Sample records for border cell separation

  1. Role of pectolytic enzymes in the programmed separation of cells from the root cap of higher plants. Final report

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

    Hawes, M.C.

    1995-03-01

    The objective of this research was to develop a model system to study border cell separation in transgenic pea roots. In addition, the hypothesis that genes encoding pectolytic enzymes in the root cap play a role in the programmed separation of root border cells from the root tip was tested. The following objectives have been accomplished: (1) the use of transgenic hairy roots to study border cell separation has been optimized for Pisum sativum; (2) a cDNA encoding a root cap pectinmethylesterase (PME) has been cloned; (3) PME and polygalacturonase activities in cell walls of the root cap have beenmore » characterized and shown to be correlated with border cell separation. A fusion gene encoding pectate lyase has also been transformed into pea hairy root cells.« less

  2. Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics1[OPEN

    PubMed Central

    2017-01-01

    The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. Border cells (in some species known as border-like cells) provide an expendable barrier between roots and the environment. Their maturation and release is an important but poorly characterized cell separation event. To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. Our findings provide a new level of detail about border cell maturation and enable us to develop a model of the separation process. We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. PMID:28400496

  3. Border cell release: Cell separation without cell wall degradation?

    PubMed

    Mravec, Jozef

    2017-07-03

    Plant border cells are specialized cells derived from the root cap with roles in the biomechanics of root growth and in forming a barrier against pathogens. The mechanism of highly localized cell separation which is essential for their release to the environment is little understood. Here I present in situ analysis of Brachypodium distachyon, a model organism for grasses which possess type II primary cell walls poor in pectin content. Results suggest similarity in spatial dynamics of pectic homogalacturonan during dicot and monocot border cell release. Integration of observations from different species leads to the hypothesis that this process most likely does not involve degradation of cell wall material but rather uses unique cell wall structural and compositional means enabling both the rigidity of the root cap as well as detachability of given cells on its surface.

  4. Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics.

    PubMed

    Mravec, Jozef; Guo, Xiaoyuan; Hansen, Aleksander Riise; Schückel, Julia; Kračun, Stjepan Krešimir; Mikkelsen, Maria Dalgaard; Mouille, Grégory; Johansen, Ida Elisabeth; Ulvskov, Peter; Domozych, David S; Willats, William George Tycho

    2017-06-01

    The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. Border cells (in some species known as border-like cells) provide an expendable barrier between roots and the environment. Their maturation and release is an important but poorly characterized cell separation event. To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea ( Pisum sativum ) root tip cell walls. Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. Our findings provide a new level of detail about border cell maturation and enable us to develop a model of the separation process. We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. © 2017 American Society of Plant Biologists. All Rights Reserved.

  5. The organization pattern of root border-like cells of Arabidopsis is dependent on cell wall homogalacturonan.

    PubMed

    Durand, Caroline; Vicré-Gibouin, Maïté; Follet-Gueye, Marie Laure; Duponchel, Ludovic; Moreau, Myriam; Lerouge, Patrice; Driouich, Azeddine

    2009-07-01

    Border-like cells are released by Arabidopsis (Arabidopsis thaliana) root tips as organized layers of several cells that remain attached to each other rather than completely detached from each other, as is usually observed in border cells of many species. Unlike border cells, cell attachment between border-like cells is maintained after their release into the external environment. To investigate the role of cell wall polysaccharides in the attachment and organization of border-like cells, we have examined their release in several well-characterized mutants defective in the biosynthesis of xyloglucan, cellulose, or pectin. Our data show that among all mutants examined, only quasimodo mutants (qua1-1 and qua2-1), which have been characterized as producing less homogalacturonan, had an altered border-like cell phenotype as compared with the wild type. Border-like cells in both lines were released as isolated cells separated from each other, with the phenotype being much more pronounced in qua1-1 than in qua2-1. Further analysis of border-like cells in the qua1-1 mutant using immunocytochemistry and a set of anti-cell wall polysaccharide antibodies showed that the loss of the wild-type phenotype was accompanied by (1) a reduction in homogalacturonan-JIM5 epitope in the cell wall of border-like cells, confirmed by Fourier transform infrared microspectrometry, and (2) the secretion of an abundant mucilage that is enriched in xylogalacturonan and arabinogalactan-protein epitopes, in which the cells are trapped in the vicinity of the root tip.

  6. The Organization Pattern of Root Border-Like Cells of Arabidopsis Is Dependent on Cell Wall Homogalacturonan12[C][W

    PubMed Central

    Durand, Caroline; Vicré-Gibouin, Maïté; Follet-Gueye, Marie Laure; Duponchel, Ludovic; Moreau, Myriam; Lerouge, Patrice; Driouich, Azeddine

    2009-01-01

    Border-like cells are released by Arabidopsis (Arabidopsis thaliana) root tips as organized layers of several cells that remain attached to each other rather than completely detached from each other, as is usually observed in border cells of many species. Unlike border cells, cell attachment between border-like cells is maintained after their release into the external environment. To investigate the role of cell wall polysaccharides in the attachment and organization of border-like cells, we have examined their release in several well-characterized mutants defective in the biosynthesis of xyloglucan, cellulose, or pectin. Our data show that among all mutants examined, only quasimodo mutants (qua1-1 and qua2-1), which have been characterized as producing less homogalacturonan, had an altered border-like cell phenotype as compared with the wild type. Border-like cells in both lines were released as isolated cells separated from each other, with the phenotype being much more pronounced in qua1-1 than in qua2-1. Further analysis of border-like cells in the qua1-1 mutant using immunocytochemistry and a set of anti-cell wall polysaccharide antibodies showed that the loss of the wild-type phenotype was accompanied by (1) a reduction in homogalacturonan-JIM5 epitope in the cell wall of border-like cells, confirmed by Fourier transform infrared microspectrometry, and (2) the secretion of an abundant mucilage that is enriched in xylogalacturonan and arabinogalactan-protein epitopes, in which the cells are trapped in the vicinity of the root tip. PMID:19448034

  7. Root Border Cells and Their Role in Plant Defense.

    PubMed

    Hawes, Martha; Allen, Caitilyn; Turgeon, B Gillian; Curlango-Rivera, Gilberto; Minh Tran, Tuan; Huskey, David A; Xiong, Zhongguo

    2016-08-04

    Root border cells separate from plant root tips and disperse into the soil environment. In most species, each root tip can produce thousands of metabolically active cells daily, with specialized patterns of gene expression. Their function has been an enduring mystery. Recent studies suggest that border cells operate in a manner similar to mammalian neutrophils: Both cell types export a complex of extracellular DNA (exDNA) and antimicrobial proteins that neutralize threats by trapping pathogens and thereby preventing invasion of host tissues. Extracellular DNases (exDNases) of pathogens promote virulence and systemic spread of the microbes. In plants, adding DNase I to root tips eliminates border cell extracellular traps and abolishes root tip resistance to infection. Mutation of genes encoding exDNase activity in plant-pathogenic bacteria (Ralstonia solanacearum) and fungi (Cochliobolus heterostrophus) results in reduced virulence. The study of exDNase activities in plant pathogens may yield new targets for disease control.

  8. The Selector Gene apterous and Notch Are Required to Locally Increase Mechanical Cell Bond Tension at the Drosophila Dorsoventral Compartment Boundary

    PubMed Central

    Michel, Marcus; Aliee, Maryam; Rudolf, Katrin; Bialas, Lisa; Jülicher, Frank; Dahmann, Christian

    2016-01-01

    The separation of cells with distinct fates and functions is important for tissue and organ formation during animal development. Regions of different fates within tissues are often separated from another along straight boundaries. These compartment boundaries play a crucial role in tissue patterning and growth by stably positioning organizers. In Drosophila, the wing imaginal disc is subdivided into a dorsal and a ventral compartment. Cells of the dorsal, but not ventral, compartment express the selector gene apterous. Apterous expression sets in motion a gene regulatory cascade that leads to the activation of Notch signaling in a few cell rows on either side of the dorsoventral compartment boundary. Both Notch and apterous mutant clones disturb the separation of dorsal and ventral cells. Maintenance of the straight shape of the dorsoventral boundary involves a local increase in mechanical tension at cell bonds along the boundary. The mechanisms by which cell bond tension is locally increased however remain unknown. Here we use a combination of laser ablation of cell bonds, quantitative image analysis, and genetic mutants to show that Notch and Apterous are required to increase cell bond tension along the dorsoventral compartment boundary. Moreover, clonal expression of the Apterous target gene capricious results in cell separation and increased cell bond tension at the clone borders. Finally, using a vertex model to simulate tissue growth, we find that an increase in cell bond tension at the borders of cell clones, but not throughout the cell clone, can lead to cell separation. We conclude that Apterous and Notch maintain the characteristic straight shape of the dorsoventral compartment boundary by locally increasing cell bond tension. PMID:27552097

  9. Consistency of Border-Ownership Cells across Artificial Stimuli, Natural Stimuli, and Stimuli with Ambiguous Contours.

    PubMed

    Hesse, Janis K; Tsao, Doris Y

    2016-11-02

    Segmentation and recognition of objects in a visual scene are two problems that are hard to solve separately from each other. When segmenting an ambiguous scene, it is helpful to already know the present objects and their shapes. However, for recognizing an object in clutter, one would like to consider its isolated segment alone to avoid confounds from features of other objects. Border-ownership cells (Zhou et al., 2000) appear to play an important role in segmentation, as they signal the side-of-figure of artificial stimuli. The present work explores the role of border-ownership cells in dorsal macaque visual areas V2 and V3 in the segmentation of natural object stimuli and locally ambiguous stimuli. We report two major results. First, compared with previous estimates, we found a smaller percentage of cells that were consistent across artificial stimuli used previously. Second, we found that the average response of those neurons that did respond consistently to the side-of-figure of artificial stimuli also consistently signaled, as a population, the side-of-figure for borders of single faces, occluding faces and, with higher latencies, even stimuli with illusory contours, such as Mooney faces and natural faces completely missing local edge information. In contrast, the local edge or the outlines of the face alone could not always evoke a significant border-ownership signal. Our results underscore that border ownership is coded by a population of cells, and indicate that these cells integrate a variety of cues, including low-level features and global object context, to compute the segmentation of the scene. To distinguish different objects in a natural scene, the brain must segment the image into regions corresponding to objects. The so-called "border-ownership" cells appear to be dedicated to this task, as they signal for a given edge on which side the object is that owns it. Here, we report that individual border-ownership cells are unreliable when tested across a battery of artificial stimuli used previously but can signal border-ownership consistently as a population. We show that these border-ownership population signals are also suited for signaling border-ownership for natural objects and at longer latency, even for stimuli without local edge information. Our results suggest that border-ownership cells integrate both local, low-level and global, high-level cues to segment the scene. Copyright © 2016 the authors 0270-6474/16/3611338-12$15.00/0.

  10. Statistical image segmentation for the detection of skin lesion borders in UV fluorescence excitation

    NASA Astrophysics Data System (ADS)

    Ortega-Martinez, Antonio; Padilla-Martinez, Juan Pablo; Franco, Walfre

    2016-04-01

    The skin contains several fluorescent molecules or fluorophores that serve as markers of structure, function and composition. UV fluorescence excitation photography is a simple and effective way to image specific intrinsic fluorophores, such as the one ascribed to tryptophan which emits at a wavelength of 345 nm upon excitation at 295 nm, and is a marker of cellular proliferation. Earlier, we built a clinical UV photography system to image cellular proliferation. In some samples, the naturally low intensity of the fluorescence can make it difficult to separate the fluorescence of cells in higher proliferation states from background fluorescence and other imaging artifacts -- like electronic noise. In this work, we describe a statistical image segmentation method to separate the fluorescence of interest. Statistical image segmentation is based on image averaging, background subtraction and pixel statistics. This method allows to better quantify the intensity and surface distributions of fluorescence, which in turn simplify the detection of borders. Using this method we delineated the borders of highly-proliferative skin conditions and diseases, in particular, allergic contact dermatitis, psoriatic lesions and basal cell carcinoma. Segmented images clearly define lesion borders. UV fluorescence excitation photography along with statistical image segmentation may serve as a quick and simple diagnostic tool for clinicians.

  11. A neural model of border-ownership from kinetic occlusion.

    PubMed

    Layton, Oliver W; Yazdanbakhsh, Arash

    2015-01-01

    Camouflaged animals that have very similar textures to their surroundings are difficult to detect when stationary. However, when an animal moves, humans readily see a figure at a different depth than the background. How do humans perceive a figure breaking camouflage, even though the texture of the figure and its background may be statistically identical in luminance? We present a model that demonstrates how the primate visual system performs figure-ground segregation in extreme cases of breaking camouflage based on motion alone. Border-ownership signals develop as an emergent property in model V2 units whose receptive fields are nearby kinetically defined borders that separate the figure and background. Model simulations support border-ownership as a general mechanism by which the visual system performs figure-ground segregation, despite whether figure-ground boundaries are defined by luminance or motion contrast. The gradient of motion- and luminance-related border-ownership signals explains the perceived depth ordering of the foreground and background surfaces. Our model predicts that V2 neurons, which are sensitive to kinetic edges, are selective to border-ownership (magnocellular B cells). A distinct population of model V2 neurons is selective to border-ownership in figures defined by luminance contrast (parvocellular B cells). B cells in model V2 receive feedback from neurons in V4 and MT with larger receptive fields to bias border-ownership signals toward the figure. We predict that neurons in V4 and MT sensitive to kinetically defined figures play a crucial role in determining whether the foreground surface accretes, deletes, or produces a shearing motion with respect to the background. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Transnational ties and the health of sub-Saharan African migrants: The moderating role of gender and family separation.

    PubMed

    Afulani, Patience A; Torres, Jacqueline M; Sudhinaraset, May; Asunka, Joseph

    2016-11-01

    Recent scholarship has focused on the role that cross-border social and economic ties play in shaping health outcomes for migrant populations. Nevertheless, the extant empirical work on this topic has paid little attention to the health impacts of cross-border separation from close family members. In this paper we examine the association between cross-border ties-and cross-border separation-with the health of sub-Saharan African (SSA) migrant adults living in metropolitan France using data from the nationally representative "Trajectoire et Origines" survey (n = 1980 SSA migrants). In logistic regression analyses we find that remitting money and having a child abroad are each associated with poor health among women, but not men. The effect of remittances on health is also modified by the location of one's children: remittance sending is associated with poor health only for SSA-migrants separated from their children. These findings underscore the importance of examining both cross-border connection and cross-border separation in studies of immigrant health, and also underscore the heterogeneous relationships between cross-border ties and health for men and women. This is the first study to our knowledge that examines the relationship between cross-border ties and health for migrants in Europe, with a focus on SSA-migrants in France. These findings have important implications for the health of the growing immigrant and refugee populations in Europe and around the globe. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. Glial Scar Borders Are Formed by Newly Proliferated, Elongated Astrocytes That Interact to Corral Inflammatory and Fibrotic Cells via STAT3-Dependent Mechanisms after Spinal Cord Injury

    PubMed Central

    Anderson, Mark A.; Song, Bingbing; Levine, Jaclynn; Fernandez, Ana; Gray-Thompson, Zachary; Ao, Yan

    2013-01-01

    Astroglial scars surround damaged tissue after trauma, stroke, infection, or autoimmune inflammation in the CNS. They are essential for wound repair, but also interfere with axonal regrowth. A better understanding of the cellular mechanisms, regulation, and functions of astroglial scar formation is fundamental to developing safe interventions for many CNS disorders. We used wild-type and transgenic mice to quantify and dissect these parameters. Adjacent to crush spinal cord injury (SCI), reactive astrocytes exhibited heterogeneous phenotypes as regards proliferation, morphology, and chemistry, which all varied with distance from lesions. Mature scar borders at 14 d after SCI consisted primarily of newly proliferated astroglia with elongated cell processes that surrounded large and small clusters of inflammatory, fibrotic, and other cells. During scar formation from 5 to 14 d after SCI, cell processes deriving from different astroglia associated into overlapping bundles that quantifiably reoriented and organized into dense mesh-like arrangements. Selective deletion of STAT3 from astroglia quantifiably disrupted the organization of elongated astroglia into scar borders, and caused a failure of astroglia to surround inflammatory cells, resulting in increased spread of these cells and neuronal loss. In cocultures, wild-type astroglia spontaneously corralled inflammatory or fibromeningeal cells into segregated clusters, whereas STAT3-deficient astroglia failed to do so. These findings demonstrate heterogeneity of reactive astroglia and show that scar borders are formed by newly proliferated, elongated astroglia, which organize via STAT3-dependent mechanisms to corral inflammatory and fibrotic cells into discrete areas separated from adjacent tissue that contains viable neurons. PMID:23904622

  14. WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate.

    PubMed

    Leung, Alan W; Murdoch, Barbara; Salem, Ahmed F; Prasad, Maneeshi S; Gomez, Gustavo A; García-Castro, Martín I

    2016-02-01

    Neural crest (NC) cells arise early in vertebrate development, migrate extensively and contribute to a diverse array of ectodermal and mesenchymal derivatives. Previous models of NC formation suggested derivation from neuralized ectoderm, via meso-ectodermal, or neural-non-neural ectoderm interactions. Recent studies using bird and amphibian embryos suggest an earlier origin of NC, independent of neural and mesodermal tissues. Here, we set out to generate a model in which to decipher signaling and tissue interactions involved in human NC induction. Our novel human embryonic stem cell (ESC)-based model yields high proportions of multipotent NC cells (expressing SOX10, PAX7 and TFAP2A) in 5 days. We demonstrate a crucial role for WNT/β-catenin signaling in launching NC development, while blocking placodal and surface ectoderm fates. We provide evidence of the delicate temporal effects of BMP and FGF signaling, and find that NC development is separable from neural and/or mesodermal contributions. We further substantiate the notion of a neural-independent origin of NC through PAX6 expression and knockdown studies. Finally, we identify a novel pre-neural border state characterized by early WNT/β-catenin signaling targets that displays distinct responses to BMP and FGF signaling from the traditional neural border genes. In summary, our work provides a fast and efficient protocol for human NC differentiation under signaling constraints similar to those identified in vivo in model organisms, and strengthens a framework for neural crest ontogeny that is separable from neural and mesodermal fates. © 2016. Published by The Company of Biologists Ltd.

  15. DE-Cadherin Is Required for Intercellular Motility during Drosophila Oogenesis

    PubMed Central

    Niewiadomska, Paulina; Godt, Dorothea; Tepass, Ulrich

    1999-01-01

    Cadherins are involved in a variety of morphogenetic movements during animal development. However, it has been difficult to pinpoint the precise function of cadherins in morphogenetic processes due to the multifunctional nature of cadherin requirement. The data presented here indicate that homophilic adhesion promoted by Drosophila E-cadherin (DE-cadherin) mediates two cell migration events during Drosophila oogenesis. In Drosophila follicles, two groups of follicle cells, the border cells and the centripetal cells migrate on the surface of germline cells. We show that the border cells migrate as an epithelial patch in which two centrally located cells retain epithelial polarity and peripheral cells are partially depolarized. Both follicle cells and germline cells express DE-cadherin, and border cells and centripetal cells strongly upregulate the expression of DE-cadherin shortly before and during their migration. Removing DE-cadherin from either the follicle cells or the germline cells blocks migration of border cells and centripetal cells on the surface of germline cells. The function of DE-cadherin in border cells appears to be specific for migration as the formation of the border cell cluster and the adhesion between border cells are not disrupted in the absence of DE-cadherin. The speed of migration depends on the level of DE-cadherin expression, as border cells migrate more slowly when DE-cadherin activity is reduced. Finally, we show that the upregulation of DE-cadherin expression in border cells depends on the activity of the Drosophila C/EBP transcription factor that is essential for border cell migration. PMID:9971747

  16. Nonuniform Effect of Carrier Separation Efficiency and Light Absorption in Type-II Perovskite Nanowire Solar Cells

    NASA Astrophysics Data System (ADS)

    Wang, Weiping; He, Jialun; Cao, Yiyan; Kong, Lijing; Zheng, Xuanli; Wu, Yaping; Chen, Xiaohong; Li, Shuping; Wu, Zhiming; Kang, Junyong

    2017-03-01

    Coaxial structures exhibit great potential for the application of high-efficiency solar cells due to the novel mechanism of radial charge separation. Here, we intensively investigate the nonuniform effect of carrier separation efficiency (CSE) and light absorption in perovskite-based type-II coaxial nanowire solar cells (ZnO/CH3NH3PbI3). Results show that the CSE rapidly decreases along the radial direction in the shell, and the value at the outer side becomes extremely low for the thick shell. Besides, the position of the main light absorption gradually moves to the outer side with the increase of the shell thickness. As a result, the external quantum efficiency shows a positional dependence with a maximal value close to the border of the nanowire. Eventually, in our case, it is found that the maximal power conversion efficiency of the solar cells reduces from 19.5 to 17.9% under the effect of the nonuniformity of CSE and light absorption. This work provides a basis for the design of high-efficiency solar cells, especially type-II nanowire solar cells.

  17. Nonuniform Effect of Carrier Separation Efficiency and Light Absorption in Type-II Perovskite Nanowire Solar Cells.

    PubMed

    Wang, Weiping; He, Jialun; Cao, Yiyan; Kong, Lijing; Zheng, Xuanli; Wu, Yaping; Chen, Xiaohong; Li, Shuping; Wu, Zhiming; Kang, Junyong

    2017-12-01

    Coaxial structures exhibit great potential for the application of high-efficiency solar cells due to the novel mechanism of radial charge separation. Here, we intensively investigate the nonuniform effect of carrier separation efficiency (CSE) and light absorption in perovskite-based type-II coaxial nanowire solar cells (ZnO/CH 3 NH 3 PbI 3 ). Results show that the CSE rapidly decreases along the radial direction in the shell, and the value at the outer side becomes extremely low for the thick shell. Besides, the position of the main light absorption gradually moves to the outer side with the increase of the shell thickness. As a result, the external quantum efficiency shows a positional dependence with a maximal value close to the border of the nanowire. Eventually, in our case, it is found that the maximal power conversion efficiency of the solar cells reduces from 19.5 to 17.9% under the effect of the nonuniformity of CSE and light absorption. This work provides a basis for the design of high-efficiency solar cells, especially type-II nanowire solar cells.

  18. Border Lakes land-cover classification

    Treesearch

    Marvin Bauer; Brian Loeffelholz; Doug Shinneman

    2009-01-01

    This document contains metadata and description of land-cover classification of approximately 5.1 million acres of land bordering Minnesota, U.S.A. and Ontario, Canada. The classification focused on the separation and identification of specific forest-cover types. Some separation of the nonforest classes also was performed. The classification was derived from multi-...

  19. Integrated Metabolomics and Transcriptomics Reveal Enhanced Specialized Metabolism in Medicago truncatula Root Border Cells1[OPEN

    PubMed Central

    Watson, Bonnie S.; Bedair, Mohamed F.; Urbanczyk-Wochniak, Ewa; Huhman, David V.; Yang, Dong Sik; Allen, Stacy N.; Li, Wensheng; Tang, Yuhong; Sumner, Lloyd W.

    2015-01-01

    Integrated metabolomics and transcriptomics of Medicago truncatula seedling border cells and root tips revealed substantial metabolic differences between these distinct and spatially segregated root regions. Large differential increases in oxylipin-pathway lipoxygenases and auxin-responsive transcript levels in border cells corresponded to differences in phytohormone and volatile levels compared with adjacent root tips. Morphological examinations of border cells revealed the presence of significant starch deposits that serve as critical energy and carbon reserves, as documented through increased β-amylase transcript levels and associated starch hydrolysis metabolites. A substantial proportion of primary metabolism transcripts were decreased in border cells, while many flavonoid- and triterpenoid-related metabolite and transcript levels were increased dramatically. The cumulative data provide compounding evidence that primary and secondary metabolism are differentially programmed in border cells relative to root tips. Metabolic resources normally destined for growth and development are redirected toward elevated accumulation of specialized metabolites in border cells, resulting in constitutively elevated defense and signaling compounds needed to protect the delicate root cap and signal motile rhizobia required for symbiotic nitrogen fixation. Elevated levels of 7,4′-dihydroxyflavone were further increased in border cells of roots exposed to cotton root rot (Phymatotrichopsis omnivora), and the value of 7,4′-dihydroxyflavone as an antimicrobial compound was demonstrated using in vitro growth inhibition assays. The cumulative and pathway-specific data provide key insights into the metabolic programming of border cells that strongly implicate a more prominent mechanistic role for border cells in plant-microbe signaling, defense, and interactions than envisioned previously. PMID:25667316

  20. Combined quantitative and qualitative two-channel optical biopsy technique for discrimination of tumor borders

    NASA Astrophysics Data System (ADS)

    Bocher, Thomas; Beuthan, Juergen; Scheller, M.; Hopf, Juergen U. G.; Linnarz, Marietta; Naber, Rolf-Dieter; Minet, Olaf; Becker, Wolfgang; Mueller, Gerhard J.

    1995-12-01

    Conventional laser-induced fluorescence spectroscopy (LIFS) of endogenous chromophores like NADH (Nicotineamide Adenine Dinucleotide, reduced form) and PP IX (Protoporphyrin IX) provides information about the relative amounts of these metabolites in the observed cells. But for diagnostic applications the concentrations of these chromophores have to be determined quantitatively to establish tissue-independent differentiation criterions. It is well- known that the individually and locally varying optical tissue parameters are major obstacles for the determination of the true chromophore concentrations by simple fluorescence spectroscopy. To overcome these problems a fiber-based, 2-channel technique including a rescaled NADH-channel (delivering quantitative values) and a relative PP IX-channel was developed. Using the accumulated information of both channels can provide good tissue state separation. Ex-vivo studies with resected and frozen samples (with LN2) of squamous cells in the histologically confirmed states: normal, tumor border, inflammation and hyperplasia were performed. Each state was represented in this series with at least 7 samples. At the identical tissue spot both, the rescaled NADH-fluorescence and the relative PP IX- fluorescence, were determined. In the first case a nitrogen laser (337 nm, 500 ps, 200 microjoule, 10 Hz) in the latter case a diode laser (633 nm, 15 mW, cw) were used as excitation sources. In this ex-vivo study a good separation between the different tissue states was achieved. With a device constructed for clinical usage one quantitative, in-vivo NADH- measurement was done recently showing similar separation capabilities.

  1. Cytopathologic differential diagnosis of low-grade urothelial carcinoma and reactive urothelial proliferation in bladder washings: a logistic regression analysis.

    PubMed

    Cakir, Ebru; Kucuk, Ulku; Pala, Emel Ebru; Sezer, Ozlem; Ekin, Rahmi Gokhan; Cakmak, Ozgur

    2017-05-01

    Conventional cytomorphologic assessment is the first step to establish an accurate diagnosis in urinary cytology. In cytologic preparations, the separation of low-grade urothelial carcinoma (LGUC) from reactive urothelial proliferation (RUP) can be exceedingly difficult. The bladder washing cytologies of 32 LGUC and 29 RUP were reviewed. The cytologic slides were examined for the presence or absence of the 28 cytologic features. The cytologic criteria showing statistical significance in LGUC were increased numbers of monotonous single (non-umbrella) cells, three-dimensional cellular papillary clusters without fibrovascular cores, irregular bordered clusters, atypical single cells, irregular nuclear overlap, cytoplasmic homogeneity, increased N/C ratio, pleomorphism, nuclear border irregularity, nuclear eccentricity, elongated nuclei, and hyperchromasia (p ˂ 0.05), and the cytologic criteria showing statistical significance in RUP were inflammatory background, mixture of small and large urothelial cells, loose monolayer aggregates, and vacuolated cytoplasm (p ˂ 0.05). When these variables were subjected to a stepwise logistic regression analysis, four features were selected to distinguish LGUC from RUP: increased numbers of monotonous single (non-umbrella) cells, increased nuclear cytoplasmic ratio, hyperchromasia, and presence of small and large urothelial cells (p = 0.0001). By this logistic model of the 32 cases with proven LGUC, the stepwise logistic regression analysis correctly predicted 31 (96.9%) patients with this diagnosis, and of the 29 patients with RUP, the logistic model correctly predicted 26 (89.7%) patients as having this disease. There are several cytologic features to separate LGUC from RUP. Stepwise logistic regression analysis is a valuable tool for determining the most useful cytologic criteria to distinguish these entities. © 2017 APMIS. Published by John Wiley & Sons Ltd.

  2. Systematic Analysis of the Transcriptional Switch Inducing Migration of Border Cells

    PubMed Central

    Borghese, Lodovica; Fletcher, Georgina; Mathieu, Juliette; Atzberger, Ann; Eades, William C.; Cagan, Ross L.; Rørth, Pernille

    2010-01-01

    Summary Cell migration within a natural context is tightly controlled, often by specific transcription factors. However, the switch from stationary to migratory behavior is poorly understood. Border cells perform a spatially and temporally controlled invasive migration during Drosophila oogenesis. Slbo, a C/EBP family transcriptional activator, is required for them to become migratory. We purified wild-type and slbo mutant border cells as well as nonmigratory follicle cells and performed comparative whole-genome expression profiling, followed by functional tests of the contributions of identified targets to migration. About 300 genes were significantly upregulated in border cells, many dependent on Slbo. Among these, the microtubule regulator Stathmin was strongly upregulated and was required for normal migration. Actin cytoskeleton regulators were also induced, including, surprisingly, a large cluster of “muscle-specific” genes. We conclude that Slbo induces multiple cytoskeletal effectors, and that each contributes to the behavioral changes in border cells. PMID:16580994

  3. Relating proton pumps with gap junctions: colocalization of ductin, the channel-forming subunit c of V-ATPase, with subunit a and with innexins 2 and 3 during Drosophila oogenesis.

