jing; Suzuki, Masato; Shimono, Ken; Nakatsuji, Norio; Kotera, Hitetoshi; Liu, Li; Chen, Yong

    2016-01-01

    Human induced pluripotent stem cell (hiPSC) derived cardiomyocytes (CMs) hold high potential for use in drug assessment and myocardial regeneration. To create tissue-like constructs of CMs for extracellular monitoring, we placed aligned fibers (AFs) on the surface of a microelectrode array and then seeded hiPSC-CMs for subsequent monitoring for 14 days. As expected, the CMs organized into anisotropic and matured tissue and the extracellular recordings showed reduced premature beating higher signal amplitude and a higher probability of T-wave detection as compared to the culture without fibers. The CMs on the aligned fibers samples also exhibited anisotropic propagation of the field potential. These results therefore suggest that the hiPSC-CMs cultured on AFs can be used more reliably for cell based assays. PMID:27446217

  4. Proteomic Analysis of Lipid Raft-Like Detergent-Resistant Membranes of Lens Fiber Cells

    PubMed Central

    Wang, Zhen; Schey, Kevin L.

    2015-01-01

    Purpose Plasma membranes of lens fiber cells have high levels of long-chain saturated fatty acids, cholesterol, and sphingolipids—key components of lipid rafts. Thus, lipid rafts are expected to constitute a significant portion of fiber cell membranes and play important roles in lens biology. The purpose of this study was to characterize the lens lipid raft proteome. Methods Quantitative proteomics, both label-free and iTRAQ methods, were used to characterize lens fiber cell lipid raft proteins. Detergent-resistant, lipid raft membrane (DRM) fractions were isolated by sucrose gradient centrifugation. To confirm protein localization to lipid rafts, protein sensitivity to cholesterol removal by methyl-β-cyclodextrin was quantified by iTRAQ analysis. Results A total of 506 proteins were identified in raft-like detergent-resistant membranes. Proteins identified support important functions of raft domains in fiber cells, including trafficking, signal transduction, and cytoskeletal organization. In cholesterol-sensitivity studies, 200 proteins were quantified and 71 proteins were strongly affected by cholesterol removal. Lipid raft markers flotillin-1 and flotillin-2 and a significant fraction of AQP0, MP20, and AQP5 were found in the DRM fraction and were highly sensitive to cholesterol removal. Connexins 46 and 50 were more abundant in nonraft fractions, but a small fraction of each was found in the DRM fraction and was strongly affected by cholesterol removal. Quantification of modified AQP0 confirmed that fatty acylation targeted this protein to membrane raft domains. Conclusions These data represent the first comprehensive profile of the lipid raft proteome of lens fiber cells and provide information on membrane protein organization in these cells. PMID:26747763

  5. Long Life Nickel Electrodes for Nickel-Hydrogen Cells: Fiber Substrates Nickel Electrodes

    NASA Technical Reports Server (NTRS)

    Rogers, Howard H.

    2000-01-01

    Samples of nickel fiber mat electrodes were investigated over a wide range of fiber diameters, electrode thickness, porosity and active material loading levels. Thickness' were 0.040, 0.060 and 0.080 inches for the plaque: fiber diameters were primarily 2, 4, and 8 micron and porosity was 85, 90, and 95%. Capacities of 3.5 in. diameter electrodes were determined in the flooded condition with both 26 and 31% potassium hydroxide solution. These capacity tests indicated that the highest capacities per unit weight were obtained at the 90% porosity level with a 4 micron diameter fiber plaque. It appeared that the thinner electrodes had somewhat better performance, consistent with sintered electrode history. Limited testing with two-positive-electrode boiler plate cells was also carried out. Considerable difficulty with constructing the cells was encountered with short circuits the major problem. Nevertheless, four cells were tested. The cell with 95% porosity electrodes failed during conditioning cycling due to high voltage during charge. Discharge showed that this cell had lost nearly all of its capacity. The other three cells after 20 conditioning cycles showed capacities consistent with the flooded capacities of the electrodes. Positive electrodes made from fiber substrates may well show a weight advantage of standard sintered electrodes, but need considerably more work to prove this statement. A major problem to be investigated is the lower strength of the substrate compared to standard sintered electrodes. Problems with welding of leads were significant and implications that the electrodes would expand more than sintered electrodes need to be investigated. Loading levels were lower than had been expected based on sintered electrode experiences and the lower loading led to lower capacity values. However, lower loading causes less expansion and contraction during cycling so that stress on the substrate is reduced.

  6. Increased Adipogenesis of Human Adipose-Derived Stem Cells on Polycaprolactone Fiber Matrices

    PubMed Central

    Brännmark, Cecilia; Paul, Alexandra; Ribeiro, Diana; Magnusson, Björn; Brolén, Gabriella; Enejder, Annika; Forslöw, Anna

    2014-01-01

    With accelerating rates of obesity and type 2 diabetes world-wide, interest in studying the adipocyte and adipose tissue is increasing. Human adipose derived stem cells - differentiated to adipocytes in vitro - are frequently used as a model system for white adipocytes, as most of their pathways and functions resemble mature adipocytes in vivo. However, these cells are not completely like in vivo mature adipocytes. Hosting the cells in a more physiologically relevant environment compared to conventional two-dimensional cell culturing on plastic surfaces, can produce spatial cues that drive the cells towards a more mature state. We investigated the adipogenesis of adipose derived stem cells on electro spun polycaprolactone matrices and compared functionality to conventional two-dimensional cultures as well as to human primary mature adipocytes. To assess the degree of adipogenesis we measured cellular glucose-uptake and lipolysis and used a range of different methods to evaluate lipid accumulation. We compared the averaged results from a whole population with the single cell characteristics – studied by coherent anti-Stokes Raman scattering microscopy - to gain a comprehensive picture of the cell phenotypes. In adipose derived stem cells differentiated on a polycaprolactone-fiber matrix; an increased sensitivity in insulin-stimulated glucose uptake was detected when cells were grown on either aligned or random matrices. Furthermore, comparing differentiation of adipose derived stem cells on aligned polycaprolactone-fiber matrixes, to those differentiated in two-dimensional cultures showed, an increase in the cellular lipid accumulation, and hormone sensitive lipase content. In conclusion, we propose an adipocyte cell model created by differentiation of adipose derived stem cells on aligned polycaprolactone-fiber matrices which demonstrates increased maturity, compared to 2D cultured cells. PMID:25419971

  7. FT-IR examination of the development of secondary cell wall in cotton fibers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The secondary cell wall development of cotton fibers harvested at 18, 20, 24, 28, 32, 36 and 40 days after flowering was examined using attenuated total reflection Fourier transform-infrared (ATR FT-IR) spectroscopy. Generally, a progressive intensity increase for bands assigned to cellulose Iß was ...

  8. Fiber-Shaped Perovskite Solar Cells with High Power Conversion Efficiency.

    PubMed

    Qiu, Longbin; He, Sisi; Yang, Jiahua; Deng, Jue; Peng, Huisheng

    2016-05-01

    A perovskite solar cell fiber is created with a high power conversion efficiency of 7.1% through a controllable deposition method. A combination of aligned TiO2 nanotubes, a uniform perovskite layer, and transparent aligned carbon nanotube sheet contributes to the high photovoltaic performance. It is flexible and stable, and can be woven into smart clothes for wearable applications. PMID:27002590

  9. Clusters of cerebellar Purkinje cells control their afferent climbing fiber discharge

    PubMed Central

    Chaumont, Joseph; Guyon, Nicolas; Valera, Antoine M.; Dugué, Guillaume P.; Popa, Daniela; Marcaggi, Paikan; Gautheron, Vanessa; Reibel-Foisset, Sophie; Dieudonné, Stéphane; Stephan, Aline; Barrot, Michel; Cassel, Jean-Christophe; Dupont, Jean-Luc; Doussau, Frédéric; Poulain, Bernard; Selimi, Fekrije; Léna, Clément; Isope, Philippe

    2013-01-01

    Climbing fibers, the projections from the inferior olive to the cerebellar cortex, carry sensorimotor error and clock signals that trigger motor learning by controlling cerebellar Purkinje cell synaptic plasticity and discharge. Purkinje cells target the deep cerebellar nuclei, which are the output of the cerebellum and include an inhibitory GABAergic projection to the inferior olive. This pathway identifies a potential closed loop in the olivo-cortico-nuclear network. Therefore, sets of Purkinje cells may phasically control their own climbing fiber afferents. Here, using in vitro and in vivo recordings, we describe a genetically modified mouse model that allows the specific optogenetic control of Purkinje cell discharge. Tetrode recordings in the cerebellar nuclei demonstrate that focal stimulations of Purkinje cells strongly inhibit spatially restricted sets of cerebellar nuclear neurons. Strikingly, such stimulations trigger delayed climbing-fiber input signals in the stimulated Purkinje cells. Therefore, our results demonstrate that Purkinje cells phasically control the discharge of their own olivary afferents and thus might participate in the regulation of cerebellar motor learning. PMID:24046366

  10. Clusters of cerebellar Purkinje cells control their afferent climbing fiber discharge.

    PubMed

    Chaumont, Joseph; Guyon, Nicolas; Valera, Antoine M; Dugué, Guillaume P; Popa, Daniela; Marcaggi, Paikan; Gautheron, Vanessa; Reibel-Foisset, Sophie; Dieudonné, Stéphane; Stephan, Aline; Barrot, Michel; Cassel, Jean-Christophe; Dupont, Jean-Luc; Doussau, Frédéric; Poulain, Bernard; Selimi, Fekrije; Léna, Clément; Isope, Philippe

    2013-10-01

    Climbing fibers, the projections from the inferior olive to the cerebellar cortex, carry sensorimotor error and clock signals that trigger motor learning by controlling cerebellar Purkinje cell synaptic plasticity and discharge. Purkinje cells target the deep cerebellar nuclei, which are the output of the cerebellum and include an inhibitory GABAergic projection to the inferior olive. This pathway identifies a potential closed loop in the olivo-cortico-nuclear network. Therefore, sets of Purkinje cells may phasically control their own climbing fiber afferents. Here, using in vitro and in vivo recordings, we describe a genetically modified mouse model that allows the specific optogenetic control of Purkinje cell discharge. Tetrode recordings in the cerebellar nuclei demonstrate that focal stimulations of Purkinje cells strongly inhibit spatially restricted sets of cerebellar nuclear neurons. Strikingly, such stimulations trigger delayed climbing-fiber input signals in the stimulated Purkinje cells. Therefore, our results demonstrate that Purkinje cells phasically control the discharge of their own olivary afferents and thus might participate in the regulation of cerebellar motor learning. PMID:24046366

  11. Cells on fibers to degrade PAH and upgrade coal

    SciTech Connect

    Clyde, R.

    1997-12-31

    There are over 2000 sites contaminated with PAH`s from coal burning plants. White rot fungus degrades phenanthrene and anthracene, but the fungus needs air to grow. When grown on old cardboard boxes and buried, air is entrapped in the corrugations for growth of the fungus. When holes are put in the valleys of the corrugations and rotated in a half full reactor, drops are formed. Mass transfer to drops is much faster than to a flat surface, as described in Patent 5,256,570, so the fungus grows faster. Low rank coal can be upgraded to more valuable products with the fungus, say some Australians, but the problem is supplying oxygen. Celite can be entrapped in the fibers to ferment coal derived synthesis gas. The paper describes these processes.

  12. Fiber-Optic SPR Immunosensors Tailored To Target Epithelial Cells through Membrane Receptors.

    PubMed

    Malachovská, Viera; Ribaut, Clotilde; Voisin, Valérie; Surin, Mathieu; Leclère, Philippe; Wattiez, Ruddy; Caucheteur, Christophe

    2015-06-16

    We report, for the first time, the use of a surface plasmon resonance (SPR) fiber-optic immunosensor for selective cellular detection through membrane protein targeting. The sensor architecture lies on gold-coated tilted fiber Bragg gratings (Au-coated TFBGs) photoimprinted in the fiber core via a laser technique. TFBGs operate in the near-infrared wavelength range at ∼1550 nm, yielding optical and SPR sensing characteristics that are advantageous for the analyses of cellular bindings and technical compatibility with relatively low-cost telecommunication-grade measurement devices. In this work, we take consider their numerous assets to figure out their ability to selectively detect intact epithelial cells as analytes in cell suspensions in the range of 2-5 × 10(6) cells mL(-1). For this, the probe was first thermally annealed to ensure a strong adhesion of the metallic coating to the fiber surface. Its surface was then functionalized with specific monoclonal antibodies via alkanethiol self-assembled monolayers (SAMs) against extracellular domain of epidermal growth factor receptors (EGFRs) and characterized by peak force tapping atomic force microscopy. A differential diagnosis has been demonstrated between two model systems. The developed immunosensors were able to monitor, in real time, the specific attachment of single intact cells in concentrations from 3 × 10(6) cells mL(-1). Such results confirm that the developed probe fits the lab-on-fiber technology and has the potential to be used as a disposable device for in situ and real-time clinical diagnosis. PMID:25962700

  13. Efficient Gene Transduction of Dispersed Islet Cells in Culture Using Fiber-Modified Adenoviral Vectors.

    PubMed

    Hanayama, Hiroyuki; Ohashi, Kazuo; Utoh, Rie; Shimizu, Hirofumi; Ise, Kazuya; Sakurai, Fuminori; Mizuguchi, Hiroyuki; Tsuchiya, Hiroyuki; Okano, Teruo; Gotoh, Mitsukazu

    2015-12-17

    To establish novel islet-based therapies, our group has recently developed technologies for creating functional neo-islet tissues in the subcutaneous space by transplanting monolithic sheets of dispersed islet cells (islet cell sheets). Improving cellular function and viability are the next important challenges for enhancing the therapeutic effects. This article describes the adenoviral vector-mediated gene transduction of dispersed islet cells under culture conditions. Purified pancreatic islets were obtained from Lewis rats and dissociated into single islet cells. Cells were plated onto laminin-5-coated temperature-responsive polymer poly(N-isopropylacrylamide)-immobilized plastic dishes. At 0 h, islet cells were infected for 1 h with either conventional type 5 adenoviral vector (Ad-CA-GFP) or fiber-modified adenoviral vector (AdK7-CA-GFP) harboring a polylysine (K7) peptide in the C terminus of the fiber knob. We investigated gene transduction efficiency at 48 h after infection and found that AdK7-CA-GFP yielded higher transduction efficiencies than Ad-CA-GFP at a multiplicity of infection (MOI) of 5 and 10. For AdK7-CA-GFP at MOI = 10, 84.4 ± 1.5% of islet cells were found to be genetically transduced without marked vector infection-related cellular damage as determined by viable cell number and lactate dehydrogenase (LDH) release assay. After AdK7-CA-GFP infection at MOI = 10, cells remained attached and expanded to nearly full confluency, showing that this adenoviral infection protocol is a feasible approach for creating islet cell sheets. We have shown that dispersed and cultured islet cells can be genetically modified efficiently using fiber-modified adenoviral vectors. Therefore, this gene therapy technique could be used for cellular modification or biological assessment of dispersed islet cells. PMID:26858906

  14. Efficient Gene Transduction of Dispersed Islet Cells in Culture Using Fiber-Modified Adenoviral Vectors

    PubMed Central

    Hanayama, Hiroyuki; Ohashi, Kazuo; Utoh, Rie; Shimizu, Hirofumi; Ise, Kazuya; Sakurai, Fuminori; Mizuguchi, Hiroyuki; Tsuchiya, Hiroyuki; Okano, Teruo; Gotoh, Mitsukazu

    2015-01-01

    To establish novel islet-based therapies, our group has recently developed technologies for creating functional neo-islet tissues in the subcutaneous space by transplanting monolithic sheets of dispersed islet cells (islet cell sheets). Improving cellular function and viability are the next important challenges for enhancing the therapeutic effects. This article describes the adenoviral vector-mediated gene transduction of dispersed islet cells under culture conditions. Purified pancreatic islets were obtained from Lewis rats and dissociated into single islet cells. Cells were plated onto laminin-5-coated temperature-responsive polymer poly(N-isopropylacrylamide)-immobilized plastic dishes. At 0 h, islet cells were infected for 1 h with either conventional type 5 adenoviral vector (Ad-CA-GFP) or fiber-modified adenoviral vector (AdK7-CA-GFP) harboring a polylysine (K7) peptide in the C terminus of the fiber knob. We investigated gene transduction efficiency at 48 h after infection and found that AdK7-CA-GFP yielded higher transduction efficiencies than Ad-CA-GFP at a multiplicity of infection (MOI) of 5 and 10. For AdK7-CA-GFP at MOI = 10, 84.4 ± 1.5% of islet cells were found to be genetically transduced without marked vector infection-related cellular damage as determined by viable cell number and lactate dehydrogenase (LDH) release assay. After AdK7-CA-GFP infection at MOI = 10, cells remained attached and expanded to nearly full confluency, showing that this adenoviral infection protocol is a feasible approach for creating islet cell sheets. We have shown that dispersed and cultured islet cells can be genetically modified efficiently using fiber-modified adenoviral vectors. Therefore, this gene therapy technique could be used for cellular modification or biological assessment of dispersed islet cells. PMID:26858906

  15. Porous, platinum nanoparticle-adsorbed carbon nanotube yarns for efficient fiber solar cells.

    PubMed

    Zhang, Sen; Ji, Chunyan; Bian, Zuqiang; Yu, Pingrong; Zhang, Luhui; Liu, Dianyi; Shi, Enzheng; Shang, Yuanyuan; Peng, Haitao; Cheng, Qiao; Wang, Dong; Huang, Chunhui; Cao, Anyuan

    2012-08-28

    Pt is a classical catalyst that has been extensively used in fuel cell and solar cell electrodes, owing to its high catalytic activity, good conductivity, and stability. In conventional fiber-shaped solar cells, solid Pt wires are usually adopted as the electrode material. Here, we report a Pt nanoparticle-adsorbed carbon nanotube yarn made by solution adsorption and yarn spinning processes, with uniformly dispersed Pt nanoparticles through the porous nanotube network. We have fabricated TiO(2)-based dye-sensitized fiber solar cells with a Pt-nanotube hybrid yarn as counter electrode and achieved a power conversion efficiency of 4.85% under standard illumination (AM1.5, 100 mW/cm(2)), comparable to the same type of fiber cells with a Pt wire electrode (4.23%). Adsorption of Pt nanoparticles within a porous nanotube yarn results in enhanced Pt-electrolyte interfacial area and significantly reduced charge-transfer resistance across the electrolyte interface, compared to a pure nanotube yarn or Pt wire. Our porous Pt-nanotube hybrid yarns have the potential to reduce the use of noble metals, lower the device weight, and improve the solar cell efficiency. PMID:22861684

  16. The development of a potassium-sulfide glass fiber cell and studies on impurities in alkali metal-sulfur cells

    NASA Technical Reports Server (NTRS)

    Tsang, F. Y.

    1977-01-01

    Potassium sulfur rechargeable cells, having as the electrolyte the thin walls of hollow glass fibers made from permeable glass, were developed. The cells had short lives, probably due to the construction materials and impurities in the potassium. The effect of the impurities in the analogous NA-S system was studied. Calcium, potassium, and NaOH/oxide impurities caused increased resistance or corrosion of the glass fibers. For long lived cell operation, the Na must contain less than 1 ppm Ca and less than a few ppm of hydroxide/oxide. Up to 150 ppm K can be tolerated. After purification of the Na anolyte, cell lifetimes in excess of 1000 deep charge-discharge cycles or over 8 months on continuous cycling at 10-30 percent depth of discharge were obtained.