    PubMed

    Lautemann, Julia; Bohrmann, Johannes

    2016-07-13

    Ion-transport mechanisms and gap junctions are known to cooperate in creating bioelectric phenomena, like pH gradients, voltage gradients and ion fluxes within single cells, tissues, organs, and whole organisms. Such phenomena have been shown to play regulatory roles in a variety of developmental and regenerative processes. Using Drosophila oogenesis as a model system, we aim at characterizing in detail the mechanisms underlying bioelectric phenomena in order to reveal their regulatory functions. We, therefore, investigated the stage-specific distribution patterns of V-ATPase components in relation to gap-junction proteins. We analysed the localization of the V-ATPase components ductin (subunit c) and subunit a, and the gap-junction components innexins 2 and 3, especially in polar cells, border cells, stalk cells and centripetally migrating cells. These types of follicle cells had previously been shown to exhibit characteristic patterns of membrane channels as well as membrane potential and intracellular pH. Stage-specifically, ductin and subunit a were found either colocalized or separately enriched in different regions of soma and germ-line cells. While ductin was often more prominent in plasma membranes, subunit a was more prominent in cytoplasmic and nuclear vesicles. Particularly, ductin was enriched in polar cells, stalk cells, and nurse-cell membranes, whereas subunit a was enriched in the cytoplasm of border cells, columnar follicle cells and germ-line cells. Comparably, ductin and both innexins 2 and 3 were either colocalized or separately enriched in different cellular regions. While ductin often showed a continuous membrane distribution, the distribution of both innexins was mostly punctate. Particularly, ductin was enriched in polar cells and stalk cells, whereas innexin 2 was enriched in the oolemma, and innexin 3 in centripetally migrating follicle cells. In lateral follicle-cell membranes, the three proteins were found colocalized as well as separately concentrated in presumed gap-junction plaques. Our results support the notion of a large variety of gap junctions existing in the Drosophila ovary. Moreover, since ductin is the channel-forming part of a proton pump and, like the innexins, is able to form junctional as well as non-junctional membrane channels, a plethora of cellular functions could be realized by using these proteins. The distribution and activity patterns of such membrane channels are expected to contribute to developmentally important bioelectric signals.

  4. Cortical Dynamics of Figure-Ground Separation in Response to 2D Pictures and 3D Scenes: How V2 Combines Border Ownership, Stereoscopic Cues, and Gestalt Grouping Rules

    PubMed Central

    Grossberg, Stephen

    2016-01-01

    The FACADE model, and its laminar cortical realization and extension in the 3D LAMINART model, have explained, simulated, and predicted many perceptual and neurobiological data about how the visual cortex carries out 3D vision and figure-ground perception, and how these cortical mechanisms enable 2D pictures to generate 3D percepts of occluding and occluded objects. In particular, these models have proposed how border ownership occurs, but have not yet explicitly explained the correlation between multiple properties of border ownership neurons in cortical area V2 that were reported in a remarkable series of neurophysiological experiments by von der Heydt and his colleagues; namely, border ownership, contrast preference, binocular stereoscopic information, selectivity for side-of-figure, Gestalt rules, and strength of attentional modulation, as well as the time course during which such properties arise. This article shows how, by combining 3D LAMINART properties that were discovered in two parallel streams of research, a unified explanation of these properties emerges. This explanation proposes, moreover, how these properties contribute to the generation of consciously seen 3D surfaces. The first research stream models how processes like 3D boundary grouping and surface filling-in interact in multiple stages within and between the V1 interblob—V2 interstripe—V4 cortical stream and the V1 blob—V2 thin stripe—V4 cortical stream, respectively. Of particular importance for understanding figure-ground separation is how these cortical interactions convert computationally complementary boundary and surface mechanisms into a consistent conscious percept, including the critical use of surface contour feedback signals from surface representations in V2 thin stripes to boundary representations in V2 interstripes. Remarkably, key figure-ground properties emerge from these feedback interactions. The second research stream shows how cells that compute absolute disparity in cortical area V1 are transformed into cells that compute relative disparity in cortical area V2. Relative disparity is a more invariant measure of an object's depth and 3D shape, and is sensitive to figure-ground properties. PMID:26858665

  5. Cortical Dynamics of Figure-Ground Separation in Response to 2D Pictures and 3D Scenes: How V2 Combines Border Ownership, Stereoscopic Cues, and Gestalt Grouping Rules.

    PubMed

    Grossberg, Stephen

    2015-01-01

    The FACADE model, and its laminar cortical realization and extension in the 3D LAMINART model, have explained, simulated, and predicted many perceptual and neurobiological data about how the visual cortex carries out 3D vision and figure-ground perception, and how these cortical mechanisms enable 2D pictures to generate 3D percepts of occluding and occluded objects. In particular, these models have proposed how border ownership occurs, but have not yet explicitly explained the correlation between multiple properties of border ownership neurons in cortical area V2 that were reported in a remarkable series of neurophysiological experiments by von der Heydt and his colleagues; namely, border ownership, contrast preference, binocular stereoscopic information, selectivity for side-of-figure, Gestalt rules, and strength of attentional modulation, as well as the time course during which such properties arise. This article shows how, by combining 3D LAMINART properties that were discovered in two parallel streams of research, a unified explanation of these properties emerges. This explanation proposes, moreover, how these properties contribute to the generation of consciously seen 3D surfaces. The first research stream models how processes like 3D boundary grouping and surface filling-in interact in multiple stages within and between the V1 interblob-V2 interstripe-V4 cortical stream and the V1 blob-V2 thin stripe-V4 cortical stream, respectively. Of particular importance for understanding figure-ground separation is how these cortical interactions convert computationally complementary boundary and surface mechanisms into a consistent conscious percept, including the critical use of surface contour feedback signals from surface representations in V2 thin stripes to boundary representations in V2 interstripes. Remarkably, key figure-ground properties emerge from these feedback interactions. The second research stream shows how cells that compute absolute disparity in cortical area V1 are transformed into cells that compute relative disparity in cortical area V2. Relative disparity is a more invariant measure of an object's depth and 3D shape, and is sensitive to figure-ground properties.

  6. Cell Geometry Guides the Dynamic Targeting of Apoplastic GPI-Linked Lipid Transfer Protein to Cell Wall Elements and Cell Borders in Arabidopsis thaliana

    PubMed Central

    Wasteneys, Geoffrey

    2013-01-01

    During cellular morphogenesis, changes in cell shape and cell junction topology are fundamental to normal tissue and organ development. Here we show that apoplastic Glycophosphatidylinositol (GPI)-anchored Lipid Transfer Protein (LTPG) is excluded from cell junctions and flat wall regions, and passively accumulates around their borders in the epidermal cells of Arabidopsis thaliana. Beginning with intense accumulation beneath highly curved cell junction borders, this enrichment is gradually lost as cells become more bulbous during their differentiation. In fully mature epidermal cells, YFP-LTPG often shows a fibrous cellulose microfibril-like pattern within the bulging outer faces. Physical contact between a flat glass surface and bulbous cell surface induces rapid and reversible evacuation from contact sites and accumulation to the curved wall regions surrounding the contact borders. Thus, LTPG distribution is dynamic, responding to changes in cell shape and wall curvature during cell growth and differentiation. We hypothesize that this geometry-based mechanism guides wax-carrying LTPG to functional sites, where it may act to “seal” the vulnerable border surrounding cell-cell junctions and assist in cell wall fortification and cuticular wax deposition. PMID:24260561

  7. Green Belt Europe - borders separate, nature unites

    Treesearch

    Uwe Friedel

    2015-01-01

    During the period of the Cold War between 1945 and 1989, a "Green Belt" of valuable pristine landscapes developed along the border line between Eastern and Western Europe, the intensively fortified and guarded so called Iron Curtain. Due to the remoteness of the border areas, a high number of national parks and other large conservation areas can be found...

  8. Dimerization and actin-bundling properties of villin and its role in the assembly of epithelial cell brush borders.

    PubMed

    George, Sudeep P; Wang, Yaohong; Mathew, Sijo; Srinivasan, Kamalakkannan; Khurana, Seema

    2007-09-07

    Villin is a major actin-bundling protein in the brush border of epithelial cells. In this study we demonstrate for the first time that villin can bundle actin filaments using a single F-actin binding site, because it has the ability to self-associate. Using fluorescence resonance energy transfer, we demonstrate villin self-association in living cells in microvilli and in growth factor-stimulated cells in membrane ruffles and lamellipodia. Using sucrose density gradient, size-exclusion chromatography, and matrix-assisted laser desorption ionization time-of-flight, the majority of villin was identified as a monomer or dimer. Villin dimers were also identified in Caco-2 cells, which endogenously express villin and Madin-Darby canine kidney cells that ectopically express villin. Using truncation mutants of villin, site-directed mutagenesis, and fluorescence resonance energy transfer, an amino-terminal dimerization site was identified that regulated villin self-association in parallel conformation as well as actin bundling by villin. This detailed analysis describes for the first time microvillus assembly by villin, redefines the actin-bundling function of villin, and provides a molecular mechanism for actin bundling by villin, which could have wider implications for other actin cross-linking proteins that share a villin-like headpiece domain. Our study also provides a molecular basis to separate the morphologically distinct actin-severing and actin-bundling properties of villin.

  9. P-selectin mediates neutrophil adhesion to endothelial cell borders.

    PubMed

    Burns, A R; Bowden, R A; Abe, Y; Walker, D C; Simon, S I; Entman, M L; Smith, C W

    1999-03-01

    During an acute inflammatory response, endothelial P-selectin (CD62P) can mediate the initial capture of neutrophils from the free flowing bloodstream. P-selectin is stored in secretory granules (Weibel-Palade bodies) and is rapidly expressed on the endothelial surface after stimulation with histamine or thrombin. Because neutrophil transmigration occurs preferentially at endothelial borders, we wished to determine whether P-selectin-dependent neutrophil capture (adhesion) occurs at endothelial cell borders. Under static or hydrodynamic flow (2 dyn/cm2) conditions, histamine (10(-4) M) or thrombin (0.2 U/mL) treatment induced preferential (> or = 75%) neutrophil adhesion to the cell borders of endothelial monolayers. Blocking antibody studies established that neutrophil adhesion was completely P-selectin dependent. P-selectin surface expression increased significantly after histamine treatment and P-selectin immunostaining was concentrated along endothelial borders. We conclude that preferential P-selectin expression along endothelial borders may be an important mechanism for targeting neutrophil migration at endothelial borders.

  10. Border patrol: insights into the unique role of perlecan/heparan sulfate proteoglycan 2 at cell and tissue borders.

    PubMed

    Farach-Carson, Mary C; Warren, Curtis R; Harrington, Daniel A; Carson, Daniel D

    2014-02-01

    The extracellular matrix proteoglycan (ECM) perlecan, also known as heparan sulfate proteoglycan 2 or HSPG2, is one of the largest (>200 nm) and oldest (>550 M years) extracellular matrix molecules. In vertebrates, perlecan's five-domain structure contains numerous independently folding modules with sequence similarities to other ECM proteins, all connected like cars into one long, diverse complex train following a unique N-terminal domain I decorated with three long glycosaminoglycan chains, and an additional glycosaminoglycan attachment site in the C-terminal domain V. In lower invertebrates, perlecan is not typically a proteoglycan, possessing the majority of the core protein modules, but lacking domain I where the attachment sites for glycosaminoglycan chains are located. This suggests that uniting the heparan sulfate binding growth factor functions of domain I and the core protein functions of the rest of the molecule in domains II-V occurred later in evolution for a new functional purpose. In this review, we surveyed several decades of pertinent literature to ask a fundamental question: Why did nature design this protein uniquely as an extraordinarily long multifunctional proteoglycan with a single promoter regulating expression, rather than separating these functions into individual proteins that could be independently regulated? We arrived at the conclusion that the concentration of perlecan at functional borders separating tissues and tissue layers is an ancient key function of the core protein. The addition of the heparan sulfate chains in domain I likely occurred as an additional means of binding the core protein to other ECM proteins in territorial matrices and basement membranes, and as a means to reserve growth factors in an on-site depot to assist with rapid repair of those borders when compromised, such as would occur during wounding. We propose a function for perlecan that extends its role from that of an extracellular scaffold, as we previously suggested, to that of a critical agent for establishing and patrolling tissue borders in complex tissues in metazoans. We also propose that understanding these unique functions of the individual portions of the perlecan molecule can provide new insights and tools for engineering of complex multi-layered tissues including providing the necessary cues for establishing neotissue borders. © 2013.

  11. Border Patrol: Insights into the Unique Role of Perlecan/Heparan Sulfate Proteoglycan2 at Cell and Tissue Borders

    PubMed Central

    Farach-Carson, Mary C.; Warren, Curtis R.; Harrington, Daniel A.; Carson, Daniel D.

    2013-01-01

    The extracellular matrix proteoglycan (ECM) perlecan, also known as heparan sulfate proteoglycan 2 or HSPG2, is one of the largest (>200 nm) and oldest (>550M years) extracellular matrix molecules. In vertebrates, perlecan’s five-domain structure contains numerous independently folding modules with sequence similarities to other ECM proteins, all connected like cars into one long, diverse complex train following a unique N-terminal domain I decorated with three long glycosaminoglycan chains, and an additional glycosaminoglycan attachment site in the C-terminal domain V. In lower invertebrates, perlecan is not typically a proteoglycan, possessing the majority of the core protein modules, but lacking domain I where the attachment sites for glycosaminoglycan chains are located. This suggests that uniting the heparan sulfate binding growth factor functions of domain I and the core protein functions of the rest of the molecule in domains II-V occurred later in evolution for a new functional purpose. In this review, we surveyed several decades of pertinent literature to ask a fundamental question: Why did nature design this protein uniquely as an extraordinarily long multifunctional proteoglycan with a single promoter regulating expression, rather than separating these functions into individual proteins that could be independently regulated? We arrived at the conclusion that the concentration of perlecan at functional borders separating tissues and tissue layers is an ancient key function of the core protein. The addition of the heparan sulfate chains in domain I likely occurred as an additional means of binding the core protein to other ECM proteins in territorial matrices and basement membranes, and as a means to reserve growth factors in an on-site depot to assist with rapid repair of those borders when compromised, such as would occur during wounding. We propose a function for perlecan that extends its role from that of an extracellular scaffold, as we previously suggested, to that of a critical agent for establishing and patrolling tissue borders in complex tissues in metazoans. We also propose that understanding these unique functions of the individual portions of the perlecan molecule can provide new insights and tools for engineering of complex multi-layered tissues including providing the necessary cues for establishing neotissue borders. PMID:24001398

  12. Teaching "Sin Fronteras" (Without Borders)

    ERIC Educational Resources Information Center

    Diaz, Marisol; Baptiste, H. Prentice

    2017-01-01

    By breaking down "fronteras" (borders) in classroom instruction, educators can interrupt and examine the interactions and discourses that create separations and alienations among people. The authors describe an effective instructional approach used in a second-grade class and modified for a class of preservice teachers.

  13. Group choreography: mechanisms orchestrating the collective movement of border cells

    PubMed Central

    Montell, Denise J.; Yoon, Wan Hee; Starz-Gaiano, Michelle

    2014-01-01

    Cell movements are essential for animal development and homeostasis but also contribute to disease. Moving cells typically extend protrusions towards a chemoattractant, adhere to the substrate, contract and detach at the rear. It is less clear how cells that migrate in interconnected groups in vivo coordinate their behaviour and navigate through natural environments. The border cells of the Drosophila melanogaster ovary have emerged as an excellent model for the study of collective cell movement, aided by innovative genetic, live imaging, and photomanipulation techniques. Here we provide an overview of the molecular choreography of border cells and its more general implications. PMID:23000794

  14. Development of Cell Analysis Software for Cultivated Corneal Endothelial Cells.

    PubMed

    Okumura, Naoki; Ishida, Naoya; Kakutani, Kazuya; Hongo, Akane; Hiwa, Satoru; Hiroyasu, Tomoyuki; Koizumi, Noriko

    2017-11-01

    To develop analysis software for cultured human corneal endothelial cells (HCECs). Software was designed to recognize cell borders and to provide parameters such as cell density, coefficient of variation, and polygonality of cultured HCECs based on phase contrast images. Cultured HCECs with high or low cell density were incubated with Ca-free and Mg-free phosphate-buffered saline for 10 minutes to reveal the cell borders and were then analyzed with software (n = 50). Phase contrast images showed that cell borders were not distinctly outlined, but these borders became more distinctly outlined after phosphate-buffered saline treatment and were recognized by cell analysis software. The cell density value provided by software was similar to that obtained using manual cell counting by an experienced researcher. Morphometric parameters, such as the coefficient of variation and polygonality, were also produced by software, and these values were significantly correlated with cell density (Pearson correlation coefficients -0.62 and 0.63, respectively). The software described here provides morphometric information from phase contrast images, and it enables subjective and noninvasive quality assessment for tissue engineering therapy of the corneal endothelium.

  15. Novel rapid genotyping assays for neuronal ceroid lipofuscinosis in Border Collie dogs and high frequency of the mutant allele in Japan.

    PubMed

    Mizukami, Keijiro; Chang, Hye-Sook; Yabuki, Akira; Kawamichi, Takuji; Kawahara, Natsuko; Hayashi, Daisuke; Hossain, Mohammad A; Rahman, Mohammad M; Uddin, Mohammad M; Yamato, Osamu

    2011-11-01

    Neuronal ceroid lipofuscinosis (NCL) constitutes a group of recessively inherited lysosomal storage diseases that primarily affect neuronal cells. Such diseases share certain clinical and pathologic features in human beings and animals. Neuronal ceroid lipofuscinosis in Border Collie dogs was first detected in Australia in the 1980s, and the pathogenic mutation was shown to be a nonsense mutation (c.619C>T) in exon 4 in canine CLN5 gene. In the present study, novel rapid genotyping assays including polymerase chain reaction (PCR)-restriction fragment length polymorphism, PCR primer-induced restriction analysis, mutagenically separated PCR, and real-time PCR with TaqMan minor groove binder probes, were developed. The utility of microchip electrophoresis was also evaluated. Furthermore, a genotyping survey was carried out in a population of Border Collies in Japan using these assays to determine the current allele frequency in Japan, providing information to control and prevent this disease in the next stage. All assays developed in the current study are available to discriminate these genotypes, and microchip electrophoresis showed a timesaving advantage over agarose gel electrophoresis. Of all assays, real-time PCR was the most suitable for large-scale examination because of its high throughput. The genotyping survey demonstrated that the carrier frequency was 8.1%. This finding suggested that the mutant allele frequency of NCL in Border Collies is high enough in Japan that measures to control and prevent the disease would be warranted. The genotyping assays developed in the present study could contribute to the prevention of NCL in Border Collies.

  16. Semi-automated discrimination of retinal pigmented epithelial cells in two-photon fluorescence images of mouse retinas.

    PubMed

    Alexander, Nathan S; Palczewska, Grazyna; Palczewski, Krzysztof

    2015-08-01

    Automated image segmentation is a critical step toward achieving a quantitative evaluation of disease states with imaging techniques. Two-photon fluorescence microscopy (TPM) has been employed to visualize the retinal pigmented epithelium (RPE) and provide images indicating the health of the retina. However, segmentation of RPE cells within TPM images is difficult due to small differences in fluorescence intensity between cell borders and cell bodies. Here we present a semi-automated method for segmenting RPE cells that relies upon multiple weak features that differentiate cell borders from the remaining image. These features were scored by a search optimization procedure that built up the cell border in segments around a nucleus of interest. With six images used as a test, our method correctly identified cell borders for 69% of nuclei on average. Performance was strongly dependent upon increasing retinosome content in the RPE. TPM image analysis has the potential of providing improved early quantitative assessments of diseases affecting the RPE.

  17. Production of non viral DNA vectors.

    PubMed

    Schleef, Martin; Blaesen, Markus; Schmeer, Marco; Baier, Ruth; Marie, Corinne; Dickson, George; Scherman, Daniel

    2010-12-01

    After some decades of research, development and first clinical approaches to use DNA vectors in gene therapy, cell therapy and DNA vaccination, the requirements for the pharmaceutical manufacturing of gene vectors has improved significantly step by step. Even the expression level and specificity of non viral DNA vectors were significantly modified and followed the success of viral vectors. The strict separation of "viral" and "non viral" gene transfer are historic borders between scientist and we will show that both fields together are able to allow the next step towards successful prevention and therapy. Here we summarize the features of producing and modifying these non-viral gene vectors to ensure the required quality to modify cells and to treat human and animals.

  18. Cross border hospital use: analysis using data linkage across four Australian states.

    PubMed

    Spilsbury, Katrina; Rosman, Diana; Alan, Janine; Boyd, James H; Ferrante, Anna M; Semmens, James B

    2015-06-15

    To determine the quality and effectiveness of national data linkage capacity by performing a proof-of-concept project investigating cross-border hospital use and hospital-related deaths. Analysis of person-level linked hospital separation and death registration data of all public and private hospital patients in New South Wales, Queensland and Western Australia and of public hospital patients in South Australia, totalling 7.7 million hospital patients from 1 July 2004 to 30 June 2009. Counts and proportions of hospital stays and patient movement patterns. 223 262 patients (3.0%) travelled across a state border to attend hospitals, in particular, far northern and western NSW patients travelling to Queensland and SA hospitals, respectively. A further 48 575 patients (0.6%) moved their place of residence interstate between hospital visits, particularly to and from areas associated with major mining and tourism industries. Over 11 000 cross-border hospital transfers were also identified. Of patients who travelled across a state border to hospital, 2800 (1.3%) died in that hospital. An additional 496 deaths recorded in one jurisdiction occurred within 30 days of hospital separation from another jurisdiction. Access to person-level data linked across jurisdictions identified geographical hot spots of cross-border hospital use and hospital-related deaths in Australia. This has implications for planning of health service delivery and for longitudinal follow-up studies, particularly those involving mobile populations.

  19. A novel role for integrin-linked kinase in epithelial sheet morphogenesis.

    PubMed

    Vespa, Alisa; D'Souza, Sudhir J A; Dagnino, Lina

    2005-09-01

    Integrin-linked kinase (ILK) is a multidomain protein involved in cell motility and cell-extracellular matrix interactions. ILK is found in integrin-containing focal adhesions in undifferentiated primary epidermal keratinocytes. Induction of keratinocyte differentiation by treatment with Ca(2+) triggers formation of cell-cell junctions, loss of focal adhesions, and ILK distribution to cell borders. We now show that Ca(2+) treatment of keratinocytes induces rapid (6 h) localization of tight junction (TJ) proteins. The kinetics of ILK movement toward the cell periphery mimics that of AJ components, suggesting that ILK plays a role in the early formation of cell-cell contacts. Whereas the N terminus in ILK mediates localization to cell borders, expression of an ILK deletion mutant incapable of localizing to the cell membrane (ILK 191-452) interferes with translocation of E-cadherin/beta-catenin to cell borders, precluding Ca(2+)-induced AJ formation. Cells expressing ILK 191-452 also fail to form TJ and sealed cell-cell borders and do not form epithelial sheets. Thus, we have uncovered a novel role for ILK in epithelial cell-cell adhesion, independent of its well-established role in integrin-mediated adhesion and migration.

  20. Dynamic transcriptional signature and cell fate analysis reveals plasticity of individual neural plate border cells

    PubMed Central

    Roellig, Daniela; Tan-Cabugao, Johanna; Esaian, Sevan; Bronner, Marianne E

    2017-01-01

    The ‘neural plate border’ of vertebrate embryos contains precursors of neural crest and placode cells, both defining vertebrate characteristics. How these lineages segregate from neural and epidermal fates has been a matter of debate. We address this by performing a fine-scale quantitative temporal analysis of transcription factor expression in the neural plate border of chick embryos. The results reveal significant overlap of transcription factors characteristic of multiple lineages in individual border cells from gastrula through neurula stages. Cell fate analysis using a Sox2 (neural) enhancer reveals that cells that are initially Sox2+ cells can contribute not only to neural tube but also to neural crest and epidermis. Moreover, modulating levels of Sox2 or Pax7 alters the apportionment of neural tube versus neural crest fates. Our results resolve a long-standing question and suggest that many individual border cells maintain ability to contribute to multiple ectodermal lineages until or beyond neural tube closure. DOI: http://dx.doi.org/10.7554/eLife.21620.001 PMID:28355135

  1. Implantation of a cone-shaped double-fixed patch increases abdominal space and prevents recurrence of large defects in congenital diaphragmatic hernia.

    PubMed

    Loff, Steffan; Wirth, Hartmut; Jester, Iwgo; Hosie, Stuart; Wollmann, Carmen; Schaible, Thomas; Ataman, Ozge; Waag, Karl-Ludwig

    2005-11-01

    Large defects in patients with congenital diaphragmatic hernia (CDH) are frequently closed with a polytetrafluoroethylene patch (PTFE). Intraoperative problems include lack of abdominal domain for the reduction of organs and closure of the abdominal wall. Main surgical postoperative complication is the recurrence of the hernia. We suggest a new and easy method of patch implantation, improving these problems, and report first follow-up results. In our clinic, 103 children with CDH were treated, and 87 children underwent reconstruction of the diaphragm in the 5 years between 1998 and 2002. In 52 patients, a patch implantation had to be performed. We have been optimizing our complete pediatric and surgical procedure and present a new standardized technique of preparation and implantation of a PTFE patch. The flat patch is folded to a 90 degrees cone. The cone is fixed in its form with few single stitches. It is implanted with an overlapping border of 1 cm circumferentially. The border is separately fixed with absorbable single stitches to keep from rolling up. The rough side of the patch points toward the rim of the diaphragm to enable ingrowth of the connective tissue. In a 1-year follow-up study, the recurrences in the 3 following groups of PTFE patches were studied: conventional implantation (simple patch without overlapping border), patch with separately fixed overlapping border, and cone-shaped patch with overlapping separately fixed border. Thirty-three patients were included in the study. After conventional PTFE-patch implantation, 6 (46%) of 13 patients developed reherniation. After PTFE-patch implantation with separately fixed overlapping border, 1 (11%) of 9 patients had a recurrent hernia. In the group with the PTFE-cone implantation, 1 (9%) of 11 patients developed a recurrence. Meanwhile, another 20 CDH patients received implantation of a cone-shaped patch, and no further recurrence occurred up to now. With the additional space (20 mL) provided by the cone-shaped patch, the closure of the abdomen was easier, and the fundus had intraoperatively a physiological position. This optimized patch implantation technique in large diaphragmatic defects offers considerable advantages especially regarding recurrence of the hernia and closure of the abdomen, which are currently the most challenging surgical problems. 1. The cone-shaped 3-dimensional patch increases abdominal capacity. 2. Redundant chest capacity is reduced, and the reconstructed diaphragm shows a physiological shape. 3. The dome of the patch allows a physiological position of the gastric fundus and a normal Hiss angle, thus preventing gastroesophageal reflux. 4. Additional safety of the implantation is achieved by separate fixation of the overlapping border of the cone, preventing recurrence.

  2. Reconsidering Privatization in Cross-Border Engagements: The Sometimes Public Nature of Private Activity

    ERIC Educational Resources Information Center

    Lane, Jason E.; Kinser, Kevin

    2011-01-01

    Privatization trends in higher education have typically been analyzed from the perspective of the institution and its relationship with the sponsoring state. The recent phenomenon of international cross-border higher education, however, represents a more complicated picture of privatization. Geographic separation from the sponsoring state is an…

  3. A Novel Role for Integrin-linked Kinase in Epithelial Sheet Morphogenesis

    PubMed Central

    Vespa, Alisa; D'Souza, Sudhir J.A.; Dagnino, Lina

    2005-01-01

    Integrin-linked kinase (ILK) is a multidomain protein involved in cell motility and cell-extracellular matrix interactions. ILK is found in integrin-containing focal adhesions in undifferentiated primary epidermal keratinocytes. Induction of keratinocyte differentiation by treatment with Ca2+ triggers formation of cell–cell junctions, loss of focal adhesions, and ILK distribution to cell borders. We now show that Ca2+ treatment of keratinocytes induces rapid (≤1 h) translocation to the cell membrane of the adherens junction (AJ) proteins E-cadherin and β-catenin. This is followed by slower (>6 h) localization of tight junction (TJ) proteins. The kinetics of ILK movement toward the cell periphery mimics that of AJ components, suggesting that ILK plays a role in the early formation of cell–cell contacts. Whereas the N terminus in ILK mediates localization to cell borders, expression of an ILK deletion mutant incapable of localizing to the cell membrane (ILK 191-452) interferes with translocation of E-cadherin/β-catenin to cell borders, precluding Ca2+-induced AJ formation. Cells expressing ILK 191-452 also fail to form TJ and sealed cell–cell borders and do not form epithelial sheets. Thus, we have uncovered a novel role for ILK in epithelial cell–cell adhesion, independent of its well-established role in integrin-mediated adhesion and migration. PMID:15975904

  4. Neuroendocrine carcinoma of the mammary gland in a dog.

    PubMed

    Nakahira, R; Michishita, M; Yoshimura, H; Hatakeyama, H; Takahashi, K

    2015-01-01

    A 10-year-old female border collie was presented with a mass (2 cm diameter) in the fifth mammary gland. The mass was located in the subcutis and the cut surface was grey-white in colour. Microscopically, the mass was composed of tumour cells arranged in nests of various sizes separated by delicate fibrovascular stroma. The tumour cells had small, round hypochromatic nuclei and abundant cytoplasm. Metastases were observed in the inguinal lymph node. Immunohistochemically, most tumour cells expressed cytokeratin (CK) 20, chromogranin A, neuron-specific enolase, synaptophysin and oestrogen receptor-β, but not low molecular weight CK (CAM5.2), p63 and insulin. Ultrastructurally, the tumour cells contained a large number of electron-dense granules corresponding to neuroendocrine granules. Based on these findings, this case was diagnosed as a neuroendocrine carcinoma of the mammary gland. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Art and cartography as a critique of borders

    NASA Astrophysics Data System (ADS)

    Melo Ribeiro, Daniel

    2018-05-01

    This study focuses on the relationship between art and cartography. The main objective is to analyze how contemporary art uses maps to criticize borders. Inspired by the arguments raised by the Critical Cartography against the false neutrality of maps, we emphasize the potential of artworks to communicate different insights about how we experience and live the contemporary space. In that sense, art plays an important role not only to discuss the articulation of power and knowledge in cartography, but also to propose other categories of thought. Considering that borders are one of the most relevant visual elements on a map, we propose the following question: how the intersection between art and cartography can improve the critical thinking about borders? By questioning borders, art underlines that physical world is characterized by liminal spaces, not by absolute or strict separations. We briefly analyzed some examples of artworks that deal with political issues regarding this topic. Our findings suggest that art could reveal the impact of imposing borders in a space, whose arbitrary delimitation reflects power relations.

  6. On the role of PDZ domain-encoding genes in Drosophila border cell migration.

    PubMed

    Aranjuez, George; Kudlaty, Elizabeth; Longworth, Michelle S; McDonald, Jocelyn A

    2012-11-01

    Cells often move as collective groups during normal embryonic development and wound healing, although the mechanisms governing this type of migration are poorly understood. The Drosophila melanogaster border cells migrate as a cluster during late oogenesis and serve as a powerful in vivo genetic model for collective cell migration. To discover new genes that participate in border cell migration, 64 out of 66 genes that encode PDZ domain-containing proteins were systematically targeted by in vivo RNAi knockdown. The PDZ domain is one of the largest families of protein-protein interaction domains found in eukaryotes. Proteins that contain PDZ domains participate in a variety of biological processes, including signal transduction and establishment of epithelial apical-basal polarity. Targeting PDZ proteins effectively assesses a larger number of genes via the protein complexes and pathways through which these proteins function. par-6, a known regulator of border cell migration, was a positive hit and thus validated the approach. Knockdown of 14 PDZ domain genes disrupted migration with multiple RNAi lines. The candidate genes have diverse predicted cellular functions and are anticipated to provide new insights into the mechanisms that control border cell movement. As a test of this concept, two genes that disrupted migration were characterized in more detail: big bang and the Dlg5 homolog CG6509. We present evidence that Big bang regulates JAK/STAT signaling, whereas Dlg5/CG6509 maintains cluster cohesion. Moreover, these results demonstrate that targeting a selected class of genes by RNAi can uncover novel regulators of collective cell migration.

  7. Solare Cell Roof Tile And Method Of Forming Same

    DOEpatents

    Hanoka, Jack I.; Real, Markus

    1999-11-16

    A solar cell roof tile includes a front support layer, a transparent encapsulant layer, a plurality of interconnected solar cells and a backskin layer. The front support layer is formed of light transmitting material and has first and second surfaces. The transparent encapsulant layer is disposed adjacent the second surface of the front support layer. The interconnected solar cells has a first surface disposed adjacent the transparent encapsulant layer. The backskin layer has a first surface disposed adjacent a second surface of the interconnected solar cells, wherein a portion of the backskin layer wraps around and contacts the first surface of the front support layer to form the border region. A portion of the border region has an extended width. The solar cell roof tile may have stand-offs disposed on the extended width border region for providing vertical spacing with respect to an adjacent solar cell roof tile.

  8. Flexibility contra Stiffness: The Phragmoplast as a Physical Barrier for Beads But Not for Vesicles[OA

    PubMed Central

    Esseling-Ozdoba, Agnieszka; Kik, Richard A.; van Lammeren, André A.M.; Kleijn, J. Mieke; Emons, Anne Mie C.

    2010-01-01

    In plant cells, Golgi vesicles are transported to the division plane to fuse with each other, forming the cell plate, the initial membrane-bordered cell wall separating daughter cells. Vesicles, but not organelles, move through the phragmoplast, which consists of two opposing cylinders of microtubules and actin filaments, interlaced with endoplasmic reticulum membrane. To study physical aspects of this transport/inhibition process, we microinjected fluorescent synthetic 1,2-dioleoyl-sn-glycero-3-phospho-rac-1-glycerol (DOPG) vesicles and polystyrene beads into Tradescantia virginiana stamen hair cells. The phragmoplast was nonselective for DOPG vesicles of a size up to 150 nm in diameter but was a physical barrier for polystyrene beads having a diameter of 20 and 40 nm and also when beads were coated with the same DOPG membrane. We conclude that stiffness is a parameter for vesicle transit through the phragmoplast and discuss that cytoskeleton configurations can physically block such transit. PMID:19939943

  9. Propagation of a transverse wave on a foam microchannel

    NASA Astrophysics Data System (ADS)

    Derec, C.; Leroy, V.; Kaurin, D.; Arbogast, L.; Gay, C.; Elias, F.

    2015-11-01

    In a dry foam, soap films meet by three in the liquid microchannels, called Plateau borders, which contain most of the liquid of the foam. We investigated here the transverse vibration of a single Plateau border isolated on a rigid frame. We measured and we computed numerically and analytically the propagation of a transverse pulse along the channel in the 20-2000 Hz frequency range. The dispersion relation shows different scaling regimes, which provide information on the role of inertial and elastic forces acting on the Plateau border. At low frequency, the dispersion relation is dominated by the vibration of the air set into motion by the transverse vibration of the adjacent soap films. The inertia of the liquid in the Plateau border plays a role at high frequency, the critical frequency separating the low-frequency and the high-frequency regimes being a decreasing function of the radius R of the Plateau border.

  10. Evaluation of possible error sources in corneal endothelial morphometry with a semiautomated noncontact specular microscope.

    PubMed

    Doughty, Michael J

    2013-09-01

    To assess the corneal endothelium, particularly the polymegethism feature, using the Topcon SP-3000P specular microscope with newer center-dot software. Forty-eight healthy, normal weight, noncontact lens wearers of Asian ethnicity were assessed. Single endothelial images from each subject were processed with center-dot software, reevaluated after correction of obvious errors, and then by manual border marking and planimetry. Endothelial cell density based on average cell area and the coefficient of variation (COV) of cell area (polymegethism) were calculated. Error sources are associated with erroneous location of cell borders (usually creating larger or smaller "cells") or failure to assign cell borders to a marked cell. On the initial application of the center-dot software, the endothelial cell density values ranged from 1822 to 3244 cells per square millimeter (mean, 2644 cells/mm); this range was reduced (eg, 1955-3054 cells/mm; mean 2690 cells/mm on editing or in manual planimetry). The COV values ranged from 17% to 39% (mean, 27.5% ± 5.5%), with one third of the endothelia yielding COV values of greater than or equal to 30%. On editing or in manual planimetry, the COV values were reduced to between 17% and 29% (mean, 24.5% ± 3.2%; P < 0.001). In the use of a center-dot endothelial analysis program with cell border identification, it is likely that at least 1 set of editing steps is required to produce reasonable results.

  11. A synthetic mammalian network to compute population borders based on engineered reciprocal cell-cell communication.

    PubMed

    Kolar, Katja; Wischhusen, Hanna M; Müller, Konrad; Karlsson, Maria; Weber, Wilfried; Zurbriggen, Matias D

    2015-12-30

    Multicellular organisms depend on the exchange of information between specialized cells. This communication is often difficult to decipher in its native context, but synthetic biology provides tools to engineer well-defined systems that allow the convenient study and manipulation of intercellular communication networks. Here, we present the first mammalian synthetic network for reciprocal cell-cell communication to compute the border between a sender/receiver and a processing cell population. The two populations communicate via L-tryptophan and interleukin-4 to highlight the population border by the production of a fluorescent protein. The sharpness of that visualized edge can be adjusted by modulating key parameters of the network. We anticipate that this network will on the one hand be a useful tool to gain deeper insights into the mechanisms of tissue formation in nature and will on the other hand contribute to our ability to engineer artificial tissues.

  12. Ion channels versus ion pumps: the principal difference, in principle.

    PubMed

    Gadsby, David C

    2009-05-01

    The incessant traffic of ions across cell membranes is controlled by two kinds of border guards: ion channels and ion pumps. Open channels let selected ions diffuse rapidly down electrical and concentration gradients, whereas ion pumps labour tirelessly to maintain the gradients by consuming energy to slowly move ions thermodynamically uphill. Because of the diametrically opposed tasks and the divergent speeds of channels and pumps, they have traditionally been viewed as completely different entities, as alike as chalk and cheese. But new structural and mechanistic information about both of these classes of molecular machines challenges this comfortable separation and forces its re-evaluation.

  13. Sovereign Challenge Conference (6th). Borders & Security: Similarities, Differences and Shared Affinities. Held in El Paso, Texas on November 7-10, 2010

    DTIC Science & Technology

    2010-11-01

    protection and to isolate threats in order to survive and develop. The cell model implies osmosis , allowing flows to take place across the borders and the...relationships to new levels, much as cell phone technology replaces the laying of copper wire for traditional telephone service. Whole-of-Government/Whole...in urban, rural, and remote environments. This practical experience with the men and women of the U.S. Border Patrol animated many of the concepts

  14. Structural specificity of mucosal-cell transport and metabolism of peptide drugs: implication for oral peptide drug delivery

    NASA Technical Reports Server (NTRS)

    Bai, J. P.; Amidon, G. L.

    1992-01-01

    The brush border membrane of intestinal mucosal cells contains a peptide carrier system with rather broad substrate specificity and various endo- and exopeptidase activities. Small peptide (di-/tripeptide)-type drugs with or without an N-terminal alpha-amino group, including beta-lactam antibiotics and angiotensin-converting enzyme (ACE) inhibitors, are transported by the peptide transporter. Polypeptide drugs are hydrolyzed by brush border membrane proteolytic enzymes to di-/tripeptides and amino acids. Therefore, while the intestinal brush border membrane has a carrier system facilitating the absorption of di-/tripeptide drugs, it is a major barrier limiting oral availability of polypeptide drugs. In this paper, the specificity of peptide transport and metabolism in the intestinal brush border membrane is reviewed.

  15. Border-Crossing Model for the Diffusive Coarsening of Wet Foams

    NASA Astrophysics Data System (ADS)

    Durian, Douglas; Schimming, Cody

    For dry foams, the transport of gas from small high-pressure bubbles to large low-pressure bubbles is dominated by diffusion across the thin soap films separating neighboring bubbles. For wetter foams, the film areas become smaller as the Plateau borders and vertices inflate with liquid. So-called ``border-blocking'' models can explain some features of wet-foam coarsening based on the presumption that the inflated borders totally block the gas flux; however, this approximation dramatically fails in the wet/unjamming limit where the bubbles become close-packed spheres. Here, we account for the ever-present border-crossing flux by a new length scale defined by the average gradient of gas concentration inside the borders. We argue that it is proportional to the geometric average of film and border thicknesses, and we verify this scaling and the numerical prefactor by numerical solution of the diffusion equation. Then we show how the dA / dt =K0 (n - 6) von Neumann law is modified by the appearance of terms that depend on bubble size and shape as well as the concentration gradient length scale. Finally, we use the modified von Neumann law to compute the growth rate of the average bubble, which is not constant.

  16. Structurally stable, thin silicon solar cells

    NASA Technical Reports Server (NTRS)

    Arndt, R. A.; Meulenberg, A.

    1984-01-01

    A fabrication process for structurally stable thin solar cell wafers that produce good power output after irradiation is described. The fabrication process is as follows. A 6 mil, circular wafer is oxidized on both sides. One side is then patterned with a rectangular array of holes in the oxide that are nominally 75 mils square and separated by 2 mil spacings. Wells are then etched into the silicon with KOH to a depth of 4 mils, leaving a 2 mil, unetched thickness. Two areas on the surface are left unetched to provide pads for bonding or testing. All oxide is then removed and the rest of the processing is normal; the unetched face is used as the illuminated face. When all other processing is complete, a 2 X 2 cm cell is sawed from the starting wafer leaving a border that is approximately 10 mils wide. The effective thickness, determined by weighing an unmetallized sample, of such a cell is about 2.4 mil.

  17. Cross-border ties as a source of risk and resilience: Do cross-border ties moderate the relationship between migration-related stress and psychological distress among Latino migrants in the United States?

    PubMed Central

    Torres, Jacqueline M.; Alcántara, Carmela; Rudolph, Kara E.; Viruell-Fuentes, Edna A.

    2017-01-01

    Few studies have examined the associations between health and the cross-border ties that migrants maintain with their family members in communities of origin. We draw on theory related to social ties, ethnic identity, and mental health to examine cross-border ties as potential moderators of the association between migration-related stress and psychological distress among Latino migrants. Using data from the National Latino and Asian American Survey, we find that remittance sending is associated with significantly lower levels of psychological distress for Cuban migrants, and difficulty visiting home is associated with significantly greater psychological distress for Puerto Rican migrants. There were significant associations between migration-related stressors and psychological distress, although these associations fell to non-significance after accounting for multiple testing. We found little evidence that cross-border ties either buffer or exacerbate the association between migration-related stressors and psychological distress. We consider the findings within the current political and historical context of cross-border ties and separation. PMID:27803264

  18. Cross-border Ties as Sources of Risk and Resilience: Do Cross-border Ties Moderate the Relationship between Migration-related Stress and Psychological Distress for Latino Migrants in the United States?

    PubMed

    Torres, Jacqueline M; Alcántara, Carmela; Rudolph, Kara E; Viruell-Fuentes, Edna A

    2016-12-01

    Few studies have examined the associations between health and the cross-border ties that migrants maintain with their family members in communities of origin. We draw on theory related to social ties, ethnic identity, and mental health to examine cross-border ties as potential moderators of the association between migration-related stress and psychological distress among Latino migrants. Using data from the National Latino and Asian American Survey, we find that remittance sending is associated with significantly lower levels of psychological distress for Cuban migrants, and difficulty visiting home is associated with significantly greater psychological distress for Puerto Rican migrants. There were significant associations between migration-related stressors and psychological distress, although these associations fell to nonsignificance after accounting for multiple testing. We found little evidence that cross-border ties either buffer or exacerbate the association between migration-related stressors and psychological distress. We consider the findings within the current political and historical context of cross-border ties and separation. © American Sociological Association 2016.

  19. In vivo antibody-mediated modulation of aminopeptidase A in mouse proximal tubular epithelial cells.

    PubMed

    Mentzel, S; Dijkman, H B; van Son, J P; Wetzels, J F; Assmann, K J

    1999-07-01

    Aminopeptidase A (APA) is one of the many renal hydrolases. In mouse kidney, APA is predominantly expressed on the brush borders and sparsely on the basolateral membranes of proximal tubular epithelial cells. However, when large amounts of monoclonal antibodies (MAbs) against APA were injected into mice, we observed strong binding of the MAbs to the basolateral membranes, whereas the MAbs bound only transiently to the brush borders of the proximal tubular epithelial cells. In parallel, APA itself disappeared from the brush borders by both endocytosis and shedding, whereas it was increasingly expressed on the basolateral sides. Using ultrastructural immunohistology, we found no evidence for transcellular transport of endocytosed APA to the basolateral side of the proximal tubular epithelial cells. The absence of transcellular transport was confirmed by experiments in which we used a low dose of the MAbs. Such a low dose did not result in binding of the MAbs to the brush borders and had no effect on the presence of APA in the brush borders of the proximal tubular epithelial cells. In these experiments we still could observe binding of the MAbs to the basolateral membranes in parallel with the local appearance of APA. In addition, treatment of mice with chlorpromazine, a calmodulin antagonist that interferes with cytoskeletal function, largely inhibited the MAb-induced modulation of APA. Our studies suggest that injection of MAbs to APA specifically interrupts the normal intracellular traffic of this enzyme in proximal tubular epithelial cells. This intracellular transport is dependent on the action of cytoskeletal proteins.

  20. Dlx proteins position the neural plate border and determine adjacent cell fates.

    PubMed

    Woda, Juliana M; Pastagia, Julie; Mercola, Mark; Artinger, Kristin Bruk

    2003-01-01

    The lateral border of the neural plate is a major source of signals that induce primary neurons, neural crest cells and cranial placodes as well as provide patterning cues to mesodermal structures such as somites and heart. Whereas secreted BMP, FGF and Wnt proteins influence the differentiation of neural and non-neural ectoderm, we show here that members of the Dlx family of transcription factors position the border between neural and non-neural ectoderm and are required for the specification of adjacent cell fates. Inhibition of endogenous Dlx activity in Xenopus embryos with an EnR-Dlx homeodomain fusion protein expands the neural plate into non-neural ectoderm tissue whereas ectopic activation of Dlx target genes inhibits neural plate differentiation. Importantly, the stereotypic pattern of border cell fates in the adjacent ectoderm is re-established only under conditions where the expanded neural plate abuts Dlx-positive non-neural ectoderm. Experiments in which presumptive neural plate was grafted to ventral ectoderm reiterate induction of neural crest and placodal lineages and also demonstrate that Dlx activity is required in non-neural ectoderm for the production of signals needed for induction of these cells. We propose that Dlx proteins regulate intercellular signaling across the interface between neural and non-neural ectoderm that is critical for inducing and patterning adjacent cell fates.

  1. Analysis of the Highly Diverse Gene Borders in Ebola Virus Reveals a Distinct Mechanism of Transcriptional Regulation

    PubMed Central

    Brauburger, Kristina; Boehmann, Yannik; Tsuda, Yoshimi; Hoenen, Thomas; Olejnik, Judith; Schümann, Michael; Ebihara, Hideki

    2014-01-01

    ABSTRACT Ebola virus (EBOV) belongs to the group of nonsegmented negative-sense RNA viruses. The seven EBOV genes are separated by variable gene borders, including short (4- or 5-nucleotide) intergenic regions (IRs), a single long (144-nucleotide) IR, and gene overlaps, where the neighboring gene end and start signals share five conserved nucleotides. The unique structure of the gene overlaps and the presence of a single long IR are conserved among all filoviruses. Here, we sought to determine the impact of the EBOV gene borders during viral transcription. We show that readthrough mRNA synthesis occurs in EBOV-infected cells irrespective of the structure of the gene border, indicating that the gene overlaps do not promote recognition of the gene end signal. However, two consecutive gene end signals at the VP24 gene might improve termination at the VP24-L gene border, ensuring efficient L gene expression. We further demonstrate that the long IR is not essential for but regulates transcription reinitiation in a length-dependent but sequence-independent manner. Mutational analysis of bicistronic minigenomes and recombinant EBOVs showed no direct correlation between IR length and reinitiation rates but demonstrated that specific IR lengths not found naturally in filoviruses profoundly inhibit downstream gene expression. Intriguingly, although truncation of the 144-nucleotide-long IR to 5 nucleotides did not substantially affect EBOV transcription, it led to a significant reduction of viral growth. IMPORTANCE Our current understanding of EBOV transcription regulation is limited due to the requirement for high-containment conditions to study this highly pathogenic virus. EBOV is thought to share many mechanistic features with well-analyzed prototype nonsegmented negative-sense RNA viruses. A single polymerase entry site at the 3′ end of the genome determines that transcription of the genes is mainly controlled by gene order and cis-acting signals found at the gene borders. Here, we examined the regulatory role of the structurally unique EBOV gene borders during viral transcription. Our data suggest that transcriptional regulation in EBOV is highly complex and differs from that in prototype viruses and further the understanding of this most fundamental process in the filovirus replication cycle. Moreover, our results with recombinant EBOVs suggest a novel role of the long IR found in all filovirus genomes during the viral replication cycle. PMID:25142600

  2. Analysis of the highly diverse gene borders in Ebola virus reveals a distinct mechanism of transcriptional regulation.

    PubMed

    Brauburger, Kristina; Boehmann, Yannik; Tsuda, Yoshimi; Hoenen, Thomas; Olejnik, Judith; Schümann, Michael; Ebihara, Hideki; Mühlberger, Elke

    2014-11-01

    Ebola virus (EBOV) belongs to the group of nonsegmented negative-sense RNA viruses. The seven EBOV genes are separated by variable gene borders, including short (4- or 5-nucleotide) intergenic regions (IRs), a single long (144-nucleotide) IR, and gene overlaps, where the neighboring gene end and start signals share five conserved nucleotides. The unique structure of the gene overlaps and the presence of a single long IR are conserved among all filoviruses. Here, we sought to determine the impact of the EBOV gene borders during viral transcription. We show that readthrough mRNA synthesis occurs in EBOV-infected cells irrespective of the structure of the gene border, indicating that the gene overlaps do not promote recognition of the gene end signal. However, two consecutive gene end signals at the VP24 gene might improve termination at the VP24-L gene border, ensuring efficient L gene expression. We further demonstrate that the long IR is not essential for but regulates transcription reinitiation in a length-dependent but sequence-independent manner. Mutational analysis of bicistronic minigenomes and recombinant EBOVs showed no direct correlation between IR length and reinitiation rates but demonstrated that specific IR lengths not found naturally in filoviruses profoundly inhibit downstream gene expression. Intriguingly, although truncation of the 144-nucleotide-long IR to 5 nucleotides did not substantially affect EBOV transcription, it led to a significant reduction of viral growth. Our current understanding of EBOV transcription regulation is limited due to the requirement for high-containment conditions to study this highly pathogenic virus. EBOV is thought to share many mechanistic features with well-analyzed prototype nonsegmented negative-sense RNA viruses. A single polymerase entry site at the 3' end of the genome determines that transcription of the genes is mainly controlled by gene order and cis-acting signals found at the gene borders. Here, we examined the regulatory role of the structurally unique EBOV gene borders during viral transcription. Our data suggest that transcriptional regulation in EBOV is highly complex and differs from that in prototype viruses and further the understanding of this most fundamental process in the filovirus replication cycle. Moreover, our results with recombinant EBOVs suggest a novel role of the long IR found in all filovirus genomes during the viral replication cycle. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  3. 47 CFR 73.807 - Minimum distance separation between stations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... and the right-hand column lists (for informational purposes only) the minimum distance necessary for...) Within 320 km of the Mexican border, LP100 stations must meet the following separations with respect to any Mexican stations: Mexican station class Co-channel (km) First-adjacent channel (km) Second-third...

  4. The placenta of the salp (Tunicata: Thaliacea).

    PubMed

    Bone, Q; Pulsford, A L; Amoroso, E C

    1985-01-01

    The morphology of the mature 'placenta' of the pelagic tunicate Salpa fusiformis is described, and it is shown that two syncytial layers, intimately connected by interdigitating microvilli, separate maternal and embryonic circulations. The central placental layer facing the maternal circulation is bordered by membrane infoldings; the cortical layer facing the embryonic circulation is bordered by extensively branching microvilli. Both layers are of maternal origin, although embryonic leucocytes pass into, and add to, the cortical layer.

  5. Impact of various color LED flashlights and different lighting source to skin distances on the manual and the computer-aided detection of basal cell carcinoma borders.

    PubMed

    Bakht, Mohamadreza K; Pouladian, Majid; Mofrad, Farshid B; Honarpisheh, Hamid

    2014-02-01

    Quantitative analysis based on digital skin image has been proven to be helpful in dermatology. Moreover, the borders of the basal cell carcinoma (BCC) lesions have been challenging borders for the automatic detection methods. In this work, a computer-aided dermatoscopy system was proposed to enhance the clinical detection of BCC lesion borders. Fifty cases of BCC were selected and 2000 pictures were taken. The lesion images data were obtained with eight colors of flashlights and in five different lighting source to skin distances (SSDs). Then, the image-processing techniques were used for automatic detection of lesion borders. Further, the dermatologists marked the lesions on the obtained photos. Considerable differences between the obtained values referring to the photographs that were taken at super blue and aqua green color lighting were observed for most of the BCC borders. It was observed that by changing the SSD, an optimum distance could be found where that the accuracy of the detection reaches to a maximum value. This study clearly indicates that by changing SSD and lighting color, manual and automatic detection of BCC lesions borders can be enhanced. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  6. Border-crossing model for the diffusive coarsening of two-dimensional and quasi-two-dimensional wet foams

    NASA Astrophysics Data System (ADS)

    Schimming, C. D.; Durian, D. J.

    2017-09-01

    For dry foams, the transport of gas from small high-pressure bubbles to large low-pressure bubbles is dominated by diffusion across the thin soap films separating neighboring bubbles. For wetter foams, the film areas become smaller as the Plateau borders and vertices inflate with liquid. So-called "border-blocking" models can explain some features of wet-foam coarsening based on the presumption that the inflated borders totally block the gas flux; however, this approximation dramatically fails in the wet or unjamming limit where the bubbles become close-packed spheres and coarsening proceeds even though there are no films. Here, we account for the ever-present border-crossing flux by a new length scale defined by the average gradient of gas concentration inside the borders. We compute that it is proportional to the geometric average of film and border thicknesses, and we verify this scaling by numerical solution of the diffusion equation. We similarly consider transport across inflated vertices and surface Plateau borders in quasi-two-dimensional foams. And we show how the d A /d t =K0(n -6 ) von Neumann law is modified by the appearance of terms that depend on bubble size and shape as well as the concentration gradient length scales. Finally, we use the modified von Neumann law to compute the growth rate of the average bubble area, which is not constant.

  7. Escaping Underground Nets: Extracellular DNases Degrade Plant Extracellular Traps and Contribute to Virulence of the Plant Pathogenic Bacterium Ralstonia solanacearum

    PubMed Central

    Tran, Tuan Minh; MacIntyre, April; Hawes, Martha; Allen, Caitilyn

    2016-01-01

    Plant root border cells have been recently recognized as an important physical defense against soil-borne pathogens. Root border cells produce an extracellular matrix of protein, polysaccharide and DNA that functions like animal neutrophil extracellular traps to immobilize pathogens. Exposing pea root border cells to the root-infecting bacterial wilt pathogen Ralstonia solanacearum triggered release of DNA-containing extracellular traps in a flagellin-dependent manner. These traps rapidly immobilized the pathogen and killed some cells, but most of the entangled bacteria eventually escaped. The R. solanacearum genome encodes two putative extracellular DNases (exDNases) that are expressed during pathogenesis, suggesting that these exDNases contribute to bacterial virulence by enabling the bacterium to degrade and escape root border cell traps. We tested this hypothesis with R. solanacearum deletion mutants lacking one or both of these nucleases, named NucA and NucB. Functional studies with purified proteins revealed that NucA and NucB are non-specific endonucleases and that NucA is membrane-associated and cation-dependent. Single ΔnucA and ΔnucB mutants and the ΔnucA/B double mutant all had reduced virulence on wilt-susceptible tomato plants in a naturalistic soil-soak inoculation assay. The ΔnucA/B mutant was out-competed by the wild-type strain in planta and was less able to stunt root growth or colonize plant stems. Further, the double nuclease mutant could not escape from root border cells in vitro and was defective in attachment to pea roots. Taken together, these results demonstrate that extracellular DNases are novel virulence factors that help R. solanacearum successfully overcome plant defenses to infect plant roots and cause bacterial wilt disease. PMID:27336156

  8. Psychological Functions of Semiotic Borders in Sense-Making: Liminality of Narrative Processes

    PubMed Central

    De Luca Picione, Raffaele; Valsiner, Jaan

    2017-01-01

    In this paper we discuss the semiotic functions of the psychological borders that structure the flow of narrative processes. Each narration is always a contextual, situated and contingent process of sensemaking, made possible by the creation of borders, such as dynamic semiotic devices that are capable of connecting the past and the future, the inside and the outside, and the me with the non-me. Borders enable us to narratively construct one’s own experiences using three inherent processes: contextualization, intersubjective positioning and setting of pertinence. The narrative process – as a subjective articulation of signs in a contingent social context – involves several functions of semiotic borders: separation, differentiation, distinction-making, connection, articulation and relation-enabling. The relevant psychological aspect highlighted here is that a border is a semiotic device which is required for both maintaining stability and inducing transformation at the same time. The peculiar dynamics and the semiotic structure of borders generate a liminal space, which is characterized by instability, by a blurred space-time distinction and by ambiguities in the semantic and syntactic processes of sensemaking. The psychological processes that occur in liminal space are strongly affectively loaded, yet it is exactly the setting and activation of liminality processes that lead to novelty and creativity and enable the creation of new narrative forms. PMID:28904600

  9. Psychological Functions of Semiotic Borders in Sense-Making: Liminality of Narrative Processes.

    PubMed

    De Luca Picione, Raffaele; Valsiner, Jaan

    2017-08-01

    In this paper we discuss the semiotic functions of the psychological borders that structure the flow of narrative processes. Each narration is always a contextual, situated and contingent process of sensemaking, made possible by the creation of borders, such as dynamic semiotic devices that are capable of connecting the past and the future, the inside and the outside, and the me with the non-me. Borders enable us to narratively construct one's own experiences using three inherent processes: contextualization, intersubjective positioning and setting of pertinence. The narrative process - as a subjective articulation of signs in a contingent social context - involves several functions of semiotic borders: separation, differentiation, distinction-making, connection, articulation and relation-enabling. The relevant psychological aspect highlighted here is that a border is a semiotic device which is required for both maintaining stability and inducing transformation at the same time. The peculiar dynamics and the semiotic structure of borders generate a liminal space, which is characterized by instability, by a blurred space-time distinction and by ambiguities in the semantic and syntactic processes of sensemaking. The psychological processes that occur in liminal space are strongly affectively loaded, yet it is exactly the setting and activation of liminality processes that lead to novelty and creativity and enable the creation of new narrative forms.