  17. Tmem2 regulates cell-matrix interactions that are essential for muscle fiber attachment.

    PubMed

    Ryckebüsch, Lucile; Hernandez, Lydia; Wang, Carole; Phan, Jenny; Yelon, Deborah

    2016-08-15

    Skeletal muscle morphogenesis depends upon interactions between developing muscle fibers and the extracellular matrix (ECM) that anchors fibers to the myotendinous junction (MTJ). The pathways that organize the ECM and regulate its engagement by cell-matrix adhesion complexes (CMACs) are therefore essential for muscle integrity. Here, we demonstrate the impact of transmembrane protein 2 (tmem2) on cell-matrix interactions during muscle morphogenesis in zebrafish. Maternal-zygotic tmem2 mutants (MZtmem2) exhibit muscle fiber detachment, in association with impaired laminin organization and ineffective fibronectin degradation at the MTJ. Similarly, disorganized laminin and fibronectin surround MZtmem2 cardiomyocytes, which could account for their hindered movement during cardiac morphogenesis. In addition to ECM defects, MZtmem2 mutants display hypoglycosylation of α-dystroglycan within the CMAC, which could contribute to the observed fiber detachment. Expression of the Tmem2 ectodomain can rescue aspects of the MZtmem2 phenotype, consistent with a possible extracellular function of Tmem2. Together, our results suggest that Tmem2 regulates cell-matrix interactions by affecting both ECM organization and CMAC activity. These findings evoke possible connections between the functions of Tmem2 and the etiologies of congenital muscular dystrophies, particularly dystroglycanopathies. PMID:27471259

  18. Evaluation of toxic agent effects on lung cells by fiber evanescent wave spectroscopy.

    PubMed

    Lucas, Pierre; Le Coq, David; Juncker, Christophe; Collier, Jayne; Boesewetter, Dianne E; Boussard-Plédel, Catherine; Bureau, Bruno; Riley, Mark R

    2005-01-01

    Biochemical changes in living cells are detected using a fiber probe system composed of a single chalcogenide fiber acting as both the sensor and transmission line for infrared optical signals. The signal is collected via evanescent wave absorption along the tapered sensing zone of the fiber. We spectroscopically monitored the effects of the surfactant Triton X-100, which serves as a toxic agent simulant on a transformed human lung carcinoma type II epithelial cell line (A549). We observe spectral changes between 2800-3000 cm(-1) in four absorptions bands, which are assigned to hydrocarbon vibrations of methylene and methyl groups in membrane lipids. Comparison of fiber and transmission spectra shows that the present technique allows one to locally probe the cell plasma membrane in the lipid spectral region. These optical responses are correlated with cellular metabolic activity measurements and LDH (lactate dehydrogenase) release assays that indicate a loss of cellular function and membrane integrity as would be expected in response to the membrane solubilizing Triton. The spectroscopic technique shows a significantly greater detection resolution in time and concentration. PMID:15720730

  19. Mesenchymal stem cells support neuronal fiber growth in an organotypic brain slice co-culture model.

    PubMed

    Sygnecka, Katja; Heider, Andreas; Scherf, Nico; Alt, Rüdiger; Franke, Heike; Heine, Claudia

    2015-04-01

    Mesenchymal stem cells (MSCs) have been identified as promising candidates for neuroregenerative cell therapies. However, the impact of different isolation procedures on the functional and regenerative characteristics of MSC populations has not been studied thoroughly. To quantify these differences, we directly compared classically isolated bulk bone marrow-derived MSCs (bulk BM-MSCs) to the subpopulation Sca-1(+)Lin(-)CD45(-)-derived MSCs(-) (SL45-MSCs), isolated by fluorescence-activated cell sorting from bulk BM-cell suspensions. Both populations were analyzed with respect to functional readouts, that are, frequency of fibroblast colony forming units (CFU-f), general morphology, and expression of stem cell markers. The SL45-MSC population is characterized by greater morphological homogeneity, higher CFU-f frequency, and significantly increased nestin expression compared with bulk BM-MSCs. We further quantified the potential of both cell populations to enhance neuronal fiber growth, using an ex vivo model of organotypic brain slice co-cultures of the mesocortical dopaminergic projection system. The MSC populations were cultivated underneath the slice co-cultures without direct contact using a transwell system. After cultivation, the fiber density in the border region between the two brain slices was quantified. While both populations significantly enhanced fiber outgrowth as compared with controls, purified SL45-MSCs stimulated fiber growth to a larger degree. Subsequently, we analyzed the expression of different growth factors in both cell populations. The results show a significantly higher expression of brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor in the SL45-MSCs population. Altogether, we conclude that MSC preparations enriched for primary MSCs promote neuronal regeneration and axonal regrowth, more effectively than bulk BM-MSCs, an effect that may be mediated by a higher BDNF secretion. PMID:25390472

  20. Design and performance of fiber optic pressure cell based on polarimetric sensing

    NASA Astrophysics Data System (ADS)

    Bock, Wojtek J.; Voet, Mark R.; Beaulieu, Mario; Chen, Jiahua

    1993-03-01

    In this paper we propose replacing a widely used but often difficult and cumbersome technique of hydraulic evaluation of stress in concrete materials with a new fiber-optic measurement device, which has all inherent advantages of fiber-optic sensors. The sensing element of the device consists of a highly birefringent (HB) polarization-maintaining optical fiber. The stress inside it induced by external pressure modulates the polarization state of the output light signal at the detection end of the system. The all-fiber instrumentation system of the sensor consists of a semiconductor pigtailed laser, input and output HB optical fibers, an analyzer and a computer-controlled synchronous detection system. A specially designed leadthrough integrated with the sensor head allowed us to insert the sensor inside a pressure pad filled with oil or alternatively with mercury. For calibration purposes, the pressure cell was placed inside a large pressure chamber designed to simulate the real environment. Characterization of the device for hysteresis, selectivity and sensitivity was performed for pressures up to 70 bar and for ambient temperatures. The described sensor is simple, cost-effective, safe in explosive environments and well adapted for stress monitoring in the large-scale structures.

  1. Composite fiber structures with antiproliferative agents exhibit advantageous drug delivery and cell growth inhibition in vitro.

    PubMed

    Kraitzer, Amir; Kloog, Yoel; Haklai, Roni; Zilberman, Meital

    2011-01-01

    Composite core/shell fiber structures loaded with the antiproliferative drugs paclitaxel or farnesylthiosalicylate (FTS) were developed and studied. The latter is a specific nontoxic Ras inhibitor with a mild hydrophobic nature, which can also be used for local cancer treatment and stent applications. The fibers were composed of a dense polyglyconate core and a porous drug-loaded poly(D,L-lactic-glycolic acid) shell, prepared using freeze drying of inverted emulsions. Our study focused on the release profile of the antiproliferative drugs from the fibers, the shell morphology and its degradation and erosion. The postfabrication antiproliferative effect of the drugs was tested in a cell culture. The process parameters were found to affect the drug-release profile via two routes: (1) direct, through water uptake and swelling of the structure leading to FTS release, or through degradation of the host polymer leading to paclitaxel release at a later stage; (2) indirect effect of the microstructure on the release profile. The fabrication process did not reduce the pharmacological activity of either paclitaxel or FTS. FTS-eluting composite fibers proved to effectively induce growth inhibition or cell death by a gradient effect and dose-dependent manner. The combined effect of the targeted mechanism of FTS as a Ras inhibitor together with the localized and controlled release characteristics of the fiber is an advantageous antiproliferative quality. It is therefore suggested that our drug-eluting fibers may be used in biomedical applications that require short release (restenosis) or prolonged release (cancer therapy). PMID:20623695

  2. Three-dimensional fiber deposition of cell-laden, viable, patterned constructs for bone tissue printing.

    PubMed

    Fedorovich, Natalja E; De Wijn, Joost R; Verbout, Abraham J; Alblas, Jacqueline; Dhert, Wouter J A

    2008-01-01

    Organ or tissue printing, a novel approach in tissue engineering, creates layered, cell-laden hydrogel scaffolds with a defined three-dimensional (3D) structure and organized cell placement. In applying the concept of tissue printing for the development of vascularized bone grafts, the primary focus lies on combining endothelial progenitors and bone marrow stromal cells (BMSCs). Here we characterize the applicability of 3D fiber deposition with a plotting device, Bioplotter, for the fabrication of spatially organized, cell-laden hydrogel constructs. The viability of printed BMSCs was studied in time, in several hydrogels, and extruded from different needle diameters. Our findings indicate that cells survive the extrusion and that their subsequent viability was not different from that of unprinted cells. The applied extrusion conditions did not affect cell survival, and BMSCs could subsequently differentiate along the osteoblast lineage. Furthermore, we were able to combine two distinct cell populations within a single scaffold by exchanging the printing syringe during deposition, indicating that this 3D fiber deposition system is suited for the development of bone grafts containing multiple cell types. PMID:18333811

  3. Secondary cell wall development in cotton fibers as examined with attenuated total reflection Fourier transform infrared spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cotton fibers harvested at 18, 20, 24, 28, 32, 36 and 40 days after flowering were examined using attenuated total reflection Fourier transform-infrared (ATR FT-IR) spectroscopy. The selected harvesting points coincide with secondary cell wall (SCW) development in the fibers. Progressive but moderat...

  4. Secreted frizzled-related protein disrupts PCP in eye lens fiber cells that have polarised primary cilia.

    PubMed

    Sugiyama, Yuki; Stump, Richard J W; Nguyen, Anke; Wen, Li; Chen, Yongjuan; Wang, Yanshu; Murdoch, Jennifer N; Lovicu, Frank J; McAvoy, John W

    2010-02-15

    Planar cell polarity (PCP) signaling polarises cells along tissue axes. Although pathways involved are becoming better understood, outstanding issues include; (i) existence/identity of cues that orchestrate global polarisation in tissues, and (ii) the generality of the link between polarisation of primary cilia and asymmetric localisation of PCP proteins. Mammalian lenses are mainly comprised of epithelial-derived fiber cells. Concentrically arranged fibers are precisely aligned as they elongate along the anterior-posterior axis and orientate towards lens poles where they meet fibers from other segments to form characteristic sutures. We show that lens exhibits PCP, with each fiber cell having an apically situated cilium and in most cases this is polarised towards the anterior pole. Frizzled and other PCP proteins are also asymmetrically localised along the equatorial-anterior axis. Mutations in core PCP genes Van Gogh-like 2 and Celsr1 perturb oriented fiber alignment and suture formation. Suppression of the PCP pathway by overexpressing Sfrp2 shows that whilst local groups of fibers are often similarly oriented, they lack global orientation; consequently when local groups of fibers with different orientations meet they form multiple, small, ectopic suture-like configurations. This indicates that this extracellular inhibitor disrupts a global polarising signal that utilises a PCP-mediated mechanism to coordinate the global alignment and orientation of fibers to lens poles. PMID:19968984

  5. Identification and characterization of plasma membrane aquaporins isolated from fiber cells of Calotropis procera

    PubMed Central

    Aslam, Usman; Khatoon, Asia; Cheema, Hafiza Masooma Naseer; Bashir, Aftab

    2013-01-01

    Calotropis procera, commonly known as “milkweed”, possesses long seed trichomes for seed dispersal and has the ability to survive under harsh conditions such as drought and salinity. Aquaporins are water channel proteins expressed in all land plants, divided into five subfamilies plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), NOD26-like proteins (NIPs), small basic intrinsic proteins (SIPs), and the unfamiliar X intrinsic proteins (XIPs). PIPs constitute the largest group of water channel proteins that are involved in different developmental and regulatory mechanisms including water permeability, cell elongation, and stomata opening. Aquaporins are also involved in abiotic stress tolerance and cell expansion mechanisms, but their role in seed trichomes (fiber cells) has never been investigated. A large number of clones isolated from C. procera fiber cDNA library showed sequence homology to PIPs. Both expressed sequence tags (ESTs) and real-time polymerase chain reaction (PCR) studies revealed that the transcript abundance of this gene family in fiber cells of C. procera is greater than that of cotton. Full-length cDNAs of CpPIP1 and CpPIP2 were isolated from C. procera fiber cDNA library and used for constructing plant expression vectors under constitutive (2×35S) and trichome-specific (GhLTP3) promoters. Transgenic tobacco plants were developed via Agrobacterium-mediated transformation. The phenotypic characteristics of the plants were observed after confirming the integration of transgene in plants. It was observed that CpPIP2 expression cassette under 2×35S and GhLTP3 promoter enhanced the numbers of stem and leave trichomes. However, 2×35S::CpPIP2 has a more amplified effect on trichome density and length than GhLTP3::CpPIP2 and other PIP constructs. These findings imply the role of C. procera PIP aquaporins in fiber cell elongation. The PIPs-derived cell expansion mechanism may be exploited through transgenic approaches

  6. Identification and characterization of plasma membrane aquaporins isolated from fiber cells of Calotropis procera.

    PubMed

    Aslam, Usman; Khatoon, Asia; Cheema, Hafiza Masooma Naseer; Bashir, Aftab

    2013-07-01

    Calotropis procera, commonly known as "milkweed", possesses long seed trichomes for seed dispersal and has the ability to survive under harsh conditions such as drought and salinity. Aquaporins are water channel proteins expressed in all land plants, divided into five subfamilies plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), NOD26-like proteins (NIPs), small basic intrinsic proteins (SIPs), and the unfamiliar X intrinsic proteins (XIPs). PIPs constitute the largest group of water channel proteins that are involved in different developmental and regulatory mechanisms including water permeability, cell elongation, and stomata opening. Aquaporins are also involved in abiotic stress tolerance and cell expansion mechanisms, but their role in seed trichomes (fiber cells) has never been investigated. A large number of clones isolated from C. procera fiber cDNA library showed sequence homology to PIPs. Both expressed sequence tags (ESTs) and real-time polymerase chain reaction (PCR) studies revealed that the transcript abundance of this gene family in fiber cells of C. procera is greater than that of cotton. Full-length cDNAs of CpPIP1 and CpPIP2 were isolated from C. procera fiber cDNA library and used for constructing plant expression vectors under constitutive (2×35S) and trichome-specific (GhLTP3) promoters. Transgenic tobacco plants were developed via Agrobacterium-mediated transformation. The phenotypic characteristics of the plants were observed after confirming the integration of transgene in plants. It was observed that CpPIP2 expression cassette under 2×35S and GhLTP3 promoter enhanced the numbers of stem and leave trichomes. However, 2×35S::CpPIP2 has a more amplified effect on trichome density and length than GhLTP3::CpPIP2 and other PIP constructs. These findings imply the role of C. procera PIP aquaporins in fiber cell elongation. The PIPs-derived cell expansion mechanism may be exploited through transgenic approaches for

  7. Cancer cell aggregate hypoxia visualized in vitro via biocompatible fiber sensors.

    PubMed

    Xue, Ruipeng; Nelson, M Tyler; Teixeira, Silvia A; Viapiano, Mariano S; Lannutti, John J

    2016-01-01

    To fully understand biological behavior in vitro often dictates that oxygen be reported at either a local or a cellular level. Oxygen sensors based on the luminescent quenching of a specific form of electrospun fiber were developed for measurement of both gaseous and dissolved oxygen concentrations. Electrospinning was used to fabricate "core-shell" fiber configurations in which oxygen-sensitive transition-metal porphyrin complexes are embedded in an optically clear, gas permeable polycarbonate polymer 'core' while polycaprolactone provided a protective yet biocompatible 'shell'. By taking advantage of the resulting high sensitivity and fast response of electrospun core-shell fiber sensors, we were able to locate and image hypoxic regions in contact with aggregates of glioblastoma cells. Nanoscale, biomimetic sensors containing oxygen-sensitive porphyrins are particularly well suited to biological applications. These 'smart' nanofiber based sensors do not consume oxygen, their mechanical and chemical characteristics can be finely tuned allowing tailoring of biocompatibility and microstructure. Core-shell nanofiber oxygen sensing fibers could provide real-time assessments of tumor cell response to pharmacological innovations designed to target hypoxic regions driving new knowledge and technological advancement. PMID:26524540

  8. Detection of low levels of Listeria monocytogenes cells by using a fiber-optic immunosensor.

    PubMed

    Geng, Tao; Morgan, Mark T; Bhunia, Arun K

    2004-10-01

    Biosensor technology has a great potential to meet the need for sensitive and nearly real-time microbial detection from foods. An antibody-based fiber-optic biosensor to detect low levels of Listeria monocytogenes cells following an enrichment step was developed. The principle of the sensor is a sandwich immunoassay where a rabbit polyclonal antibody was first immobilized on polystyrene fiber waveguides through a biotin-streptavidin reaction to capture Listeria cells on the fiber. Capture of cells on the fibers was confirmed by scanning electron microscopy. A cyanine 5-labeled murine monoclonal antibody, C11E9, was used to generate a specific fluorescent signal, which was acquired by launching a 635-nm laser light from an Analyte 2000 and collected by a photodetector at 670 to 710 nm. This immunosensor was specific for L. monocytogenes and showed a significantly higher signal strength than for other Listeria species or other microorganisms, including Escherichia coli, Enterococcus faecalis, Salmonella enterica, Lactobacillus plantarum, Carnobacterium gallinarum, Hafnia alvei, Corynebacterium glutamicum, Enterobacter aerogenes, Pseudomonas aeruginosa, and Serratia marcescens, in pure or in mixed-culture setup. Fiber-optic results could be obtained within 2.5 h of sampling. The sensitivity threshold was about 4.3 x 10(3) CFU/ml for a pure culture of L. monocytogenes grown at 37 degrees C. When L. monocytogenes was mixed with lactic acid bacteria or grown at 10 degrees C with 3.5% NaCl, the detection threshold was 4.1 x 10(4) or 2.8 x 10(7) CFU/ml, respectively. In less than 24 h, this method could detect L. monocytogenes in hot dog or bologna naturally contaminated or artificially inoculated with 10 to 1,000 CFU/g after enrichment in buffered Listeria enrichment broth. PMID:15466560

  9. Assaying Human Myogenic Progenitor Cell Activity by Reconstitution of Muscle Fibers and Satellite Cells in Immunodeficient Mice.

    PubMed

    Parker, Maura H

    2016-01-01

    Comparing the functional myogenic potential of various human cell populations is an important step in the preclinical evaluation of cell transplantation as a means to treat human muscle disease and degeneration. Culture systems allow one to gage the potential of cell populations to proliferate and undergo myogenic differentiation under specific conditions. An in vivo assay evaluates the ability of cells to differentiate and generate muscle fibers within a natural environment, and importantly, evaluates the potential of donor cells to reconstitute the satellite cell niche. In this chapter, we describe a technique for isolating mononuclear cells from human muscle samples, and a method of xenotransplantation for assessing functional myogenic potential in vivo. Briefly, cell populations are injected into the pre-irradiated and regenerating muscle of immunodeficient mice. The injected muscle is frozen at specific time points after injection and cryosections analyzed by immunostaining. The number of human dystrophin-expressing fibers and the number of Pax7(+) human lamin A/C(+) nuclei are determined, which provides a quantitative method of comparing the in vivo functional potential of cell populations. PMID:27492175

  10. Fibroblast growth factor receptor signaling is essential for lens fiber cell differentiation

    PubMed Central

    Zhao, Haotian; Yang, Tianyu; Madakashira, Bhavani P.; Thiels, Cornelius A.; Bechtle, Chad A.; Garcia, Claudia M.; Zhang, Huiming; Yu, Kai; Ornitz, David M.; Beebe, David C.; Robinson, Michael L.

    2008-01-01

    The vertebrate lens provides an excellent model to study the mechanisms that regulate terminal differentiation. Although fibroblast growth factors (FGFs) are thought to be important for lens cell differentiation, it is unclear which FGF receptors mediate these processes during different stages of lens development. Deletion of three FGF receptors (Fgfr1-3) early in lens development demonstrated that expression of only a single allele of Fgfr2 or Fgfr3 was sufficient for grossly normal lens development, while mice possessing only a single Fgfr1 allele developed cataracts and microphthalmia. Profound defects were observed in lenses lacking all three Fgfrs. These included lack of fiber cell elongation, abnormal proliferation in prospective lens fiber cells, reduced expression of the cell cycle inhibitors p27kip1 and p57kip2, increased apoptosis and aberrant or reduced expression of Prox1, Pax6, c-Maf, E-cadherin and α-, β- and γ-crystallins. Therefore, while signaling by FGF receptors is essential for lens fiber differentiation, different FGF receptors function redundantly. PMID:18455718

  11. Increased epidermal growth factor-receptor protein in a human mesothelial cell line in response to long asbestos fibers.

    PubMed Central

    Pache, J. C.; Janssen, Y. M.; Walsh, E. S.; Quinlan, T. R.; Zanella, C. L.; Low, R. B.; Taatjes, D. J.; Mossman, B. T.

    1998-01-01

    Epidermal growth factor (EGF) is a potent mitogen for human mesothelial cells, and autophosphorylation of the EGF receptor (EGF-R) occurs in these cell types after exposure to asbestos, a carcinogen associated with the development of mesothelioma. Here, the intensity and distribution of EGF-R protein was documented by immunocytochemistry in a human mesothelial cell line (MET5A) exposed to various concentrations of crocidolite asbestos and man-made vitreous fibers (MMVF-10). Whereas cells in contact with or phagocytizing shorter asbestos fibers (<60 microm length) or MMVF-10 at a range of concentrations showed no increase in EGF-R protein as determined by immunofluorescence, elongated cells phagocytizing and surrounding longer fibers (> or =60 microm) showed intense staining for EGF-R. In contrast, human A549 lung carcinoma cells showed neither elongation nor increased accumulation of EGF-R protein in response to long fibers. Patterns of aggregation and increases in EGF-R protein in mesothelial cells phagocytizing long asbestos fibers were distinct from diffuse staining of phosphotyrosine residues observed in asbestos-exposed cultures. These studies indicate that aggregation of EGF-R by long fibers may initiate cell signaling cascades important in asbestos-induced mitogenesis and carcinogenesis. Images Figure 1 Figure 2 Figure 3 PMID:9466557

  12. Proteomic Studies of a Single CNS Synapse Type: The Parallel Fiber/Purkinje Cell Synapse

    PubMed Central

    Selimi, Fekrije; Cristea, Ileana M; Heller, Elizabeth; Chait, Brian T; Heintz, Nathaniel

    2009-01-01

    Precise neuronal networks underlie normal brain function and require distinct classes of synaptic connections. Although it has been shown that certain individual proteins can localize to different classes of synapses, the biochemical composition of specific synapse types is not known. Here, we have used a combination of genetically engineered mice, affinity purification, and mass spectrometry to profile proteins at parallel fiber/Purkinje cell synapses. We identify approximately 60 candidate postsynaptic proteins that can be classified into 11 functional categories. Proteins involved in phospholipid metabolism and signaling, such as the protein kinase MRCKγ, are major unrecognized components of this synapse type. We demonstrate that MRCKγ can modulate maturation of dendritic spines in cultured cortical neurons, and that it is localized specifically to parallel fiber/Purkinje cell synapses in vivo. Our data identify a novel synapse-specific signaling pathway, and provide an approach for detailed investigations of the biochemical complexity of central nervous system synapse types. PMID:19402746

  13. Proteomic studies of a single CNS synapse type: the parallel fiber/purkinje cell synapse.

    PubMed

    Selimi, Fekrije; Cristea, Ileana M; Heller, Elizabeth; Chait, Brian T; Heintz, Nathaniel

    2009-04-14

    Precise neuronal networks underlie normal brain function and require distinct classes of synaptic connections. Although it has been shown that certain individual proteins can localize to different classes of synapses, the biochemical composition of specific synapse types is not known. Here, we have used a combination of genetically engineered mice, affinity purification, and mass spectrometry to profile proteins at parallel fiber/Purkinje cell synapses. We identify approximately 60 candidate postsynaptic proteins that can be classified into 11 functional categories. Proteins involved in phospholipid metabolism and signaling, such as the protein kinase MRCKgamma, are major unrecognized components of this synapse type. We demonstrate that MRCKgamma can modulate maturation of dendritic spines in cultured cortical neurons, and that it is localized specifically to parallel fiber/Purkinje cell synapses in vivo. Our data identify a novel synapse-specific signaling pathway, and provide an approach for detailed investigations of the biochemical complexity of central nervous system synapse types. PMID:19402746

  14. Viscoelastic Retraction of Single Living Stress Fibers and Its Impact on Cell Shape, Cytoskeletal Organization, and Extracellular Matrix Mechanics

    PubMed Central

    Kumar, Sanjay; Maxwell, Iva Z.; Heisterkamp, Alexander; Polte, Thomas R.; Lele, Tanmay P.; Salanga, Matthew; Mazur, Eric; Ingber, Donald E.