  10. Late oligocene and miocene faulting and sedimentation, and evolution of the southern Rio Grande rift, New Mexico, USA

    NASA Astrophysics Data System (ADS)

    Mack, Greg H.; Seager, William R.; Kieling, John

    1994-08-01

    The distribution of nonmarine lithofacies, paleocurrents, and provenance data are used to define the evolution of late Oligocene and Miocene basins and complementary uplifts in the southern Rio Grande rift in the vicinity of Hatch, New Mexico, USA. The late Oligocene-middle Miocene Hayner Ranch Formation, which consists of a maximum of 1000 m of alluvial-fan, alluvial-flat, and lacustrine-carbonate lithofacies, was deposited in a narrow (12 km), northwest-trending, northeast-tilted half graben, whose footwall was the Caballo Mountains block. Stratigraphic separation on the border faults of the Caballo Mountains block was approximately 1615 m. An additional 854 m of stratigraphic separation along the Caballo Mountains border faults occurred during deposition of the middle-late Miocene Rincon Valley Formation, which is composed of up to 610 m of alluvial-fan, alluvial-flat, braided-fluvial, and gypsiferous playa lithofacies. Two new, north-trending fault blocks (Sierra de las Uvas and Dona Ana Mountains) and complementary west-northwest-tilted half graben also developed during Rincon Valley time, with approximately 549 m of stratigraphic separation along the border fault of the Sierra de las Uvas block. In latest Miocene and early Pliocene time, following deposition of the Rincon Valley Formation, movement continued along the border faults of the Caballo Mountains, Dona Ana Mountains, and Sierra de las Uvas blocks, and large parts of the Hayner Ranch and Rincon Valley basins were segmented into smaller fault blocks and basins by movement along new, largely north-trending faults. Analysis of the Hayner Ranch and Rincon Valley Formations, along with previous studies of the early Oligocene Bell Top Formation and late Pliocene-early Pleistocene Camp Rice Formation, indicate that the traditional two-stage model for development of the southern Rio Grande rift should be abandoned in favor of at least four episodes of block faulting beginning 35 Ma ago. With the exception of two northwest-trending border faults of the Caballo Mountains block that may be reactivated along Eocene compressional structures, the majority of border faults and complementary basins throughout the history of the southern Rio Grande rift were north-trending, which challenges the conventional idea of a clockwise change in stress through time.

  11. Crossing Boundaries in Literacy Research: Challenges and Opportunities

    ERIC Educational Resources Information Center

    Almasi, Janice F.

    2016-01-01

    The concept of boundaries, as borders that separate two entities, can be problematic in that those who "belong" within a boundary in a given social world or entity are separated from those who do not, creating binaries. The field of literacy research has had a long history of binarism in which instructional methods, epistemological…

  12. Demographic Trends in the United States: A Review of Research in the 2000s

    ERIC Educational Resources Information Center

    Cherlin, Andrew J.

    2010-01-01

    Demographic trends in the 2000s showed the continuing separation of family and household because of factors such as childbearing among single parents, the dissolution of cohabiting unions, divorce, repartnering, and remarriage. The transnational families of many immigrants also displayed this separation, as families extended across borders. In…

  13. Corneal squamous cell carcinoma in a Border Collie.

    PubMed

    Busse, Claudia; Sansom, Jane; Dubielzig, R R; Hayes, Alison

    2008-01-01

    A 6-year-old, female, spayed Border Collie was presented to the Unit of Comparative Ophthalmology at the Animal Health Trust with a 6-month history of a progressive nonpainful opacity of the left cornea. A keratectomy was performed and the tissue submitted for histopathology. The diagnosis was squamous cell carcinoma. There has been no recurrence of the neoplasm to date (5 months). Canine corneal squamous cell carcinoma (SCC) has not been reported previously in the UK.

  14. Dlx proteins position the neural plate border and determine adjacent cell fates

    PubMed Central

    Woda, Juliana M.; Pastagia, Julie; Mercola, Mark; Artinger, Kristin Bruk

    2014-01-01

    Summary The lateral border of the neural plate is a major source of signals that induce primary neurons, neural crest cells and cranial placodes as well as provide patterning cues to mesodermal structures such as somites and heart. Whereas secreted BMP, FGF and Wnt proteins influence the differentiation of neural and non-neural ectoderm, we show here that members of the Dlx family of transcription factors position the border between neural and non-neural ectoderm and are required for the specification of adjacent cell fates. Inhibition of endogenous Dlx activity in Xenopus embryos with an EnR-Dlx homeodomain fusion protein expands the neural plate into non-neural ectoderm tissue whereas ectopic activation of Dlx target genes inhibits neural plate differentiation. Importantly, the stereotypic pattern of border cell fates in the adjacent ectoderm is re-established only under conditions where the expanded neural plate abuts Dlx-positive non-neural ectoderm. Experiments in which presumptive neural plate was grafted to ventral ectoderm reiterate induction of neural crest and placodal lineages and also demonstrate that Dlx activity is required in non-neural ectoderm for the production of signals needed for induction of these cells. We propose that Dlx proteins regulate intercellular signaling across the interface between neural and non-neural ectoderm that is critical for inducing and patterning adjacent cell fates. PMID:12466200

  15. Optoelectronic stereoscopic device for diagnostics, treatment, and developing of binocular vision

    NASA Astrophysics Data System (ADS)

    Pautova, Larisa; Elkhov, Victor A.; Ovechkis, Yuri N.

    2003-08-01

    Operation of the device is based on alternative generation of pictures for left and right eyes on the monitor screen. Controller gives pulses on LCG so that shutter for left or right eye opens synchronously with pictures. The device provides frequency of switching more than 100 Hz, and that is why the flickering is absent. Thus, a separate demonstration of images to the left eye or to the right one in turn is obtained for patients being unaware and creates the conditions of binocular perception clsoe to natural ones without any additional separation of vision fields. LC-cell transfer characteristic coodination with time parameters of monitor screen has enabled to improve stereo image quality. Complicated problem of computer stereo images with LC-glasses is so called 'ghosts' - noise images that come to blocked eye. We reduced its influence by adapting stereo images to phosphor and LC-cells characteristics. The device is intended for diagnostics and treatment of stabismus, amblyopia and other binocular and stereoscopic vision impairments, for cultivating, training and developing of stereoscopic vision, for measurements of horizontal and vertical phoria, phusion reserves, the stereovision acuity and some else, for fixing central scotoma borders, as well as suppression scotoma in strabismus too.

  16. Environmental cues and symbiont microbe-associated molecular patterns function in concert to drive the daily remodelling of the crypt-cell brush border of the Euprymna scolopes light organ

    PubMed Central

    Heath-Heckman, Elizabeth A.C.; Foster, Jamie; Apicella, Michael A.; Goldman, William E.; McFall-Ngai, Margaret

    2016-01-01

    Summary Recent research has shown that the microbiota affects the biology of associated host epithelial tissues, including their circadian rhythms, although few data are available on how such influences shape the microarchitecture of the brush border. The squid-vibrio system exhibits two modifications of the brush border that supports the symbionts: effacement and repolarization. Together these occur on a daily rhythm in adult animals, at the dawn expulsion of symbionts into the environment, and symbiont colonization of the juvenile host induces an increase in microvillar density. Here we sought to define how these processes are related and the roles of both symbiont colonization and environmental cues. Ultrastructural analyses showed that the juvenile-organ brush borders also efface concomitantly with daily dawn-cued expulsion of symbionts. Manipulation of the environmental light cue and juvenile symbiotic state demonstrated that this behaviour requires the light cue, but not colonization. In contrast, symbionts were required for the observed increase in microvillar density that accompanies post dawn brush-border repolarization; this increase was induced solely by host exposure to phosphorylated lipid A of symbiont cells. These data demonstrate that a partnering of environmental and symbiont cues shapes the brush border and that microbe-associated molecular patterns play a role in the regulation of brush-border microarchitecture. PMID:27062511

  17. Paraquat initially damages cochlear support cells leading to anoikis-like hair cell death.

    PubMed

    Zhang, Jianhui; Sun, Hong; Salvi, Richard; Ding, Dalian

    2018-07-01

    Paraquat (PQ), one of the most widely used herbicides, is extremely dangerous because it generates the highly toxic superoxide radical. When paraquat was applied to cochlear organotypic cultures, it not only damaged the outer hair cells (OHCs) and inner hair cells (IHCs), but also caused dislocation of the hair cell rows. We hypothesized that the dislocation arose from damage to the support cells (SCs) that anchors hair cells within the epithelium. To test this hypothesis, rat postnatal cochlear cultures were treated with PQ. Shortly after PQ treatment, the rows of OHCs separated from one another and migrated radially away from IHCs suggesting loss of cell-cell adhesion that hold the hair cells in proper alignment. Hair cells dislocation was associated with extensive loss of SCs in the organ of Corti, loss of tympanic border cells (TBCs) beneath the basilar membrane, the early appearance of superoxide staining and caspase-8 labeling in SCs below the OHCs and disintegration of E-cadherin and β-catenin in the organ of Corti. Damage to the TBCs and SCs occurred prior to loss of OHC or IHC loss suggesting a form of detachment-induced apoptosis referred to as anoikis. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Effect of arabinogalactan proteins from the root caps of pea and Brassica napus on Aphanomyces euteiches zoospore chemotaxis and germination.

    PubMed

    Cannesan, Marc Antoine; Durand, Caroline; Burel, Carole; Gangneux, Christophe; Lerouge, Patrice; Ishii, Tadashi; Laval, Karine; Follet-Gueye, Marie-Laure; Driouich, Azeddine; Vicré-Gibouin, Maïté

    2012-08-01

    Root tips of many plant species release a number of border, or border-like, cells that are thought to play a major role in the protection of root meristem. However, little is currently known on the structure and function of the cell wall components of such root cells. Here, we investigate the sugar composition of the cell wall of the root cap in two species: pea (Pisum sativum), which makes border cells, and Brassica napus, which makes border-like cells. We find that the cell walls are highly enriched in arabinose and galactose, two major residues of arabinogalactan proteins. We confirm the presence of arabinogalactan protein epitopes on root cap cell walls using immunofluorescence microscopy. We then focused on these proteoglycans by analyzing their carbohydrate moieties, linkages, and electrophoretic characteristics. The data reveal (1) significant structural differences between B. napus and pea root cap arabinogalactan proteins and (2) a cross-link between these proteoglycans and pectic polysaccharides. Finally, we assessed the impact of root cap arabinogalactan proteins on the behavior of zoospores of Aphanomyces euteiches, an oomycetous pathogen of pea roots. We find that although the arabinogalactan proteins of both species induce encystment and prevent germination, the effects of both species are similar. However, the arabinogalactan protein fraction from pea attracts zoospores far more effectively than that from B. napus. This suggests that root arabinogalactan proteins are involved in the control of early infection of roots and highlights a novel role for these proteoglycans in root-microbe interactions.

  19. Effect of Arabinogalactan Proteins from the Root Caps of Pea and Brassica napus on Aphanomyces euteiches Zoospore Chemotaxis and Germination12[C][W

    PubMed Central

    Cannesan, Marc Antoine; Durand, Caroline; Burel, Carole; Gangneux, Christophe; Lerouge, Patrice; Ishii, Tadashi; Laval, Karine; Follet-Gueye, Marie-Laure; Driouich, Azeddine; Vicré-Gibouin, Maïté

    2012-01-01

    Root tips of many plant species release a number of border, or border-like, cells that are thought to play a major role in the protection of root meristem. However, little is currently known on the structure and function of the cell wall components of such root cells. Here, we investigate the sugar composition of the cell wall of the root cap in two species: pea (Pisum sativum), which makes border cells, and Brassica napus, which makes border-like cells. We find that the cell walls are highly enriched in arabinose and galactose, two major residues of arabinogalactan proteins. We confirm the presence of arabinogalactan protein epitopes on root cap cell walls using immunofluorescence microscopy. We then focused on these proteoglycans by analyzing their carbohydrate moieties, linkages, and electrophoretic characteristics. The data reveal (1) significant structural differences between B. napus and pea root cap arabinogalactan proteins and (2) a cross-link between these proteoglycans and pectic polysaccharides. Finally, we assessed the impact of root cap arabinogalactan proteins on the behavior of zoospores of Aphanomyces euteiches, an oomycetous pathogen of pea roots. We find that although the arabinogalactan proteins of both species induce encystment and prevent germination, the effects of both species are similar. However, the arabinogalactan protein fraction from pea attracts zoospores far more effectively than that from B. napus. This suggests that root arabinogalactan proteins are involved in the control of early infection of roots and highlights a novel role for these proteoglycans in root-microbe interactions. PMID:22645070

  20. Intervention of ginger or propolis ameliorates methotrexate-induced ileum toxicity.

    PubMed

    Abdul-Hamid, Manal; Salah, Marwa

    2016-02-01

    The long-term clinical use of methotrexate (MTX) is restricted due to its severe intestinal toxicity. The protective effect of ginger or propolis on the toxicity induced by MTX is relatively less understood, so the possible protective effect of ginger or propolis, used separately, was investigated. A total of 60 male albino rats were divided into six groups as follows: (1) control group; (2) ginger group; (3) propolis group; (4) MTX group; (5) ginger + MTX group; and (6) propolis + MTX group. The present results show that MTX caused ileum injury, including shortening and fusion of the villi, inflammatory cell infiltration and goblet cell depletion. Administration of ginger or propolis ameliorated the MTX-induced ileum injury as shown by histological, immunohistochemical and ultrastructural investigations and statistical analysis. This is revealed by intact villi, which shows marked increase in brown colouration of proliferating cell nuclear antigen positive nuclei in the crypts region, improvement in the number of goblet cells and brush border length of ileum. The current results conclude the efficacy and safety of ginger and propolis, which may be due to their antioxidant properties. © The Author(s) 2013.

  1. Ceroid-lipofuscinosis in border collie dogs.

    PubMed

    Taylor, R M; Farrow, B R

    1988-01-01

    Five Border Collie dogs with ceroid-lipofuscinosis developed progressive neurological disease between 18 and 22 months of age. These dogs had behavioural abnormalities, gait and visual deficits and became progressively demented. All dogs examined had common ancestors. Light microscopic examination of tissues demonstrated extensive accumulation of granular, sudan black-staining autofluorescent material in the cytoplasm of neurones, retinal ganglion cells and some visceral cells. At ultrastructural examination inclusions of variable morphology were observed.

  2. Induction of a gradual, reversible morphogenesis of its host's epithelial brush border by Vibrio fischeri.

    PubMed

    Lamarcq, L H; McFall-Ngai, M J

    1998-02-01

    Bacteria exert a variety of influences on the morphology and physiology of animal cells whether they are pathogens or cooperative partners. The association between the luminous bacterium Vibrio fischeri and the sepiolid squid Euprymna scolopes provides an experimental model for the study of the influence of extracellular bacteria on the development of host epithelia. In this study, we analyzed bacterium-induced changes in the brush borders of the light organ crypt epithelia during the initial hours following colonization of this tissue. Transmission electron microscopy of the brush border morphology in colonized and uncolonized hosts revealed that the bacteria effect a fourfold increase in microvillar density over the first 4 days of the association. Estimates of the proportions of bacterial cells in contact with host microvilli showed that the intimacy of the bacterial cells with animal cell surfaces increases significantly during this time. Antibiotic curing of the organ following colonization showed that sustained interaction with bacteria is essential for the retention of the induced morphological changes. Bacteria that are defective in either light production or colonization efficiency produced changes similar to those by the parent strain. Conventional fluorescence and confocal scanning laser microscopy revealed that the brush border is supported by abundant filamentous actin. However, in situ hybridization with beta-actin probes did not show marked bacterium-induced increases in beta-actin gene expression. These experiments demonstrate that the E. scolopes-V. fischeri system is a viable model for the experimental study of bacterium-induced changes in host brush border morphology.

  3. Induction of a Gradual, Reversible Morphogenesis of Its Host’s Epithelial Brush Border by Vibrio fischeri

    PubMed Central

    Lamarcq, Laurence H.; McFall-Ngai, Margaret J.

    1998-01-01

    Bacteria exert a variety of influences on the morphology and physiology of animal cells whether they are pathogens or cooperative partners. The association between the luminous bacterium Vibrio fischeri and the sepiolid squid Euprymna scolopes provides an experimental model for the study of the influence of extracellular bacteria on the development of host epithelia. In this study, we analyzed bacterium-induced changes in the brush borders of the light organ crypt epithelia during the initial hours following colonization of this tissue. Transmission electron microscopy of the brush border morphology in colonized and uncolonized hosts revealed that the bacteria effect a fourfold increase in microvillar density over the first 4 days of the association. Estimates of the proportions of bacterial cells in contact with host microvilli showed that the intimacy of the bacterial cells with animal cell surfaces increases significantly during this time. Antibiotic curing of the organ following colonization showed that sustained interaction with bacteria is essential for the retention of the induced morphological changes. Bacteria that are defective in either light production or colonization efficiency produced changes similar to those by the parent strain. Conventional fluorescence and confocal scanning laser microscopy revealed that the brush border is supported by abundant filamentous actin. However, in situ hybridization with β-actin probes did not show marked bacterium-induced increases in β-actin gene expression. These experiments demonstrate that the E. scolopes-V. fischeri system is a viable model for the experimental study of bacterium-induced changes in host brush border morphology. PMID:9453641

  4. Trafficking Ion Transporters to the Apical Membrane of Polarized Intestinal Enterocytes.

    PubMed

    Engevik, Amy Christine; Goldenring, James R

    2018-01-02

    Epithelial cells lining the gastrointestinal tract require distinct apical and basolateral domains to function properly. Trafficking and insertion of enzymes and transporters into the apical brush border of intestinal epithelial cells is essential for effective digestion and absorption of nutrients. Specific critical ion transporters are delivered to the apical brush border to facilitate fluid and electrolyte uptake. Maintenance of these apical transporters requires both targeted delivery and regulated membrane recycling. Examination of altered apical trafficking in patients with Microvillus Inclusion disease caused by inactivating mutations in MYO5B has led to insights into the regulation of apical trafficking by elements of the apical recycling system. Modeling of MYO5B loss in cell culture and animal models has led to recognition of Rab11a and Rab8a as critical regulators of apical brush border function. All of these studies show the importance of apical membrane trafficking dynamics in maintenance of polarized epithelial cell function. Copyright © 2018 Cold Spring Harbor Laboratory Press; all rights reserved.

  5. Cerebellar abiotrophy in a family of Border Collie dogs.

    PubMed

    Sandy, J R; Slocombe, R E; Mitten, R W; Jedwab, D

    2002-11-01

    Cerebellar abiotrophies have a nonsex-linked, autosomal, recessively inherited basis in a number of species, and lesions typically reflect profound and progressive loss of Purkinje cells. In this report, an unusual form of abiotrophy is described for two sibling Border Collies. Extensive loss of the cerebellar granular cell layer was present with relative sparing of Purkinje cells of two female pups. The biochemical basis for this form of cerebellar abiotrophy is unknown, but the lack of disease in other siblings supports an autosomal recessive mode of inheritance.

  6. HIV Transmission Networks in the San Diego–Tijuana Border Region

    PubMed Central

    Mehta, Sanjay R.; Wertheim, Joel O.; Brouwer, Kimberly C.; Wagner, Karla D.; Chaillon, Antoine; Strathdee, Steffanie; Patterson, Thomas L.; Rangel, Maria G.; Vargas, Mlenka; Murrell, Ben; Garfein, Richard; Little, Susan J.; Smith, Davey M.

    2015-01-01

    Background HIV sequence data can be used to reconstruct local transmission networks. Along international borders, like the San Diego–Tijuana region, understanding the dynamics of HIV transmission across reported risks, racial/ethnic groups, and geography can help direct effective prevention efforts on both sides of the border. Methods We gathered sociodemographic, geographic, clinical, and viral sequence data from HIV infected individuals participating in ten studies in the San Diego–Tijuana border region. Phylogenetic and network analysis was performed to infer putative relationships between HIV sequences. Correlates of identified clusters were evaluated and spatiotemporal relationships were explored using Bayesian phylogeographic analysis. Findings After quality filtering, 843 HIV sequences with associated demographic data and 263 background sequences from the region were analyzed, and 138 clusters were inferred (2–23 individuals). Overall, the rate of clustering did not differ by ethnicity, residence, or sex, but bisexuals were less likely to cluster than heterosexuals or men who have sex with men (p = 0.043), and individuals identifying as white (p ≤ 0.01) were more likely to cluster than other races. Clustering individuals were also 3.5 years younger than non-clustering individuals (p < 0.001). Although the sampled San Diego and Tijuana epidemics were phylogenetically compartmentalized, five clusters contained individuals residing on both sides of the border. Interpretation This study sampled ~ 7% of HIV infected individuals in the border region, and although the sampled networks on each side of the border were largely separate, there was evidence of persistent bidirectional cross-border transmissions that linked risk groups, thus highlighting the importance of the border region as a “melting pot” of risk groups. Funding NIH, VA, and Pendleton Foundation. PMID:26629540

  7. HIV Transmission Networks in the San Diego-Tijuana Border Region.

    PubMed

    Mehta, Sanjay R; Wertheim, Joel O; Brouwer, Kimberly C; Wagner, Karla D; Chaillon, Antoine; Strathdee, Steffanie; Patterson, Thomas L; Rangel, Maria G; Vargas, Mlenka; Murrell, Ben; Garfein, Richard; Little, Susan J; Smith, Davey M

    2015-10-01

    HIV sequence data can be used to reconstruct local transmission networks. Along international borders, like the San Diego-Tijuana region, understanding the dynamics of HIV transmission across reported risks, racial/ethnic groups, and geography can help direct effective prevention efforts on both sides of the border. We gathered sociodemographic, geographic, clinical, and viral sequence data from HIV infected individuals participating in ten studies in the San Diego-Tijuana border region. Phylogenetic and network analysis was performed to infer putative relationships between HIV sequences. Correlates of identified clusters were evaluated and spatiotemporal relationships were explored using Bayesian phylogeographic analysis. After quality filtering, 843 HIV sequences with associated demographic data and 263 background sequences from the region were analyzed, and 138 clusters were inferred (2-23 individuals). Overall, the rate of clustering did not differ by ethnicity, residence, or sex, but bisexuals were less likely to cluster than heterosexuals or men who have sex with men (p = 0.043), and individuals identifying as white (p ≤ 0.01) were more likely to cluster than other races. Clustering individuals were also 3.5 years younger than non-clustering individuals (p < 0.001). Although the sampled San Diego and Tijuana epidemics were phylogenetically compartmentalized, five clusters contained individuals residing on both sides of the border. This study sampled ~ 7% of HIV infected individuals in the border region, and although the sampled networks on each side of the border were largely separate, there was evidence of persistent bidirectional cross-border transmissions that linked risk groups, thus highlighting the importance of the border region as a "melting pot" of risk groups. NIH, VA, and Pendleton Foundation.

  8. The Neural Border: Induction, Specification and Maturation of the territory that generates Neural Crest cells.

    PubMed

    Pla, Patrick; Monsoro-Burq, Anne H

    2018-05-28

    The neural crest is induced at the edge between the neural plate and the nonneural ectoderm, in an area called the neural (plate) border, during gastrulation and neurulation. In recent years, many studies have explored how this domain is patterned, and how the neural crest is induced within this territory, that also participates to the prospective dorsal neural tube, the dorsalmost nonneural ectoderm, as well as placode derivatives in the anterior area. This review highlights the tissue interactions, the cell-cell signaling and the molecular mechanisms involved in this dynamic spatiotemporal patterning, resulting in the induction of the premigratory neural crest. Collectively, these studies allow building a complex neural border and early neural crest gene regulatory network, mostly composed by transcriptional regulations but also, more recently, including novel signaling interactions. Copyright © 2018. Published by Elsevier Inc.

  9. The border-to-border distribution method for analysis of cytoplasmic particles and organelles.

    PubMed

    Yacovone, Shalane K; Ornelles, David A; Lyles, Douglas S

    2016-02-01

    Comparing the distribution of cytoplasmic particles and organelles between different experimental conditions can be challenging due to the heterogeneous nature of cell morphologies. The border-to-border distribution method was created to enable the quantitative analysis of fluorescently labeled cytoplasmic particles and organelles of multiple cells from images obtained by confocal microscopy. The method consists of four steps: (1) imaging of fluorescently labeled cells, (2) division of the image of the cytoplasm into radial segments, (3) selection of segments of interest, and (4) population analysis of fluorescence intensities at the pixel level either as a function of distance along the selected radial segments or as a function of angle around an annulus. The method was validated using the well-characterized effect of brefeldin A (BFA) on the distribution of the vesicular stomatitis virus G protein, in which intensely labeled Golgi membranes are redistributed within the cytoplasm. Surprisingly, in untreated cells, the distribution of fluorescence in Golgi membrane-containing radial segments was similar to the distribution of fluorescence in other G protein-containing segments, indicating that the presence of Golgi membranes did not shift the distribution of G protein towards the nucleus compared to the distribution of G protein in other regions of the cell. Treatment with BFA caused only a slight shift in the distribution of the brightest G protein-containing segments which had a distribution similar to that in untreated cells. Instead, the major effect of BFA was to alter the annular distribution of G protein in the perinuclear region.

  10. Restoring effect of selenium on the molecular content, structure and fluidity of diabetic rat kidney brush border cell membrane.

    PubMed

    Gurbanov, Rafig; Bilgin, Mehmet; Severcan, Feride

    2016-04-01

    Diabetic kidney disease (DKD) is a dominant factor standing for kidney impairments during diabetes. In this study, attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy was used to disclose the diabetes-induced structural changes in the kidney and evaluate the effects of selenium on diabetes. The increase in the area of the olefinic band indicated increased amount of lipid peroxidation end products in diabetic kidney brush border cell membrane. Moreover, saturated lipid content of this cell membrane considerably diminished. DKD was found to disrupt lipid order and cause a decrease in membrane dynamics. However, the administration of selenium at low and medium doses was shown to improve these conditions by changing the lipid contents toward control values, restoring the ordered structure of the lipids and membrane dynamics. Curve-fitting and artificial neural network (ANN) analyses of secondary structures of proteins demonstrated a relative increase in α-helix and reduction in the β-sheet during diabetes in comparison to the control group, which were ameliorated following selenium treatment at low and medium doses. These findings were further confirmed by applying hierarchical cluster analysis (HCA) and principal component analysis (PCA). A clear separation of the experimental groups was obtained with high heterogeneity in the lipid and protein regions. These chemometric analyses showed that the low and medium doses of selenium-treated diabetic groups are successfully segregated from the diabetic group and clustered closer to the control. The study suggests that medium and, more predominantly, low-dose selenium treatment can be efficient in eliminating diabetes-induced structural alterations. Copyright © 2016 Elsevier B.V. All rights reserved

  11. Los Angeles Basin

    NASA Image and Video Library

    2009-06-29

    The Los Angeles Basin is bordered on the north by the San Gabriel Mountains. Other smaller basins are separated by smaller mountain ranges, like the Verdugo Hills, and the Santa Monica Mountains in this image from NASA Terra spacecraft.

  12. Humane Enforcement and Legal Protections for Separated Children Act

    THOMAS, 111th Congress

    Rep. Woolsey, Lynn C. [D-CA-6

    2009-07-31

    House - 09/14/2009 Referred to the Subcommittee on Immigration, Citizenship, Refugees, Border Security, and International Law. (All Actions) Tracker: This bill has the status IntroducedHere are the steps for Status of Legislation:

  13. Regional photochemical air quality modeling in the Mexico-US border area

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

    Mendoza, A.; Russell, A.G.; Mejia, G.M.

    1998-12-31

    The Mexico-United States border area has become an increasingly important region due to its commercial, industrial and urban growth. As a result, environmental concerns have risen. Treaties like the North American Free Trade Agreement (NAFTA) have further motivated the development of environmental impact assessment in the area. Of particular concern are air quality, and how the activities on both sides of the border contribute to its degradation. This paper presents results of applying a three-dimensional photochemical airshed model to study air pollution dynamics along the Mexico-United States border. In addition, studies were conducted to assess how size resolution impacts themore » model performance. The model performed within acceptable statistic limits using 12.5 x 12.5 km{sup 2} grid cells, and the benefits using finer grids were limited. Results were further used to assess the influence of grid-cell size on the modeling of control strategies, where coarser grids lead to significant loss of information.« less

  14. Post-repolarization refractoriness increases vulnerability to block and initiation of reentrant impulses in heterogeneous infarcted myocardium.