    2006-01-01

    Cells change their form and function by assembling actin stress fibers at their base and exerting traction forces on their extracellular matrix (ECM) adhesions. Individual stress fibers are thought to be actively tensed by the action of actomyosin motors and to function as elastic cables that structurally reinforce the basal portion of the cytoskeleton; however, these principles have not been directly tested in living cells, and their significance for overall cell shape control is poorly understood. Here we combine a laser nanoscissor, traction force microscopy, and fluorescence photobleaching methods to confirm that stress fibers in living cells behave as viscoelastic cables that are tensed through the action of actomyosin motors, to quantify their retraction kinetics in situ, and to explore their contribution to overall mechanical stability of the cell and interconnected ECM. These studies reveal that viscoelastic recoil of individual stress fibers after laser severing is partially slowed by inhibition of Rho-associated kinase and virtually abolished by direct inhibition of myosin light chain kinase. Importantly, cells cultured on stiff ECM substrates can tolerate disruption of multiple stress fibers with negligible overall change in cell shape, whereas disruption of a single stress fiber in cells anchored to compliant ECM substrates compromises the entire cellular force balance, induces cytoskeletal rearrangements, and produces ECM retraction many microns away from the site of incision; this results in large-scale changes of cell shape (> 5% elongation). In addition to revealing fundamental insight into the mechanical properties and cell shape contributions of individual stress fibers and confirming that the ECM is effectively a physical extension of the cell and cytoskeleton, the technologies described here offer a novel approach to spatially map the cytoskeletal mechanics of living cells on the nanoscale. PMID:16500961

  15. A Raman cell based on hollow optical fibers for breath analysis

    NASA Astrophysics Data System (ADS)

    Okita, Y.; Katagiri, T.; Matsuura, Y.

    2010-02-01

    A compact Raman cell based on the hollow optical fiber for highly sensitive breath analysis is reported. A polycarbonate tube-based hollow optical fiber with inner coating of silver is used for both a gas cell and a Stokes collector. An excitation laser light at 532 nm is launched into the cell filled with analytes and the Stokes light collected in the cell is detected by the multichannel Raman spectrometer. A high-reflectivity mirror was placed at the output end of the cell for the effective excitation of trace gases. The Raman spectrum of major breath molecule (oxygen, carbon dioxide, and water) is obtained without a serious decrease of the signal-to-noise ratio even if the cell is coiled into a multiple loop with a 3.8 cm radius. Because the cell examined in this report needs very small volume of only 0.4 ml or less, it has great potential for gas analyses that need fast response such as in critical care and operating rooms.

  16. Accumulation and diffusion of crystallin inside single fiber cells in intact chicken embryo lenses.

    PubMed Central

    Peetermans, J A; Foy, B D; Tanaka, T

    1987-01-01

    The use of microscope laser light-scattering spectroscopy allows for the measurement of dynamic properties of intracellular particles inside single fiber cells at different locations in the intact chicken embryo lens. Profiles of the diffusive properties of the delta-crystallin proteins across the lens are reported for developing chickens from day 5 to day 37. A clear decrease of the diffusion is observed in the lens nucleus relative to the cortex beginning with day 10. Images PMID:3470754

  17. Fiber and fabric solar cells by directly weaving carbon nanotube yarns with CdSe nanowire-based electrodes

    NASA Astrophysics Data System (ADS)

    Zhang, Luhui; Shi, Enzheng; Ji, Chunyan; Li, Zhen; Li, Peixu; Shang, Yuanyuan; Li, Yibin; Wei, Jinquan; Wang, Kunlin; Zhu, Hongwei; Wu, Dehai; Cao, Anyuan

    2012-07-01

    Electrode materials are key components for fiber solar cells, and when combined with active layers (for light absorption and charge generation) in appropriate ways, they enable design and fabrication of efficient and innovative device structures. Here, we apply carbon nanotube yarns as counter electrodes in combination with CdSe nanowire-grafted primary electrodes (Ti wire) for making fiber and fabric-shaped photoelectrochemical cells with power conversion efficiencies in the range 1% to 2.9%. The spun-twist long nanotube yarns possess both good electrical conductivity and mechanical flexibility compared to conventional metal wires or carbon fibers, which facilitate fabrication of solar cells with versatile configurations. A unique feature of our process is that instead of making individual fiber cells, we directly weave single or multiple nanotube yarns with primary electrodes into a functional fabric. Our results demonstrate promising applications of semiconducting nanowires and carbon nanotubes in woven photovoltaics.Electrode materials are key components for fiber solar cells, and when combined with active layers (for light absorption and charge generation) in appropriate ways, they enable design and fabrication of efficient and innovative device structures. Here, we apply carbon nanotube yarns as counter electrodes in combination with CdSe nanowire-grafted primary electrodes (Ti wire) for making fiber and fabric-shaped photoelectrochemical cells with power conversion efficiencies in the range 1% to 2.9%. The spun-twist long nanotube yarns possess both good electrical conductivity and mechanical flexibility compared to conventional metal wires or carbon fibers, which facilitate fabrication of solar cells with versatile configurations. A unique feature of our process is that instead of making individual fiber cells, we directly weave single or multiple nanotube yarns with primary electrodes into a functional fabric. Our results demonstrate promising applications

  18. Proliferative Dynamics and the Role of FGF2 During Myogenesis of Rat Satellite Cells on Isolated Fibers

    PubMed Central

    Yablonka-Reuveni, Zipora; Rivera, Anthony J.

    2015-01-01

    Myogenic precursors in adult skeletal muscle (satellite cells) are mitotically quiescent but can proliferate in response to a variety of stresses including muscle injury. To gain further understanding of adult myoblasts, we are analyzing myogenesis of satellite cells on fibers isolated from adult rat muscle. In this culture model, satellite cells are maintained in their in situ position underneath the fiber basement membrane. Employing two different approaches to monitor proliferation of satellite cells on isolated fibers (autoradiography following 3H-thymidine incorporation and immunofluorescence of cells positive for proliferating cell nuclear antigen (PCNA)), we show in the present study that satellite cells initiate cell proliferation at 12 to 24 hours following fiber culture establishment and that cell proliferation is reduced to minimal levels by 60 to 72 hours in culture. Maximal number of proliferating cells is seen at 36 to 48 hours in culture. These PCNA+ satellite cells transit into the differentiated, myogenin+ state following about 24 hours in the proliferative state. Continuous exposure of the fiber culture to FGF2 (basic FGF; added at the time of culture establishment) leads to a 2 fold increase in the number of PCNA+ cells by 48 hours in culture but the overall schedule of proliferation and transition into the myogenin+ state is not affected. Delaying the addition of FGF2 until 15 to 18 hours following the initiation of the fiber culture does not reduce its effect. However, the addition of FGF2 at 24 hours or later results in a progressive reduction in the number of proliferating satellite cells. Exposure of fiber cultures to transforming growth factor β (TGFβ1) leads to a reduction in the number of proliferating cells in both the absence or presence of FGF2. We propose that FGF2 enhances the number of proliferating cells by facilitating the recruitment of additional satellite cells from the quiescent state. However, satellite cells on isolated

  19. Low optical insertion-loss and vacuum-pressure all-fiber acetylene cell based on hollow-core photonic crystal fiber.

    PubMed

    Light, P S; Couny, F; Benabid, F

    2006-09-01

    We report a novel and easy-to-implement hollow-core photonic crystal fiber cell fabrication technique based on helium diffusion through silica. The formed gas cells combine low optical insertion loss (1.8 dB) and vacuum acetylene pressure (microbar regime). The estimates of the final gas pressure, using both Voigt interpolation and electromagnetically induced transparency, show a good match with the initial fitting pressure. PMID:16902611

  20. Pax7 Reveals a Greater Frequency and Concentration of Satellite Cells at the Ends of Growing Skeletal Muscle Fibers

    PubMed Central

    Allouh, Mohammed Z.; Yablonka-Reuveni, Zipora; Rosser, Benjamin W.C.

    2008-01-01

    The main sites of longitudinal growth in skeletal muscle are the ends of the fibers. This study tests the hypothesis that satellite cells (SCs) are at a greater frequency (#SC nuclei/all nuclei within basal laminae) and concentration (closer together) within growing fiber ends of posthatch chicken pectoralis. SCs were localized by their Pax7 expression, and fiber ends were identified by their retention of neonatal myosin heavy chains and small cross-sectional profiles. Whereas SC frequency decreased from about 20% at 9 days posthatch to <5% at 115 days, fiber ends retained a frequency of ∼16%. Calculated mean area of sarcolemma per SC revealed higher concentrations of SCs at fiber ends. There was also a strong inverse correlation between SC frequency and fiber profile cross-sectional size throughout development. This study suggests that SCs at fiber ends play a key role in the longitudinal growth of muscle fibers, and that fiber profile size may impact SC distribution. (J Histochem Cytochem 56:77–87, 2008) PMID:17938281

  1. Hollow core photonic crystal fiber for monitoring leukemia cells using surface enhanced Raman scattering (SERS).

    PubMed

    Khetani, Altaf; Momenpour, Ali; Alarcon, Emilio I; Anis, Hanan

    2015-11-01

    The present paper demonstrates an antibody-free, robust, fast, and portable platform for detection of leukemia cells using Raman spectroscopy with a 785-nm laser diode coupled to a hollow core photonic crystal (HC-PCF) containing silver nanoparticles. Acute myeloid leukemia is one of the most common bone marrow cancers in children and youths. Clinical studies suggest that early diagnosis and remission evaluation of myoblasts in the bone marrow are pivotal for improving patient survival. However, the current protocols for leukemic cells detection involve the use of expensive antibodies and flow cytometers. Thus, we have developed a new technology for detection of leukemia cells up to 300 cells/ml using a compact fiber HC-PCF, which offers a novel alternative to existing clinical standards. Furthermore, we were also able to accurately distinguish live, apoptotic and necrotic leukemic cells. PMID:26601021

  2. Hollow core photonic crystal fiber for monitoring leukemia cells using surface enhanced Raman scattering (SERS)

    PubMed Central

    Khetani, Altaf; Momenpour, Ali; Alarcon, Emilio I.; Anis, Hanan

    2015-01-01

    The present paper demonstrates an antibody-free, robust, fast, and portable platform for detection of leukemia cells using Raman spectroscopy with a 785-nm laser diode coupled to a hollow core photonic crystal (HC-PCF) containing silver nanoparticles. Acute myeloid leukemia is one of the most common bone marrow cancers in children and youths. Clinical studies suggest that early diagnosis and remission evaluation of myoblasts in the bone marrow are pivotal for improving patient survival. However, the current protocols for leukemic cells detection involve the use of expensive antibodies and flow cytometers. Thus, we have developed a new technology for detection of leukemia cells up to 300 cells/ml using a compact fiber HC-PCF, which offers a novel alternative to existing clinical standards. Furthermore, we were also able to accurately distinguish live, apoptotic and necrotic leukemic cells. PMID:26601021

  3. Columnar deformation of human red blood cell by highly localized fiber optic Bessel beam stretcher

    PubMed Central

    Lee, Sungrae; Joo, Boram; Jeon, Pyo Jin; Im, Seongil; Oh, Kyunghwan

    2015-01-01

    A single human red blood cell was optically stretched along two counter-propagating fiber-optic Bessel-like beams in an integrated lab-on-a-chip structure. The beam enabled highly localized stretching of RBC, and it induced a nonlinear mechanical deformation to finally reach an irreversible columnar shape that has not been reported. We characterized and systematically quantified this optically induced mechanical deformation by the geometrical aspect ratio of stretched RBC and the irreversible stretching time. The proposed RBC mechanism can realize a versatile and compact opto-mechanical platform for optical diagnosis of biological substances in the single cell level. PMID:26601005

  4. Columnar deformation of human red blood cell by highly localized fiber optic Bessel beam stretcher.

    PubMed

    Lee, Sungrae; Joo, Boram; Jeon, Pyo Jin; Im, Seongil; Oh, Kyunghwan

    2015-11-01

    A single human red blood cell was optically stretched along two counter-propagating fiber-optic Bessel-like beams in an integrated lab-on-a-chip structure. The beam enabled highly localized stretching of RBC, and it induced a nonlinear mechanical deformation to finally reach an irreversible columnar shape that has not been reported. We characterized and systematically quantified this optically induced mechanical deformation by the geometrical aspect ratio of stretched RBC and the irreversible stretching time. The proposed RBC mechanism can realize a versatile and compact opto-mechanical platform for optical diagnosis of biological substances in the single cell level. PMID:26601005

  5. Dynamics of myosin II organization into contractile networks and fibers at the medial cell cortex

    NASA Astrophysics Data System (ADS)

    Nie, Wei

    The cellular morphology of adhered cells depends crucially on the formation of a contractile meshwork of parallel and cross-linked stress fibers along the contacting surface. The motor activity and mini-filament assembly of non-muscle myosin II is an important component of cell-level cytoskeletal remodeling during mechanosensing. To monitor the dynamics of non-muscle myosin II, we used confocal microscopy to image cultured HeLa cells that stably express myosin regulatory light chain tagged with GFP (MRLC-GFP). MRLC-GFP was monitored in time-lapse movies at steady state and during the response of cells to varying concentrations of blebbistatin (which disrupts actomyosin stress fibers). Using image correlation spectroscopy analysis, we quantified the kinetics of disassembly and reassembly of actomyosin networks and compared to studies by other groups. This analysis suggested the following processes: myosin minifilament assembly and disassembly; aligning and contraction; myosin filament stabilization upon increasing contractile tension. Numerical simulations that include those processes capture some of the main features observed in the experiments. This study provides a framework to help interpret how different cortical myosin remodeling kinetics may contribute to different cell shape and rigidity depending on substrate stiffness. We discuss methods to monitor myosin reorganization using non-linear imaging methods.

  6. Asbestos fibers mediate transformation of monkey cells by exogenous plasmid DNA

    SciTech Connect

    Appel, J.D.; Fasy, T.M.; Kohtz, D.S.; Kohtz, J.D.; Johnson, E.M. )

    1988-10-01

    The authors have tested the ability of chrysotile asbestos fibers to introduce plasmid DNA into monkey COS-7 cells and the ability of this DNA to function in both replication and gene expression. Chrysotile fibers are at least as effective as calcium phosphate in standard transfection assays at optimal ratios of asbestos to DNA. After transfection with chrysotile, a minor percentage of introduced plasmid DNA bearing a simian virus 40 origin of replication replicates after 24 hr. Fragmentation of entering DNA is more prominent with asbestos than with calcium phosphate, and after 72 hr most DNA introduced by asbestos is associated with chromosomal DNA. Cells transfected with plasmid p11-4, bearing the p53 protooncogene, express this gene. Cells transfected with pSV2-neo express a gene conferring resistance of antibiotic G418, allowing isolation of colonies of transformed cells after 18 days. The introduction of exogenous DNA into eukaryotic cells could cause mutations in several ways and thus contribute to asbestos-induced oncogenesis.

  7. Characterization of Palladin, a Novel Protein Localized to Stress Fibers and Cell Adhesions

    PubMed Central

    Parast, Mana M.; Otey, Carol A.

    2000-01-01

    Here, we describe the identification of a novel phosphoprotein named palladin, which colocalizes with α-actinin in the stress fibers, focal adhesions, cell–cell junctions, and embryonic Z-lines. Palladin is expressed as a 90–92-kD doublet in fibroblasts and coimmunoprecipitates in a complex with α-actinin in fibroblast lysates. A cDNA encoding palladin was isolated by screening a mouse embryo library with mAbs. Palladin has a proline-rich region in the NH2-terminal half of the molecule and three tandem Ig C2 domains in the COOH-terminal half. In Northern and Western blots of chick and mouse tissues, multiple isoforms of palladin were detected. Palladin expression is ubiquitous in embryonic tissues, and is downregulated in certain adult tissues in the mouse. To probe the function of palladin in cultured cells, the Rcho-1 trophoblast model was used. Palladin expression was observed to increase in Rcho-1 cells when they began to assemble stress fibers. Antisense constructs were used to attenuate expression of palladin in Rcho-1 cells and fibroblasts, and disruption of the cytoskeleton was observed in both cell types. At longer times after antisense treatment, fibroblasts became fully rounded. These results suggest that palladin is required for the normal organization of the actin cytoskeleton and focal adhesions. PMID:10931874

  8. Use of fiber-optic-based flow cells and probes in the chemical and petroleum industries

    NASA Astrophysics Data System (ADS)

    Ponstingl, Mike; Vetter, Hans

    1992-08-01

    Fiber-optic-based, on-line photometric/spectrophotometric analytic methods are becoming increasingly more important in the process control industries because of unique benefits such as: safety, real-time data capture, immunity to EMI/RFI, and simplicity of installation. End- users employing fiber-optic methods are experiencing increased production yields, less waster, and greater product consistency all because the process can be controlled more efficiently. Extractive and in-situ flow cells have proven to be valuable means of 'looking' at the process stream. These techniques are currently being offered by several manufacturers of on-line fiber-optic photometric/spectrophotometric instruments. Custom Sensors & Technology has developed practical techniques for optical energy transmission in the 250 - 2000 nm wavelength range. In addition to discussing extractive and in-situ methods of sampling, various design considerations are addressed which relate to the efficiency of coupling light energy into and out of extractive flow cells and in-situ probes. In-situ probes can be of the transmission, turbidity, attenuated total reflection, or diffuse reflection types; and can be installed in a sanitary or threaded pipe fitting.

  9. Dietary fiber and short-chain fatty acids affect cell proliferation and protein synthesis in isolated rat colonocytes.

    PubMed

    Marsman, K E; McBurney, M I

    1996-05-01

    Colonic metabolism may be affected by dietary fiber and short-chain fatty acids, the products of fiber fermentation. The aim of this study was to assess the effects of fiber supplementation (150 g/kg diet) on dynamic measurements of metabolism in isolated rat colonic epithelial cells. Additionally, we investigated the effect of in vitro short-chain fatty acid and glutamine concentrations and media osmolarity on oxygen uptake, protein synthesis, cell proliferation and anaplerotic flux. Colonocyte oxygen consumption did not differ due to fiber supplementation or the inclusion of short -chain fatty acids in incubation media. Cell proliferation (3H-thymidine uptake) was increased by fiber consumption (P fiber supplementation but was decreased when short-chain fatty acids were present in incubation media (P fiber supplementation increases in vitro colonocyte proliferation, the unchanged oxygen uptake rate indicates that there was no concurrent increase in energy expenditure. PMID:8618140

  10. Plastic fiber optics for micro-imaging of fluorescence signals in living cells

    NASA Astrophysics Data System (ADS)

    Sakurai, Takashi; Natsume, Mitsuo; Koida, Kowa

    2015-03-01

    The fiber-coupled microscope (FCM) enables in vivo imaging at deep sites in the tissues or organs that other optical techniques are unable to reach. To develop FCM-based intravital imaging, we employed a plastic optical fiber (POF) bundle that included more than 10,000-units of polystyrene core and polymethyl methacrylate cladding. Each POF had a diameter of less than 5 μm the tip of the bundle was less than 0.5 mm wide, and the flexible wire had a length of 1,000 mm. The optical performance of the plastic FCM was sufficient for detection of significant signal changes in an acinus of rat pancreas labeled with a calcium ion-sensitive fluorescent dye. In the future, the potential power of plastic FCM is expected to increase, enabling analysis of structure and organization of specific functions in live cells within vulnerable organs.

  11. Miniaturized ascorbic acid fuel cells with flexible electrodes made of graphene-coated carbon fiber cloth

    NASA Astrophysics Data System (ADS)

    Hoshi, Kazuki; Muramatsu, Kazuo; Sumi, Hisato; Nishioka, Yasushiro

    2016-04-01

    Ascorbic acid (AA) is a biologically friendly compound and exists in many products such as sports drinks, fruit, and even in human blood. Thus, a miniaturized and flexible ascorbic acid fuel cell (AAFC) is expected be a power source for portable or implantable electric devices. In this study, we fabricated an AAFC with anode and cathode dimensions of 3 × 10 mm2 made of a graphene-coated carbon fiber cloth (GCFC) and found that GCFC electrodes significantly improve the power generated by the AAFC. This is because the GCFC has more than two times the effective surface area of a conventional carbon fiber cloth and it can contain more enzymes. The power density of the AAFC in a phosphate buffer solution containing 100 mM AA at room temperature was 34.1 µW/cm2 at 0.46 V. Technical issues in applying the AAFC to portable devices are also discussed.

  12. GMP-Compliant Expansion of Clinical-Grade Human Mesenchymal Stromal/Stem Cells Using a Closed Hollow Fiber Bioreactor.

    PubMed

    Barckhausen, Christina; Rice, Brent; Baila, Stefano; Sensebé, Luc; Schrezenmeier, Hubert; Nold, Philipp; Hackstein, Holger; Rojewski, Markus Thomas

    2016-01-01

    This chapter describes a method for GMP-compliant expansion of human mesenchymal stromal/stem cells (hMSC) from bone marrow aspirates, using the Quantum(®) Cell Expansion System from Terumo BCT. The Quantum system is a functionally closed, automated hollow fiber bioreactor system designed to reproducibly grow cells in either GMP or research laboratory environments. The chapter includes protocols for preparation of media, setup of the Quantum system, coating of the hollow fiber bioreactor, as well as loading, feeding, and harvesting of cells. We suggest a panel of quality controls for the starting material, the interim product, as well as the final product. PMID:27236685

  13. Sialylation of vitronectin regulates stress fiber formation and cell spreading of dermal fibroblasts via a heparin-binding site.

    PubMed

    Miyamoto, Yasunori; Tanabe, Mio; Date, Kimie; Sakuda, Kanoko; Sano, Kotone; Ogawa, Haruko

    2016-04-01

    Vitronectin (VN) plays an important role in tissue regeneration. We previously reported that VN from partial hepatectomized (PH) rats results in a decrease of sialylation of VN and de-sialylation of VN decreases the cell spreading of hepatic stellate cells. In this study, we analyzed the mechanism how sialylation of VN regulates the properties of mouse primary cultured dermal fibroblasts (MDF) and a dermal fibroblast cell line, Swiss 3T3 cells. At first, we confirmed that VN from PH rats or de-sialylated VN also decreased cell spreading in MDF and Swiss 3T3 cells. The de-sialylation suppressed stress fiber formation in Swiss 3T3 cells. Next, we analyzed the effect of the de-sialylation of VN on stress fiber formation in Swiss 3T3 cells. RGD peptide, an inhibitor for a cell binding site of VN, did not affect the cell attachment of Swiss 3T3 cells on untreated VN but significantly decreased it on de-sialylated VN, suggesting that the de-sialylation attenuates the binding activity of an RGD-independent binding site in VN. To analyze a candidate RGD-independent binding site, an inhibition experiment of stress fiber formation for a heparin binding site was performed. The addition of heparin and treatment of cells with heparinase decreased stress fiber formation in Swiss 3T3 cells. Furthermore, de-sialylation increased the binding activity of VN to heparin, as detected by surface plasmon resonance (SPR). These results demonstrate that sialylation of VN glycans regulates stress fiber formation and cell spreading of dermal fibroblast cells via a heparin binding site. PMID:26979432

  14. Influence of exercise contraction mode and protein supplementation on human skeletal muscle satellite cell content and muscle fiber growth

    PubMed Central

    Farup, Jean; Rahbek, Stine Klejs; Riis, Simon; Vendelbo, Mikkel Holm; de Paoli, Frank

    2014-01-01

    Skeletal muscle satellite cells (SCs) are involved in remodeling and hypertrophy processes of skeletal muscle. However, little knowledge exists on extrinsic factors that influence the content of SCs in skeletal muscle. In a comparative human study, we investigated the muscle fiber type-specific association between emergence of satellite cells (SCs), muscle growth, and remodeling in response to 12 wk unilateral resistance training performed as eccentric (Ecc) or concentric (Conc) resistance training ± whey protein (Whey, 19.5 g protein + 19.5 g glucose) or placebo (Placebo, 39 g glucose) supplementation. Muscle biopsies (vastus lateralis) were analyzed for fiber type-specific SCs, myonuclei, and fiber cross-sectional area (CSA). Following training, SCs increased with Conc in both type I and type II fibers (P < 0.01) and exhibited a group difference from Ecc (P < 0.05), which did not increase. Myonuclei content in type I fibers increased in all groups (P < 0.01), while a specific accretion of myonuclei in type II fibers was observed in the Whey-Conc (P < 0.01) and Placebo-Ecc (P < 0.01) groups. Similarly, whereas type I fiber CSA increased independently of intervention (P < 0.001), type II fiber CSA increased exclusively with Whey-Conc (P < 0.01) and type II fiber hypertrophy correlated with whole muscle hypertrophy exclusively following Conc training (P < 0.01). In conclusion, isolated concentric knee extensor resistance training appears to constitute a stronger driver of SC content than eccentric resistance training while type II fiber hypertrophy was accentuated when combining concentric resistance training with whey protein supplementation. PMID:25103976

  15. High Intensity Training May Reverse the Fiber Type Specific Decline in Myogenic Stem Cells in Multiple Sclerosis Patients.

    PubMed

    Farup, Jean; Dalgas, Ulrik; Keytsman, Charly; Eijnde, Bert O; Wens, Inez

    2016-01-01

    Multiple sclerosis (MS) is associated with loss of skeletal muscle mass and function. The myogenic stem cells (satellite cells-SCs) are instrumental to accretion of myonuclei, but remain to be investigated in MS. The present study aimed to compare the SC and myonuclei content between MS patients (n = 23) and age matched healthy controls (HC, n = 18). Furthermore, the effects of 12 weeks of high intensity training on SC and myonuclei content were explored in MS. Muscle biopsies were obtained from m. Vastus Lateralis at baseline (MS and HC) and following 12 weeks of training (MS only). Frozen biopsies were sectioned followed by immunohistochemical analysis for fiber type specific SCs (Pax7(+)), myonuclei (MN) and central nuclei content and fiber cross-sectional area (fCSA) was quantified using ATPase histochemistry. At baseline the SCs per fiber was lower in type II compared to type I fibers in both MS (119%, p < 0.01) and HC (69%, p < 0.05), whereas the SCs per fCSA was lower in type II fibers compared to type I only in MS (72%, p < 0.05). No differences were observed in MN or central nuclei between MS and HC. Following training the type II fiber SCs per fiber and per fCSA in MS patients increased by 165% (p < 0.05) and 135% (p < 0.05), respectively. Furthermore, the type II fiber MN content tended (p = 0.06) to be increased by 35% following training. In conclusion, the SC content is lower in type II compared to type I fibers in both MS and HC. Furthermore, high intensity training was observed to selectively increase the SC and myonuclei content in type II fibers in MS patients. PMID:27303309

  16. Intraluminal acid activates esophageal nodose C fibers after mast cell activation.

    PubMed

    Zhang, Shizhong; Liu, Zhenyu; Heldsinger, Andrea; Owyang, Chung; Yu, Shaoyong

    2014-02-01

    Acid reflux in the esophagus can induce esophageal painful sensations such as heartburn and noncardiac chest pain. The mechanisms underlying acid-induced esophageal nociception are not clearly understood. In our previous studies, we characterized esophageal vagal nociceptive afferents and defined their responses to noxious mechanical and chemical stimulation. In the present study, we aim to determine their responses to intraluminal acid infusion. Extracellular single-unit recordings were performed in nodose ganglion neurons with intact nerve endings in the esophagus using ex vivo esophageal-vagal preparations. Action potentials evoked by esophageal intraluminal acid perfusion were compared in naive and ovalbumin (OVA)-challenged animals, followed by measurements of transepithelial electrical resistance (TEER) and the expression of tight junction proteins (zona occludens-1 and occludin). In naive guinea pigs, intraluminal infusion with either acid (pH = 2-3) or capsaicin did not evoke an action potential discharge in esophageal nodose C fibers. In OVA-sensitized animals, following esophageal mast cell activation by in vivo OVA inhalation, intraluminal acid infusion for about 20 min started to evoke action potential discharges. This effect is further confirmed by selective mast cell activation using in vitro tissue OVA challenge in esophageal-vagal preparations. OVA inhalation leads to decreased TEER and zona occludens-1 expression, suggesting an impaired esophageal epithelial barrier function after mast cell activation. These data for the first time provide direct evidence of intraluminal acid-induced activation of esophageal nociceptive C fibers and suggest that mast cell activation may make esophageal epithelium more permeable to acid, which subsequently may increase esophageal vagal nociceptive C fiber activation. PMID:24264049

  17. Application of differential fiber Bragg grating displacement cell in bridge crack monitoring

    NASA Astrophysics Data System (ADS)

    Yue, Li-na; Huang, Jun; Jiang, De-sheng; Wang, Jun-jie

    2008-12-01

    During the construction and service period of concrete bridges, the cracks often influence the quality of the project even the safety of the structure. An effective and long-term crack monitoring of concrete bridges with the appropriate choice of displacement sensors is imperative under the situation. The differential fiber bragg grating displacement cell is based on the composite structure which consisted of pulling spring and cantilever. It has realized differential measure of normal FBG displacement sensor and has solved the serious problem of temperature disturbance. The differential fiber bragg grating displacement cell has the advantages of high accuracy, anti-interference, long distance transmitting and good durability etc. In this paper, the differential fiber bragg grating displacement cells were applied to monitor the cracks of web slabs during the tension process of external prestressed tendons when the continuous prestressed concrete box girder bridges and continuous concrete rigid frame bridges were maintained and reinforced. A group of typical cracks of web slabs was selected respectively in the continuous prestressed concrete box girder bridge and the continuous concrete rigid frame bridge. And a group of three sensors were installed across the three cracks. The external prestressed tendons had been tensioned by four grades. Then the widths of these cracks were recorded in accordance with the four tension grades of the external prestressed tendons: before tension, 10% tension, 80% tension, 100% tension. The results of the differential FBG displacement cells used during the process of tension of external prestressed tendons show that the cracks monitoring data are accurate and in accordance with the cracks changing rule.

  18. Miniature fiber optic spectrometer-based quantitative fluorescence resonance energy transfer measurement in single living cells

    NASA Astrophysics Data System (ADS)

    Chai, Liuying; Zhang, Jianwei; Zhang, Lili; Chen, Tongsheng

    2015-03-01

    Spectral measurement of fluorescence resonance energy transfer (FRET), spFRET, is a widely used FRET quantification method in living cells today. We set up a spectrometer-microscope platform that consists of a miniature fiber optic spectrometer and a widefield fluorescence microscope for the spectral measurement of absolute FRET efficiency (E) and acceptor-to-donor concentration ratio (RC) in single living cells. The microscope was used for guiding cells and the spectra were simultaneously detected by the miniature fiber optic spectrometer. Moreover, our platform has independent excitation and emission controllers, so different excitations can share the same emission channel. In addition, we developed a modified spectral FRET quantification method (mlux-FRET) for the multiple donors and multiple acceptors FRET construct (mD˜nA) sample, and we also developed a spectra-based 2-channel acceptor-sensitized FRET quantification method (spE-FRET). We implemented these modified FRET quantification methods on our platform to measure the absolute E and RC values of tandem constructs with different acceptor/donor stoichiometries in single living Huh-7 cells.

  19. ADF and Cofilin1 Control Actin Stress Fibers, Nuclear Integrity, and Cell Survival

    PubMed Central

    Kanellos, Georgios; Zhou, Jing; Patel, Hitesh; Ridgway, Rachel A.; Huels, David; Gurniak, Christine B.; Sandilands, Emma; Carragher, Neil O.; Sansom, Owen J.; Witke, Walter; Brunton, Valerie G.; Frame, Margaret C.

    2015-01-01

    Summary Genetic co-depletion of the actin-severing proteins ADF and CFL1 triggers catastrophic loss of adult homeostasis in multiple tissues. There is impaired cell-cell adhesion in skin keratinocytes with dysregulation of E-cadherin, hyperproliferation of differentiated cells, and ultimately apoptosis. Mechanistically, the primary consequence of depleting both ADF and CFL1 is uncontrolled accumulation of contractile actin stress fibers associated with enlarged focal adhesions at the plasma membrane, as well as reduced rates of membrane protrusions. This generates increased intracellular acto-myosin tension that promotes nuclear deformation and physical disruption of the nuclear lamina via the LINC complex that normally connects regulated actin filaments to the nuclear envelope. We therefore describe a pathway involving the actin-severing proteins ADF and CFL1 in regulating the dynamic turnover of contractile actin stress fibers, and this is vital to prevent the nucleus from being damaged by actin contractility, in turn preserving cell survival and tissue homeostasis. PMID:26655907

  20. Muscle cell membranes from early degeneration muscle cell fibers in Solenopsis are leaky to lanthanum: electron microscopy and X-ray analysis

    SciTech Connect

    Jones, R.G.; Davis, W.L.

    1985-06-01

    Lanthanum infusion techniques, transmission electron microscopy, and X-ray microanalysis were utilized to compare the permeability of muscle cell membranes from normal and degenerating muscle fibers of Solenopsis spp. In normal fibers, the electron-dense tracer was limited to components of the sarcotubular system. However, the insemination-induced degeneration of muscle fibers was characterized by the presence of an electron-dense precipitate within the myofibrils and mitochondria as well as in the extramyofibrillar spaces. The electron-dense material was subsequently identified by elemental analysis to be lanthanum. Such data indicate that one of the earliest stages of muscle degeneration involves an alteration in cell membrane permeability.

  1. The effect of various denier capillary channel polymer fibers on the alignment of NHDF cells and type I collagen.

    PubMed

    Sinclair, Kristofer D; Webb, Ken; Brown, Philip J

    2010-12-15

    If tissue engineers are to successfully repair and regenerate native tendons and ligaments, it will be essential to implement contact guidance to induce cellular and type I collagen alignment to replicate the native structure. Capillary channel polymer (CC-P) fibers fabricated by melt-extrusion have aligned micrometer scale surface channels that may serve the goal of achieving biomimetic, physical templates for ligament growth and regeneration. Previous work characterizing the behavior of normal human dermal fibroblasts (NHDF), on the 19 denier per filament (dpf) CC-P fibers, demonstrated a need for improved cellular and type I collagen alignment. Therefore, 5 and 9 dpf CC-P fibers were manufactured to determine whether their channel dimensions would achieve greater alignment. A 29 dpf CC-P fiber was also examined to determine whether cellular guidance could still be achieved within the larger dimensions of the fiber's channels. The 9 dpf CC-P fiber appeared to approach the topographical constraints necessary to induce the cellular and type I collagen architecture that most closely mirrored that of native ACL tissue. This work demonstrated that the novel cross-section of the CC-P fiber geometry could approach the necessary surface topography to align NHDF cells along the longitudinal axis of each fiber. PMID:20925084

  2. Identification and Ultrastructural Characterization of a Novel Nuclear Degradation Complex in Differentiating Lens Fiber Cells.

    PubMed

    Costello, M Joseph; Brennan, Lisa A; Mohamed, Ashik; Gilliland, Kurt O; Johnsen, Sönke; Kantorow, Marc

    2016-01-01

    An unresolved issue in structural biology is how the encapsulated lens removes membranous organelles to carry out its role as a transparent optical element. In this ultrastructural study, we establish a mechanism for nuclear elimination in the developing chick lens during the formation of the organelle-free zone. Day 12-15 chick embryo lenses were examined by high-resolution confocal light microscopy and thin section transmission electron microscopy (TEM) following fixation in 10% formalin and 4% paraformaldehyde, and then processing for confocal or TEM as described previously. Examination of developing fiber cells revealed normal nuclei with dispersed chromatin and clear nucleoli typical of cells in active ribosome production to support protein synthesis. Early signs of nuclear degradation were observed about 300 μm from the lens capsule in Day 15 lenses where the nuclei display irregular nuclear stain and prominent indentations that sometimes contained a previously undescribed macromolecular aggregate attached to the nuclear envelope. We have termed this novel structure the nuclear excisosome. This complex by confocal is closely adherent to the nuclear envelope and by TEM appears to degrade the outer leaflet of the nuclear envelope, then the inner leaflet up to 500 μm depth. The images suggest that the nuclear excisosome separates nuclear membrane proteins from lipids, which then form multilamellar assemblies that stain intensely in confocal and in TEM have 5 nm spacing consistent with pure lipid bilayers. The denuded nucleoplasm then degrades by condensation and loss of structure in the range 600 to 700 μm depth producing pyknotic nuclear remnants. None of these stages display any classic autophagic vesicles or lysosomes associated with nuclei. Uniquely, the origin of the nuclear excisosome is from filopodial-like projections of adjacent lens fiber cells that initially contact, and then appear to fuse with the outer nuclear membrane. These filopodial

  3. Identification and Ultrastructural Characterization of a Novel Nuclear Degradation Complex in Differentiating Lens Fiber Cells

    PubMed Central

    Costello, M. Joseph; Brennan, Lisa A.; Gilliland, Kurt O.; Johnsen, Sönke; Kantorow, Marc

    2016-01-01

    An unresolved issue in structural biology is how the encapsulated lens removes membranous organelles to carry out its role as a transparent optical element. In this ultrastructural study, we establish a mechanism for nuclear elimination in the developing chick lens during the formation of the organelle-free zone. Day 12–15 chick embryo lenses were examined by high-resolution confocal light microscopy and thin section transmission electron microscopy (TEM) following fixation in 10% formalin and 4% paraformaldehyde, and then processing for confocal or TEM as described previously. Examination of developing fiber cells revealed normal nuclei with dispersed chromatin and clear nucleoli typical of cells in active ribosome production to support protein synthesis. Early signs of nuclear degradation were observed about 300 μm from the lens capsule in Day 15 lenses where the nuclei display irregular nuclear stain and prominent indentations that sometimes contained a previously undescribed macromolecular aggregate attached to the nuclear envelope. We have termed this novel structure the nuclear excisosome. This complex by confocal is closely adherent to the nuclear envelope and by TEM appears to degrade the outer leaflet of the nuclear envelope, then the inner leaflet up to 500 μm depth. The images suggest that the nuclear excisosome separates nuclear membrane proteins from lipids, which then form multilamellar assemblies that stain intensely in confocal and in TEM have 5 nm spacing consistent with pure lipid bilayers. The denuded nucleoplasm then degrades by condensation and loss of structure in the range 600 to 700 μm depth producing pyknotic nuclear remnants. None of these stages display any classic autophagic vesicles or lysosomes associated with nuclei. Uniquely, the origin of the nuclear excisosome is from filopodial-like projections of adjacent lens fiber cells that initially contact, and then appear to fuse with the outer nuclear membrane. These filopodial

  4. Smooth Muscle Cell Alignment and Phenotype Control by Melt Spun Polycaprolactone Fibers for Seeding of Tissue Engineered Blood Vessels

    PubMed Central

    Agrawal, Animesh; Lee, Bae Hoon; Irvine, Scott A.; An, Jia; Bhuthalingam, Ramya; Singh, Vaishali; Low, Kok Yao; Chua, Chee Kai; Venkatraman, Subbu S.

    2015-01-01

    A method has been developed to induce and retain a contractile phenotype for vascular smooth muscle cells, as the first step towards the development of a biomimetic blood vessel construct with minimal compliance mismatch. Melt spun PCL fibers were deposited on a mandrel to form aligned fibers of 10 μm in diameter. The fibers were bonded into aligned arrangement through dip coating in chitosan solution. This formed a surface of parallel grooves, 10 μm deep by 10 μm across, presenting a surface layer of chitosan to promote cell surface interactions. The aligned fiber surface was used to culture cells present in the vascular wall, in particular fibroblasts and smooth muscle cells. This topography induced “surface guidance” over the orientation of the cells, which adopted an elongated spindle-like morphology, whereas cells on the unpatterned control surface did not show such orientation, assuming more rhomboid shapes. The preservation of VSMC contractile phenotype on the aligned scaffold was demonstrated by the retention of α-SMA expression after several days of culture. The effect was assessed on a prototype vascular graft prosthesis fabricated from polylactide caprolactone; VSMCs aligned longitudinally along a fiberless tube, whereas, for the aligned fiber coated tubes, the VSMCs aligned in the required circumferential orientation. PMID:26413093

  5. Short-term modulation of cerebellar Purkinje cell activity after spontaneous climbing fiber input.

    PubMed

    Sato, Y; Miura, A; Fushiki, H; Kawasaki, T

    1992-12-01

    1. There are two opposite points of view concerning the way climbing fiber input in a Purkinje cell modifies simple spike (SS) activity transiently: depression versus enhancement of SS activity. The different groups of investigators favored one effect predominating over the other. In the decerebrate unanesthetized cat, we recorded spontaneous activity of single Purkinje cells and investigated time course of SS activity after the complex spike (CS). 2. In the peri-CS time histogram, there was a SS pause lasting, on average, 10.8 ms after onset of the CS in all of the 316 cells recorded. The pause was followed by a rapid increase in SS activity to a maximum, which was on average 175.6% of a pre-CS control level, and a gradual return to around the control level in the majority of the cells recorded (pause-facilitation type, 71.2%). The increase in SS activity was significant (P < 0.01, t test) during 20-100 ms. The SS activity during the 20-100 ms was, on average, 163.7% of the control level. In some cells (pure-pause type, 25.3%), no significant changes were found (P > 0.01) in the post-pause SS firing. In contrast, only 3.5% of the cells (pause-reduction type) showed a significant (P < 0.01) firing decrease (average 54.0% of the control level) lasting 20-60 ms after the pause period. 3. Analysis of the pre-CS time histogram revealed no significant differences (P > 0.01) in the SS activity between pre-CS periods in all of the cells recorded, suggesting that the SS activity enhancement is not due to a coactivated mossy fiber input just preceding the activation of the climbing fiber input. 4. Analysis of the raster diagram revealed variability of individual SS responses after the CS. The probability of occurrence of the increase in SS number during a post-CS period of 0-100 ms with respect to that during a pre-CS period of -100-0 ms in individual raster traces was high (on average 78.2%), medium (57.3%), and low (36.3%) in the pause-facilitation, pure-pause, and pause

  6. Enhanced performance of electrospun carbon fibers modified with carbon nanotubes: promising electrodes for enzymatic biofuel cells

    NASA Astrophysics Data System (ADS)

    Both Engel, A.; Cherifi, A.; Tingry, S.; Cornu, D.; Peigney, A.; Laurent, Ch

    2013-06-01

    New nanostructured electrodes, promising for the production of clean and renewable energy in biofuel cells, were developed with success. For this purpose, carbon nanofibers were produced by the electrospinning of polyacrylonitrile solution followed by convenient thermal treatments (stabilization followed by carbonization at 1000, 1200 and 1400° C), and carbon nanotubes were adsorbed on the surfaces of the fibers by a dipping method. The morphology of the developed electrodes was characterized by several techniques (SEM, Raman spectroscopy, electrical conductivity measurement). The electrochemical properties were evaluated through cyclic voltammetry, where the influence of the carbonization temperature of the fibers and the beneficial contribution of the carbon nanotubes were observed through the reversibility and size of the redox peaks of K3Fe(CN)6 versus Ag/AgCl. Subsequently, redox enzymes were immobilized on the electrodes and the electroreduction of oxygen to water was realized as a test of their efficiency as biocathodes. Due to the fibrous and porous structure of these new electrodes, and to the fact that carbon nanotubes may have the ability to promote electron transfer reactions of redox biomolecules, the new electrodes developed were capable of producing higher current densities than an electrode composed only of electrospun carbon fibers.