    PubMed

    Cabo, Candido

    2015-10-01

    Myocardial infarction causes remodeling of the tissue structure and the density and kinetics of several ion channels in the cell membrane. Heterogeneities in refractory period (ERP) have been shown to occur in the infarct border zone and have been proposed to lead to initiation of arrhythmias. The purpose of this study is to quantify the window of vulnerability (WV) to block and initiation of reentrant impulses in myocardium with ERP heterogeneities using computer simulations. We found that ERP transitions at the border between normal ventricular cells (NZ) with different ERPs are smooth, whereas ERP transitions between NZ and infarct border zone cells (IZ) are abrupt. The profile of the ERP transitions is a combination of electrotonic interaction between NZ and IZ cells and the characteristic post-repolarization refractoriness (PRR) of IZ cells. ERP heterogeneities between NZ and IZ cells are more vulnerable to block and initiation of reentrant impulses than ERP heterogeneities between NZ cells. The relationship between coupling intervals of premature impulses (V1V2) and coupling intervals between premature and first reentrant impulses (V2T1) at NZ/NZ and NZ/IZ borders is inverse (i.e. the longer the coupling intervals of premature impulses the shorter the coupling interval between the premature and first reentrant impulses); this is in contrast with the reported V1V2/V2T1 relationship measured during initiation of reentrant impulses in canine infarcted hearts which is direct. (1) ERP transitions at the NZ-IZ border are abrupt as a consequence of PRR; (2) PRR increases the vulnerability to block and initiation of reentrant impulses in heterogeneous myocardium; (3) V1V2/V2T1 relationships measured at ERP heterogeneities in the computer model and in experimental canine infarcts are not consistent. Therefore, it is likely that other mechanisms like micro and/or macro structural heterogeneities also contribute to initiation of reentrant impulses in infarcted hearts. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Alcohol Use Patterns and DSM-5 Alcohol Use Disorder on Both Sides of the US-Mexico Border

    PubMed Central

    Greenfield, Thomas K.; Ye, Yu; Lown, E. Anne; Cherpitel, Cheryl J.; Zemore, Sarah; Borges, Guilherme

    2017-01-01

    Background Alcohol consumption patterns on the US-Mexico border and their relationships with DSM-5 Alcohol Use Disorders (AUDs) have been understudied. Yet, effects of drinking by Mexican-origin individuals may differ between cities on vs. off the border both in the US and Mexico. We characterize prior 12-months drinking patterns, and examine their relationships with AUD, in border and off-border cities of Texas and adjacent Mexican states. Methods Data come from the US-Mexico Study of Alcohol and Related Conditions (UMSARC) involving 2,336 Mexican Americans in Texas and 2,460 Mexicans in bordering states of Nuevo Leon and Tamaulipas in Mexico. Drinking pattern was defined as an interaction between volume and maximum amount, or intensity (never vs. ever 5+/4+ [men/women], 8+, and 12+ drinks in a day). DSM-5 AUD was assessed using an adaptation of the Alcohol Section of the WHO Composite International Diagnostic Interview (CIDI) core. Separately by gender, five logistic regressions models controlling for age were estimated predicting symptoms in 2 or more AUD criteria domains from volume, heavy pattern and, successively, effects of country, and (by country) residing on vs. off the border, or in each of 3 cities/country. Results A segmentation analysis for Texas males based on rate of experiencing AUD generated several distinct volume-groups, each partitioned by an empirically selected maximum, helped identify a drinking pattern typology. In gender-stratified models of AUD rates using this typology, adjusting for age, significant volume and intensity effects were seen, more strongly in the US. Border vs. interior differences implied more AUD for given patterns at the border in the US and the reverse in Mexico, with some city differences also evident. Conclusion Drinking pattern analyses confirm that border proximity may affect drinking problems but in opposite directions in the US and Mexico, possibly related to economic and psychological stresses specific to respective communities. PMID:28207949

  16. [Population in the northern border area. Urban dynamism and binational interrelation].

    PubMed

    Ham Chande, R

    1988-01-01

    The 3300 km border between Mexico and the US constitutes the geopolitical separation between an underdeveloped country on the 1 hand and 1 of the most technologically and economically powerful countries in the world on the other. The border region is characterized by the contrasts on either side of the border and by the strong interrelation between both sides. Vast streams of persons, merchandise, money, services, communications, and cultural influences flow from 1 side to the other. The border region as a seat of population has a recent history. The border was defined in near current form only in the mid-19th century, when the expansionist tendencies of the US encountered a vast area of very sparse population. In 1900, the principal localities of the border zone had only about 39,000 inhabitants, of whom fewer than 5000 lived west of Ciudad Juarez. Between 1910-20, the population of the border region increased from 53,000 to 96,000 as a result of migrants fleeing the ravages of the revolution. The population of the border region was estimated at 3.826 million in 1988, resulting from rates of growth above Mexico's national average. Settlement in the area has depended on events and conditions in Mexico and on such US occurrences as Prohibition, the Great Depression, the 2nd World War, the Bracero program, and the Program of Border Industrialization. 82% of the border population lives in urban zones, partly because of lack of water. 80% of the urban population is concentrated in 6 cities, Juarez, Tijuana, Mexicali, Nuevo Laredo, Reynosa, and Matamoros. Much of the population of the 6 cities is composed of persons born elsewhere. The border area also has a large floating population of undocumented migrants in transit to or from the US. The high rates of urbanization and of binational interaction are reflected in demographic dynamics. In 1979, 71% of women in union in the border area vs 54% in the rest of Mexico had used contraception, and the infant mortality rate was 10% below the national average. In the 20-29 cohort there were 88 men per 100 women, reflecting male emigration and female inmigration. The rate of economic participation was higher than the national average, per capita production was greater, and unemployment rates were lower.

  17. Ceroid lipofuscinosis in the border collie dog: retinal lesions in an animal model of juvenile Batten disease.

    PubMed

    Taylor, R M; Farrow, B R

    1992-02-15

    Ceroid lipofuscinosis, an inherited disorder of lipopigment accumulation, was identified in a group of Border Collie dogs. The dogs developed mental, motor, and visual signs between age 15 and 22 months and progressed rapidly to severe neurological disease. The principal signs were blindness and gait and behavioural abnormalities with progressive dementia. Lipopigment accumulation was severe in neurones and glial cells of the central nervous system and was present in some visceral cells. Inclusions with variable ultrastructure were common in all cells of the retina, but the pigment accumulation did not damage the retinal architecture. The cytoplasmic inclusions were granular, sudanophilic, eosinophilic, and autofluorescent. Ultrastructural morphology varied, but fingerprint and curvilinear patterns predominated. The retinal lesions in the Border Collies were similar to those in English Setters with ceroid lipofuscinosis, but were much less severe than in juvenile human ceroid lipofuscinosis. This disorder bears a close resemblance to ceroid lipofuscinosis in English Setters and is another useful model for Batten's disease.

  18. Replication landscape of the human genome

    PubMed Central

    Petryk, Nataliya; Kahli, Malik; d'Aubenton-Carafa, Yves; Jaszczyszyn, Yan; Shen, Yimin; Silvain, Maud; Thermes, Claude; Chen, Chun-Long; Hyrien, Olivier

    2016-01-01

    Despite intense investigation, human replication origins and termini remain elusive. Existing data have shown strong discrepancies. Here we sequenced highly purified Okazaki fragments from two cell types and, for the first time, quantitated replication fork directionality and delineated initiation and termination zones genome-wide. Replication initiates stochastically, primarily within non-transcribed, broad (up to 150 kb) zones that often abut transcribed genes, and terminates dispersively between them. Replication fork progression is significantly co-oriented with the transcription. Initiation and termination zones are frequently contiguous, sometimes separated by regions of unidirectional replication. Initiation zones are enriched in open chromatin and enhancer marks, even when not flanked by genes, and often border ‘topologically associating domains' (TADs). Initiation zones are enriched in origin recognition complex (ORC)-binding sites and better align to origins previously mapped using bubble-trap than λ-exonuclease. This novel panorama of replication reveals how chromatin and transcription modulate the initiation process to create cell-type-specific replication programs. PMID:26751768

  19. An Apical MRCK-driven Morphogenetic Pathway Controls Epithelial Polarity

    PubMed Central

    Zihni, Ceniz; Vlassaks, Evi; Terry, Stephen; Carlton, Jeremy; Leung, Thomas King Chor; Olson, Michael; Pichaud, Franck; Balda, Maria Susana; Matter, Karl

    2017-01-01

    Polarized epithelia develop distinct cell surface domains, with the apical membrane acquiring characteristic morphological features such as microvilli. Cell polarization is driven by polarity determinants including the evolutionarily conserved partitioning defective (PAR) proteins that are separated into distinct cortical domains. PAR protein segregation is thought to be a consequence of asymmetric actomyosin contractions. The mechanism of activation of apically polarized actomyosin contractility is unknown. Here we show that the Cdc42 effector MRCK activates Myosin-II at the apical pole to segregate aPKC-Par6 from junctional Par3, defining the apical domain. Apically polarized MRCK-activated actomyosin contractility is reinforced by cooperation with aPKC-Par6 downregulating antagonistic RhoA-driven junctional actomyosin contractility, and drives polarization of cytosolic brush border determinants and apical morphogenesis. MRCK-activated polarized actomyosin contractility is required for apical differentiation and morphogenesis in vertebrate epithelia and Drosophila photoreceptors. Our results identify an apical origin of actomyosin-driven morphogenesis that couples cytoskeletal reorganization to PAR polarity signalling. PMID:28825699

  20. Jejunal brush border microvillous alterations in Giardia muris-infected mice: role of T lymphocytes and interleukin-6.

    PubMed

    Scott, K G; Logan, M R; Klammer, G M; Teoh, D A; Buret, A G

    2000-06-01

    Intestinal colonization with the protozoan Giardia causes diffuse brush border microvillous alterations and disaccharidase deficiencies, which in turn are responsible for intestinal malabsorption and maldigestion. The role of T cells and/or cytokines in the pathogenesis of Giardia-induced microvillous injury remains unclear. The aim of this study was to assess the role of T cells and interleukin-6 (IL-6) in the brush border pathophysiology of acute murine giardiasis in vivo. Athymic nude (nu(-)/nu(-)) CD-1 mice and isogenic immunocompetent (nu(+)/nu(+)) CD-1 mice (4 weeks old) received an axenic Giardia muris trophozoite inoculum or vehicle (control) via orogastric gavage. Weight gain and food intake were assessed daily. On day 6, segments of jejunum were assessed for parasite load, brush border ultrastructure, IL-6 content, maltase and sucrase activities, villus-crypt architecture, and intraepithelial lymphocyte (IEL) infiltration. Despite similar parasitic loads on day 6, infected immunocompetent animals, but not infected nude mice, showed a diffuse loss of brush border microvillous surface area, which was correlated with a significant reduction in maltase and sucrase activities and a decrease in jejunal IL-6 concentration. In both athymic control and infected mice, jejunal brush border surface area and disaccharidases were high, but levels of tissue IL-6 were low and comparable to the concentration measured in immunocompetent infected animals. In both immunocompetent and nude mice, infection caused a small but significant increase in the numbers of IELs. These findings suggest that the enterocyte brush border injury and malfunction seen in giardiasis is, at least in part, mediated by thymus-derived T lymphocytes and that suppressed jejunal IL-6 does not necessarily accompany microvillous shortening.

  1. Cell tracing reveals a dorsoventral lineage restriction plane in the mouse limb bud mesenchyme.

    PubMed

    Arques, Carlos G; Doohan, Roisin; Sharpe, James; Torres, Miguel

    2007-10-01

    Regionalization of embryonic fields into independent units of growth and patterning is a widespread strategy during metazoan development. Compartments represent a particular instance of this regionalization, in which unit coherence is maintained by cell lineage restriction between adjacent regions. Lineage compartments have been described during insect and vertebrate development. Two common characteristics of the compartments described so far are their occurrence in epithelial structures and the presence of signaling regions at compartment borders. Whereas Drosophila compartmental organization represents a background subdivision of embryonic fields that is not necessarily related to anatomical structures, vertebrate compartment borders described thus far coincide with, or anticipate, anatomical or cell-type discontinuities. Here, we describe a general method for clonal analysis in the mouse and use it to determine the topology of clone distribution along the three limb axes. We identify a lineage restriction boundary at the limb mesenchyme dorsoventral border that is unrelated to any anatomical discontinuity, and whose lineage restriction border is not obviously associated with any signaling center. This restriction is the first example in vertebrates of a mechanism of primordium subdivision unrelated to anatomical boundaries. Furthermore, this is the first lineage compartment described within a mesenchymal structure in any organism, suggesting that lineage restrictions are fundamental not only for epithelial structures, but also for mesenchymal field patterning. No lineage compartmentalization was found along the proximodistal or anteroposterior axes, indicating that patterning along these axes does not involve restriction of cell dispersion at specific axial positions.

  2. Robust nuclei segmentation in cyto-histopathological images using statistical level set approach with topology preserving constraint

    NASA Astrophysics Data System (ADS)

    Taheri, Shaghayegh; Fevens, Thomas; Bui, Tien D.

    2017-02-01

    Computerized assessments for diagnosis or malignancy grading of cyto-histopathological specimens have drawn increased attention in the field of digital pathology. Automatic segmentation of cell nuclei is a fundamental step in such automated systems. Despite considerable research, nuclei segmentation is still a challenging task due noise, nonuniform illumination, and most importantly, in 2D projection images, overlapping and touching nuclei. In most published approaches, nuclei refinement is a post-processing step after segmentation, which usually refers to the task of detaching the aggregated nuclei or merging the over-segmented nuclei. In this work, we present a novel segmentation technique which effectively addresses the problem of individually segmenting touching or overlapping cell nuclei during the segmentation process. The proposed framework is a region-based segmentation method, which consists of three major modules: i) the image is passed through a color deconvolution step to extract the desired stains; ii) then the generalized fast radial symmetry transform is applied to the image followed by non-maxima suppression to specify the initial seed points for nuclei, and their corresponding GFRS ellipses which are interpreted as the initial nuclei borders for segmentation; iii) finally, these nuclei border initial curves are evolved through the use of a statistical level-set approach along with topology preserving criteria for segmentation and separation of nuclei at the same time. The proposed method is evaluated using Hematoxylin and Eosin, and fluorescent stained images, performing qualitative and quantitative analysis, showing that the method outperforms thresholding and watershed segmentation approaches.

  3. Understanding the role of the cytoskeleton in wood formation in angiosperm trees: hybrid aspen (Populus tremula x P. tremuloides) as the model species.

    PubMed

    Chaffey, Nigel; Barlow, Peter; Sundberg, Björn

    2002-03-01

    The involvement of microfilaments and microtubules in the development of the radial and axial components of secondary xylem (wood) in hybrid aspen (Populus tremula L. x P. tremuloides Michx.) was studied by indirect immunofluorescent localization techniques. In addition to cambial cells, the differentiated cell types considered were early- and late-wood vessel elements, axial parenchyma, normal-wood fibers and gelatinous fibers, and contact and isolation ray cells. Microfilaments were rare in ray cambial cells, but were abundant and axially arranged in their derivatives once cell elongation had begun, and persisted in that orientation in mature ray cells. Microfilaments were axially arranged in fusiform cambial cells and persisted in that orientation in all xylem derivatives of those cells. Microtubules were randomly oriented in ray and fusiform cells of the cambial zone. Dense arrays of parallel-aligned microtubules were oriented near axially in the developing gelatinous fibers, but at a wide range of angles in normal-wood fibers. Ellipses of microfilaments were associated with pit development in fiber cells and isolation ray cells. Rings of co-localized microtubules and microfilaments were associated with developing inter-vessel bordered pits and vessel-contact ray cell contact pits, and, in the case of bordered pits, these rings decreased in diameter as the over-arching pit border increased in size. Although only microtubules were seen at the periphery of the perforation plate of vessel elements, a prominent meshwork of microfilaments overlaid the perforation plate itself. A consensus view of the roles of the cytoskeleton during wood formation in angiosperm trees is presented.

  4. Border Forces and Friction Control Epithelial Closure Dynamics

    PubMed Central

    Cochet-Escartin, Olivier; Ranft, Jonas; Silberzan, Pascal; Marcq, Philippe

    2014-01-01

    We study the closure dynamics of a large number of well-controlled circular apertures within an epithelial monolayer, where the collective cell migration responsible for epithelization is triggered by the removal of a spatial constraint rather than by scratching. Based on experimental observations, we propose a physical model that takes into account border forces, friction with the substrate, and tissue rheology. Border protrusive activity drives epithelization despite the presence of a contractile actomyosin cable at the periphery of the wound. The closure dynamics is quantified by an epithelization coefficient, defined as the ratio of protrusive stress to tissue-substrate friction, that allows classification of different phenotypes. The same analysis demonstrates a distinct signature for human cells bearing the oncogenic RasV12 mutation, demonstrating the potential of the approach to quantitatively characterize metastatic transformations. PMID:24411238

  5. Optical coherence tomography in guided surgery of GI cancer

    NASA Astrophysics Data System (ADS)

    Zagaynova, Elena V.; Abelevich, Alexander I.; Zagaynov, Vladimir E.; Gladkova, Natalia D.; Denisenko, Arkady N.; Feldchtein, Felix I.; Snopova, Ludmila B.; Kutis, Irina S.

    2005-04-01

    Optical Coherence Tomography (OCT) is a new high spatial resolution, real-time optical imaging modality, known from prior pilot studies for its high sensitivity to invasive cancer. We reported our results in an OCT feasibility study for accurate determination of the proximal border for esophageal carcinoma and the distal border for rectal carcinoma. The OCT study enrolled 19 patients with rectal adenocarcinoma and 24 patients with distal esophageal carcinoma (14 squamous cell carcinomas, 10 adenocarcinomas). During pre-surgery planning endoscopy we performed in vivo OCT imaging of the tumor border at four dial clock axes (12, 3, 6 and 9 o"clock). The OCT border then was marked by an electrocoagulator, or by a methylene blue tattoo. A cold biopsy (from the esophagus) was performed at visual and OCT borders and compared with visual and OCT readings. 27 post-surgery excised specimens were analyzed. OCT borders matched the histopathology in 94% cases in the rectum and 83.3% in the esophagus. In the cases of a mismatch between the OCT and histology borders, a deep tumor invasion occurred in the muscle layer (esophagus, rectum). Because of its high sensitivity to mucosal cancer, OCT can be used for pre-surgery planning and surgery guidance of the proximal border for esophageal carcinoma and the distal border for rectal carcinoma. However, deep invasion in the rectum or esophageal wall has to be controlled by alternative diagnostic modalities.

  6. Coral disease physiology: the impact of Acroporid white syndrome on Symbiodinium

    NASA Astrophysics Data System (ADS)

    Roff, G.; Kvennefors, E. C. E.; Ulstrup, K. E.; Fine, M.; Hoegh-Guldberg, O.

    2008-06-01

    Acroporid white syndrome, a disease-like syndrome from the Great Barrier Reef, results from degenerative host tissue at lesion borders. Tissue preceding lesion borders appears visually healthy, but it is currently unclear whether the endosymbiotic zooxanthellae ( Symbiodinium) are physiologically impacted. Compared to healthy colonies, this study found no significant differences in symbiont density, mitotic index or chlorophyll a content in tissue bordering (0 cm), and 8 cm away from white syndrome lesions. Using chlorophyll a fluorescence techniques, the border tissue did not appear to be photosynthetically compromised, and Symbiodinium extracted from this area were photosynthetically competent. Transmission electron microscopy revealed extensive degeneration of host tissues surrounding symbionts in affected areas, however, Symbiodinium cells were structurally intact with no sign of in situ degradation. Collectively, these results suggest that Symbiodinium at white syndrome lesion borders exist in a dynamic intra-cellular state during active host tissue loss, yet remain physiologically uncompromised.

  7. Proteomic analysis of the enterocyte brush border

    PubMed Central

    McConnell, Russell E.; Benesh, Andrew E.; Mao, Suli; Tabb, David L.

    2011-01-01

    The brush border domain at the apex of intestinal epithelial cells is the primary site of nutrient absorption in the intestinal tract and the primary surface of interaction with microbes that reside in the lumen. Because the brush border is positioned at such a critical physiological interface, we set out to create a comprehensive list of the proteins that reside in this domain using shotgun mass spectrometry. The resulting proteome contains 646 proteins with diverse functions. In addition to the expected collection of nutrient processing and transport components, we also identified molecules expected to function in the regulation of actin dynamics, membrane bending, and extracellular adhesion. These results provide a foundation for future studies aimed at defining the molecular mechanisms underpinning brush border assembly and function. PMID:21330445

  8. Astronomers Without Borders: A Global Astronomy Community

    NASA Astrophysics Data System (ADS)

    Simmons, M.

    2011-10-01

    Astronomers Without Borders (AWB) brings together astronomy enthusiasts of all types - amateur astronomers, educators, professionals and "armchair" astronomers for a variety of online and physicalworld programs. The AWB web site provides social networking and a base for online programs that engage people worldwide in astronomy activities that transcend geopolitical and cultural borders. There is universal interest in astronomy, which has been present in all cultures throughout recorded history. Astronomy is also among the most accessible of sciences with the natural laboratory of the sky being available to people worldwide. There are few other interests for which people widely separated geographically can engage in activities involving the same objects. AWB builds on those advantages to bring people together. AWB also provides a platform where projects can reach a global audience. AWB also provides unique opportunities for multidisciplinary collaboration in EPO programs. Several programs including The World at Night, Global Astronomy Month and others will be described along with lessons learned.

  9. A new double right border binary vector for producing marker-free transgenic plants

    PubMed Central

    2013-01-01

    Background Once a transgenic plant is developed, the selectable marker gene (SMG) becomes unnecessary in the plant. In fact, the continued presence of the SMG in the transgenic plant may cause unexpected pleiotropic effects as well as environmental or biosafety issues. Several methods for removal of SMGs that have been reported remain inaccessible due to protection by patents, while development of new ones is expensive and cost prohibitive. Here, we describe the development of a new vector for producing marker-free plants by simply adapting an ordinary binary vector to the double right border (DRB) vector design using conventional cloning procedures. Findings We developed the DRB vector pMarkfree5.0 by placing the bar gene (representing genes of interest) between two copies of T-DNA right border sequences. The β-glucuronidase (gus) and nptII genes (representing the selectable marker gene) were cloned next followed by one copy of the left border sequence. When tested in a model species (tobacco), this vector system enabled the generation of 55.6% kanamycin-resistant plants by Agrobacterium-mediated transformation. The frequency of cotransformation of the nptII and bar transgenes using the vector was 66.7%. Using the leaf bleach and Basta assays, we confirmed that the nptII and bar transgenes were coexpressed and segregated independently in the transgenic plants. This enable separation of the transgenes in plants cotransformed using pMarkfree5.0. Conclusions The results suggest that the DRB system developed here is a practical and effective approach for separation of gene(s) of interest from a SMG and production of SMG-free plants. Therefore this system could be instrumental in production of “clean” plants containing genes of agronomic importance. PMID:24207020

  10. Evidence for tension-based regulation of Drosophila MAL and SRF during invasive cell migration.

    PubMed

    Somogyi, Kálmán; Rørth, Pernille

    2004-07-01

    Cells migrating through a tissue exert force via their cytoskeleton and are themselves subject to tension, but the effects of physical forces on cell behavior in vivo are poorly understood. Border cell migration during Drosophila oogenesis is a useful model for invasive cell movement. We report that this migration requires the activity of the transcriptional factor serum response factor (SRF) and its cofactor MAL-D and present evidence that nuclear accumulation of MAL-D is induced by cell stretching. Border cells that cannot migrate lack nuclear MAL-D but can accumulate it if they are pulled by other migrating cells. Like mammalian MAL, MAL-D also responds to activated Diaphanous, which affects actin dynamics. MAL-D/SRF activity is required to build a robust actin cytoskeleton in the migrating cells; mutant cells break apart when initiating migration. Thus, tension-induced MAL-D activity may provide a feedback mechanism for enhancing cytoskeletal strength during invasive migration.

  11. Alcohol Use Patterns and DSM-5 Alcohol Use Disorder on Both Sides of the U.S.-Mexico Border.

    PubMed

    Greenfield, Thomas K; Ye, Yu; Lown, E Anne; Cherpitel, Cheryl J; Zemore, Sarah; Borges, Guilherme

    2017-04-01

    Alcohol consumption patterns on the U.S.-Mexico border and their relationships with DSM-5 alcohol use disorders (AUD) have been understudied. Yet, the effects of drinking by Mexican-origin individuals may differ between cities on versus off the border both in the United States and in Mexico. We characterize prior 12-month drinking patterns and examine their relationships with AUD, in border and off-border cities of Texas and adjacent Mexican states. Data come from the U.S.-Mexico Study of Alcohol and Related Conditions involving 2,336 Mexican Americans in Texas and 2,460 Mexicans in bordering states of Nuevo Leon and Tamaulipas in Mexico. Drinking pattern was defined as an interaction between volume and maximum amount, or intensity (never vs. ever 5+/4+ [men/women], 8+, and 12+ drinks in a day). DSM-5 AUD was assessed using an adaptation of the Alcohol Section of the World Health Organization Composite International Diagnostic Interview core. Separately by gender, 5 logistic regressions models controlling for age were estimated predicting symptoms in 2 or more AUD criteria domains from volume, heavy pattern and, successively, effects of country, and (by country) residing on vs. off the border, or in each of 3 cities/country. A segmentation analysis for Texas males based on rate of experiencing AUD generated several distinct volume groups, each partitioned by an empirically selected maximum, and helped identify a drinking-pattern typology. In gender-stratified models of AUD rates using this typology, adjusting for age, significant volume and intensity effects were seen, more strongly in the United States. Border versus interior differences implied more AUD for given patterns at the border in the United States and the reverse in Mexico, with some city differences also evident. Drinking-pattern analyses confirm that border proximity may affect drinking problems but in opposite directions in the United States and Mexico, possibly related to economic and psychological stresses specific to respective communities. Copyright © 2017 by the Research Society on Alcoholism.

  12. DREW-UCLA Breast Cancer Research and Training Program: Molecular/Cellular Pathogenesis Model

    DTIC Science & Technology

    2007-03-01

    system in bovine renal brush- border [13] and in a bovine renal epithelial cell line NBL -1 [14]. In order to highlight the substrate difference with an... NBL -1 expresses a broad specificity Na(+)-dependent neu- tral amino acid transport system (System B0) similar to that in bovine renal brush border

  13. Multi-frequency inversion-charge pumping for charge separation and mobility analysis in high-k/InGaAs metal-oxide-semiconductor field-effect transistors

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

    Djara, V.; Cherkaoui, K.; Negara, M. A.

    2015-11-28

    An alternative multi-frequency inversion-charge pumping (MFICP) technique was developed to directly separate the inversion charge density (N{sub inv}) from the trapped charge density in high-k/InGaAs metal-oxide-semiconductor field-effect transistors (MOSFETs). This approach relies on the fitting of the frequency response of border traps, obtained from inversion-charge pumping measurements performed over a wide range of frequencies at room temperature on a single MOSFET, using a modified charge trapping model. The obtained model yielded the capture time constant and density of border traps located at energy levels aligned with the InGaAs conduction band. Moreover, the combination of MFICP and pulsed I{sub d}-V{sub g}more » measurements enabled an accurate effective mobility vs N{sub inv} extraction and analysis. The data obtained using the MFICP approach are consistent with the most recent reports on high-k/InGaAs.« less

  14. Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three-dimensional organization and protein composition.

    PubMed

    Fey, E G; Wan, K M; Penman, S

    1984-06-01

    Madin-Darby canine kidney (MDCK) cells grow as differentiated, epithelial colonies that display tissue-like organization. We examined the structural elements underlying the colony morphology in situ using three consecutive extractions that produce well-defined fractions for both microscopy and biochemical analysis. First, soluble proteins and phospholipid were removed with Triton X-100 in a physiological buffer. The resulting skeletal framework retained nuclei, dense cytoplasmic filament networks, intercellular junctional complexes, and apical microvillar structures. Scanning electron microscopy showed that the apical cell morphology is largely unaltered by detergent extraction. Residual desmosomes, as can be seen in thin sections, were also well-preserved. The skeletal framework was visualized in three dimensions as an unembedded whole mount that revealed the filament networks that were masked in Epon-embedded thin sections of the same preparation. The topography of cytoskeletal filaments was relatively constant throughout the epithelial sheet, particularly across intercellular borders. This ordering of epithelial skeletal filaments across contiguous cell boundaries was in sharp contrast to the more independent organization of networks in autonomous cells such as fibroblasts. Further extraction removed the proteins of the salt-labile cytoskeleton and the chromatin as separate fractions, and left the nuclear matrix-intermediate filament (NM-IF) scaffold. The NM-IF contained only 5% of total cellular protein, but whole mount transmission electron microscopy and immunofluorescence showed that this scaffold was organized as in the intact epithelium. Immunoblots demonstrate that vimentin, cytokeratins, desmosomal proteins, and a 52,000-mol-wt nuclear matrix protein were found almost exclusively in the NM-IF scaffold. Vimentin was largely perinuclear while the cytokeratins were localized at the cell borders. The 52,000-mol-wt nuclear matrix protein was confined to the chromatin-depleted matrix and the desmosomal proteins were observed in punctate polygonal arrays at intercellular junctions. The filaments of the NM-IF were seen to be interconnected, via the desmosomes, over the entire epithelial colony. The differentiated epithelial morphology was reflected in both the cytoskeletal framework and the NM-IF scaffold.

  15. Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three-dimensional organization and protein composition

    PubMed Central

    1984-01-01

    Madin-Darby canine kidney (MDCK) cells grow as differentiated, epithelial colonies that display tissue-like organization. We examined the structural elements underlying the colony morphology in situ using three consecutive extractions that produce well-defined fractions for both microscopy and biochemical analysis. First, soluble proteins and phospholipid were removed with Triton X-100 in a physiological buffer. The resulting skeletal framework retained nuclei, dense cytoplasmic filament networks, intercellular junctional complexes, and apical microvillar structures. Scanning electron microscopy showed that the apical cell morphology is largely unaltered by detergent extraction. Residual desmosomes, as can be seen in thin sections, were also well- preserved. The skeletal framework was visualized in three dimensions as an unembedded whole mount that revealed the filament networks that were masked in Epon-embedded thin sections of the same preparation. The topography of cytoskeletal filaments was relatively constant throughout the epithelial sheet, particularly across intercellular borders. This ordering of epithelial skeletal filaments across contiguous cell boundaries was in sharp contrast to the more independent organization of networks in autonomous cells such as fibroblasts. Further extraction removed the proteins of the salt-labile cytoskeleton and the chromatin as separate fractions, and left the nuclear matrix-intermediate filament (NM-IF) scaffold. The NM-IF contained only 5% of total cellular protein, but whole mount transmission electron microscopy and immunofluorescence showed that this scaffold was organized as in the intact epithelium. Immunoblots demonstrate that vimentin, cytokeratins, desmosomal proteins, and a 52,000-mol-wt nuclear matrix protein were found almost exclusively in the NM-IF scaffold. Vimentin was largely perinuclear while the cytokeratins were localized at the cell borders. The 52,000-mol-wt nuclear matrix protein was confined to the chromatin- depleted matrix and the desmosomal proteins were observed in punctate polygonal arrays at intercellular junctions. The filaments of the NM-IF were seen to be interconnected, via the desmosomes, over the entire epithelial colony. The differentiated epithelial morphology was reflected in both the cytoskeletal framework and the NM-IF scaffold. PMID:6202700

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

    Herring, G. M.; Vaughan, Janet; Williamson, Margaret

    Characteristics of bone surfaces are discussed in relation to the uptake of bone-seeking isotopes. The alkaline earths are concentrated behind the osteoid border of active surfaces. Yttrium, americium, and plutonium are concentrated on quiescent and resorbing surfaces; traces only occur in and beneath the osteoid border. In view of evidence of mucoproteins at sites where the latter elemerts are found in concentration, a separation of the mucoproteins in cortical bone was undertaken. Mucosubstances have been isolated which give the same reaction with certain reagents as the bone surfaces in question. One of these, a sialoprotein, might be expected, in viewmore » of its acidic nature, to be capable of metal binding.« less

  17. 19 CFR 10.102 - Duty-free entries.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY... on a separate paper attached to the entry or withdrawal form filed by the Government agency or office... withdrawal. (1) Articles for military departments, subheading 9808.00.30, HTSUS. I certify that the...