  7. Independent replication of mitochondrial genes supports the transcriptional program in developing fiber cells of cotton (Gossypium hirsutum L.).

    PubMed

    Thyssen, Gregory N; Song, Xianliang; Naoumkina, Marina; Kim, Hee-Jin; Fang, David D

    2014-07-01

    The mitochondrial genomes of flowering plants exist both as a "master circle" chromosome and as numerous subgenomic sublimons that are generated by intramolecular recombination. Differential stability or replication of these sublimons allows individual mitochondrial gene copy numbers to vary independently between different cell types and developmental stages. Our objective was to determine the relationship between mitochondrial gene copy number and transcript abundance in the elongating fiber cells of Upland cotton (Gossypium hirsutum L.). We compared RNA and DNA from cotton fiber cells at five developmental time points from early elongation through secondary cell wall thickening from the Ligon-lintless 2 (Li2) short fiber mutant and its wild type near isogenic line (NIL) DP5690. Mitochondrial gene copy number decreased from 3 to 8-DPA in the developing cotton fiber cells while transcript levels remained low. As secondary cell wall biosynthesis began in developing fibers, the expression levels and copy numbers of mitochondrial genes involved in energy production and respiration were up-regulated in wild type cotton DP5690. However, the short fiber mutant Li2, failed to increase expression of these genes, which include three subunits of ATP synthase, atp1, atp8 and atp9 and two cytochrome genes cox1 and cob. At the same time, Li2 failed to increase the copy numbers of these highly expressed genes. Surprisingly, we found that when mitochondrial genes were highly transcribed, they also had very high copy numbers. This observation suggests that in developing cotton fibers, increased mitochondrial sublimon replication may support increases in gene transcription. PMID:24768176

  8. Development of a micro-fiber nickel electrode for nickel-hydrogen cell

    NASA Technical Reports Server (NTRS)

    Britton, Doris L.

    1995-01-01

    Development of a high specific energy nickel electrode is the main goal of the lightweight nickel electrode program at the NASA Lewis Research Center. The approach has been to improve the nickel electrode by continuing combined in-house and contract efforts to develop a more efficient and lighter weight electrode for the nickel-hydrogen cell. Small fiber diameter nickel plaques are used as conductive supports for the nickel hydroxide active material. These plaques are commercial products and have an advantage of increased surface area available for the deposition of active material. Initial tests include activation and capacity measurements at different discharge levels followed by half-cell cycle testing at 80 percent depth-of-discharge in a low-Earth-orbit regime. The electrodes that pass the initial tests are life cycle-tested in a boiler plate nickel-hydrogen cell before flightweight designs are built and tested.

  9. Development of a Micro-Fiber Nickel Electrode for Nickel-Hydrogen Cell

    NASA Technical Reports Server (NTRS)

    Britton, Doris L.

    1996-01-01

    The development of a high specific energy battery is one of the objectives of the lightweight nickel-hydrogen (NiH2) program at the NASA Lewis Research Center. The approach has been to improve the nickel electrode by continuing combined in-house and contract efforts to develop a more efficient and lighter weight electrode for the nickel-hydrogen fuel cell. Small fiber diameter nickel plaques are used as conductive supports for the nickel hydroxide active material. These plaques are commercial products and have an advantage of increased surface area available for the deposition of active materials. Initial tests include activation and capacity measurements at different discharge levels followed by half-cell cycle testing at 80 percent depth-of-discharge in a low Earth orbit regime. The electrodes that pass the initial tests are life cycle tested in a boiler plate nickel-hydrogen cell before flightweight designs are built and tested.

  10. Direct Tracking of Amyloid and Tu Dynamics in Neuroblastoma Cells Using Nanoplasmonic Fiber Tip Probes.

    PubMed

    Liang, Feng; Zhang, Yiying; Hong, Wooyoung; Dong, Yuanlin; Xie, Zhongcong; Quan, Qimin

    2016-07-13

    Amyloid plaques and neurofibrillary tangles are the pathological hallmarks of Alzheimer's disease. However, there has been a long-standing discussion on the dynamic relations between Aβ and tau proteins, partially due to the lack of a tool to track protein dynamics in individual live neurons at the early stage of Aβ generation and tau phosphorylation. Here, we developed nanoplasmonic fiber tip probe (nFTP) technology to simultaneously monitor Aβ42 generation and tau phosphorylation (at serine 262) in living, single neuroblastoma cells over 12 h. We observed that Aβ42 generation, under clinically relevant anesthetic treatment, preceded tau phosphorylation, which then facilitated Aβ42 generation. This observation is also supported by measuring proteins in cell lysates using the ultrasensitive label-free photonic crystal nanosensors. nFTP therefore provides an advanced method to investigate protein expression and post-translational modification in live cells and determine outcomes of intervention of Alzheimer's disease and other neurodegenerative disorders. PMID:27266855

  11. Ultrafine polybenzimidazole (PBI) fibers. [separators for alkaline batteries and dfuel cells

    NASA Technical Reports Server (NTRS)

    Chenevey, E. C.

    1979-01-01

    Mats were made from ultrafine polybenzimidazole (PBI) fibers to provide an alternate to the use of asbestos as separators in fuel cells and alkaline batteries. To minimize distortion during mat drying, a process to provide a dry fibrid was developed. Two fibrid types were developed: one coarse, making mats for battery separators; the other fine, making low permeability matrices for fuel cells. Eventually, it was demonstrated that suitable mat fabrication techniques yielded fuel cell separators from the coarser alkaline battery fibrids. The stability of PBI mats to 45% KOH at 123 C can be increased by heat treatment at high temperatures. Weight loss data to 1000 hours exposure show the alkali resistance of the mats to be superior to that of asbestos.

  12. Registration of Charged Particles by Scintillating Fibers Coupled with μ-CELL SI APDG

    NASA Astrophysics Data System (ADS)

    Basharuli, N.; Bondarenko, G.; Bekenov, B.; Golovin, V.; Petrov, V.; Ponomarev, N.; Grigoriev, E.

    2002-11-01

    Silicon μ-cell Avalanche Photodiode operating in Geiger mode (APDG) was used to detect light produced in scintillating fibers of 1 mm diameter by electrons from a 90Sr-source and by α-particles from a 238Pu-source. This recently developed in mesa-technology square 1 mm2 APDG, consisting of 1370 μ-cells, has enhanced inter-cell optical isolation and individual quenching resistors. It showed at room temperature and low biasing voltages (45-47 V) very high gain (up to 106), low dark counting rates (below 3 × 105sec-1) and high detection efficiency for photons of green light (> 35%). Basic characteristics - internal gain, dark counting rate and average number of detected photoelectrons as a function of bias voltage were measured.

  13. Monitoring glutamine in animal cell cultures using a chemiluminescence fiber optic biosensor.

    PubMed

    Cattaneo, M V; Luong, J H

    1993-03-15

    Together with flow injection analysis (FIA), a chemiluminescence (CL) fiber optic biosensor system has been developed for determining glutamine in animal cell cultures. Glutaminase (GAH) and glutamate oxidase (GLO) were onto separate porous aminopropyl glass beads via glutaraldehyde activation and packed to form an enzyme column. These two enzymes acted in sequence on glutamine to produce hydrogen peroxide, which was then reacted with luminol in the presence of ferricyanide to produce a light signal. An anion exchanger was introduced on-line to eliminate interfering endogenous glutamate in view of its negative charge at pH above 3.22 (isoelectric pH). Among several resins tested, the acetate form was most effective, and this type of ion exchanger also effectively adsorbed uric acid, acetaminophen, and aspartic acid.There was an excellent linear relationship between the CL response and standard glutamine concentration in the range 1 to 100 muM. A complete analysis could be performed in 2 min, including sampling and washing with a good reproducibility (+/- 4.4%). Both the bi-enzymic and ion exchange columns were useful for at least 500 analyses when the biosensor system was applied for the glutamine determination in murine hybridoma cell cultures and insect cell cultures. The values obtained compared well with those of HPLC, thus validating the applicability of the CL fiber optic system. PMID:18609602

  14. Carbon nano-chain and carbon nano-fibers based gas diffusion layers for proton exchange membrane fuel cells

    NASA Astrophysics Data System (ADS)

    Kannan, Arunachala M.; Munukutla, Lakshmi

    Gas diffusion layers (GDL) for proton exchange membrane fuel cell have been developed using a partially ordered graphitized nano-carbon chain (Pureblack ® carbon) and carbon nano-fibers. The GDL samples' characteristics such as, surface morphology, surface energy, bubble-point pressure and pore size distribution were characterized using electron microscope, inverse gas chromatograph, gas permeability and mercury porosimetry, respectively. Fuel cell performance of the GDLs was evaluated using single cell with hydrogen/air at ambient pressure, 70 °C and 100% RH. The GDLs with combination of vapor grown carbon nano-fibers with Pureblack carbon showed significant improvement in mechanical robustness as well as fuel cell performance. The micro-porous layer of the GDLs as seen under scanning electron microscope showed excellent surface morphology showing the reinforcement with nano-fibers and the surface homogeneity without any cracks.

  15. High Intensity Training May Reverse the Fiber Type Specific Decline in Myogenic Stem Cells in Multiple Sclerosis Patients

    PubMed Central

    Farup, Jean; Dalgas, Ulrik; Keytsman, Charly; Eijnde, Bert O.; Wens, Inez

    2016-01-01

    Multiple sclerosis (MS) is associated with loss of skeletal muscle mass and function. The myogenic stem cells (satellite cells—SCs) are instrumental to accretion of myonuclei, but remain to be investigated in MS. The present study aimed to compare the SC and myonuclei content between MS patients (n = 23) and age matched healthy controls (HC, n = 18). Furthermore, the effects of 12 weeks of high intensity training on SC and myonuclei content were explored in MS. Muscle biopsies were obtained from m. Vastus Lateralis at baseline (MS and HC) and following 12 weeks of training (MS only). Frozen biopsies were sectioned followed by immunohistochemical analysis for fiber type specific SCs (Pax7+), myonuclei (MN) and central nuclei content and fiber cross-sectional area (fCSA) was quantified using ATPase histochemistry. At baseline the SCs per fiber was lower in type II compared to type I fibers in both MS (119%, p < 0.01) and HC (69%, p < 0.05), whereas the SCs per fCSA was lower in type II fibers compared to type I only in MS (72%, p < 0.05). No differences were observed in MN or central nuclei between MS and HC. Following training the type II fiber SCs per fiber and per fCSA in MS patients increased by 165% (p < 0.05) and 135% (p < 0.05), respectively. Furthermore, the type II fiber MN content tended (p = 0.06) to be increased by 35% following training. In conclusion, the SC content is lower in type II compared to type I fibers in both MS and HC. Furthermore, high intensity training was observed to selectively increase the SC and myonuclei content in type II fibers in MS patients. PMID:27303309

  16. Genes Involved in Osmoregulation during Turgor-Driven Cell Expansion of Developing Cotton Fibers Are Differentially Regulated1

    PubMed Central

    Smart, Lawrence B.; Vojdani, Fakrieh; Maeshima, Masayoshi; Wilkins, Thea A.

    1998-01-01

    Cotton (Gossypium hirsutum L.) fibers are single-celled trichomes that synchronously undergo a phase of rapid cell expansion, then a phase including secondary cell wall deposition, and finally maturation. To determine if there is coordinated regulation of gene expression during fiber expansion, we analyzed the expression of components involved in turgor regulation and a cytoskeletal protein by measuring levels of mRNA and protein accumulation and enzyme activity. Fragments of the genes for the plasma membrane proton-translocating ATPase, vacuole-ATPase, proton-translocating pyrophosphatase (PPase), phosphoenolpyruvate carboxylase, major intrinsic protein, and α-tubulin were amplified by polymerase chain reaction and used as probes in ribonuclease protection assays of RNA from a fiber developmental series, revealing two discrete patterns of mRNA accumulation. Transcripts of all but the PPase accumulated to highest levels during the period of peak expansion (+12–15 d postanthesis [dpa]), then declined with the onset of secondary cell wall synthesis. The PPase was constitutively expressed through fiber development. Activity of the two proton-translocating-ATPases peaked at +15 dpa, whereas PPase activity peaked at +20 dpa, suggesting that all are involved in the process of cell expansion but with varying roles. Patterns of protein accumulation and enzyme activity for some of the proteins examined suggest posttranslational regulation through fiber development. PMID:9536073

  17. A method to integrate patterned electrospun fibers with microfluidic systems to generate complex microenvironments for cell culture applications.

    PubMed

    Wallin, Patric; Zandén, Carl; Carlberg, Björn; Hellström Erkenstam, Nina; Liu, Johan; Gold, Julie

    2012-06-01

    The properties of a cell's microenvironment are one of the main driving forces in cellular fate processes and phenotype expression invivo. The ability to create controlled cell microenvironments invitro becomes increasingly important for studying or controlling phenotype expression in tissue engineering and drug discovery applications. This includes the capability to modify material surface properties within well-defined liquid environments in cell culture systems. One successful approach to mimic extra cellular matrix is with porous electrospun polymer fiber scaffolds, while microfluidic networks have been shown to efficiently generate spatially and temporally defined liquid microenvironments. Here, a method to integrate electrospun fibers with microfluidic networks was developed in order to form complex cell microenvironments with the capability to vary relevant parameters. Spatially defined regions of electrospun fibers of both aligned and random orientation were patterned on glass substrates that were irreversibly bonded to microfluidic networks produced in poly-dimethyl-siloxane. Concentration gradients obtained in the fiber containing channels were characterized experimentally and compared with values obtained by computational fluid dynamic simulations. Velocity and shear stress profiles, as well as vortex formation, were calculated to evaluate the influence of fiber pads on fluidic properties. The suitability of the system to support cell attachment and growth was demonstrated with a fibroblast cell line. The potential of the platform was further verified by a functional investigation of neural stem cell alignment in response to orientation of electrospun fibers versus a microfluidic generated chemoattractant gradient of stromal cell-derived factor 1 alpha. The described method is a competitive strategy to create complex microenvironments invitro that allow detailed studies on the interplay of topography, substrate surface properties, and soluble

  18. Transgenic Expression of AQP1 in the Fiber Cells of AQP0 Knockout Mouse: Effects on Lens Transparency

    PubMed Central

    Varadaraj, K.; Kumari, S.S.; Mathias, R.T.

    2010-01-01

    Mutations and knockout of aquaporin 0 (AQP0) result in dominant lens cataract. To date, several functions have been proposed for AQP0; however, two functions, water permeability and cell-to-cell adhesion have been supported by several investigators and only water channel function has been readily authenticated by in vitro and ex vivo studies. Lens shifts protein expression from the more efficient AQP1 in the equatorial epithelial cells to the less efficient water channel, AQP0, in the differentiating secondary fiber cells; perhaps, AQP0 performs a distinctive function. If AQP0 has only water permeability function, can the more efficient water channel AQP1 transgenically expressed in the fiber cells compensate and restore lens transparency in the AQP0 knockout (AQP0-/-) mouse? To investigate, we generated a transgenic wild type mouse line expressing AQP1 in the fiber cells using αA-crystallin promoter. These transgenic mice (TgAQP1+/+) showed increase in fiber cell membrane water permeability without any morphological, anatomical or physiological defects compared to the wild type indicating that the main purpose of the shift in expression from AQP1 to AQP0 may not be to lessen the membrane water permeability. Further, we transgenically expressed AQP1 in the lens fiber cells of AQP0 knockout mouse (TgAQP1+/+/AQP0-/-) to determine whether AQP1 could restore AQP0 water channel function and regain lens transparency. Fiber cells of these mice showed 2.6 times more water permeability than the wild type. Transgene AQP1 reduced the severity of lens cataract and prevented dramatic acceleration of cataractogenesis. However, lens fiber cells showed deformities and lack of compact cellular architecture. Loss of lens transparency due to the absence of AQP0 was not completely restored indicating an additional function for AQP0. In vitro studies showed that AQP0 is capable of cell-to-cell adhesion while AQP1 is not. To our knowledge, this is the first report which uses an animal

  19. Sonic hedgehog enhances somite cell viability and formation of primary slow muscle fibers in avian segmented mesoderm.

    PubMed

    Cann, G M; Lee, J W; Stockdale, F E

    1999-09-01

    Primary skeletal muscle fibers first form in the segmented portions of paraxial mesoderm called somites. Although the neural tube and notochord are recognized as crucial in patterning myogenic cell lineages during avian and mammalian somitic myogenesis, the source, identities, and actions of the signals governing this process remain controversial. It has been shown that signals emanating from the ventral neural tube and/or notochord alone or Shh alone serve to activate MyoD expression in somites. However, beyond a role in initiating MyoD expression, little is known about the effects of Shh on primary muscle fiber formation in somites of higher vertebrates. The studies reported here investigate how the ventral neural tube promotes myogenesis and compare the effects of the ventral neural tube with those of purified Shh protein on fiber formation in somites. We show that purified Shh protein mimics actions of the ventral neural tube on somites including initiation of muscle fiber formation, enhancement of numbers of primary muscle fibers, and particularly, the formation of primary fibers that express slow myosin. There is a marked increase in slow myosin expression in fibers in response to Shh as somites mature. The effects of ventral neural tube on fiber formation can be blocked by disrupting the Shh signaling pathway by increasing the activity of somitic cyclic AMP-dependent protein kinase A. Furthermore, it was demonstrated that apoptosis is a dominant fate of somite cells, but not somitic muscle fibers, when cultured in the absence of the neural tube, and that application of Shh protein to somites reduced apoptosis. The block to apoptosis by Shh is a manifestation of the maturity of the somite with a progressive increase in the block as somites are displaced rostrally from somite III forward. We conclude that purified Shh protein in mimicking the effects of the ventral neural tube on segmented mesoderm can exert pleiotropic effects during primary myogenesis

  20. Phagosomal pH and glass fiber dissolution in cultured nasal epithelial cells and alveolar macrophages: a preliminary study.

    PubMed Central

    Johnson, N F

    1994-01-01

    The dissolution rate of glass fibers has been shown to be pH sensitive using in vitro lung fluid simulant models. The current study investigated whether there is a difference in phagosomal pH (ppH) between rat alveolar macrophages (AM) and rat nasal epithelial cells (RNEC) and whether such a difference would influence the dissolution of glass fibers. The ppH was measured in cultured AM and RNEC using flow cytometric, fluorescence-emission rationing techniques with fluorescein-labeled, amorphous silica particles. Glass fiber dissolution was determined in AM and RNEC cultured for 3 weeks with fast dissolving glass fibers (GF-A) or slow dissolving ones (GF-B). The mean diameters of GF-A were 2.7 microns and of GF-B, 2.6 microns, the average length of both fibers was approximately 22 to 25 microns. Dissolution was monitored by measuring the length and diameter of intracellular fibers and estimating the volume, assuming a cylindrical morphology. The ppH of AM was 5.2 to 5.8, and the ppH of RNEC was 7.0 to 7.5. The GF-A dissolved more slowly in RNEC than in AM, and no dissolution was evident in either cell type with GF-B. The volume loss with GF-A after a 3-week culture with AM was 66% compared to 45% for cultured RNEC. These results are different from those obtained using in vitro lung fluid-simulant models where dissolution is faster at higher pH. This difference suggests that dissolution rates of glass fibers in AM should not be applied to the dissolution of fibers in epithelial cells. Images Figure 1. a Figure 1. b Figure 2. a Figure 2. b Figure 3. a Figure 3. b PMID:7882965

  1. Proteomics analysis of nasopharyngeal carcinoma cell secretome using a hollow fiber culture system and mass spectrometry.