  18. 19 CFR 10.102 - Duty-free entries.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY... on a separate paper attached to the entry or withdrawal form filed by the Government agency or office... withdrawal. (1) Articles for military departments, subheading 9808.00.30, HTSUS. I certify that the...

  19. Border preserving skin lesion segmentation

    NASA Astrophysics Data System (ADS)

    Kamali, Mostafa; Samei, Golnoosh

    2008-03-01

    Melanoma is a fatal cancer with a growing incident rate. However it could be cured if diagnosed in early stages. The first step in detecting melanoma is the separation of skin lesion from healthy skin. There are particular features associated with a malignant lesion whose successful detection relies upon accurately extracted borders. We propose a two step approach. First, we apply K-means clustering method (to 3D RGB space) that extracts relatively accurate borders. In the second step we perform an extra refining step for detecting the fading area around some lesions as accurately as possible. Our method has a number of novelties. Firstly as the clustering method is directly applied to the 3D color space, we do not overlook the dependencies between different color channels. In addition, it is capable of extracting fine lesion borders up to pixel level in spite of the difficulties associated with fading areas around the lesion. Performing clustering in different color spaces reveals that 3D RGB color space is preferred. The application of the proposed algorithm to an extensive data-base of skin lesions shows that its performance is superior to that of existing methods both in terms of accuracy and computational complexity.

  20. The Brassicaceae species Heliophila coronopifolia produces root border-like cells that protect the root tip and secrete defensin peptides.

    PubMed

    Weiller, Florent; Moore, John P; Young, Philip; Driouich, Azeddine; Vivier, Melané A

    2017-03-01

    Root border cells and border-like cells (BLCs), the latter originally described in Arabidopsis thaliana , have been described as cells released at the root tips of the species in which they occur. BLCs are thought to provide protection to root meristems similar to classical root border cells. In addition, four defensin peptides (Hc-AFP1-4) have previously been characterized from Heliophila coronopifolia , a South African semi-desert flower, and found to be strongly antifungal. This provided an opportunity to evaluate if the BLCs of H. coronopifolia indeed produce these defensins, which would provide evidence towards a defence role for BLCs. Fluorescence microscopy, using live-cell-imaging technology, was used to characterize the BLCs of H. coronopifolia . Quantitative real-time PCR (qRT-PCR) analysis and immunofluorescence microscopy was used to characterize these defensin peptides. BLCs originated at the root apical meristem and formed a protective sheath at the tip and along the sides as the root elongated in solid medium. BLCs have a cellulose-enriched cell wall, intact nuclei and are embedded in a layer of pectin-rich mucilage. Pectinase treatments led to the dissolution of the sheath and dissociation of the root BLCs. Hc-AFP1-4 genes were all expressed in root tissues, but Hc-AFP3 transcripts were the most abundant in these tissues as measured by qRT-PCR. A polyclonal antibody that was cross-reactive with all four defensins, and probably recognizing a general plant defensin epitope, was used in fluorescence microscopy analysis to examine the presence of the peptides in the root tip and BLCs. Data confirmed the peptides present in the root tip tissues, the mucilage sheath and the BLCs. This study provides a link between defensin peptides and BLCs, both embedded in a protective pectin mucilage sheath, during normal plant growth and development. The presence of the Hc-AFP3 defensin peptides in the BLCs suggests a role for these cells in root protection. © The Author 2016. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com

  1. The Brassicaceae species Heliophila coronopifolia produces root border-like cells that protect the root tip and secrete defensin peptides

    PubMed Central

    Weiller, Florent; Young, Philip; Driouich, Azeddine; Vivier, Melané A.

    2017-01-01

    Background and Aims Root border cells and border-like cells (BLCs), the latter originally described in Arabidopsis thaliana, have been described as cells released at the root tips of the species in which they occur. BLCs are thought to provide protection to root meristems similar to classical root border cells. In addition, four defensin peptides (Hc-AFP1–4) have previously been characterized from Heliophila coronopifolia, a South African semi-desert flower, and found to be strongly antifungal. This provided an opportunity to evaluate if the BLCs of H. coronopifolia indeed produce these defensins, which would provide evidence towards a defence role for BLCs. Methods Fluorescence microscopy, using live-cell-imaging technology, was used to characterize the BLCs of H. coronopifolia. Quantitative real-time PCR (qRT-PCR) analysis and immunofluorescence microscopy was used to characterize these defensin peptides. Key Results BLCs originated at the root apical meristem and formed a protective sheath at the tip and along the sides as the root elongated in solid medium. BLCs have a cellulose-enriched cell wall, intact nuclei and are embedded in a layer of pectin-rich mucilage. Pectinase treatments led to the dissolution of the sheath and dissociation of the root BLCs. Hc-AFP1–4 genes were all expressed in root tissues, but Hc-AFP3 transcripts were the most abundant in these tissues as measured by qRT-PCR. A polyclonal antibody that was cross-reactive with all four defensins, and probably recognizing a general plant defensin epitope, was used in fluorescence microscopy analysis to examine the presence of the peptides in the root tip and BLCs. Data confirmed the peptides present in the root tip tissues, the mucilage sheath and the BLCs. Conclusions This study provides a link between defensin peptides and BLCs, both embedded in a protective pectin mucilage sheath, during normal plant growth and development. The presence of the Hc-AFP3 defensin peptides in the BLCs suggests a role for these cells in root protection. PMID:27481828

  2. An amphioxus winged helix/forkhead gene, AmphiFoxD: insights into vertebrate neural crest evolution

    NASA Technical Reports Server (NTRS)

    Yu, Jr-Kai; Holland, Nicholas D.; Holland, Linda Z.

    2002-01-01

    During amphioxus development, the neural plate is bordered by cells expressing many genes with homologs involved in vertebrate neural crest induction. However, these amphioxus cells evidently lack additional genetic programs for the cell delaminations, migrations, and differentiations characterizing definitive vertebrate neural crest. We characterize an amphioxus winged helix/forkhead gene (AmphiFoxD) closely related to vertebrate FoxD genes. Phylogenetic analysis indicates that the AmphiFoxD is basal to vertebrate FoxD1, FoxD2, FoxD3, FoxD4, and FoxD5. One of these vertebrate genes (FoxD3) consistently marks neural crest during development. Early in amphioxus development, AmphiFoxD is expressed medially in the anterior neural plate as well as in axial (notochordal) and paraxial mesoderm; later, the gene is expressed in the somites, notochord, cerebral vesicle (diencephalon), and hindgut endoderm. However, there is never any expression in cells bordering the neural plate. We speculate that an AmphiFoxD homolog in the common ancestor of amphioxus and vertebrates was involved in histogenic processes in the mesoderm (evagination and delamination of the somites and notochord); then, in the early vertebrates, descendant paralogs of this gene began functioning in the presumptive neural crest bordering the neural plate to help make possible the delaminations and cell migrations that characterize definitive vertebrate neural crest. Copyright 2002 Wiley-Liss, Inc.

  3. Lamellipodin and the Scar/WAVE complex cooperate to promote cell migration in vivo

    PubMed Central

    Law, Ah-Lai; Vehlow, Anne; Kotini, Maria; Dodgson, Lauren; Soong, Daniel; Theveneau, Eric; Bodo, Cristian; Taylor, Eleanor; Navarro, Christel; Perera, Upamali; Michael, Magdalene; Dunn, Graham A.; Bennett, Daimark; Mayor, Roberto

    2013-01-01

    Cell migration is essential for development, but its deregulation causes metastasis. The Scar/WAVE complex is absolutely required for lamellipodia and is a key effector in cell migration, but its regulation in vivo is enigmatic. Lamellipodin (Lpd) controls lamellipodium formation through an unknown mechanism. Here, we report that Lpd directly binds active Rac, which regulates a direct interaction between Lpd and the Scar/WAVE complex via Abi. Consequently, Lpd controls lamellipodium size, cell migration speed, and persistence via Scar/WAVE in vitro. Moreover, Lpd knockout mice display defective pigmentation because fewer migrating neural crest-derived melanoblasts reach their target during development. Consistently, Lpd regulates mesenchymal neural crest cell migration cell autonomously in Xenopus laevis via the Scar/WAVE complex. Further, Lpd’s Drosophila melanogaster orthologue Pico binds Scar, and both regulate collective epithelial border cell migration. Pico also controls directed cell protrusions of border cell clusters in a Scar-dependent manner. Taken together, Lpd is an essential, evolutionary conserved regulator of the Scar/WAVE complex during cell migration in vivo. PMID:24247431

  4. Developmental and cell-specific expression of thyroid hormone transporters in the mouse cochlea.

    PubMed

    Sharlin, David S; Visser, Theo J; Forrest, Douglas

    2011-12-01

    Thyroid hormone is essential for the development of the cochlea and auditory function. Cochlear response tissues, which express thyroid hormone receptor β (encoded by Thrb), include the greater epithelial ridge and sensory epithelium residing inside the bony labyrinth. However, these response tissues lack direct blood flow, implying that mechanisms exist to shuttle hormone from the circulation to target tissues. Therefore, we investigated expression of candidate thyroid hormone transporters L-type amino acid transporter 1 (Lat1), monocarboxylate transporter (Mct)8, Mct10, and organic anion transporting polypeptide 1c1 (Oatp1c1) in mouse cochlear development by in situ hybridization and immunofluorescence analysis. L-type amino acid transporter 1 localized to cochlear blood vessels and transiently to sensory hair cells. Mct8 localized to the greater epithelial ridge, tympanic border cells underlying the sensory epithelium, spiral ligament fibrocytes, and spiral ganglion neurons, partly overlapping with the Thrb expression pattern. Mct10 was detected in a highly restricted pattern in the outer sulcus epithelium and weakly in tympanic border cells and hair cells. Organic anion transporting polypeptide 1c1 localized primarily to fibrocytes in vascularized tissues of the spiral limbus and spiral ligament and to tympanic border cells. Investigation of hypothyroid Tshr(-/-) mice showed that transporter expression was delayed consistent with retardation of cochlear tissue maturation but not with compensatory responses to hypothyroidism. The results demonstrate specific expression of thyroid hormone transporters in the cochlea and suggest that a network of thyroid hormone transport underlies cochlear development.

  5. Conserved gene regulatory module specifies lateral neural borders across bilaterians

    PubMed Central

    Li, Yongbin; Zhao, Di; Horie, Takeo; Chen, Geng; Bao, Hongcun; Chen, Siyu; Liu, Weihong; Horie, Ryoko; Liang, Tao; Dong, Biyu; Feng, Qianqian; Tao, Qinghua

    2017-01-01

    The lateral neural plate border (NPB), the neural part of the vertebrate neural border, is composed of central nervous system (CNS) progenitors and peripheral nervous system (PNS) progenitors. In invertebrates, PNS progenitors are also juxtaposed to the lateral boundary of the CNS. Whether there are conserved molecular mechanisms determining vertebrate and invertebrate lateral neural borders remains unclear. Using single-cell-resolution gene-expression profiling and genetic analysis, we present evidence that orthologs of the NPB specification module specify the invertebrate lateral neural border, which is composed of CNS and PNS progenitors. First, like in vertebrates, the conserved neuroectoderm lateral border specifier Msx/vab-15 specifies lateral neuroblasts in Caenorhabditis elegans. Second, orthologs of the vertebrate NPB specification module (Msx/vab-15, Pax3/7/pax-3, and Zic/ref-2) are significantly enriched in worm lateral neuroblasts. In addition, like in other bilaterians, the expression domain of Msx/vab-15 is more lateral than those of Pax3/7/pax-3 and Zic/ref-2 in C. elegans. Third, we show that Msx/vab-15 regulates the development of mechanosensory neurons derived from lateral neural progenitors in multiple invertebrate species, including C. elegans, Drosophila melanogaster, and Ciona intestinalis. We also identify a novel lateral neural border specifier, ZNF703/tlp-1, which functions synergistically with Msx/vab-15 in both C. elegans and Xenopus laevis. These data suggest a common origin of the molecular mechanism specifying lateral neural borders across bilaterians. PMID:28716930

  6. Conserved gene regulatory module specifies lateral neural borders across bilaterians.

    PubMed

    Li, Yongbin; Zhao, Di; Horie, Takeo; Chen, Geng; Bao, Hongcun; Chen, Siyu; Liu, Weihong; Horie, Ryoko; Liang, Tao; Dong, Biyu; Feng, Qianqian; Tao, Qinghua; Liu, Xiao

    2017-08-01

    The lateral neural plate border (NPB), the neural part of the vertebrate neural border, is composed of central nervous system (CNS) progenitors and peripheral nervous system (PNS) progenitors. In invertebrates, PNS progenitors are also juxtaposed to the lateral boundary of the CNS. Whether there are conserved molecular mechanisms determining vertebrate and invertebrate lateral neural borders remains unclear. Using single-cell-resolution gene-expression profiling and genetic analysis, we present evidence that orthologs of the NPB specification module specify the invertebrate lateral neural border, which is composed of CNS and PNS progenitors. First, like in vertebrates, the conserved neuroectoderm lateral border specifier Msx/vab-15 specifies lateral neuroblasts in Caenorhabditis elegans Second, orthologs of the vertebrate NPB specification module ( Msx/vab-15 , Pax3/7/pax-3 , and Zic/ref-2 ) are significantly enriched in worm lateral neuroblasts. In addition, like in other bilaterians, the expression domain of Msx/vab-15 is more lateral than those of Pax3/7/pax-3 and Zic/ref- 2 in C. elegans Third, we show that Msx/vab-15 regulates the development of mechanosensory neurons derived from lateral neural progenitors in multiple invertebrate species, including C. elegans , Drosophila melanogaster , and Ciona intestinalis We also identify a novel lateral neural border specifier, ZNF703/tlp-1 , which functions synergistically with Msx/vab- 15 in both C. elegans and Xenopus laevis These data suggest a common origin of the molecular mechanism specifying lateral neural borders across bilaterians.

  7. Drawing lines and borders: how the dehiscent fruit of Arabidopsis is patterned.

    PubMed

    Dinneny, José R; Yanofsky, Martin F

    2005-01-01

    The advent of fruits marked a key innovation in the evolution of flowering plants and helped generate a diverse array of mechanisms for seed dispersal. In the model plant, Arabidopsis thaliana, seed dispersal occurs through a process known as "pod-shatter" in which the fruit structure falls to pieces upon light mechanical pressures. This dispersal mechanism is dependent on the careful patterning of tissues in the fruit, which perform diverse functions that enable the fruit to open at maturation. Using the genetic power of Arabidopsis, many of the molecular components that help specify these tissues have been identified. Studies of the interactions among these genes have revealed a regulatory network that limits processes such as cell-cell separation and lignification to discreet regions of the fruit. Knowledge of these processes in a model fruit creates a foundation on which to build an understanding of the evolution of fruit form in other species and provides tools to engineer shatter-resistant seed pods to prevent crop loss in plants of agronomic importance such as canola. Copyright 2004 Wiley Periodicals, Inc.

  8. Canine Mesenchymal Stem Cell Potential and the Importance of Dog Breed: Implication for Cell-Based Therapies.

    PubMed

    Bertolo, Alessandro; Steffen, Frank; Malonzo-Marty, Cherry; Stoyanov, Jivko

    2015-01-01

    The study of canine bone marrow-derived mesenchymal stem cells (MSCs) has a prominent position in veterinary cell-based applications. Yet the plethora of breeds, their different life spans, and interbreed variations provide unclearness on what can be achieved specifically by such therapies. In this study, we compared a set of morphological, physiological, and genetic markers of MSCs derived from large dog breeds, namely, Border collie, German shepherd, Labrador, Malinois, Golden retriever, and Hovawart. We compared colony-forming units (CFUs) assay, population doubling time (PDT), senescence-associated β-galactosidase (SA-β-gal) activity, telomere length, and gene expression of MSCs, as well as the ability of cells to differentiate to osteogenic, adipogenic, and chondrogenic phenotypes. The influence of the culture media α-MEM, low-glucose DMEM, and high-glucose DMEM, used in cell isolation and expansion, was investigated in the presence and absence of basic fibroblast growth factor (bFGF). Initial cell yield was not affected by culturing medium, but MSCs expanded best in α-MEM supplemented with bFGF. After isolation, the number of MSCs was similar among breeds--as shown by equivalent CFUs--except in the Hovawart samples, which had fivefold less CFU. Telomere lengths were similar among breeds. MSCs divided actively only for 4 weeks in culture (PDT = ∼50 h/division), except Border collie cells divided for a longer time than cells from other groups. The percentage of senescent cells increased linearly in all breeds with time, with a faster rate in German shepherd, Labrador, and Golden retriever. Border collie cells underwent efficient osteogenic differentiation, Hovawart cells performed the best in chondrogenic differentiation, and Labrador cells in both, while German shepherd cells had the lower differentiation potential. MSCs from all breeds preserved the same adipogenic differentiation potential. In conclusion, despite variations, isolated MSCs can be expanded and differentiated in vitro, and all breeds are eligible for MSC-based therapies.

  9. [Calcium distribution in the central cell of lettuce (Lactuca sativa L.) before and after pollination].

    PubMed

    Qiu, Yi Lan; Liu, Ru Shi; Ye, Lv; Tian, Hui

    2008-02-01

    Potassium antimonite precipitation was used to locate calcium in the central cell of lettuce (Lactuca sativa L.) before and after pollination. At 3d before anthesis, two polar nuclei of central cell separately located at two polarity of the cell, and few calcium precipitates (ppts) appeared in the polar nuclei and cytoplasm, but some ppts in its small vacuoles. At 2d before anthesis, two polar nuclei moved toward the middle of the cell and fused to form a secondary nucleus, and the ppts evidently increased in the nucleus and cytoplasm. At 1d before anthesis, secondary nucleus again moved toward micropylar end and located near the egg to prepare for fertilization. Calcium precipitates were mainly accumulated in the secondary nucleus. After pollination and before fertilization, the distribution of calcium ppts was similar to that before pollination. At 4h after pollination, the central cell was fertilized, and calcium ppts evidently increased in the cell and numerous were accumulated in its nucleus and cytoplasm. At 6h after pollination, the primary endosperm nucleus completed its first division and formed two dissociate endosperm nuclei, and still many calcium precipitates appeared in the nucleus and cytoplasm. With endosperm development, calcium ppts decreased in the endosperm cell. At 1d after emasculated and without pollination, the secondary nucleus of the cell still bordered on the egg and some calcium ppts appeared in the secondary nucleus. The results indicated that the temporal and spatial changes of calcium in the central cell may play an important physiological role during the development of the central cell and endosperm.

  10. Dynamic signatures of quiet sun magnetic fields

    NASA Technical Reports Server (NTRS)

    Martin, S. F.

    1983-01-01

    The collision and disappearance of opposite polarity fields is observed most frequently at the borders of network cells. Due to observational limitations, the frequency, magnitude, and spatial distribution of magnetic flux loss have not yet been quantitatively determined at the borders or within the interiors of the cells. However, in agreement with published hypotheses of other authors, the disapperance of magnetic flux is speculated to be a consequence of either gradual or rapid magnetic reconnection which could be the means of converting magnetic energy into the kinetic, thermal, and nonthermal sources of energy for microflares, spicules, the solar wind, and the heating of the solar corona.

  11. 77 FR 17439 - Crystalline Silicon Photovoltaic Cells, Whether or Not Assembled Into Modules, From the People's...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-26

    ... silicon photovoltaic cells, whether or not assembled into modules (solar cells) from the People's Republic... History The Department initiated a countervailing duty (CVD) investigation of solar cells from the PRC on.... Customs and Border Protection (CBP) entry data for U.S. imports of solar cells from the PRC for the period...

  12. Effect of cephalosporins on organic ion transport in renal membrane vesicles from rat and rabbit kidney cortex.

    PubMed

    Williams, P D; Hitchcock, M J; Hottendorf, G H

    1985-03-01

    The effects of cephaloridine and cephalothin on prototypical organic anion (p-aminohippurate, PAH) and cation (N-methylnicotinamide, NMN) transport were observed in brush border and basolateral membrane vesicles prepared from rat and rabbit renal cortex. The cephalosporins interacted with both the cationic and anionic transport systems. Cephalothin inhibited PAH transport in basolateral and brush border membrane in both rats and rabbits. Cephaloridine on the other hand inhibited PAH and NMN transport across rabbit basolateral membranes while it showed a lack of interaction with transport systems in rat basolateral membranes. Conversely, cephaloridine inhibited brush border transport of PAH and NMN in the rat but not in the rabbit. These results provide indirect evidence that cephalothin may be secreted across the renal tubule cell in rats and rabbits while cephaloridine may not accumulate in the rat kidney and becomes trapped in rabbit renal tubule cells. The differences in transport effects observed may explain intra- and interspecies differences in susceptibility to cephalosporin nephrotoxicity.

  13. Aluminum-induced cell death of barley-root border cells is correlated with peroxidase- and oxalate oxidase-mediated hydrogen peroxide production.

    PubMed

    Tamás, L; Budíková, S; Huttová, J; Mistrík, I; Simonovicová, M; Siroká, B

    2005-06-01

    The function of root border cells (RBC) during aluminum (Al) stress and the involvement of oxalate oxidase, peroxidase and H(2)O(2) generation in Al toxicity were studied in barley roots. Our results suggest that RBC effectively protect the barley root tip from Al relative to the situation in roots cultivated in hydroponics where RBC are not sustained in the area surrounding the root tip. The removal of RBC from Al-treated roots increased root growth inhibition, Al and Evans blue uptake, inhibition of RBC production, the level of dead RBC, peroxidase and oxalate oxidase activity and the production of H(2)O(2). Our results suggest that even though RBC actively produce active oxygen species during Al stress, their role in the protection of root tips against Al toxicity is to chelate Al in their dead cell body.

  14. Escuela de la Frontera (Border School)

    ERIC Educational Resources Information Center

    Kilman, Carrie

    2006-01-01

    This article features Lourdes Catholic School, a small Catholic school where students grapple with life and learning in the shadow of two nations. The school, which is governed by the Minim Daughters of Mary Immaculate, is located in Nogales, a desert mountain town straddling the line that separates Arizona from the Mexican state of Sonora. Though…

  15. Embryonic Stem Cell Specific “Master” Replication Origins at the Heart of the Loss of Pluripotency

    PubMed Central

    Julienne, Hanna; Audit, Benjamin; Arneodo, Alain

    2015-01-01

    Epigenetic regulation of the replication program during mammalian cell differentiation remains poorly understood. We performed an integrative analysis of eleven genome-wide epigenetic profiles at 100 kb resolution of Mean Replication Timing (MRT) data in six human cell lines. Compared to the organization in four chromatin states shared by the five somatic cell lines, embryonic stem cell (ESC) line H1 displays (i) a gene-poor but highly dynamic chromatin state (EC4) associated to histone variant H2AZ rather than a HP1-associated heterochromatin state (C4) and (ii) a mid-S accessible chromatin state with bivalent gene marks instead of a polycomb-repressed heterochromatin state. Plastic MRT regions (≲ 20% of the genome) are predominantly localized at the borders of U-shaped timing domains. Whereas somatic-specific U-domain borders are gene-dense GC-rich regions, 31.6% of H1-specific U-domain borders are early EC4 regions enriched in pluripotency transcription factors NANOG and OCT4 despite being GC poor and gene deserts. Silencing of these ESC-specific “master” replication initiation zones during differentiation corresponds to a loss of H2AZ and an enrichment in H3K9me3 mark characteristic of late replicating C4 heterochromatin. These results shed a new light on the epigenetically regulated global chromatin reorganization that underlies the loss of pluripotency and lineage commitment. PMID:25658386

  16. SPACELAB (SL)-1 - INSIGNIAS

    NASA Image and Video Library

    1981-10-01

    S82-31408 (May 1983) --- The Spacelab 2 emblem is a symbolic representation of the scientific objectives of the mission. The emblem is in the shape of a triangular shield with convexly curved edges. Across the top of a black out border are the words ?SPACELAB 2?. Within the black border is a sky blue border carryhing the words: ?ASTRONOMY?, ON TOP? ?PHYSICS?, on the left; and ?BIOLOGY?, on the right. Within the blue border is a schematic view of the sun, the earth, and the orbiter with Spacelab 2. The sun appears in the upper right background as a white disc surrounded by six concentric rings ranging grom bright yellow near the disc through yellow-red to a dark red out ring. A sector of the earth with blue ocean and a black portion of North America is in the upper left corner. The black and white Orbiter is seen from directly overhead in the foreground, the right side illuminated by the sun, the left side in shadow. Although the payload bay doors are not open, the Spacelab 2 payload is seen as if the doors were open. In black on white are seen the three pallets, and the separately mounted cosmic ray experiment at the aft end of the bay.

  17. The Border Ranges fault system in Glacier Bay National Park, Alaska: Evidence for major early Cenozoic dextral strike-slip motion

    USGS Publications Warehouse

    Smart, K.J.; Pavlis, T.L.; Sisson, V.B.; Roeske, S.M.; Snee, L.W.

    1996-01-01

    The Border Ranges fault system of southern Alaska, the fundamental break between the arc basement and the forearc accretionary complex, is the boundary between the Peninsular-Alexander-Wrangellia terrane and the Chugach terrane. The fault system separates crystalline rocks of the Alexander terrane from metamorphic rocks of the Chugach terrane in Glacier Bay National Park. Mylonitic rocks in the zone record abundant evidence for dextral strike-slip motion along north-northwest-striking subvertical surfaces. Geochronologic data together with regional correlations of Chugach terrane rocks involved in the deformation constrain this movement between latest Cretaceous and Early Eocene (???50 Ma). These findings are in agreement with studies to the northwest and southeast along the Border Ranges fault system which show dextral strike-slip motion occurring between 58 and 50 Ma. Correlations between Glacier Bay plutons and rocks of similar ages elsewhere along the Border Ranges fault system suggest that as much as 700 km of dextral motion may have been accommodated by this structure. These observations are consistent with oblique convergence of the Kula plate during early Cenozoic and forearc slivering above an ancient subduction zone following late Mesozoic accretion of the Peninsular-Alexander-Wrangellia terrane to North America.

  18. Local pH domains regulate NHE3-mediated Na+ reabsorption in the renal proximal tubule

    PubMed Central

    Burford, James L.; McDonough, Alicia A.; Holstein-Rathlou, Niels-Henrik; Peti-Peterdi, Janos

    2014-01-01

    The proximal tubule Na+/H+ exchanger 3 (NHE3), located in the apical dense microvilli (brush border), plays a major role in the reabsorption of NaCl and water in the renal proximal tubule. In response to a rise in blood pressure NHE3 redistributes in the plane of the plasma membrane to the base of the brush border, where NHE3 activity is reduced. This NHE3 redistribution is assumed to provoke pressure natriuresis; however, it is unclear how NHE3 redistribution per se reduces NHE3 activity. To investigate if the distribution of NHE3 in the brush border can change the reabsorption rate, we constructed a spatiotemporal mathematical model of NHE3-mediated Na+ reabsorption across a proximal tubule cell and compared the model results with in vivo experiments in rats. The model predicts that when NHE3 is localized exclusively at the base of the brush border, it creates local pH microdomains that reduce NHE3 activity by >30%. We tested the model's prediction experimentally: the rat kidney cortex was loaded with the pH-sensitive fluorescent dye BCECF, and cells of the proximal tubule were imaged in vivo using confocal fluorescence microscopy before and after an increase of blood pressure by ∼50 mmHg. The experimental results supported the model by demonstrating that a rise of blood pressure induces the development of pH microdomains near the bottom of the brush border. These local changes in pH reduce NHE3 activity, which may explain the pressure natriuresis response to NHE3 redistribution. PMID:25298526

  19. Developmental and Cell-Specific Expression of Thyroid Hormone Transporters in the Mouse Cochlea

    PubMed Central

    Sharlin, David S.; Visser, Theo J.