    PubMed

    Wu, Hsin-Yi; Chang, Ying-Hwa; Chang, Yu-Chen; Liao, Pao-Chi

    2009-01-01

    Secreted proteins, referred to as the secretome, are known to regulate a variety of biological functions and are involved in a multitude of pathological processes. However, some secreted proteins from cell cultures are difficult to detect because of their intrinsic low abundance. They are frequently masked by proteins shed from lysed cells and the substantial amounts of serum proteins used in culture medium. We have proposed an analytical platform for sensitive detection of secreted proteins by utilizing a hollow fiber culture (HFC) system coupled with proteomic approaches. The HFC system enables culture of high-density cells in a small volume where secreted proteins can be accumulated. In addition, cell lysis rates can be greatly reduced, which alleviates the contamination from lysed cells. In this study, nasopharyngeal carcinoma (NPC) cells were utilized to evaluate the efficiency of this system in the collection and analysis of the cell secretome. Cells were adapted to serum-free medium and inoculated into the HFC system. The cell lysis rate in the culture system was estimated to be 0.001-0.022%, as determined by probing four intracellular proteins in the conditioned medium (CM), while a cell lysis rate of 0.32-1.84% was observed in dish cultures. Proteins in the CM were analyzed using SDS-PAGE and liquid chromatography tandem mass spectrometry (LC-MS/MS). A total of 134 proteins were identified in 62 gel bands, of which 61% possess a signal peptide and/or a transmembrane domain. In addition, 37% of the identified secretome were classified as extracellular or membrane proteins, whereas 98% of the lysate proteins were identified as intracellular proteins. We suggest that the HFC system may be used to collect secreted proteins efficiently and facilitate comprehensive characterization of cell secretome. PMID:19012429

  2. A method to integrate patterned electrospun fibers with microfluidic systems to generate complex microenvironments for cell culture applications

    PubMed Central

    Wallin, Patric; Zandén, Carl; Carlberg, Björn; Hellström Erkenstam, Nina; Liu, Johan; Gold, Julie

    2012-01-01

    The properties of a cell’s microenvironment are one of the main driving forces in cellular fate processes and phenotype expression invivo. The ability to create controlled cell microenvironments invitro becomes increasingly important for studying or controlling phenotype expression in tissue engineering and drug discovery applications. This includes the capability to modify material surface properties within well-defined liquid environments in cell culture systems. One successful approach to mimic extra cellular matrix is with porous electrospun polymer fiber scaffolds, while microfluidic networks have been shown to efficiently generate spatially and temporally defined liquid microenvironments. Here, a method to integrate electrospun fibers with microfluidic networks was developed in order to form complex cell microenvironments with the capability to vary relevant parameters. Spatially defined regions of electrospun fibers of both aligned and random orientation were patterned on glass substrates that were irreversibly bonded to microfluidic networks produced in poly-dimethyl-siloxane. Concentration gradients obtained in the fiber containing channels were characterized experimentally and compared with values obtained by computational fluid dynamic simulations. Velocity and shear stress profiles, as well as vortex formation, were calculated to evaluate the influence of fiber pads on fluidic properties. The suitability of the system to support cell attachment and growth was demonstrated with a fibroblast cell line. The potential of the platform was further verified by a functional investigation of neural stem cell alignment in response to orientation of electrospun fibers versus a microfluidic generated chemoattractant gradient of stromal cell-derived factor 1 alpha. The described method is a competitive strategy to create complex microenvironments invitro that allow detailed studies on the interplay of topography, substrate surface properties, and soluble

  3. Asbestos-induced endothelial cell activation and injury. Demonstration of fiber phagocytosis and oxidant-dependent toxicity.

    PubMed

    Garcia, J G; Gray, L D; Dodson, R F; Callahan, K S

    1988-10-01

    Vascular endothelial cell injury is important in the development of a variety of chronic interstitial lung disorders. However, the involvement of such injury in the inflammatory response associated with the inhalation of asbestos fibers is unclear and the mechanism of asbestos fiber cytotoxicity remains unknown. In the present study, human umbilical vein endothelial cells were challenged with amosite asbestos and several parameters of cellular function were examined. Electron microscopic examination revealed that endothelial cell exposure to asbestos resulted in active phagocytosis of these particulates. Biochemical evidence of dose-dependent asbestos-mediated endothelial cell activation was indicated by increased metabolism of arachidonic acid. For example, amosite asbestos (500 micrograms/ml) produced a ninefold increase in prostacyclin (PGI2) levels over those levels in non-exposed cells. Incubation of human endothelial cells with asbestos fibers induced specific 51Cr release in both a dose- and time-dependent fashion indicative of cellular injury. Injury induced by amosite asbestos was not significantly attenuated by treatment of the endothelial cell monolayer with either the iron chelator deferoxamine, which prevents hydroxyl radical (.OH) formation, or by the superoxide anion (O2-) scavenger, superoxide dismutase. However, significant dose-dependent protection was observed with the hydrogen peroxide (H2O2) scavenger, catalase. Chelation of elemental iron present within amosite asbestos fibers by deferoxamine produced a 33% reduction in asbestos cytotoxicity, suggesting a potential role for hydroxyl radical-mediated injury via the iron-catalyzed Haber-Weiss reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:3202471

  4. A versatile micro-mechanical tester for actin stress fibers isolated from cells.

    PubMed

    Matsui, Tsubasa S; Deguchi, Shinji; Sakamoto, Naoya; Ohashi, Toshiro; Sato, Masaaki

    2009-01-01

    Conventional atomic force microscopy is one of the major techniques to evaluate mechanical properties of cells and subcellular components. The use of a cantilever probe for sample manipulation within the vertical plane often makes absolute positioning of the probe, subject to thermal drift, difficult. In addition, the vertical test is unable to observe changes in the sample structure responsible for mechanical behavior detected by the probe. In the present study, an alternative mechanical tester was developed that incorporated a pair of micro-needles to manipulate a sample in a project plane, allowing acquisition of the accurate probe position and entire sample image. Using a vision-based feedback control, a micro-needle driven by a piezo actuator is moved to give user-defined displacements or forces to sample. To show its usefulness and versatility, three types of viscoelastic measurements on actin stress fibers isolated from smooth muscle cells were demonstrated: strain rate-controlled tensile tests, relaxation tests and creep tests. Fluorescence imaging of the stress fibers using Qdots over the course of the measurements, obtained through multiple image detectors, was also carried out. The technique described here is useful for examining the quantitative relationship between mechanical behavior and related structural changes of biomaterials. PMID:19940356

  5. Thickness-controllable electrospun fibers promote tubular structure formation by endothelial progenitor cells

    PubMed Central

    Hong, Jong Kyu; Bang, Ju Yup; Xu, Guan; Lee, Jun-Hee; Kim, Yeon-Ju; Lee, Ho-Jun; Kim, Han Seong; Kwon, Sang-Mo

    2015-01-01

    Controlling the thickness of an electrospun nanofibrous scaffold by altering its pore size has been shown to regulate cell behaviors such as cell infiltration into a three-dimensional (3D) scaffold. This is of great importance when manufacturing tissue-engineering scaffolds using an electrospinning process. In this study, we report the development of a novel process whereby additional aluminum foil layers were applied to the accumulated electrospun fibers of an existing aluminum foil collector, effectively reducing the incidence of charge buildup. Using this process, we fabricated an electrospun scaffold with a large pore (pore size >40 μm) while simultaneously controlling the thickness. We demonstrate that the large pore size triggered rapid infiltration (160 μm in 4 hours of cell culture) of individual endothelial progenitor cells (EPCs) and rapid cell colonization after seeding EPC spheroids. We confirmed that the 3D, but not two-dimensional, scaffold structures regulated tubular structure formation by the EPCs. Thus, incorporation of stem cells into a highly porous 3D scaffold with tunable thickness has implications for the regeneration of vascularized thick tissues and cardiac patch development. PMID:25709441

  6. Thickness-controllable electrospun fibers promote tubular structure formation by endothelial progenitor cells.

    PubMed

    Hong, Jong Kyu; Bang, Ju Yup; Xu, Guan; Lee, Jun-Hee; Kim, Yeon-Ju; Lee, Ho-Jun; Kim, Han Seong; Kwon, Sang-Mo

    2015-01-01

    Controlling the thickness of an electrospun nanofibrous scaffold by altering its pore size has been shown to regulate cell behaviors such as cell infiltration into a three-dimensional (3D) scaffold. This is of great importance when manufacturing tissue-engineering scaffolds using an electrospinning process. In this study, we report the development of a novel process whereby additional aluminum foil layers were applied to the accumulated electrospun fibers of an existing aluminum foil collector, effectively reducing the incidence of charge buildup. Using this process, we fabricated an electrospun scaffold with a large pore (pore size >40 μm) while simultaneously controlling the thickness. We demonstrate that the large pore size triggered rapid infiltration (160 μm in 4 hours of cell culture) of individual endothelial progenitor cells (EPCs) and rapid cell colonization after seeding EPC spheroids. We confirmed that the 3D, but not two-dimensional, scaffold structures regulated tubular structure formation by the EPCs. Thus, incorporation of stem cells into a highly porous 3D scaffold with tunable thickness has implications for the regeneration of vascularized thick tissues and cardiac patch development. PMID:25709441

  7. Hollow fiber integrated microfluidic platforms for in vitro Co-culture of multiple cell types.

    PubMed

    Huang, Jen-Huang; Harris, Jennifer F; Nath, Pulak; Iyer, Rashi

    2016-10-01

    This study demonstrates a rapid prototyping approach for fabricating and integrating porous hollow fibers (HFs) into microfluidic device. Integration of HF can enhance mass transfer and recapitulate tubular shapes for tissue-engineered environments. We demonstrate the integration of single or multiple HFs, which can give the users the flexibility to control the total surface area for tissue development. We also present three microfluidic designs to enable different co-culture conditions such as the ability to co-culture multiple cell types simultaneously on a flat and tubular surface, or inside the lumen of multiple HFs. Additionally, we introduce a pressurized cell seeding process that can allow the cells to uniformly adhere on the inner surface of HFs without losing their viabilities. Co-cultures of lung epithelial cells and microvascular endothelial cells were demonstrated on the different platforms for at least five days. Overall, these platforms provide new opportunities for co-culturing of multiple cell types in a single device to reconstruct native tissue micro-environment for biomedical and tissue engineering research. PMID:27613401

  8. Passively mode-locked fiber laser by a cell-type WS2 nanosheets saturable absorber

    PubMed Central

    Yan, Peiguang; Liu, Aijiang; Chen, Yushan; Wang, JinZhang; Ruan, Shuangchen; Chen, Hao; Ding, Jinfei

    2015-01-01

    A cell-type saturable absorber has been demonstrated by filling the single mode photonic crystal fiber (SMPCF) with tungsten disulfide (WS2) nanosheets. The modulation depth, saturable intensity, and non-saturable loss of this SA are measured to be 3.53%, 159 MW/cm2 and 23.2%, respectively. Based on this SA, a passively mode-locked EDF laser has been achieved with pulse duration of 808 fs and repetition rate of 19.57 MHz, and signal-noise-ratio (SNR) of 60.5 dB. Our results demonstrate that the cell-type WS2 nanosheets SA can serve as a good candidate for short-pulse mode locker. PMID:26213180

  9. "allometry" Deterministic Approaches in Cell Size, Cell Number and Crude Fiber Content Related to the Physical Quality of Kangkong (Ipomoea reptans) Grown Under Different Plant Density Pressures

    NASA Astrophysics Data System (ADS)

    Selamat, A.; Atiman, S. A.; Puteh, A.; Abdullah, N. A. P.; Mohamed, M. T. M.; Zulkeefli, A. A.; Othman, S.

    Kangkong, especially the upland type (Ipomoea reptans) is popularly consumed as a vegetable dish in the South East Asian countries for its quality related to Vitamins (A and C) and crude fiber contents. Higher fiber contents would prevent from the occurrence of colon cancer and diverticular disease. With young stem edible portion, its cell number and size contribute to the stem crude fiber content. The mathematical approach of allometry of cell size, number, and fiber content of stem could be used in determining the 'best' plant density pressure in producing the quality young stem to be consumed. Basically, allometry is the ratio of relative increment (growth or change) rates of two parameters, or the change rate associated to the log of measured variables relationship. Kangkog grown equal or lower than 55 plants m-2 produced bigger individual plant and good quality (physical) kangkong leafy vegetable, but with lower total yield per unit area as compared to those grown at higher densities.

  10. Fractionation of human bone marrow cell suspensions in nylon fiber columns: an efficient method for the removal of cells that produce colony stimulating factor (CSF).

    PubMed

    Sullivan, R; Gans, P J

    1980-09-01

    In this report, we describe an efficient technique for the extraction of CSF-producing cells from human marrow suspensions. Prior to plating in agar cultures, we incubated buoyant human marrow cells for 45 min in columns packed with nylon fiber or subjected the cells to two one-hour incubations in glass petri dishes. Recoveries of total cells, differential marrow elements, and committed granulocyte-monocyte progenitor cells (CFUc) were similar after each separative procedure. However, spontaneous CFUc proliferation was more effectively eliminated when cells were fractionated in nylon fiber columns. After the removal of cells which were adherent to glass, spontaneous CFUc proliferation in cultures containing no exogenous CSF accounted for 2.1% of total CFUc at a plating concentration of 10(5) cells/ml and 7.8% at a concentration of 3 X 10(5) cells/ml. After the fractionation of marrow cell suspensions in nylon fiber columns, spontaneous CFUc growth was completely obliterated at a plating concentration of 10(5) cells/ml, and at a concentration of 3 X 10(5) cells/ml accounted for only 0.09% of total CFUc. Further experiments were undertaken which demonstrated that buoyant marrow cells after incubation in nylon fiber columns may be employed to assay CSF in extremely dilute concentrations. Because of the simplicity and efficiency of this procedure, nylon fiber chromatography appears to be a highly useful technique for the rapid semi-purification of marrow suspensions for use in the assay of human CSF. PMID:6970676

  11. Cotton GhMYB7 is predominantly expressed in developing fibers and regulates secondary cell wall biosynthesis in transgenic Arabidopsis.

    PubMed

    Huang, Junfeng; Chen, Feng; Wu, Siyu; Li, Juan; Xu, Wenliang

    2016-02-01

    The secondary cell wall in mature cotton fibers contains over 90% cellulose with low quantities of xylan and lignin. However, little is known regarding the regulation of secondary cell wall biosynthesis in cotton fibers. In this study, we characterized an R2R3-MYB transcription factor, GhMYB7, in cotton. GhMYB7 is expressed at a high level in developing fibers and encodes a MYB protein that is targeted to the cell nucleus and has transcriptional activation activity. Ectopic expression of GhMYB7 in Arabidopsis resulted in small, curled, dark green leaves and also led to shorter inflorescence stems. A cross-sectional assay of basal stems revealed that cell wall thickness of vessels and interfascicular fibers was higher in transgenic lines overexpressing GhMYB7 than in the wild type. Constitutive expression of GhMYB7 in Arabidopsis activated the expression of a suite of secondary cell wall biosynthesis-related genes (including some secondary cell wall-associated transcription factors), leading to the ectopic deposition of cellulose and lignin. The ectopic deposition of secondary cell walls may have been initiated before the cessation of cell expansion. Moreover, GhMYB7 was capable of binding to the promoter regions of AtSND1 and AtCesA4, suggesting that GhMYB7 may function upstream of NAC transcription factors. Collectively, these findings suggest that GhMYB7 is a potential transcriptional activator, which may participate in regulating secondary cell wall biosynthesis of cotton fibers. PMID:26803299

  12. Muscle fiber characteristics, satellite cells and soccer performance in young athletes.

    PubMed

    Metaxas, Thomas I; Mandroukas, Athanasios; Vamvakoudis, Efstratios; Kotoglou, Kostas; Ekblom, Björn; Mandroukas, Konstantinos

    2014-09-01

    This study is aimed to examine the muscle fiber type, composition and satellite cells in young male soccer players and to correlate them to cardiorespiratory indices and muscle strength. The participants formed three Groups: Group A (n = 13), 11.2 ± 0.4yrs, Group B (n=10), 13.1 ± 0.5yrs and Group C (n = 9), 15.2 ± 0.6yrs. Muscle biopsies were obtained from the vastus lateralis. Peak torque values of the quadriceps and hamstrings were recorded and VO2max was measured on the treadmill. Group C had lower type I percentage distribution compared to A by 21.3% (p < 0.01), while the type IIA relative percentage was higher by 18.1% and 18.4% than in Groups A and B (p < 0.05). Groups B and C had higher cross-sectional area (CSA) values in all fiber types than in Group A (0.05 < p < 0.001). The number of satellite cells did not differ between the groups. Groups B and C had higher peak torque at all angular velocities and absolute VO2max in terms of ml·min(-1) than Group A (0.05 < p < 0.001). It is concluded that the increased percentage of type IIA muscle fibers noticed in Group C in comparison to the Groups A and B should be mainly attributed to the different workload exercise and training programs. The alteration of myosin heavy chain (MHC) isoforms composition even in children is an important mechanism for skeletal muscle characteristics. Finally, CSA, isokinetic muscle strength and VO2max values seems to be expressed according to age. Key PointsFifteen years old soccer players have higher IIA percentage distribution than the younger players by approximately 18%.The age and the training status play a crucial role in muscle fibers co-expression.Specific training in young athletes seems to alter significantly the muscular metabolic profile. PMID:25177173

  13. Muscle Fiber Characteristics, Satellite Cells and Soccer Performance in Young Athletes

    PubMed Central

    Metaxas, Thomas I.; Mandroukas, Athanasios; Vamvakoudis, Efstratios; Kotoglou, Kostas; Ekblom, Björn; Mandroukas, Konstantinos

    2014-01-01

    This study is aimed to examine the muscle fiber type, composition and satellite cells in young male soccer players and to correlate them to cardiorespiratory indices and muscle strength. The participants formed three Groups: Group A (n = 13), 11.2 ± 0.4yrs, Group B (n=10), 13.1 ± 0.5yrs and Group C (n = 9), 15.2 ± 0.6yrs. Muscle biopsies were obtained from the vastus lateralis. Peak torque values of the quadriceps and hamstrings were recorded and VO2max was measured on the treadmill. Group C had lower type I percentage distribution compared to A by 21.3% (p < 0.01), while the type IIA relative percentage was higher by 18.1% and 18.4% than in Groups A and B (p < 0.05). Groups B and C had higher cross-sectional area (CSA) values in all fiber types than in Group A (0.05 < p < 0.001). The number of satellite cells did not differ between the groups. Groups B and C had higher peak torque at all angular velocities and absolute VO2max in terms of ml·min-1 than Group A (0.05 < p < 0.001). It is concluded that the increased percentage of type IIA muscle fibers noticed in Group C in comparison to the Groups A and B should be mainly attributed to the different workload exercise and training programs. The alteration of myosin heavy chain (MHC) isoforms composition even in children is an important mechanism for skeletal muscle characteristics. Finally, CSA, isokinetic muscle strength and VO2max values seems to be expressed according to age. Key Points Fifteen years old soccer players have higher IIA percentage distribution than the younger players by approximately 18%. The age and the training status play a crucial role in muscle fibers co-expression. Specific training in young athletes seems to alter significantly the muscular metabolic profile. PMID:25177173

  14. Isolation of functional giant smooth muscle cells from an invertebrate: structural features of relaxed and contracted fibers.

    PubMed Central

    Hernandez-Nicaise, M L; Bilbaut, A; Malaval, L; Nicaise, G

    1982-01-01

    The giant smooth muscle fibers of a ctenophore were isolated by enzymatic digestion. These fibers are multinucleated cells, up to 50 micrometers in diameter and 2 cm in length. Their ultrastructure and membrane electrical properties are similar to those of in situ fibers. Relaxed, coiled (partially contracted), and fully shortened states were distinguished in isolated cells and studied by scanning and transmission electron microscopy. Calcium-containing mitochondrial granules were found in the coiled cells but not in either the relaxed or the fully shortened cells. The relaxed cell is characterized in cross section by the density of myosin filaments (457 +/- 15 per micrometer2) and the thin-to-thick filament ratio (5.2 +/- 0.2). In the coiled cell, the muscle lattice does not expand uniformly, as shown by the variability of myosin spacing, and the thin-to-thick filament ratio decreases. Both clockwise and counterclockwise coiling occur along the same fiber. The implications of these findings with respect to the structure of the contractile apparatus are discussed. Images PMID:6952237

  15. Dual-modality fiber-optic imager (DFOI) for intracellular gene delivery in human cervical cancer cell

    NASA Astrophysics Data System (ADS)

    Cha, Jaepyeong; Zhang, Jing; Gurbani, Saumya; Li, Min; Kang, Jin U.

    2013-03-01

    The most common optical method to validate intracellular gene delivery in cancer is to detect tagged fluorescence signals from the cells. However, fluorescent detection is usually performed in vitro due to the limitation of standard microscopes. Herein, we propose a highly sensitive dual-modality fiber-optic imager (DFOI), which enables in vivo fluorescence imaging. Our system uses a coherent fiber bundle based imager capable of simultaneously performing both confocal reflectance and fluorescent microscopy. Non-viral vectors targeting human cervical cancer cells (HeLa) were used to evaluate the performance. Preliminary results demonstrated the DFOI is promising for in vivo evaluation of intracellular gene delivery.

  16. Regeneration of skeletal muscle fibers from autologous satellite cells multiplied in vitro. An experimental model for testing cultured cell myogenicity

    SciTech Connect

    Alameddine, H.S.; Dehaupas, M.; Fardeau, M. )

    1989-07-01

    An experimental model used to test in vivo myogenicity of autologous satellite cells multiplied in vitro is described. Free muscle autotransplantation served as the basis and was combined with x-irradiation. Administration of 1500, 2500, and 3500 rad doses 24 hours before or after ischemia showed that inhibition of spontaneous regeneration is dose dependent and more efficient when irradiation was applied before injury. A single dose of 2500 rad before injury resulted in the formation of a cystic structure ideal for cell implantation. FITC-latex beads and/or carbocyanine dyes were internalized by mononucleated satellite cells in vitro. Labeling did not affect survival or development of these cells. No sign of marker release or spreading from labeled to unlabeled cells was detectable unless by the fusion process. These labels were retained for several weeks. Grafting of labeled dense cellular suspensions into x-irradiated ischemic muscles indicated that satellite cells retain their myogenic characteristic and are able to reform fully differentiated muscle fibers. 55 references.