    2011-01-01

    Thyroid hormone is essential for the development of the cochlea and auditory function. Cochlear response tissues, which express thyroid hormone receptor β (encoded by Thrb), include the greater epithelial ridge and sensory epithelium residing inside the bony labyrinth. However, these response tissues lack direct blood flow, implying that mechanisms exist to shuttle hormone from the circulation to target tissues. Therefore, we investigated expression of candidate thyroid hormone transporters L-type amino acid transporter 1 (Lat1), monocarboxylate transporter (Mct)8, Mct10, and organic anion transporting polypeptide 1c1 (Oatp1c1) in mouse cochlear development by in situ hybridization and immunofluorescence analysis. L-type amino acid transporter 1 localized to cochlear blood vessels and transiently to sensory hair cells. Mct8 localized to the greater epithelial ridge, tympanic border cells underlying the sensory epithelium, spiral ligament fibrocytes, and spiral ganglion neurons, partly overlapping with the Thrb expression pattern. Mct10 was detected in a highly restricted pattern in the outer sulcus epithelium and weakly in tympanic border cells and hair cells. Organic anion transporting polypeptide 1c1 localized primarily to fibrocytes in vascularized tissues of the spiral limbus and spiral ligament and to tympanic border cells. Investigation of hypothyroid Tshr−/− mice showed that transporter expression was delayed consistent with retardation of cochlear tissue maturation but not with compensatory responses to hypothyroidism. The results demonstrate specific expression of thyroid hormone transporters in the cochlea and suggest that a network of thyroid hormone transport underlies cochlear development. PMID:21878515

  20. Phenotypic Tfh development promoted by CXCR5-controlled re-localization and IL-6 from radiation-resistant cells.

    PubMed

    Chen, Xin; Ma, Weiwei; Zhang, Tingxin; Wu, Longyan; Qi, Hai

    2015-11-01

    How follicular T-helper (Tfh) cells develop is incompletely understood. We find that, upon antigen exposure in vivo, both naïve and antigen-experienced T cells sequentially upregulate CXCR5 and Bcl6 within the first 24 h, relocate to the T-B border, and give rise to phenotypic Bcl6(+)CXCR5(+) Tfh cells before the first cell division. CXCR5 upregulation is more dependent on ICOS costimulation than that of Bcl6, and early Bcl6 induction requires T-cell expression of CXCR5 and, presumably, relocation toward the follicle. This early and rapid upregulation of CXCR5 and Bcl6 depends on IL-6 produced by radiation-resistant cells. These results suggest that a Bcl6(hi)CXCR5(hi) phenotype does not automatically define a Tfh lineage but might reflect a state of antigen exposure and non-commitment to terminal effector fates and that niches in the T-B border and/or the follicle are important for optimal Bcl6 induction and maintenance.

  1. Mechanical analysis of the strains generated by water tension in plant stems. Part I: stress transmission from the water to the cell walls.

    PubMed

    Alméras, Tancrède; Gril, Joseph

    2007-11-01

    Plant tissues shrink and swell in response to changes in water pressure. These strains can be easily measured, e.g., at the surface of tree stems, to obtain indirect information about plant water status and other physiological parameters. We developed a mechanical model to clarify how water pressure is transmitted to cell walls and causes shrinkage of plant tissues, particularly in the case of thick-walled cells such as wood fibers. Our analysis shows that the stress inside the fiber cell walls is lower than the water tension. The difference is accounted for by a stress transmission factor that depends on two main effects. The first effect is the dilution of the stress through the cell wall, because water acts at the lumen border and is transmitted to the outer border of the cell, which has a larger circumference. The second effect is the partial conversion of radial stress into tangential stress. Both effects are quantified as functions of parameters of the cell wall structure and its mechanical properties.

  2. Differing strategies of patterning of follicular cells in higher and lower brachycerans (Diptera: Brachycera).

    PubMed

    Tworzydlo, Waclaw; Jablonska, Anna; Kisiel, Elzbieta; Bilinski, Szczepan M

    2005-10-01

    In all higher dipterans (Brachycera), including the fruitfly, Drosophila melanogaster, each egg chamber (ovarian follicle) consists of a group (clone) of germ cells (one oocyte and 15 accompanying nurse cells) that is surrounded by a layer of somatic mesodermal follicular cells (FCs). As oogenesis progresses the initially uniform FCs diversify into several morphologically and functionally distinct subpopulations. In D. melanogaster some of these subpopulations, e.g., border, centripetal, and dorsolateral cells, undertake coordinated migration or rearrangement over the surface of the germ cells. During the final stages of oogenesis these subpopulations participate in the formation of a complex, regionally specialized eggshell. In representatives of lower brachycerans (Orthorrhapha), only FCs that undertake active, directed migration are the border cells. These cells originate at the anterior pole of the ovarian follicle and migrate between the nurse cells to the anterior pole of the oocyte. Reduced motility of FCs in lower brachycerans results in the absence of certain FC subpopulations in their egg chambers and subsequent simplicity of their eggshells. We found that the lack of some FC subpopulations coincided with the appearance of lamellipodium-like protrusions of the oocyte. These protrusions penetrated between the apposing membranes of nurse and FCs and partially enveloped the nurse cell compartment. Analysis of whole-mount preparations stained with rhodamine-conjugated phalloidin revealed that the protrusions contained microfilaments and that their tips were equipped with actin-rich filopodium-like processes. We also found that in some lower brachycerans (representatives of the family Rhagionidae), the FCs located at the posterior pole of the oocyte, became enlarged and morphologically similar to the anterior border cells. These findings indicate that in higher dipterans the processes leading to the formation of a functional egg are variable and often markedly different from those in the model organism, D. melanogaster. genesis 43:49-58, 2005. (c) 2005 Wiley-Liss, Inc.

  3. The Transient Intermediate Plexiform Layer, a Plexiform Layer-like Structure Temporarily Existing in the Inner Nuclear Layer in Developing Rat Retina.

    PubMed

    Park, Hyung Wook; Kim, Hong-Lim; Park, Yong Soo; Kim, In-Beom

    2018-02-01

    The retina is a highly specialised part of the brain responsible for visual processing. It is well-laminated; three layers containing five different types of neurons are compartmentalised by two synaptic layers. Among the retinal layers, the inner nuclear layer (INL) is composed of horizontal, bipolar, and amacrine cell types. Bipolar cells form one sublayer in the distal half of the IPL, while amacrine cells form another sublayer in the proximal half, without any border-like structure. Here, we report that a plexiform layer-like structure exists temporarily in the border between the bipolar and amacrine sublayers in the INL in the rat retina during retinal development. This transient intermediate plexiform layer (TIPL) appeared at postnatal day (PD) 7 and then disappeared around PD 12. Most apoptotic cells in the INL were found near the TIPL. These results suggest that the TIPL may contribute to the formation of sublayers and the cell number limit in the INL.

  4. Does C₁ fracture displacement correlate with transverse ligament integrity?

    PubMed

    Radcliff, Kristen E; Sonagli, Marcos A; Rodrigues, Luciano M; Sidhu, Gursukhman S; Albert, Todd J; Vaccaro, Alexander R

    2013-05-01

    The Rule of Spence states that displacement of the C₁ lateral masses by >6.9-8.1 mm suggests loss of transverse ligament integrity. The purpose of this study was to establish the thresholds of C₁ displacement on CT scans that correspond to transverse ligament disruption. Over four years, consecutive patients with acute C₁ fractures with at least three fracture lines were analyzed. CT measurements and MRI were assessed by blinded observers for bony displacement in the axial (internal and external lateral mass separation), coronal and sagittal planes and transverse ligament integrity. Eighteen patients were studied. Mean CT bony measurements were as follows: internal border lateral mass separation (ILM) 23.3 ± 3.4 mm, external border lateral mass separation (ELM) 50.3 ± 4.3 mm, total C₁ lateral mass overhang over the C₂ superior process (LMO) 5.4 ± 1.3 mm. Twelve patients were identified as having intact transverse ligament and six had transverse ligament disruption. There was no difference in mean normalized ILM, ELM, or LMO between patients with or without transverse ligament integrity (P > 0.05). There was no correlation between bony displacement and transverse ligament integrity. CT scans post-injury may not show the position of maximal displacement. If there is clinical concern about a possible transverse ligament injury, MRI should be performed. © 2013 Chinese Orthopaedic Association and Wiley Publishing Asia Pty Ltd.

  5. Bathrooms without Borders: Transgender Students Argue Separate Is Not Equal

    ERIC Educational Resources Information Center

    Watkins, Paul J.; Moreno, Edward

    2017-01-01

    Title IX guidelines governing equal access to collide with state legislation around gender identity and multiple use bathrooms and changing facilities. This policy review of literature argues for a stronger voice from Washington, protecting the rights of all students to feel safe using private spaces at school. The many court rulings offer a fair…

  6. New state record for Culiseta impatiens in Maryland.

    PubMed

    Pagac, B B; Harlan, H J; Doran, S D; Brosnihan, M A

    1992-06-01

    Culiseta impatiens is reported for the first time from Maryland. Collections from the southwestern border of Fort George G. Meade extend the range of this species over 400 km farther south than previous records. Multiple collections from 4 separate traps during 2 years indicate a population of Cs. impatiens is probably established in the collection area.

  7. Evaluating the compatibility of American and Mexican national forest inventory data

    Treesearch

    Todd A. Schroeder; Sean P. Healey; Gretchen G. Moisen

    2012-01-01

    The international border region between the United States and Mexico represents a point of discontinuity in forest policy, land use management and resource utilization practices. These differences along with physical barriers which separate the two countries can interact to alter the structure and functioning of forest vegetation. One valuable source of information for...

  8. Teaching Students to Write across a Border: A Writing Curriculum for Inner-City College Access Programs

    ERIC Educational Resources Information Center

    Dobbs, Jennifer Kwon

    2012-01-01

    This article explores the development of the Summer Tools, Information, Motivation, and Education (SummerTIME) Writing Program, the only program of its kind in Los Angeles that conducts self-assessment. The author describes the geographical and political boundaries separating inner-city Los Angeles high school graduates from higher education,…

  9. Historically Black Land-Grant Institutions and the Development of Agriculture and Home Economics, 1890-1990.

    ERIC Educational Resources Information Center

    Neyland, Leedell W.

    Since 1890, historically black land-grant colleges and universities have delivered quality teaching, research, and extension service primarily to black people in Southern and border states. The Second Morrill Act of 1890 required that all land-grant funds be equitably divided in states that maintained separate schools for races. Tuskeegee…

  10. Luminance, Colour, Viewpoint and Border Enhanced Disparity Energy Model

    PubMed Central

    Martins, Jaime A.; Rodrigues, João M. F.; du Buf, Hans

    2015-01-01

    The visual cortex is able to extract disparity information through the use of binocular cells. This process is reflected by the Disparity Energy Model, which describes the role and functioning of simple and complex binocular neuron populations, and how they are able to extract disparity. This model uses explicit cell parameters to mathematically determine preferred cell disparities, like spatial frequencies, orientations, binocular phases and receptive field positions. However, the brain cannot access such explicit cell parameters; it must rely on cell responses. In this article, we implemented a trained binocular neuronal population, which encodes disparity information implicitly. This allows the population to learn how to decode disparities, in a similar way to how our visual system could have developed this ability during evolution. At the same time, responses of monocular simple and complex cells can also encode line and edge information, which is useful for refining disparities at object borders. The brain should then be able, starting from a low-level disparity draft, to integrate all information, including colour and viewpoint perspective, in order to propagate better estimates to higher cortical areas. PMID:26107954

  11. A Sharp Cadherin-6 Gene Expression Boundary in the Developing Mouse Cortical Plate Demarcates the Future Functional Areal Border

    PubMed Central

    Terakawa, Youhei W.; Inoue, Yukiko U.; Asami, Junko; Hoshino, Mikio; Inoue, Takayoshi

    2013-01-01

    The mammalian cerebral cortex can be tangentially subdivided into tens of functional areas with distinct cyto-architectures and neural circuitries; however, it remains elusive how these areal borders are genetically elaborated during development. Here we establish original bacterial artificial chromosome transgenic mouse lines that specifically recapitulate cadherin-6 (Cdh6) mRNA expression profiles in the layer IV of the somatosensory cortex and by detailing their cortical development, we show that a sharp Cdh6 gene expression boundary is formed at a mediolateral coordinate along the cortical layer IV as early as the postnatal day 5 (P5). By further applying mouse genetics that allows rigid cell fate tracing with CreERT2 expression, it is demonstrated that the Cdh6 gene expression boundary set at around P4 eventually demarcates the areal border between the somatosensory barrel and limb field at P20. In the P6 cortical cell pellet culture system, neurons with Cdh6 expression preferentially form aggregates in a manner dependent on Ca2+ and electroporation-based Cdh6 overexpression limited to the postnatal stages perturbs area-specific cell organization in the barrel field. These results suggest that Cdh6 expression in the nascent cortical plate may serve solidification of the protomap for cortical functional areas. PMID:22875867

  12. Mechanisms of atrial tachyarrhythmias associated with coronary artery occlusion in a chronic canine model.

    PubMed

    Nishida, Kunihiro; Qi, Xiao Yan; Wakili, Reza; Comtois, Philippe; Chartier, Denis; Harada, Masahide; Iwasaki, Yu-ki; Romeo, Philippe; Maguy, Ange; Dobrev, Dobromir; Michael, Georghia; Talajic, Mario; Nattel, Stanley

    2011-01-18

    Coronary artery disease predisposes to atrial fibrillation (AF), but the effects of chronic atrial ischemia/infarction on AF-related substrates are unknown. Regional right atrial myocardial infarction (MI) was created in 40 dogs by ligating an artery that supplies the right atrial free wall and not the ventricles; 35 sham dogs with the same artery isolated but not ligated were controls. Dogs were observed 8 days after MI and subjected to open-chest study, in vitro optical mapping, and/or cell isolation for patch-clamp and Ca(2+) imaging on day 8. Holter ECGs showed more spontaneous atrial ectopy in MI dogs (eg, 662±281 on day 7 versus 34±25 ectopic complexes per day at baseline; 52±21 versus 1±1 atrial tachycardia episodes per day). Triggered activity was increased in MI border zone cells, which had faster decay of caffeine-evoked Ca(2+) transients and enhanced (by ≈73%) Na(+)-Ca(2+) exchange current. Spontaneous Ca(2+) sparks (confocal microscopy) occurred under β-adrenergic stimulation in more MI dog cells (66±9%) than in control cells (29±4%; P<0.01). Burst pacing induced long-lasting AF in MI dogs (1146±259 versus 30±14 seconds in shams). Increased border zone conduction heterogeneity was confirmed by both bipolar electrode mapping in vivo and optical mapping. Optical mapping demonstrated stable border zone reentry in all 9 MI preparations but in none of 6 shams. Border zone tissue showed increased fibrous tissue content. Chronic atrial ischemia/infarction creates substrates for both spontaneous ectopy (Ca(2+)-release events, increased Na(+)-Ca(2+) exchange current) and sustained reentry (conduction abnormalities that anchor reentry). Thus, chronic atrial infarction in dogs promotes both AF triggers and the substrate for AF maintenance. These results provide novel insights into potential AF mechanisms in patients with coronary artery disease.

  13. Mineralogy and origin of coarse-grained segregations in the pyrometallurgical Zn-Pb slags from Katowice-Wełnowiec (Poland)

    NASA Astrophysics Data System (ADS)

    Warchulski, R.; Gawęda, A.; Janeczek, J.; Kądziołka-Gaweł, M.

    2016-10-01

    The unique among pyrometallurgical slags, coarse-grained (up to 2.5 cm) segregations (up to 40 cm long) rimmed by "aplitic" border zones occur within holocrystalline historical Zn-smelting slag in Katowice, S Poland. Slag surrounding the segregations consists of olivine, spinel series, melilite, clinopyroxene, leucite, nepheline and sulphides. Ca-olivines, kalsilite and mica compositionally similar to oxykinoshitalite occur in border zones in addition to olivine, spinel series and melilite. Miarolitic and massive pegmatite-like segregations are built of subhedral crystals of melilite, leucite, spinel series, clinopyroxene and hematite. Melilite, clinopyroxenes and spinels in the segregations are enriched in Zn relatively to original slag and to fine-grained border zones. The segregations originated as a result of crystallization from residual melt rich in volatiles (presumably CO2). The volatile-rich melt was separated during fractional crystallization of molten slag under the cover of the overlying hot (ca. 1250 °C) vesicular slag, preventing the escape of volatiles. That unique slag system is analogous to natural magmatic systems.

  14. Transcriptional Activation by NFκB Increases Perlecan/HSPG2 Expression in the Desmoplastic Prostate Tumor Microenvironment

    PubMed Central

    Warren, Curtis R.; Grindel, Brian J.; Francis, Lewis; Carson, Daniel D.; Farach-Carson, Mary C.

    2014-01-01

    Perlecan/HSPG2, a heparan sulfate proteoglycan typically found at tissue borders including those separating epithelia and connective tissue, increases near sites of invasion of primary prostatic tumors as previously shown for other proteins involved in desmoplastic tissue reaction. Studies of prostate cancer cells and stromal cells from both prostate and bone, the major site for prostate cancer metastasis, showed that cancer cells and a subset of stromal cells increased production of perlecan in response to cytokines present in the tumor microenvironment. In silico analysis of the HSPG2 promoter revealed two conserved NFκB binding sites, in addition to the previously reported SMAD3 binding sites. By systematically transfecting cells with a variety of reporter constructs including sequences up to 2.6 kb from the start site of transcription, we identified an active cis element in the distal region of the HSPG2 promoter, and showed that it functions in regulating transcription of HSPG2. Treatment with TNF-α and/or TGFβ1 identified TNF-α as a major cytokine regulator of perlecan production. TNF-α treatment also triggered p65 nuclear translocation and binding to the HSPG2 regulatory region in stromal cells and cancer cells. In addition to stromal induction of perlecan production in the prostate, we identified a matrix-secreting bone marrow stromal cell type that may represent the source for increases in perlecan in the metastatic bone marrow environment. These studies implicate perlecan in cytokine-mediated, innate tissue responses to cancer cell invasion, a process we suggest reflects a modified wound healing tissue response co-opted by prostate cancer cells. PMID:24700612

  15. The late and dual origin of cerebrospinal fluid-contacting neurons in the mouse spinal cord

    PubMed Central

    Petracca, Yanina L.; Sartoretti, Maria Micaela; Di Bella, Daniela J.; Marin-Burgin, Antonia; Carcagno, Abel L.; Schinder, Alejandro F.; Lanuza, Guillermo M.

    2016-01-01

    Considerable progress has been made in understanding the mechanisms that control the production of specialized neuronal types. However, how the timing of differentiation contributes to neuronal diversity in the developing spinal cord is still a pending question. In this study, we show that cerebrospinal fluid-contacting neurons (CSF-cNs), an anatomically discrete cell type of the ependymal area, originate from surprisingly late neurogenic events in the ventral spinal cord. CSF-cNs are identified by the expression of the transcription factors Gata2 and Gata3, and the ionic channels Pkd2l1 and Pkd1l2. Contrasting with Gata2/3+ V2b interneurons, differentiation of CSF-cNs is independent of Foxn4 and takes place during advanced developmental stages previously assumed to be exclusively gliogenic. CSF-cNs are produced from two distinct dorsoventral regions of the mouse spinal cord. Most CSF-cNs derive from progenitors circumscribed to the late-p2 and the oligodendrogenic (pOL) domains, whereas a second subset of CSF-cNs arises from cells bordering the floor plate. The development of these two subgroups of CSF-cNs is differentially controlled by Pax6, they adopt separate locations around the postnatal central canal and they display electrophysiological differences. Our results highlight that spatiotemporal mechanisms are instrumental in creating neural cell diversity in the ventral spinal cord to produce distinct classes of interneurons, motoneurons, CSF-cNs, glial cells and ependymal cells. PMID:26839365

  16. Brush border membrane vesicle and Caco-2 cell line: Two experimental models for evaluation of absorption enhancing effects of saponins, bile salts, and some synthetic surfactants

    PubMed Central

    Moghimipour, Eskandar; Tabassi, Sayyed Abolghassem Sajadi; Ramezani, Mohammad; Handali, Somayeh; Löbenberg, Raimar

    2016-01-01

    The aim of this study was to investigate the influence of absorption enhancers in the uptake of hydrophilic compounds. The permeation of the two hydrophilic drug models gentamicin and 5 (6)-carboxyfluorescein (CF) across the brush border membrane vesicles and Caco-2 cell lines were evaluated using total saponins of Acanthophyllum squarrosum, Quillaja saponaria, sodium lauryl sulfate, sodium glycocholate, sodium taurodeoxycholate, and Tween 20 as absorption enhancers. Transepithelial electrical resistance (TEER) measurement was utilized to assess the paracellular permeability of cell lines. Confocal laser scanning microscopy (CLSM) was performed to obtain images of the distribution of CF in Caco-2 cells. These compounds were able to loosen tight junctions, thus increasing paracellular permeability. CLSM confirmed the effect of these absorption enhancers on CF transport across Caco-2 lines and increased the Caco-2 permeability via transcellular route. It was also confirmed that the decrease in TEER was transient and reversible after removal of permeation enhancers. PMID:27429925

  17. Molecular, biological, and antigenic characterization of a Border disease virus isolated from a pig during classical swine fever surveillance in Japan.

    PubMed

    Nagai, Makoto; Aoki, Hiroshi; Sakoda, Yoshihiro; Kozasa, Takashi; Tominaga-Teshima, Kaho; Mine, Junki; Abe, Yuri; Tamura, Tomokazu; Kobayashi, Tsubasa; Nishine, Kaoru; Tateishi, Kentaro; Suzuki, Yudai; Fukuhara, Mai; Ohmori, Keitaro; Todaka, Reiko; Katayama, Kazuhiko; Mizutani, Tetsuya; Nakamura, Shigeyuki; Kida, Hiroshi; Shirai, Junsuke

    2014-07-01

    In the current study, molecular, biological, and antigenic analyses were performed to characterize Border disease virus (BDV) strain FNK2012-1 isolated from a pig in 2012 in Japan. The complete genome comprises 12,327 nucleotides (nt), including a large open reading frame of 11,685 nt. Phylogenetic analysis revealed that FNK2012-1 was clustered into BDV genotype 1 with ovine strains. FNK2012-1 grew in porcine, bovine, and ovine primary cells and cell lines, but grew better in bovine and ovine cells than in porcine cells. Specific pathogen-free pigs inoculated with FNK2012-1 did not show any clinical signs. Noninoculated contact control pigs also did not show clinical signs and did not seroconvert. The results suggest that FNK2012-1 may be of ruminant origin and is poorly adapted to pigs. Such observations can provide important insights into evidence for infection and transmission of BDV, which may be of ruminant origin, among pigs.

  18. Paper SERS chromatography for detection of trace analytes in complex samples

    NASA Astrophysics Data System (ADS)

    Yu, Wei W.; White, Ian M.

    2013-05-01

    We report the application of paper SERS substrates for the detection of trace quantities of multiple analytes in a complex sample in the form of paper chromatography. Paper chromatography facilitates the separation of different analytes from a complex sample into distinct sections in the chromatogram, which can then be uniquely identified using SERS. As an example, the separation and quantitative detection of heroin in a highly fluorescent mixture is demonstrated. Paper SERS chromatography has obvious applications, including law enforcement, food safety, and border protection, and facilitates the rapid detection of chemical and biological threats at the point of sample.

  19. Stochastic models for tumoral growth

    NASA Astrophysics Data System (ADS)

    Escudero, Carlos

    2006-02-01

    Strong experimental evidence has indicated that tumor growth belongs to the molecular beam epitaxy universality class. This type of growth is characterized by the constraint of cell proliferation to the tumor border and the surface diffusion of cells at the growing edge. Tumor growth is thus conceived as a competition for space between the tumor and the host, and cell diffusion at the tumor border is an optimal strategy adopted for minimizing the pressure and helping tumor development. Two stochastic partial differential equations are reported in this paper in order to correctly model the physical properties of tumoral growth in (1+1) and (2+1) dimensions. The advantage of these models is that they reproduce the correct geometry of the tumor and are defined in terms of polar variables. An analysis of these models allows us to quantitatively estimate the response of the tumor to an unfavorable perturbation during growth.

  20. Regulation of Polysaccharide- and Protein- Specific Antibody Responses to Intact Extracellular Bacteria

    DTIC Science & Technology

    2016-03-11

    50, Immunology. 26 | P a g e blood-borne antigens. The white pulp consists of the periarteriolar lymphoid sheath (PALS) which contain T cells ...and CD8α+ dendritic (DC), and adjacent lymphoid follicles containing mainly circulating B cells , known as follicular B cells (FB). The outer boundary...complexes, for initial priming within the T cell zones of secondary lymphoid organs followed by migration of T cells to the T cell -B cell border. B

  1. Insulator function and topological domain border strength scale with architectural protein occupancy

    PubMed Central

    2014-01-01

    Background Chromosome conformation capture studies suggest that eukaryotic genomes are organized into structures called topologically associating domains. The borders of these domains are highly enriched for architectural proteins with characterized roles in insulator function. However, a majority of architectural protein binding sites localize within topological domains, suggesting sites associated with domain borders represent a functionally different subclass of these regulatory elements. How topologically associating domains are established and what differentiates border-associated from non-border architectural protein binding sites remain unanswered questions. Results By mapping the genome-wide target sites for several Drosophila architectural proteins, including previously uncharacterized profiles for TFIIIC and SMC-containing condensin complexes, we uncover an extensive pattern of colocalization in which architectural proteins establish dense clusters at the borders of topological domains. Reporter-based enhancer-blocking insulator activity as well as endogenous domain border strength scale with the occupancy level of architectural protein binding sites, suggesting co-binding by architectural proteins underlies the functional potential of these loci. Analyses in mouse and human stem cells suggest that clustering of architectural proteins is a general feature of genome organization, and conserved architectural protein binding sites may underlie the tissue-invariant nature of topologically associating domains observed in mammals. Conclusions We identify a spectrum of architectural protein occupancy that scales with the topological structure of chromosomes and the regulatory potential of these elements. Whereas high occupancy architectural protein binding sites associate with robust partitioning of topologically associating domains and robust insulator function, low occupancy sites appear reserved for gene-specific regulation within topological domains. PMID:24981874

  2. Ozone formation along the California-Mexican border region during Cal-Mex 2010 field campaign

    NASA Astrophysics Data System (ADS)

    Li, Guohui; Bei, Naifang; Zavala, Miguel; Molina, Luisa T.

    2014-05-01

    The purpose of this study is to evaluate the ozone (O3) formation along the California-Mexico border region using the WRF-CHEM model in association with the Cal-Mex 2010 field campaign. Four two-day episodes in 2010 are chosen based on plume transport patterns: 1) May 15-16 (plume north), 2) May 29-30 (plume southwest), 3) June 4-5 (plume east), and 4) June 13-14 (plume southeast). Generally, the predicted O3 spatial patterns and temporal variations agree well with the observations at the ambient monitoring sites in the San Diego-Tijuana region, but in the Calexico-Mexicali region, the model frequently underestimates the observation. In the San Diego-Tijuana region, the morning anthropogenic precursor emissions in the urbanized coastal plain are carried inland and mixed with the local biogenic emissions during transport, causing the high O3 level over the mountain region. Biogenic emissions enhance the O3 concentrations by up to 40 ppb over the mountain region in the afternoon. The factor separation approach is used to evaluate the contributions of trans-boundary transport of emissions from California and Baja California to the O3 level in the California-Mexico border region. The Baja California emissions play a minor role in the O3 formation in the San Diego region and do not seem to contribute to the O3 exceedances in the region, but have large potential to cause O3 exceedances in the Calexico region. The California emissions can considerably enhance the O3 level in the Tijuana region. Generally, the California emissions play a more important role than the Baja California emissions on O3 formation in the border region (within 40 km to the California-Mexico border). On average, the O3 concentrations in the border region are decreased by 2-4 ppb in the afternoon due to the interactions of emissions from California and Baja California. Further studies need to be conducted to improve the sea breeze simulations in the border region for evaluating O3 formation.

  3. Transcriptional Regulation in Ebola Virus: Effects of Gene Border Structure and Regulatory Elements on Gene Expression and Polymerase Scanning Behavior

    PubMed Central

    Brauburger, Kristina; Boehmann, Yannik; Krähling, Verena

    2015-01-01

    ABSTRACT The highly pathogenic Ebola virus (EBOV) has a nonsegmented negative-strand (NNS) RNA genome containing seven genes. The viral genes either are separated by intergenic regions (IRs) of variable length or overlap. The structure of the EBOV gene overlaps is conserved throughout all filovirus genomes and is distinct from that of the overlaps found in other NNS RNA viruses. Here, we analyzed how diverse gene borders and noncoding regions surrounding the gene borders influence transcript levels and govern polymerase behavior during viral transcription. Transcription of overlapping genes in EBOV bicistronic minigenomes followed the stop-start mechanism, similar to that followed by IR-containing gene borders. When the gene overlaps were extended, the EBOV polymerase was able to scan the template in an upstream direction. This polymerase feature seems to be generally conserved among NNS RNA virus polymerases. Analysis of IR-containing gene borders showed that the IR sequence plays only a minor role in transcription regulation. Changes in IR length were generally well tolerated, but specific IR lengths led to a strong decrease in downstream gene expression. Correlation analysis revealed that these effects were largely independent of the surrounding gene borders. Each EBOV gene contains exceptionally long untranslated regions (UTRs) flanking the open reading frame. Our data suggest that the UTRs adjacent to the gene borders are the main regulators of transcript levels. A highly complex interplay between the different cis-acting elements to modulate transcription was revealed for specific combinations of IRs and UTRs, emphasizing the importance of the noncoding regions in EBOV gene expression control. IMPORTANCE Our data extend those from previous analyses investigating the implication of noncoding regions at the EBOV gene borders for gene expression control. We show that EBOV transcription is regulated in a highly complex yet not easily predictable manner by a set of interacting cis-active elements. These findings are important not only for the design of recombinant filoviruses but also for the design of other replicon systems widely used as surrogate systems to study the filovirus replication cycle under low biosafety levels. Insights into the complex regulation of EBOV transcription conveyed by noncoding sequences will also help to interpret the importance of mutations that have been detected within these regions, including in isolates of the current outbreak. PMID:26656691

  4. Transcriptional Regulation in Ebola Virus: Effects of Gene Border Structure and Regulatory Elements on Gene Expression and Polymerase Scanning Behavior.