  17. Anisotropic poly (glycerol sebacate)-poly (ϵ-caprolactone) electrospun fibers promote endothelial cell guidance.

    PubMed

    Gaharwar, Akhilesh K; Nikkhah, Mehdi; Sant, Shilpa; Khademhosseini, Ali

    2015-01-01

    Topographical cell guidance is utilized to engineer highly organized and aligned cellular constructs for numerous tissue engineering applications. Recently, electrospun scaffolds fabricated using poly(glycerol sebacate) (PGS) and poly(ϵ-caprolactone) (PCL) have shown a great promise to support valvular interstitial cell functions for the development of tissue engineered heart valves. However, one of the major drawbacks of PGS-PCL scaffolds is the lack of control over cellular alignment. In this work, we investigate the role of scaffold architecture on the endothelial cell alignment, proliferation and formation of organized cellular structures. In particular, PGS-PCL scaffolds with randomly oriented and highly aligned fibers with tunable mechanical properties were fabricated using electrospinning technique. After one week of culture, endothelial cells on the aligned scaffolds exhibited higher proliferation compared to those cultures on randomly oriented fibrous scaffolds. Furthermore, the endothelial cells reorganized in response to the topographical features of aligned scaffolds forming highly organized cellular constructs. Thus, topographical contact guidance, provided by aligned PGS-PCL scaffolds, is envisioned to be useful in developing cellular structures for vascular tissue engineering. PMID:25516556

  18. Long-distance laser propulsion and deformation- monitoring of cells in optofluidic photonic crystal fiber.

    PubMed

    Unterkofler, Sarah; Garbos, Martin K; Euser, Tijmen G; St J Russell, Philip

    2013-09-01

    We introduce a unique method for laser-propelling individual cells over distances of 10s of cm through stationary liquid in a microfluidic channel. This is achieved by using liquid-filled hollow-core photonic crystal fiber (HC-PCF). HC-PCF provides low-loss light guidance in a well-defined single mode, resulting in highly uniform optical trapping and propulsive forces in the core which at the same time acts as a microfluidic channel. Cells are trapped laterally at the center of the core, typically several microns away from the glass interface, which eliminates adherence effects and external perturbations. During propagation, the velocity of the cells is conveniently monitored using a non-imaging Doppler velocimetry technique. Dynamic changes in velocity at constant optical powers up to 350 mW indicate stress-induced changes in the shape of the cells, which is confirmed by bright-field microscopy. Our results suggest that HC-PCF will be useful as a new tool for the study of single-cell biomechanics. PMID:23281270

  19. Anisotropic Poly (glycerol sebacate)-Poly (ε-caprolactone) Electrospun Fibers Promote Endothelial Cell Guidance

    PubMed Central

    Gaharwar, Akhilesh K.; Nikkhah, Mehdi; Sant, Shilpa; Khademhosseini, Ali

    2015-01-01

    Topographical cell guidance is utilized to engineer highly organized and aligned cellular constructs for numerous tissue engineering applications. Recently, electrospun scaffolds fabricated using poly(glycerol sebacate) (PGS) and poly(ε-caprolactone) (PCL) have shown a great promise to support valvular interstitial cell functions for the development of tissue engineered heart valves. However, one of the major drawbacks of PGS-PCL scaffolds is the lack of control over cellular alignment. In this work we investigate the role of scaffold architecture on the endothelial cell alignment, proliferation and formation of organized cellular structures. In particular, PGS-PCL scaffolds with randomly oriented and highly aligned fibers with tunable mechanical properties were fabricated using electrospinning technique. After one week of culture, endothelial cells on the aligned scaffolds exhibit higher proliferation compared to those cultures on randomly oriented fibrous scaffolds. Furthermore, the endothelial cells reorganize in response to the topographical features of anisotropic scaffolds forming highly organize cellular constructs. Thus, the topographical contact guidance, provided by aligned PGS-PCL scaffolds, is envisioned to be useful in developing aligned cellular structures for vascular tissue engineering. PMID:25516556

  20. [Changes in cell respiration of postural muscle fibers under long-term gravitational unloading after dietary succinate supplementation].

    PubMed

    Ogneva, I V; Veselova, O M; Larina, I M

    2011-01-01

    The intensity of cell respiration of the rat m. soleus, m. gastrocnemius c.m. and tibialis anterior fibers during 35-day gravitational unloading, with the addition of succinate in the diet at a dosage rate of 50 mg per 1 kg animal weight has been investigated. The gravitational unloading was modeled by antiorthostatic hindlimb suspension. The intensity of cell respiration was estimated by polarography. It was shown that the rate of oxygen consumption by soleus and gastrocnemius fibers on endogenous and exogenous substrates and with the addition of ADP decreases after the discharge. This may be associated with the transition to the glycolytic energy path due to a decrease in the EMG-activity. At the same time, the respiration rate after the addition of exogenous substrates in soleus fibers did not increase, indicating a disturbance in the function of the NCCR-section of the respiratory chain and more pronounced changes in the structure of muscle fibers. In tibialis anterior fibers, no changes in oxygen consumption velocity were observed. The introduction of succinate to the diet of rats makes it possible to prevent the negative effects of hypokinesia, although it reduces the basal level of intensity of cell respiration. PMID:21442893

  1. Progressive Failure of a Unidirectional Fiber-Reinforced Composite Using the Method of Cells: Discretization Objective Computational Results

    NASA Technical Reports Server (NTRS)

    Pineda, Evan J.; Bednarcyk, Brett A.; Waas, Anthony M.; Arnold, Steven M.

    2012-01-01

    The smeared crack band theory is implemented within the generalized method of cells and high-fidelity generalized method of cells micromechanics models to capture progressive failure within the constituents of a composite material while retaining objectivity with respect to the size of the discretization elements used in the model. An repeating unit cell containing 13 randomly arranged fibers is modeled and subjected to a combination of transverse tension/compression and transverse shear loading. The implementation is verified against experimental data (where available), and an equivalent finite element model utilizing the same implementation of the crack band theory. To evaluate the performance of the crack band theory within a repeating unit cell that is more amenable to a multiscale implementation, a single fiber is modeled with generalized method of cells and high-fidelity generalized method of cells using a relatively coarse subcell mesh which is subjected to the same loading scenarios as the multiple fiber repeating unit cell. The generalized method of cells and high-fidelity generalized method of cells models are validated against a very refined finite element model.

  2. Transcriptome Profiling, Molecular Biological, and Physiological Studies Reveal a Major Role for Ethylene in Cotton Fiber Cell Elongation[W][OA

    PubMed Central

    Shi, Yong-Hui; Zhu, Sheng-Wei; Mao, Xi-Zeng; Feng, Jian-Xun; Qin, Yong-Mei; Zhang, Liang; Cheng, Jing; Wei, Li-Ping; Wang, Zhi-Yong; Zhu, Yu-Xian

    2006-01-01

    Upland cotton (Gossypium hirsutum) produces the most widely used natural fibers, yet the regulatory mechanisms governing fiber cell elongation are not well understood. Through sequencing of a cotton fiber cDNA library and subsequent microarray analysis, we found that ethylene biosynthesis is one of the most significantly upregulated biochemical pathways during fiber elongation. The 1-Aminocyclopropane-1-Carboxylic Acid Oxidase1-3 (ACO1-3) genes responsible for ethylene production were expressed at significantly higher levels during this growth stage. The amount of ethylene released from cultured ovules correlated with ACO expression and the rate of fiber growth. Exogenously applied ethylene promoted robust fiber cell expansion, whereas its biosynthetic inhibitor l-(2-aminoethoxyvinyl)-glycine (AVG) specifically suppressed fiber growth. The brassinosteroid (BR) biosynthetic pathway was modestly upregulated during this growth stage, and treatment with BR or its biosynthetic inhibitor brassinazole (BRZ) also promoted or inhibited, respectively, fiber growth. However, the effect of ethylene treatment was much stronger than that of BR, and the inhibitory effect of BRZ on fiber cells could be overcome by ethylene, but the AVG effect was much less reversed by BR. These results indicate that ethylene plays a major role in promoting cotton fiber elongation. Furthermore, ethylene may promote cell elongation by increasing the expression of sucrose synthase, tubulin, and expansin genes. PMID:16461577

  3. Fluorescence image-guided photodynamic therapy of cancer cells using a scanning fiber endoscope

    NASA Astrophysics Data System (ADS)

    Woldetensae, Mikias H.; Kirshenbaum, Mark R.; Kramer, Greg M.; Zhang, Liang; Seibel, Eric J.

    2013-03-01

    A scanning fiber endoscope (SFE) and the cancer biomarker 5-aminolevulinic acid (5-ALA) were used to fluorescently detect and destroy superficial cancerous lesions, while experimenting with different dosimetry levels for concurrent or sequential imaging and laser therapy. The 1.6-mm diameter SFE was used to fluorescently image a confluent monolayer of A549 human lung cancer cells from culture, previously administered with 5 mM solution of 5-ALA for 4 hours. Twenty hours after therapy, cell cultures were stained to distinguish between living and dead cells using a laser scanning confocal microscope. To determine relative dosimetry for photodynamic therapy (PDT), 405-nm laser illumination was varied from 1 to 5 minutes with power varying from 5 to 18 mW, chosen to compare equal amounts of energy delivered to the cell culture. The SFE produced 500-line images of fluorescence at 15 Hz using the red detection channel centered at 635 nm. The results show that PDT of A549 cancer cell monolayers using 405nm light for imaging and 5-ALAinduced PpIX therapy was possible using the same SFE system. Increased duration and power of laser illumination produced an increased area of cell death upon live/dead staining. The ultrathin and flexible SFE was able to direct PDT using wide-field fluorescence imaging of a monolayer of cultured cancer cells after uptaking 5-ALA. The correlation between light intensity and duration of PDT was measured. Increased length of exposure and decreased light intensity yields larger areas of cell death than decreased length of exposure with increased light intensity.

  4. Mechanism of Ad5 Vaccine Immunity and Toxicity: Fiber Shaft Targeting of Dendritic Cells

    PubMed Central

    Kong, Wing-pui; Sheets, Rebecca L; Gomez, Phillip L; King, C. Richter; Nabel, Gary J

    2007-01-01

    Recombinant adenoviral (rAd) vectors elicit potent cellular and humoral immune responses and show promise as vaccines for HIV-1, Ebola virus, tuberculosis, malaria, and other infections. These vectors are now widely used and have been generally well tolerated in vaccine and gene therapy clinical trials, with many thousands of people exposed. At the same time, dose-limiting adverse responses have been observed, including transient low-grade fevers and a prior human gene therapy fatality, after systemic high-dose recombinant adenovirus serotype 5 (rAd5) vector administration in a human gene therapy trial. The mechanism responsible for these effects is poorly understood. Here, we define the mechanism by which Ad5 targets immune cells that stimulate adaptive immunity. rAd5 tropism for dendritic cells (DCs) was independent of the coxsackievirus and adenovirus receptor (CAR), its primary receptor or the secondary integrin RGD receptor, and was mediated instead by a heparin-sensitive receptor recognized by a distinct segment of the Ad5 fiber, the shaft. rAd vectors with CAR and RGD mutations did not infect a variety of epithelial and fibroblast cell types but retained their ability to transfect several DC types and stimulated adaptive immune responses in mice. Notably, the pyrogenic response to the administration of rAd5 also localized to the shaft region, suggesting that this interaction elicits both protective immunity and vector-induced fevers. The ability of replication-defective rAd5 viruses to elicit potent immune responses is mediated by a heparin-sensitive receptor that interacts with the Ad5 fiber shaft. Mutant CAR and RGD rAd vectors target several DC and mononuclear subsets and induce both adaptive immunity and toxicity. Understanding of these interactions facilitates the development of vectors that target DCs through alternative receptors that can improve safety while retaining the immunogenicity of rAd vaccines. PMID:17319743

  5. Identification of global gene expression differences between human lens epithelial and cortical fiber cells reveals specific genes and their associated pathways important for specialized lens cell functions

    PubMed Central

    Hawse, John R.; DeAmicis-Tress, Candida; Cowell, Tracy L.; Kantorow, Marc

    2005-01-01

    Purpose: In order to identify specific genes that may play important roles in maintaining the specialized functions of lens epithelial and fiber cells, we have analyzed the global gene expression profiles of these two cell types in the human lens. This analysis will also reveal those genes that are exclusively expressed in the epithelial and cortical fiber cells and those genes that may play important roles in the differentiation of epithelial cells to mature fiber cells. Methods: Oligonucleotide microarray hybridization was used to analyze the expression profiles of 22,215 genes between adult (average age greater than 56 years) human lens epithelial and cortical fiber cells. The expression levels of selected genes were further compared by semi-quantitative RT-PCR and selected genes were functionally clustered into common categories using the EASE bioinformatics software package. Results: Analysis of three separate microarray hybridizations revealed 1,196 transcripts that exhibit increased expression and 1,278 transcripts that exhibit decreased expression at the 2 fold or greater level between lens epithelial cells and cortical fiber cells on all three of the arrays analyzed. Of these, 222 transcripts exhibited increased expression and 135 transcripts exhibited decreased expression by an average of 5 fold or greater levels on all three arrays. Semi-quantitative RT-PCR analysis of 21 randomly selected genes revealed identical expression patterns as those detected by microarray hybridization indicating that the microarray data are accurate. Functional clustering of the identified gene expression patterns using the EASE program revealed a wide variety of biological pathways that exhibited altered expression patterns between the two cell types including mRNA processing, cell adhesion, cell proliferation, translation, protein folding, oxidative phosphorylation, and apoptosis, among others. Conclusions: These data reveal novel and previously identified gene expression

  6. Polyphosphazene functionalized polyester fiber matrices for tendon tissue engineering: in vitro evaluation with human mesenchymal stem cells.

    PubMed

    Peach, M Sean; James, Roshan; Toti, Udaya S; Deng, Meng; Morozowich, Nicole L; Allcock, Harry R; Laurencin, Cato T; Kumbar, Sangamesh G

    2012-08-01

    Poly[(ethyl alanato)(1)(p-methyl phenoxy)(1)] phosphazene (PNEA-mPh) was used to modify the surface of electrospun poly(ε-caprolactone) (PCL) nanofiber matrices having an average fiber diameter of 3000 ± 1700 nm for the purpose of tendon tissue engineering and augmentation. This study reports the effect of polyphosphazene surface functionalization on human mesenchymal stem cell (hMSC) adhesion, cell-construct infiltration, proliferation and tendon differentiation, as well as long term cellular construct mechanical properties. PCL fiber matrices functionalized with PNEA-mPh acquired a rougher surface morphology and led to enhanced cell adhesion as well as superior cell-construct infiltration when compared to smooth PCL fiber matrices. Long-term in vitro hMSC cultures on both fiber matrices were able to produce clinically relevant moduli. Both fibrous constructs expressed scleraxis, an early tendon differentiation marker, and a bimodal peak in expression of the late tendon differentiation marker tenomodulin, a pattern that was not observed in PCL thin film controls. Functionalized matrices achieved a more prominent tenogenic differentiation, possessing greater tenomodulin expression and superior phenotypic maturity according to the ratio of collagen I to collagen III expression. These findings indicate that PNEA-mPh functionalization is an efficient method for improving cell interactions with electrospun PCL matrices for the purpose of tendon repair. PMID:22736077

  7. Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro.

    PubMed

    Chal, Jérome; Al Tanoury, Ziad; Hestin, Marie; Gobert, Bénédicte; Aivio, Suvi; Hick, Aurore; Cherrier, Thomas; Nesmith, Alexander P; Parker, Kevin K; Pourquié, Olivier

    2016-10-01

    Progress toward finding a cure for muscle diseases has been slow because of the absence of relevant cellular models and the lack of a reliable source of muscle progenitors for biomedical investigation. Here we report an optimized serum-free differentiation protocol to efficiently produce striated, millimeter-long muscle fibers together with satellite-like cells from human pluripotent stem cells (hPSCs) in vitro. By mimicking key signaling events leading to muscle formation in the embryo, in particular the dual modulation of Wnt and bone morphogenetic protein (BMP) pathway signaling, this directed differentiation protocol avoids the requirement for genetic modifications or cell sorting. Robust myogenesis can be achieved in vitro within 1 month by personnel experienced in hPSC culture. The differentiating culture can be subcultured to produce large amounts of myogenic progenitors amenable to numerous downstream applications. Beyond the study of myogenesis, this differentiation method offers an attractive platform for the development of relevant in vitro models of muscle dystrophies and drug screening strategies, as well as providing a source of cells for tissue engineering and cell therapy approaches. PMID:27583644

  8. Graphene-coated carbon fiber cloth for flexible electrodes of glucose fuel cells

    NASA Astrophysics Data System (ADS)

    Hoshi, Kazuki; Muramatsu, Kazuo; Sumi, Hisato; Nishioka, Yasushiro

    2016-02-01

    In this work, we fabricated flexible electrodes for a miniaturized, simple structured, and flexible glucose biofuel cell (BFC) using a graphene-coated carbon fiber cloth (GCFC). The areas of the anode and cathode electrodes were 3 × 10 mm2. The anode area was coated with the enzyme glucose oxidase, and the cathode area was coated with the enzyme bilirubin oxidase. No ion-exchange film was needed because glucose oxidase selectively oxidizes glucose and bilirubin oxidase selectively reduces oxygen. The power density of the BFC with GCFC electrodes in a phosphate buffer solution of 200 mM glucose solution at room temperature was 34.3 µW/cm2 at 0.43 V. The power density of a BFC using carbon fiber cloth (CFC) without graphene modification was 18.5 µW/cm2 at 0.13 V. The BFC with the GCFC electrode continued to function longer than 24 h with a power density higher than 5 µW/cm2. These effects were attributed to the much larger effective surface areas of the GCFC electrodes that maintain more enzymes than those of the CFC electrodes.

  9. Defined topologically-complex protein matrices to manipulate cell shape via three-dimensional fiber-like patterns.

    PubMed

    Moraes, Christopher; Kim, Byoung Choul; Zhu, Xiaoyue; Mills, Kristen L; Dixon, Angela R; Thouless, M D; Takayama, Shuichi

    2014-07-01

    Culturing cells in three-dimensional (3D) environments has been shown to significantly influence cell function, and may provide a more physiologically relevant environment within which to study the behavior of specific cell types. 3D tissues typically present a topologically complex fibrous adhesive environment, which is technically challenging to replicate in a controlled manner. Micropatterning technologies have provided significant insights into cell-biomaterial interactions, and can be used to create fiber-like adhesive structures, but are typically limited to flat culture systems; the methods are difficult to apply to topologically-complex surfaces. In this work, we utilize crack formation in multilayered microfabricated materials under applied strain to rapidly generate well-controlled and topologically complex 'fiber-like' adhesive protein patterns, capable of supporting cell culture and controlling cell shape on three-dimensional patterns. We first demonstrate that the features of the generated adhesive environments such as width, spacing and topology can be controlled, and that these factors influence cell morphology. The patterning technique is then applied to examine the influence of fiber structure on the nuclear morphology and actin cytoskeletal structure of cells cultured in a nanofibrous biomaterial matrix. PMID:24632936

  10. Defined topologically-complex protein matrices to manipulate cell shape via three-dimensional fiber-like patterns

    PubMed Central

    Moraes, Christopher; Kim, Byoung Choul; Zhu, Xiaoyue; Mills, Kristen L.; Dixon, Angela R.; Thouless; Takayama, Shuichi

    2014-01-01

    Culturing cells in three-dimensional (3D) environments has been shown to significantly influence cell function, and may provide a more physiologically relevant environment within which to study the behavior of specific cell types. 3D tissues typically present a topologically complex fibrous adhesive environment, which is technically challenging to replicate in a controlled manner. Micropatterning technologies have provided significant insights into cell-biomaterial interactions, and can be used to create fiber-like adhesive structures, but are typically limited to flat culture systems; the methods are difficult to apply to topologically-complex surfaces. In this work, we utilize crack formation in multilayered microfabricated materials under applied strain to rapidly generate well-controlled and topologically complex ‘fiber-like’ adhesive protein patterns, capable of supporting cell culture and controlling cell shape on three-dimensional patterns. We first demonstrate that the features of the generated adhesive environments such as width, spacing and topology can be controlled, and that these factors influence cell morphology. The patterning technique is then applied to examine the influence of fiber structure on the nuclear morphology and actin cytoskeletal structure of cells cultured in a nanofibrous biomaterial matrix. PMID:24632936

  11. 3D cell culture and osteogenic differentiation of human bone marrow stromal cells plated onto jet-sprayed or electrospun micro-fiber scaffolds.

    PubMed

    Brennan, Meadhbh Á; Renaud, Audrey; Gamblin, Anne-Laure; D'Arros, Cyril; Nedellec, Steven; Trichet, Valerie; Layrolle, Pierre

    2015-08-01

    A major limitation of the 2D culture systems is that they fail to recapitulate the in vivo 3D cellular microenvironment whereby cell-cell and cell-extracellular matrix (ECM) interactions occur. In this paper, a biomaterial scaffold that mimics the structure of collagen fibers was produced by jet-spraying. This micro-fiber polycaprolactone (PCL) scaffold was evaluated for 3D culture of human bone marrow mesenchymal stromal cells (MSCs) in comparison with a commercially available electrospun scaffold. The jet-sprayed scaffolds had larger pore diameters, greater porosity, smaller diameter fibers, and more heterogeneous fiber diameter size distribution compared to the electrospun scaffolds. Cells on jet-sprayed constructs exhibited spread morphology with abundant cytoskeleton staining, whereas MSCs on electrospun scaffolds appeared less extended with fewer actin filaments. MSC proliferation and cell infiltration occurred at a faster rate on jet-sprayed compared to electrospun scaffolds. Osteogenic differentiation of MSCs and ECM production as measured by ALP, collagen and calcium deposition was superior on jet-sprayed compared to electrospun scaffolds. The jet-sprayed scaffold which mimics the native ECM and permits homogeneous cell infiltration is important for 3D in vitro applications such as bone cellular interaction studies or drug testing, as well as bone tissue engineering strategies. PMID:26238732

  12. Cytotoxicity, oxidative stress and genotoxicity induced by glass fibers on human alveolar epithelial cell line A549.

    PubMed

    Rapisarda, Venerando; Loreto, Carla; Ledda, Caterina; Musumeci, Giuseppe; Bracci, Massimo; Santarelli, Lory; Renis, Marcella; Ferrante, Margherita; Cardile, Venera

    2015-04-01

    Man-made vitreous fibers have been widely used as insulation material as asbestos substitutes; however their morphology and composition raises concerns. In 1988 the International Agency for Research on Cancer classified fiberglass, rock wool, slag wool, and ceramic fibers as Group 2B, i.e. possibly carcinogenic to humans. In 2002 it reassigned fiberglass, rock and slag wool, and continuous glass filaments to Group 3, not classifiable as carcinogenic to humans. The aim of this study was to verify the cytotoxic and genotoxic effects and oxidative stress production induced by in vitro exposure of human alveolar epithelial cells A549 to glass fibers with a predominant diameter <3 μm (97%) and length >5 μm (93%). A549 cells were incubated with 5, 50, or 100 μg/ml (2.1, 21, and 42 μg/cm(2), respectively) of glass fibers for 72 h. Cytotoxicity and DNA damage were tested by the MTT and the Comet assay, respectively. Oxidative stress was determined by measuring inducible nitric oxide synthase (iNOS) expression by Western blotting, production of nitric oxide (NO) with Griess reagent, and concentration of reactive oxygen species by fluorescent quantitative analysis with 2',7'-dichlorofluorescein-diacetate (DCFH-DA). The results showed that glass fiber exposure significantly reduced cell viability and increased DNA damage and oxidative stress production in a concentration-dependent manner, demonstrating that glass fibers exert cytotoxic and genotoxic effects related to increased oxidative stress on the human alveolar cell line A549. PMID:25620604

  13. A Novel Isoform of Sucrose Synthase Is Targeted to the Cell Wall during Secondary Cell Wall Synthesis in Cotton Fiber[C][W][OA

    PubMed Central

    Brill, Elizabeth; van Thournout, Michel; White, Rosemary G.; Llewellyn, Danny; Campbell, Peter M.; Engelen, Steven; Ruan, Yong-Ling; Arioli, Tony; Furbank, Robert T.

    2011-01-01

    Sucrose (Suc) synthase (Sus) is the major enzyme of Suc breakdown for cellulose biosynthesis in cotton (Gossypium hirsutum) fiber, an important source of fiber for the textile industry. This study examines the tissue-specific expression, relative abundance, and temporal expression of various Sus transcripts and proteins present in cotton. A novel isoform of Sus (SusC) is identified that is expressed at high levels during secondary cell wall synthesis in fiber and is present in the cell wall fraction. The phylogenetic relationships of the deduced amino acid sequences indicate two ancestral groups of Sus proteins predating the divergence of monocots and dicots and that SusC sequences form a distinct branch in the phylogeny within the dicot-specific clade. The subcellular location of the Sus isoforms is determined, and it is proposed that cell wall-localized SusC may provide UDP-glucose for cellulose and callose synthesis from extracellular sugars. PMID:21757635

  14. Ultrastructural and Chemical Evidence That the Cell Wall of Green Cotton Fiber Is Suberized 1

    PubMed Central

    Yatsu, L. Y.; Espelie, Karl E.; Kolattukudy, P. E.

    1983-01-01

    Green cotton (Gossypium hirsutum L.) fibers were shown by electron microscopy to have numerous thin concentric rings around the lumen of the cell. These rings possessed a lamellar fine structure characteristic of suberin. LiA1D4 depolymerization and gas chromatography-mass spectrometry analysis showed the presence of a suberin polymer in the green cotton with the major aliphatic monomers being ω-hydroxydocosanoic acid (70%) and docosanedoic acid (25%). Ordinary white cotton was shown by chemical and ultrastructural examination to be encircled by a thin cuticular polymer containing less than 0.5% of the aliphatic components found in green cotton. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:16663251

  15. Study of the Peripheral Nerve Fibers Myelin Structure Changes during Activation of Schwann Cell Acetylcholine Receptors

    PubMed Central

    Verdiyan, Ekaterina E.; Allakhverdiev, Elvin S.; Maksimov, Georgy V.

    2016-01-01

    In the present paper we consider a new type of mechanism by which neurotransmitter acetylcholine (ACh) regulates the properties of peripheral nerve fibers myelin. Our data show the importance of the relationship between the changes in the number of Schwann cell (SC) acetylcholine receptors (AChRs) and the axon excitation (different intervals between action potentials (APs)). Using Raman spectroscopy, an effect of activation of SC AChRs on the myelin membrane fluidity was investigated. It was found, that ACh stimulates an increase in lipid ordering degree of the myelin lipids, thus providing evidence for specific role of the “axon-SC” interactions at the axon excitation. It was proposed, that during the axon excitation, the SC membrane K+- depolarization and the Ca2+—influx led to phospholipase activation or exocytosis of intracellular membrane vesicles and myelin structure reorganization. PMID:27455410

  16. Retinal nerve fiber layer and ganglion cell-inner plexiform layer thickness in children with obesity

    PubMed Central

    Demir, Selim; Özer, Samet; Alim, Sait; Güneş, Alper; Ortak, Hüseyin; Yılmaz, Resul

    2016-01-01

    AIM To evaluate retinal nerve fiber layer (RNFL) thickness analysis of peripapillary optic nerve head (PONH) and macula as well as ganglion cell-inner plexiform layer (GCIPL) thickness in obese children. METHODS Eighty-five children with obesity and 30 controls were included in the study. The thicknesses of the PONH and macula of each subject's right eye were measured by high-resolution spectral-domain optic coherence tomography (OCT). RESULTS The RNFL thicknesses of central macular and PONH were similar between the groups (all P>0.05). The GCIPL thickness was also similar between the groups. However, the RNFL thickness of temporal outer macula were 261.7±13.7 and 268.9±14.3 µm for the obesity and the control group, respectively (P=0.034). CONCLUSION Obesity may cause a reduction in temporal outer macular RNFL thickness. PMID:27158616

  17. Wet-laid soy fiber reinforced hydrogel scaffold: Fabrication, mechano-morphological and cell studies.

    PubMed

    Wood, Andrew T; Everett, Dominique; Budhwani, Karim I; Dickinson, Brenna; Thomas, Vinoy

    2016-06-01

    Among materials used in biomedical applications, hydrogels have received consistent linear growth in interest over the past decade due to their large water volume and saliency to the natural extracellular matrix. These materials are often limited due to their sub-optimal mechanical properties which are typically improved via chemical or physical crosslinking. Chemical crosslinking forms strong inter-polymer bonds but typically uses reagents that are cytotoxic while physical crosslinking is more temperamental to environmental changes but can be formed without these toxic reagents. In this study, we added a fiber-reinforcement phase to a poly(vinyl alcohol) (PVA) hydrogel formed through successive freezing-thawing cycles by incorporating a non-woven microfiber mat formed by the wet-lay process. By reinforcing the hydrogel with a wet-laid fibrous mat, the ultimate tensile strength and modulus increased from 0.11±0.01MPa and 0.17±0.02kPa to 0.24±0.02MPa and 5.76±1.12kPa, respectively. An increase in toughness and elongation was also found increasing from 2.52±0.37MPa to 25.6±3.84 and 51.89±5.16% to 111.16±9.68%, respectively. The soy fibers were also found to induce minimal cytotoxicity with endothelial cell viability showing 96.51%±1.91 living cells after a 48h incubation. This approach to hydrogel-reinforcement presents a rapid, tunable method by which hydrogels can attain increased mechanical properties without sacrificing their inherent biologically favorable properties. PMID:27040224

  18. Role of stress fibers and focal adhesions as a mediator for mechano-signal transduction in endothelial cells in situ

    PubMed Central

    Katoh, Kazuo; Kano, Yumiko; Ookawara, Shigeo

    2008-01-01

    Fluid shear stress is the mechanical force generated by the blood flow which is applied over the apical surface of endothelial cells in situ. The findings of a recent study suggest that stress fibers and its associated focal adhesions play roles in mechano-signal transduction mechanism. Stress fibers are present along the apical and the basal portion of the endothelial cells. Endothelial cells respond to fluid shear stress and change their morphological characteristics in both their cell shape and cytoskeletal organization. Atherosclerosis is a common disease of the arteries and it occurs in areas around the branching site of blood vessels where the cells are exposed to low fluid shear stress. The organization of stress fibers and focal adhesions are strongly influenced by shear stress, and therefore the generation of atherosclerotic lesions seem to be associated with the cytoskeletal components of endothelial cells. This review describes the possible role of the cytoskeleton as a mechano-transducer in endothelial cells in situ. PMID:19337541

  19. Vascularized Bone Tissue Formation Induced by Fiber-Reinforced Scaffolds Cultured with Osteoblasts and Endothelial Cells

    PubMed Central

    Liu, Xinhui; Zhang, Guoping; Hou, Chuanyong; Wang, Hua; Yang, Yelin; Guan, Guoping; Dong, Wei; Gao, Hongyang

    2013-01-01

    The repair of the damaged bone tissue caused by damage or bone disease was still a problem. Current strategies including the use of autografts and allografts have the disadvantages, namely, diseases transmission, tissue availability and donor morbidity. Bone tissue engineering has been developed and regarded as a new way of regenerating bone tissues to repair or substitute damaged or diseased ones. The main limitation in engineering in vitro tissues is the lack of a sufficient blood vessel system, the vascularization. In this paper, a new-typed hydroxyapatite/collagen composite scaffold which was reinforced by chitosan fibers and cultured with osteoblasts and endothelial cells was fabricated. General observation, histological observation, detection of the degree of vascularization, and X-ray examination had been done to learn the effect of vascularized bone repair materials on the regeneration of bone. The results show that new vessel and bone formed using implant cultured with osteoblasts and endothelial cells. Nanofiber-reinforced scaffold cultured with osteoblasts and endothelial cells can induce vascularized bone tissue formation. PMID:24369019

  20. Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations.

    PubMed

    Jansen, J; De Napoli, I E; Fedecostante, M; Schophuizen, C M S; Chevtchik, N V; Wilmer, M J; van Asbeck, A H; Croes, H J; Pertijs, J C; Wetzels, J F M; Hilbrands, L B; van den Heuvel, L P; Hoenderop, J G; Stamatialis, D; Masereeuw, R

    2015-01-01

    The bioartificial kidney (BAK) aims at improving dialysis by developing 'living membranes' for cells-aided removal of uremic metabolites. Here, unique human conditionally immortalized proximal tubule epithelial cell (ciPTEC) monolayers were cultured on biofunctionalized MicroPES (polyethersulfone) hollow fiber membranes (HFM) and functionally tested using microfluidics. Tight monolayer formation was demonstrated by abundant zonula occludens-1 (ZO-1) protein expression along the tight junctions of matured ciPTEC on HFM. A clear barrier function of the monolayer was confirmed by limited diffusion of FITC-inulin. The activity of the organic cation transporter 2 (OCT2) in ciPTEC was evaluated in real-time using a perfusion system by confocal microscopy using 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (ASP(+)) as a fluorescent substrate. Initial ASP(+) uptake was inhibited by a cationic uremic metabolites mixture and by the histamine H2-receptor antagonist, cimetidine. In conclusion, a 'living membrane' of renal epithelial cells on MicroPES HFM with demonstrated active organic cation transport was successfully established as a first step in BAK engineering. PMID:26567716

  1. Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations

    PubMed Central

    Jansen, J.; De Napoli, I. E; Fedecostante, M.; Schophuizen, C. M. S.; Chevtchik, N. V.; Wilmer, M. J.; van Asbeck, A. H.; Croes, H. J.; Pertijs, J. C.; Wetzels, J. F. M.; Hilbrands, L. B.; van den Heuvel, L. P.; Hoenderop, J. G.; Stamatialis, D.; Masereeuw, R.

    2015-01-01

    The bioartificial kidney (BAK) aims at improving dialysis by developing ‘living membranes’ for cells-aided removal of uremic metabolites. Here, unique human conditionally immortalized proximal tubule epithelial cell (ciPTEC) monolayers were cultured on biofunctionalized MicroPES (polyethersulfone) hollow fiber membranes (HFM) and functionally tested using microfluidics. Tight monolayer formation was demonstrated by abundant zonula occludens-1 (ZO-1) protein expression along the tight junctions of matured ciPTEC on HFM. A clear barrier function of the monolayer was confirmed by limited diffusion of FITC-inulin. The activity of the organic cation transporter 2 (OCT2) in ciPTEC was evaluated in real-time using a perfusion system by confocal microscopy using 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (ASP+) as a fluorescent substrate. Initial ASP+ uptake was inhibited by a cationic uremic metabolites mixture and by the histamine H2-receptor antagonist, cimetidine. In conclusion, a ‘living membrane’ of renal epithelial cells on MicroPES HFM with demonstrated active organic cation transport was successfully established as a first step in BAK engineering. PMID:26567716

  2. The Use of Fiber-Reinforced Scaffolds Cocultured with Schwann Cells and Vascular Endothelial Cells to Repair Rabbit Sciatic Nerve Defect with Vascularization

    PubMed Central

    Gao, Hongyang; You, Yang; Zhang, Guoping; Zhao, Feng; Sha, Ziyi; Shen, Yong

    2013-01-01

    To explore the feasibility of biodegradable fiber-reinforced 3D scaffolds with satisfactory mechanical properties for the repair of long-distance sciatic nerve defect in rabbits and effects of vascularized graft in early stage on the recovery of neurological function, Schwann cells and vascular endothelial cells were cocultured in the fiber-reinforced 3D scaffolds. Experiment group which used prevascularized nerve complex for the repair of sciatic nerve defect and control group which only cultured with Schwann cells were set. The animals in both groups underwent electromyography to show the status of the neurological function recovery at 4, 8, and 16 weeks after the surgery. Sciatic nerve regeneration and myelination were observed under the light microscope and electron microscope. Myelin sheath thickness, axonal diameter, and number of myelinated nerve fiber were quantitatively analyzed using image analysis system. The recovery of foot ulcer, the velocity of nerve conduction, the number of regenerating nerve fiber, and the recovery of ultrastructure were increased in the experimental group than those in the control group. Prevascularized tissue engineered fiber-reinforced 3D scaffolds for the repair of sciatic nerve defects in rabbits can effectively promote the recovery of neurological function. PMID:24490158

  3. The use of fiber-reinforced scaffolds cocultured with Schwann cells and vascular endothelial cells to repair rabbit sciatic nerve defect with vascularization.

    PubMed

    Gao, Hongyang; You, Yang; Zhang, Guoping; Zhao, Feng; Sha, Ziyi; Shen, Yong

    2013-01-01

    To explore the feasibility of biodegradable fiber-reinforced 3D scaffolds with satisfactory mechanical properties for the repair of long-distance sciatic nerve defect in rabbits and effects of vascularized graft in early stage on the recovery of neurological function, Schwann cells and vascular endothelial cells were cocultured in the fiber-reinforced 3D scaffolds. Experiment group which used prevascularized nerve complex for the repair of sciatic nerve defect and control group which only cultured with Schwann cells were set. The animals in both groups underwent electromyography to show the status of the neurological function recovery at 4, 8, and 16 weeks after the surgery. Sciatic nerve regeneration and myelination were observed under the light microscope and electron microscope. Myelin sheath thickness, axonal diameter, and number of myelinated nerve fiber were quantitatively analyzed using image analysis system. The recovery of foot ulcer, the velocity of nerve conduction, the number of regenerating nerve fiber, and the recovery of ultrastructure were increased in the experimental group than those in the control group. Prevascularized tissue engineered fiber-reinforced 3D scaffolds for the repair of sciatic nerve defects in rabbits can effectively promote the recovery of neurological function. PMID:24490158

  4. Mast Cell-Derived Tumor Necrosis Factor Can Promote Nerve Fiber Elongation in the Skin during Contact Hypersensitivity in Mice

    PubMed Central

    Kakurai, Maki; Monteforte, Rossella; Suto, Hajime; Tsai, Mindy; Nakae, Susumu; Galli, Stephen J.

    2006-01-01

    In humans, lesions of contact eczema or atopic dermatitis can exhibit increases in epidermal nerves, but the mechanism resulting in such nerve elongation are not fully understood. We found that contact hypersensitivity reactions to oxazolone in mice were associated with significant increases in the length of nerves in the epidermis and dermis. Using genetically mast cell-deficient c-kit mutant mice selectively repaired of their dermal mast cell deficiency with either wild-type or tumor necrosis factor (TNF)-deficient mast cells, we found that mast cells, and mast cell-derived TNF, significantly contributed to the elongation of epidermal and dermal PGP 9.5+ nerves and dermal CGRP+ nerves, as well as to the inflammation observed at sites of contact hypersensitivity in response to oxazolone. Moreover, the percentage of mast cells in close proximity to dermal PGP 9.5+ nerve fibers was significantly higher in wild-type mice and in c-kit mutant mice repaired of their dermal mast cell deficiency by the adoptive transfer of wild-type mast cells than in TNF-deficient mice or in TNF−/− mast cell-engrafted c-kit mutant mice. These observations show that mast cells, and mast cell-derived TNF, can promote the elongation of cutaneous nerve fibers during contact hypersensitivity in the mouse. PMID:17071594

  5. A Specialized Outer Layer of the Primary Cell Wall Joins Elongating Cotton Fibers into Tissue-Like Bundles1[W][OA

    PubMed Central

    Singh, Bir; Avci, Utku; Eichler Inwood, Sarah E.; Grimson, Mark J.; Landgraf, Jeff; Mohnen, Debra; Sørensen, Iben; Wilkerson, Curtis G.; Willats, William G.T.; Haigler, Candace H.

    2009-01-01

    Cotton (Gossypium hirsutum) provides the world's dominant renewable textile fiber, and cotton fiber is valued as a research model because of its extensive elongation and secondary wall thickening. Previously, it was assumed that fibers elongated as individual cells. In contrast, observation by cryo-field emission-scanning electron microscopy of cotton fibers developing in situ within the boll demonstrated that fibers elongate within tissue-like bundles. These bundles were entrained by twisting fiber tips and consolidated by adhesion of a cotton fiber middle lamella (CFML). The fiber bundles consolidated via the CFML ultimately formed a packet of fiber around each seed, which helps explain how thousands of cotton fibers achieve their great length within a confined space. The cell wall nature of the CFML was characterized using transmission electron microscopy, including polymer epitope labeling. Toward the end of elongation, up-regulation occurred in gene expression and enzyme activities related to cell wall hydrolysis, and targeted breakdown of the CFML restored fiber individuality. At the same time, losses occurred in certain cell wall polymer epitopes (as revealed by comprehensive microarray polymer profiling) and sugars within noncellulosic matrix components (as revealed by gas chromatography-mass spectrometry analysis of derivatized neutral and acidic glycosyl residues). Broadly, these data show that adhesion modulated by an outer layer of the primary wall can coordinate the extensive growth of a large group of cells and illustrate dynamic changes in primary wall structure and composition occurring during the differentiation of one cell type that spends only part of its life as a tissue. PMID:19369592

  6. Actin stress fiber disruption and tropomysin isoform switching in normal thyroid epithelial cells stimulated by thyrotropin and phorbol esters

    SciTech Connect

    Roger, P.P.; Rickaert, F.; Lamy, F.; Authelet, M.; Dumont, J.E. )

    1989-05-01

    Thyrotropin (TSH), through cyclic AMP, promotes both proliferation and differentiation expression in dog thyroid epithelial cells in primary culture, whereas the tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA) also stimulates proliferation but antagonizes differentiating effects of TSH. In this study, within 20 min both factors triggered the disruption of actin-containing stress fibers. This process preceded distinct morphological changes: cytoplasmic retraction and arborization in response to TSH and cyclic AMP, cell shape distortion, and increased motility in response to TPA and diacylglycerol. TSH and TPA also induced a marked decrease in the synthesis of three high M{sub r} tropomyosin isoforms, which were not present in dog thyroid tissue but appeared in culture during cell spreading and stress fiber formation. The tropomyosin isoform switching observed here closely resembled similar processes in various cells transformed by oncogenic viruses. However, it did not correlate with differentiation or mitogenic activation. Contrasting with current hypothesis on this process in transformed cells, tropomyosin isoform switching in normal thyroid cells was preceded and thus might be caused by early disruption of stress fibers.

  7. Catalytic Improvement on Counter Electrode of Dye-Sensitized Solar Cells Using Electrospun Pt Nano-Fibers.

    PubMed

    Seol, Hyunwoong; Shiratani, Masaharu; Seneekatima, Kannanut; Pornprasertsuk, Rojana

    2016-04-01

    A dye-sensitized solar cell is one of cost-competitive photovoltaic devices. For higher performance, all components have been actively studied and improved. However, Pt is still a dominant catalyst since first development although some catalytic materials were studied so far. Catalytic materials of counter electrode play an important role in the performance because it supplies electrons from counter electrode to electrolyte. Therefore, the catalytic activation of counter electrode is closely connected with the performance enhancement. In this work, Pt nano-fiber was fabricated by electrospinning and applied for the counter electrode. Its wide surface area is advantageous for good conductivity and catalytic activation. Morphological characteristics of nano-fibers were analyzed according to electrospinning conditions. Photovoltaic properties, cyclic voltammetry, impedance analysis verified the catalytic activation. Consequently, dye-sensitized solar cell with Pt nano-fiber electrospun at 5.0 kV of applied voltage had higher performance than conventional dye-sensitized solar cell with Pt thin film. This work is significant for related researches because all nano-fibers counter electrode material proposed so far never exceeded the performance of conventional Pt counter electrode. PMID:27451627

  8. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions

    NASA Astrophysics Data System (ADS)

    Fraley, Stephanie I.; Wu, Pei-Hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D.; Wirtz, Denis

    2015-10-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility.

  9. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions.

    PubMed

    Fraley, Stephanie I; Wu, Pei-Hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D; Wirtz, Denis

    2015-01-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility. PMID:26423227

  10. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions

    PubMed Central

    Fraley, Stephanie I.; Wu, Pei-hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D.; Wirtz, Denis

    2015-01-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility. PMID:26423227