    PubMed

    Brauburger, Kristina; Boehmann, Yannik; Krähling, Verena; Mühlberger, Elke

    2016-02-15

    The highly pathogenic Ebola virus (EBOV) has a nonsegmented negative-strand (NNS) RNA genome containing seven genes. The viral genes either are separated by intergenic regions (IRs) of variable length or overlap. The structure of the EBOV gene overlaps is conserved throughout all filovirus genomes and is distinct from that of the overlaps found in other NNS RNA viruses. Here, we analyzed how diverse gene borders and noncoding regions surrounding the gene borders influence transcript levels and govern polymerase behavior during viral transcription. Transcription of overlapping genes in EBOV bicistronic minigenomes followed the stop-start mechanism, similar to that followed by IR-containing gene borders. When the gene overlaps were extended, the EBOV polymerase was able to scan the template in an upstream direction. This polymerase feature seems to be generally conserved among NNS RNA virus polymerases. Analysis of IR-containing gene borders showed that the IR sequence plays only a minor role in transcription regulation. Changes in IR length were generally well tolerated, but specific IR lengths led to a strong decrease in downstream gene expression. Correlation analysis revealed that these effects were largely independent of the surrounding gene borders. Each EBOV gene contains exceptionally long untranslated regions (UTRs) flanking the open reading frame. Our data suggest that the UTRs adjacent to the gene borders are the main regulators of transcript levels. A highly complex interplay between the different cis-acting elements to modulate transcription was revealed for specific combinations of IRs and UTRs, emphasizing the importance of the noncoding regions in EBOV gene expression control. Our data extend those from previous analyses investigating the implication of noncoding regions at the EBOV gene borders for gene expression control. We show that EBOV transcription is regulated in a highly complex yet not easily predictable manner by a set of interacting cis-active elements. These findings are important not only for the design of recombinant filoviruses but also for the design of other replicon systems widely used as surrogate systems to study the filovirus replication cycle under low biosafety levels. Insights into the complex regulation of EBOV transcription conveyed by noncoding sequences will also help to interpret the importance of mutations that have been detected within these regions, including in isolates of the current outbreak. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  5. Localization of the action of cholera toxin on adenyl cyclase in mucosal epithelial cells of rabbit intestine

    PubMed Central

    Parkinson, David K.; Ebel, Hans; DiBona, Donald R.; Sharp, Geoffrey W. G.

    1972-01-01

    Brush borders and plasma membranes have been purified from mucosal epithelial cells of rabbit ileum under control conditions and after treatment for 3 hr with cholera toxin in vivo. The activity of several enzymes in these preparations was measured. It was concluded that adenyl cyclase, like NaK-ATPase, seems not to be a normal constituent of brush borders. Both these enzymes are present in plasma membrane preparations derived largely from the basal and lateral margins of the epithelial cells, both may be phospholipid dependent enzymes and both are affected by cholera toxin. Adenyl cyclase activity is increased while NaK-ATPase is decreased. The activities of alkaline phosphatase, leucineaminopeptidase, 5′-nucleotidase, glucose-6-phosphatase, and Mg-ATPase were not found to be affected by the toxin. Cholera toxin, which makes contact with the luminal side of the epithelial cells, in the natural disease and in the experimental model, would appear to exert its pathologic effect on adenyl cyclase at the opposite (basal and lateral) side of the cells. Images PMID:4344729

  6. Coding of Border Ownership in Monkey Visual Cortex

    PubMed Central

    Zhou, Hong; Friedman, Howard S.; von der Heydt, Rüdiger

    2016-01-01

    Areas V1 and V2 of the visual cortex have traditionally been conceived as stages of local feature representations. We investigated whether neural responses carry information about how local features belong to objects. Single-cell activity was recorded in areas V1, V2, and V4 of awake behaving monkeys. Displays were used in which the same local feature (contrast edge or line) could be presented as part of different figures. For example, the same light–dark edge could be the left side of a dark square or the right side of a light square. Each display was also presented with reversed contrast. We found significant modulation of responses as a function of the side of the figure in >50% of neurons of V2 and V4 and in 18% of neurons of the top layers of V1. Thus, besides the local contrast border information, neurons were found to encode the side to which the border belongs (“border ownership coding”). A majority of these neurons coded border ownership and the local polarity of luminance–chromaticity contrast. The others were insensitive to contrast polarity. Another 20% of the neurons of V2 and V4, and 48% of top layer V1, coded local contrast polarity, but not border ownership. The border ownership-related response differences emerged soon (<25 msec) after the response onset. In V2 and V4, the differences were found to be nearly independent of figure size up to the limit set by the size of our display (21°). Displays that differed only far outside the conventional receptive field could produce markedly different responses. When tested with more complex displays in which figure-ground cues were varied, some neurons produced invariant border ownership signals, others failed to signal border ownership for some of the displays, but neurons that reversed signals were rare. The influence of visual stimulation far from the receptive field center indicates mechanisms of global context integration. The short latencies and incomplete cue invariance suggest that the border-ownership effect is generated within the visual cortex rather than projected down from higher levels. PMID:10964965

  7. Migrant and Refugee Patient Perspectives on Travel and Tuberculosis along the Thailand-Myanmar Border: A Qualitative Study.

    PubMed

    Tschirhart, Naomi; Sein, Tabitha; Nosten, Francois; Foster, Angel M

    2016-01-01

    The Thailand-Myanmar border separates two very different health systems. The healthcare system in eastern Myanmar remains underdeveloped as a result of decades of instability. Comparatively, Tak province, Thailand has more healthcare resources. In this Thai border province government hospitals and non-governmental organizations provide tuberculosis (TB) treatment to migrants and refugees. Our overall study aimed to explore accessibility of TB treatment, TB surveillance and health system responsiveness specific to migrant and refugee populations in Tak province. In this paper, we focus on the perspectives of migrant and refugee TB patients with respect to travel and treatment in Tak province. In 2014 we conducted focus group discussions with 61 TB, Tuberculosis and Human Immunodeficiency Virus co-infection, and multidrug-resistant TB patients in Tak province. We analyzed the data for content and themes and documented individual travel trajectories. Migrants are travelling with active TB within the country and between Thailand and Myanmar. Migrants primarily travelled to obtain treatment but two participants reported travelling home to seek family care in Myanmar before returning to Thailand for treatment. Travel, while expensive and arduous, is an adaptive strategy that migrants use to access healthcare. Migrant's need for travel points to larger difficulties associated with healthcare access in the border region. Long distance travel with an infectious disease can be seen as an indicator that local healthcare is not available or affordable. These findings suggest that public health officials from both sides of the border should discuss the factors that contribute to travel with active TB and explore potential solutions to mitigate disease transmission in migrant populations.

  8. Migrant and Refugee Patient Perspectives on Travel and Tuberculosis along the Thailand-Myanmar Border: A Qualitative Study

    PubMed Central

    Sein, Tabitha; Nosten, Francois; Foster, Angel M.

    2016-01-01

    Background The Thailand-Myanmar border separates two very different health systems. The healthcare system in eastern Myanmar remains underdeveloped as a result of decades of instability. Comparatively, Tak province, Thailand has more healthcare resources. In this Thai border province government hospitals and non-governmental organizations provide tuberculosis (TB) treatment to migrants and refugees. Objectives Our overall study aimed to explore accessibility of TB treatment, TB surveillance and health system responsiveness specific to migrant and refugee populations in Tak province. In this paper, we focus on the perspectives of migrant and refugee TB patients with respect to travel and treatment in Tak province. Methods In 2014 we conducted focus group discussions with 61 TB, Tuberculosis and Human Immunodeficiency Virus co-infection, and multidrug-resistant TB patients in Tak province. We analyzed the data for content and themes and documented individual travel trajectories. Results and Discussion Migrants are travelling with active TB within the country and between Thailand and Myanmar. Migrants primarily travelled to obtain treatment but two participants reported travelling home to seek family care in Myanmar before returning to Thailand for treatment. Travel, while expensive and arduous, is an adaptive strategy that migrants use to access healthcare. Conclusions Migrant’s need for travel points to larger difficulties associated with healthcare access in the border region. Long distance travel with an infectious disease can be seen as an indicator that local healthcare is not available or affordable. These findings suggest that public health officials from both sides of the border should discuss the factors that contribute to travel with active TB and explore potential solutions to mitigate disease transmission in migrant populations. PMID:27509036

  9. “Agrolistic” transformation of plant cells: Integration of T-strands generated in planta

    PubMed Central

    Hansen, Geneviève; Chilton, Mary-Dell

    1996-01-01

    We describe a novel plant transformation technique, termed “agrolistic,” that combines the advantages of the Agrobacterium transformation system with the high efficiency of biolistic DNA delivery. Agrolistic transformation allows integration of the gene of interest without undesired vector sequence. The virulence genes virD1 and virD2 from Agrobacterium tumefaciens that are required in bacteria for excision of T-strands from the tumor-inducing plasmid were placed under the control of the CaMV35S promoter and codelivered with a target plasmid containing border sequences flanking the gene of interest. Transient expression assays in tobacco and in maize cells indicated that vir gene products caused strand-specific nicking in planta at the right border sequence, similar to VirD1/VirD2-catalyzed T-strand excision observed in Agrobacterium. Agrolistically transformed tobacco calli were obtained after codelivery of virD1 and virD2 genes together with a selectable marker flanked by border sequences. Some inserts exhibited right junctions with plant DNA that corresponded precisely to the sequence expected for T-DNA (portion of the tumor-inducing plasmid that is transferred to plant cells) insertion events. We designate these as “agrolistic” inserts, as distinguished from “biolistic” inserts. Both types of inserts were found in some transformed lines. The frequency of agrolistic inserts was 20% that of biolistic inserts. PMID:8962167

  10. Sinus venosus syndrome: atrial septal defect or anomalous venous connection? A multiplane transoesophageal approach.

    PubMed

    Oliver, J M; Gallego, P; Gonzalez, A; Dominguez, F J; Aroca, A; Mesa, J M

    2002-12-01

    To discuss the anatomical features of sinus venosus atrial defect on the basis of a comprehensive transoesophageal echocardiography (TOE) examination and its relation to surgical data. 24 patients (13 men, 11 women, mean (SD) age 37 (17) years, range 17-73 years) with a posterior interatrial communication closely related to the entrance of the superior (SVC) or inferior vena cava (IVC) who underwent TOE before surgical repair. Records of these patients were retrospectively reviewed and compared with surgical assessments. In 13 patients, TOE showed a deficiency in the extraseptal wall that normally separates the left atrium and right upper pulmonary vein from the SVC and right atrium. This deficiency unroofed the right upper pulmonary vein, compelling it to drain into the SVC, which overrode the intact atrial septum. In three patients, TOE examination showed a defect in the wall of the IVC, which continued directly into the posterior border of the left atrium. Thus, the intact muscular border of the atrial septum was overridden by the mouth of the IVC, which presented a biatrial connection. In the remaining eight patients, the defect was located in the muscular posterior border of the fossa ovalis. A residuum of atrial septum was visualised in the superior margin of the defect. Neither caval vein overriding nor anomalous pulmonary vein drainage was present. Sinus venosus syndrome should be regarded as an anomalous venous connection with an interatrial communication outside the confines of the atrial septum, in the unfolding wall that normally separates the left atrium from either caval vein. It results in overriding of the caval veins across the intact atrial septum and partial pulmonary vein anomalous drainage. It should be differentiated from posterior atrial septal defect without overriding or anomalous venous connections.

  11. Sinus venosus syndrome: atrial septal defect or anomalous venous connection? A multiplane transoesophageal approach

    PubMed Central

    Oliver, J M; Gallego, P; Gonzalez, A; Dominguez, F J; Aroca, A; Mesa, J M

    2002-01-01

    Objective: To discuss the anatomical features of sinus venosus atrial defect on the basis of a comprehensive transoesophageal echocardiography (TOE) examination and its relation to surgical data. Methods: 24 patients (13 men, 11 women, mean (SD) age 37 (17) years, range 17–73 years) with a posterior interatrial communication closely related to the entrance of the superior (SVC) or inferior vena cava (IVC) who underwent TOE before surgical repair. Records of these patients were retrospectively reviewed and compared with surgical assessments. Results: In 13 patients, TOE showed a deficiency in the extraseptal wall that normally separates the left atrium and right upper pulmonary vein from the SVC and right atrium. This deficiency unroofed the right upper pulmonary vein, compelling it to drain into the SVC, which overrode the intact atrial septum. In three patients, TOE examination showed a defect in the wall of the IVC, which continued directly into the posterior border of the left atrium. Thus, the intact muscular border of the atrial septum was overridden by the mouth of the IVC, which presented a biatrial connection. In the remaining eight patients, the defect was located in the muscular posterior border of the fossa ovalis. A residuum of atrial septum was visualised in the superior margin of the defect. Neither caval vein overriding nor anomalous pulmonary vein drainage was present. Conclusions: Sinus venosus syndrome should be regarded as an anomalous venous connection with an interatrial communication outside the confines of the atrial septum, in the unfolding wall that normally separates the left atrium from either caval vein. It results in overriding of the caval veins across the intact atrial septum and partial pulmonary vein anomalous drainage. It should be differentiated from posterior atrial septal defect without overriding or anomalous venous connections. PMID:12433899

  12. The histopathological effect of intracameral ropivacaine in different concentrations on corneal endothelium.

    PubMed

    Caça, Ihsan; Kavak, Vatan; Unlü, Kaan; Ari, Seyhmus; Nergis, Yusuf; Take, Gülnür

    2006-01-01

    We evaluated the histopathological changes occurring in corneal endothelium after intracameral injection ropivacaine into rats. Intracamerally administered ropivacaine in 1, 0.5, and 0.1% concentrations resulted in impairment of hexagonal structure of corneal endothelial cells and intercellular junctions, destruction of microvilli on the cell surface, roughness of cell borders, picnotic nucleus, diffuse vacuolization, and crystalysis in mitochondria.

  13. Nickel-metal hydride battery development. Final technical report

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

    NONE

    1995-06-01

    Rechargeable batteries are used as the power source for a broad range of portable equipment. Key battery selection criteria typically are weight, volume, first cost, life cycle cost, and environmental impact. Rechargeable batteries are favored from a life cycle cost and environmental impact standpoint over primary batteries. The nickel-metal hydride (Ni-MH) battery system has emerged as the battery of choice for many applications based on its superior characteristics when judged on the above criteria against other battery types. In most cases commercial Ni-MH batteries are constructed with coiled electrodes in cylindrical metal containers. Electro Energy, Inc. (EEI) has been developingmore » a novel flat bipolar configuration of the Ni-MH system that offers weight, volume, and cost advantages when compared to cylindrical cells. The unique bipolar approach consists of fabricating individual flat wafer cells in conductive, carbon-filled, plastic face plates. The individual cells contain a nonconductive plastic border which is heat sealed around the perimeter to make a totally sealed unit cell. Multi-cell batteries are fabricated by stacking the individual wafer cells in such a way that the positive face of one cell contacts the negative face of the adjacent cell. The stack is then contained in an outer housing with end contacts. The purpose of this program was to develop, evaluate, and demonstrate the capabilities of the EEI Ni-MH battery system for consumer applications. The work was directed at the development and evaluation of the compact bipolar construction for its potential advantages of high power and energy density. Experimental investigations were performed on various nickel electrode types, hydride electrode formulations, and alternate separator materials. Studies were also directed at evaluating various oxygen recombination techniques for low pressure operation during charge and overcharge.« less

  14. Dual mechanism controls asymmetric spindle position in ascidian germ cell precursors.

    PubMed

    Prodon, François; Chenevert, Janet; Hébras, Céline; Dumollard, Rémi; Faure, Emmanuel; Gonzalez-Garcia, Jose; Nishida, Hiroki; Sardet, Christian; McDougall, Alex

    2010-06-01

    Mitotic spindle orientation with respect to cortical polarity cues generates molecularly distinct daughter cells during asymmetric cell division (ACD). However, during ACD it remains unknown how the orientation of the mitotic spindle is regulated by cortical polarity cues until furrowing begins. In ascidians, the cortical centrosome-attracting body (CAB) generates three successive unequal cleavages and the asymmetric segregation of 40 localized postplasmic/PEM RNAs in germ cell precursors from the 8-64 cell stage. By combining fast 4D confocal fluorescence imaging with gene-silencing and classical blastomere isolation experiments, we show that spindle repositioning mechanisms are active from prometaphase until anaphase, when furrowing is initiated in B5.2 cells. We show that the vegetal-most spindle pole/centrosome is attracted towards the CAB during prometaphase, causing the spindle to position asymmetrically near the cortex. Next, during anaphase, the opposite spindle pole/centrosome is attracted towards the border with neighbouring B5.1 blastomeres, causing the spindle to rotate (10 degrees /minute) and migrate (3 microm/minute). Dynamic 4D fluorescence imaging of filamentous actin and plasma membrane shows that precise orientation of the cleavage furrow is determined by this second phase of rotational spindle displacement. Furthermore, in pairs of isolated B5.2 blastomeres, the second phase of rotational spindle displacement was lost. Finally, knockdown of PEM1, a protein localized in the CAB and required for unequal cleavage in B5.2 cells, completely randomizes spindle orientation. Together these data show that two separate mechanisms active during mitosis are responsible for spindle positioning, leading to precise orientation of the cleavage furrow during ACD in the cells that give rise to the germ lineage in ascidians.

  15. Electrotonic coupling of excitable and nonexcitable cells in the heart revealed by optogenetics

    PubMed Central

    Quinn, T. Alexander; Camelliti, Patrizia; Rog-Zielinska, Eva A.; Siedlecka, Urszula; Poggioli, Tommaso; O'Toole, Eileen T.; Knöpfel, Thomas; Kohl, Peter

    2016-01-01

    Electrophysiological studies of excitable organs usually focus on action potential (AP)-generating cells, whereas nonexcitable cells are generally considered as barriers to electrical conduction. Whether nonexcitable cells may modulate excitable cell function or even contribute to AP conduction via direct electrotonic coupling to AP-generating cells is unresolved in the heart: such coupling is present in vitro, but conclusive evidence in situ is lacking. We used genetically encoded voltage-sensitive fluorescent protein 2.3 (VSFP2.3) to monitor transmembrane potential in either myocytes or nonmyocytes of murine hearts. We confirm that VSFP2.3 allows measurement of cell type-specific electrical activity. We show that VSFP2.3, expressed solely in nonmyocytes, can report cardiomyocyte AP-like signals at the border of healed cryoinjuries. Using EM-based tomographic reconstruction, we further discovered tunneling nanotube connections between myocytes and nonmyocytes in cardiac scar border tissue. Our results provide direct electrophysiological evidence of heterocellular electrotonic coupling in native myocardium and identify tunneling nanotubes as a possible substrate for electrical cell coupling that may be in addition to previously discovered connexins at sites of myocyte–nonmyocyte contact in the heart. These findings call for reevaluation of cardiac nonmyocyte roles in electrical connectivity of the heterocellular heart. PMID:27930302

  16. Fermented soya bean (tempe) extracts reduce adhesion of enterotoxigenic Escherichia coli to intestinal epithelial cells.

    PubMed

    Roubos-van den Hil, P J; Nout, M J R; Beumer, R R; van der Meulen, J; Zwietering, M H

    2009-03-01

    This study aimed to investigate the effect of processed soya bean, during the successive stages of tempe fermentation and different fermentation times, on adhesion of enterotoxigenic Escherichia coli (ETEC) K88 to intestinal brush border cells as well as Caco-2 intestinal epithelial cells; and to clarify the mechanism of action. Tempe was prepared at controlled laboratory scale using Rhizopus microsporus var. microsporus as the inoculum. Extracts of raw, soaked and cooked soya beans reduced ETEC adhesion to brush border cells by 40%. Tempe extracts reduced adhesion by 80% or more. ETEC adhesion to Caco-2 cells reduced by 50% in the presence of tempe extracts. ETEC K88 bacteria were found to interact with soya bean extracts, and this may contribute to the observed decrease of ETEC adhesion to intestinal epithelial cells. Fermented soya beans (tempe) reduce the adhesion of ETEC to intestinal epithelial cells of pig and human origin. This reduced adhesion is caused by an interaction between ETEC K88 bacteria and soya bean compounds. The results strengthen previous observations on the anti-diarrhoeal effect of tempe. This effect indicates that soya-derived compounds may reduce adhesion of ETEC to intestinal cells in pigs as well as in humans and prevent against diarrhoeal diseases.

  17. Cell cycle progression is required for zebrafish somite morphogenesis but not segmentation clock function

    PubMed Central

    Zhang, Lixia; Kendrick, Christina; Jülich, Dörthe; Holley, Scott A.

    2010-01-01

    Summary Cell division, differentiation and morphogenesis are coordinated during embryonic development and frequently in disarray in pathologies such as cancer. Here, we present a zebrafish mutant that ceases mitosis at the beginning of gastrulation, but undergoes axis elongation and develops blood, muscle and a beating heart. We identify the mutation as being in early mitotic inhibitor 1 (emi1), a negative regulator of the Anaphase Promoting Complex, and utilize the mutant to examine the role of the cell cycle in somitogenesis. The mutant phenotype indicates that axis elongation during the segmentation period is substantially driven by cell migration. We find that the segmentation clock, which regulates somitogenesis, functions normally in the absence of cell cycle progression and observe that mitosis is a modest source of noise for the clock. Somite morphogenesis involves the epithelialization of the somite border cells around a core of mesenchyme. As in wild-type embryos, somite boundary cells are polarized along a Fibronectin matrix in emi1−/−. The mutants also display evidence of segment polarity. However, in the absence of a normal cell cycle, somites appear to hyper-epithelialize as the internal mesenchymal cells exit the core of the somite after initial boundary formation. Thus, cell cycle progression is not required during the segmentation period for segmentation clock function but is necessary for normal segmental arrangement of epithelial borders and internal mesenchymal cells. PMID:18480162

  18. Effects of myocardial infarction on the distribution and transport of nutrients and oxygen in porcine myocardium.

    PubMed

    Davis, Bryce H; Morimoto, Yoshihisa; Sample, Chris; Olbrich, Kevin; Leddy, Holly A; Guilak, Farshid; Taylor, Doris A

    2012-10-01

    One of the primary limitations of cell therapy for myocardial infarction is the low survival of transplanted cells, with a loss of up to 80% of cells within 3 days of delivery. The aims of this study were to investigate the distribution of nutrients and oxygen in infarcted myocardium and to quantify how macromolecular transport properties might affect cell survival. Transmural myocardial infarction was created by controlled cryoablation in pigs. At 30 days post-infarction, oxygen and metabolite levels were measured in the peripheral skeletal muscle, normal myocardium, the infarct border zone, and the infarct interior. The diffusion coefficients of fluorescein or FITC-labeled dextran (0.3-70 kD) were measured in these tissues using fluorescence recovery after photobleaching. The vascular density was measured via endogenous alkaline phosphatase staining. To examine the influence of these infarct conditions on cells therapeutically used in vivo, skeletal myoblast survival and differentiation were studied in vitro under the oxygen and glucose concentrations measured in the infarct tissue. Glucose and oxygen concentrations, along with vascular density were significantly reduced in infarct when compared to the uninjured myocardium and infarct border zone, although the degree of decrease differed. The diffusivity of molecules smaller than 40 kD was significantly higher in infarct center and border zone as compared to uninjured heart. Skeletal myoblast differentiation and survival were decreased stepwise from control to hypoxia, starvation, and ischemia conditions. Although oxygen, glucose, and vascular density were significantly reduced in infarcted myocardium, the rate of macromolecular diffusion was significantly increased, suggesting that diffusive transport may not be inhibited in infarct tissue, and thus the supply of nutrients to transplanted cells may be possible. in vitro studies mimicking infarct conditions suggest that increasing nutrients available to transplanted cells may significantly increase their ability to survive in infarct.

  19. BMP signaling protects telencephalic fate by repressing eye identity and its Cxcr4-dependent morphogenesis.

    PubMed

    Bielen, Holger; Houart, Corinne

    2012-10-16

    Depletion of Wnt signaling is a major requirement for the induction of the anterior prosencephalon. However, the molecular events driving the differential regionalization of this area into eye-field and telencephalon fates are still unknown. Here we show that the BMP pathway is active in the anterior neural ectoderm during late blastula to early gastrula stage in zebrafish. Bmp2b mutants and mosaic loss-of-function experiments reveal that BMP acts as a repressor of eye-field fate through inhibition of its key transcription factor Rx3, thereby protecting the future telencephalon from acquiring eye identity. This BMP-driven mechanism initiates the establishment of the telencephalon prior to the involvement of Wnt antagonists from the anterior neural border. Furthermore, we demonstrate that Rx3 and BMP are respectively required to maintain and restrict the chemokine receptor cxcr4a, which in turn contributes to the morphogenetic separation of eye-field and telencephalic cells during early neurulation. Copyright © 2012 Elsevier Inc. All rights reserved.

  20. [Anatomy of fractures of the inferior scapular angle].

    PubMed

    Bartoníček, J; Tuček, M; Malík, J

    2018-01-01

    The aim of this study is to describe the anatomy of fractures of the inferior angle and the adjacent part of the scapular body, based on 3D CT reconstructions. In a series of 375 scapular fractures, we identified a total of 20 fractures of the inferior angle of the scapular body (13 men, 7 women), with a mean patient age of 50 years (range 3373). In all fractures, 3D CT reconstructions were obtained, allowing an objective evaluation of the fracture pattern with a focus on the size and shape of the inferior angle fragment, propagation of the fracture line to the lateral and medial borders of the infraspinous part of the scapular body, fragment displacement and any additional fracture of the ipsilateral scapula and the shoulder girdle. We identified a total of 5 types of fracture involving the distal half of the infraspinous part of the scapular body. The first type, recorded in 5 cases, affected only the apex of the inferior angle, with a small part of the adjacent medial border. The second type, occurring in 4 cases, involved fractures separating the entire inferior angle. The third type, represented by 4 cases, was characterized by a fracture line starting medially close above the inferior angle and passing proximolaterally. The separated fragment had a shape of a big drop, carrying also the distal half of the lateral pillar in addition to the inferior angle. In the fourth type identified in 5 fractures, the separated fragment was formed both by the inferior angle and a variable part of the medial border. The fifth type, being by its nature a transition to the fracture of the infraspinous part of the body, was recorded in 2 cases, with the same V-shaped fragment. Fractures of the inferior angle and the adjacent part of the scapular body are groups of fractures differing from other infraspinous fractures of the scapular body. Although these fractures are highly variable in terms of shape, they have the same course of fracture line and the manner of displacement.Key words: scapula scapula fractures scapular body fractures inferior angle classification of scapular body fractures.

  1. Limits, excesses and pleasures: practices and discourses of sex work in a Mexican rural community.

    PubMed

    Parrini, Rodrigo; Amuchástegui, Ana; Garibi, Cecilia

    2017-10-01

    In Tomatlán, a small city on the Mexican Pacific Coast there is a tolerance zone in which a group of sex workers, separated from the rest of the city by a garbage dump, offer their services to local men. The women present themselves materially and symbolically as agents of regulation with respect to male sexuality, itself constructed as overwhelming and insatiable, which, were it not for services provided by the sex workers, would threaten the family-based social order. Through in-depth interviews with 19 sex workers and 5 municipal officials, this paper discusses how the sex gender system results in a series of borders that are both geographical and subjective. Such borders have to do with periphery and control, body and intimacy, and the boundaries of animality.

  2. Somitic origin of the medial border of the mammalian scapula and its homology to the avian scapula blade.

    PubMed

    Valasek, Petr; Theis, Susanne; Krejci, Eliska; Grim, Milos; Maina, Flavio; Shwartz, Yulia; Otto, Anthony; Huang, Ruijin; Patel, Ketan

    2010-04-01

    The scapula is the main skeletal element of the pectoral girdle allowing muscular fixation of the forelimb to the axial skeleton. The vertebrate limb skeleton has traditionally been considered to develop from the lateral plate mesoderm, whereas the musculature originates from the axial somites. However, in birds, the scapular blade has been shown to develop from the somites. We investigated whether a somitic contribution was also present in the mammalian scapula. Using genetic lineage-tracing techniques, we show that the medial border of the mammalian scapula develops from somitic cells. The medial scapula border serves as the attachment site of girdle muscles (serratus anterior, rhomboidei and levator scapulae). We show that the development of these muscles is independent of the mechanism that controls the formation of all other limb muscles. We suggest that these muscles be specifically referred to as medial girdle muscles. Our results establish the avian scapular blade and medial border of the mammalian scapula as homologous structures as they share the same developmental origin.

  3. Somitic origin of the medial border of the mammalian scapula and its homology to the avian scapula blade

    PubMed Central

    Valasek, Petr; Theis, Susanne; Krejci, Eliska; Grim, Milos; Maina, Flavio; Shwartz, Yulia; Otto, Anthony; Huang, Ruijin; Patel, Ketan

    2010-01-01

    The scapula is the main skeletal element of the pectoral girdle allowing muscular fixation of the forelimb to the axial skeleton. The vertebrate limb skeleton has traditionally been considered to develop from the lateral plate mesoderm, whereas the musculature originates from the axial somites. However, in birds, the scapular blade has been shown to develop from the somites. We investigated whether a somitic contribution was also present in the mammalian scapula. Using genetic lineage-tracing techniques, we show that the medial border of the mammalian scapula develops from somitic cells. The medial scapula border serves as the attachment site of girdle muscles (serratus anterior, rhomboidei and levator scapulae). We show that the development of these muscles is independent of the mechanism that controls the formation of all other limb muscles. We suggest that these muscles be specifically referred to as medial girdle muscles. Our results establish the avian scapular blade and medial border of the mammalian scapula as homologous structures as they share the same developmental origin. PMID:20136669

  4. Automatic Approach for Lung Segmentation with Juxta-Pleural Nodules from Thoracic CT Based on Contour Tracing and Correction.

    PubMed

    Wang, Jinke; Guo, Haoyan

    2016-01-01

    This paper presents a fully automatic framework for lung segmentation, in which juxta-pleural nodule problem is brought into strong focus. The proposed scheme consists of three phases: skin boundary detection, rough segmentation of lung contour, and pulmonary parenchyma refinement. Firstly, chest skin boundary is extracted through image aligning, morphology operation, and connective region analysis. Secondly, diagonal-based border tracing is implemented for lung contour segmentation, with maximum cost path algorithm used for separating the left and right lungs. Finally, by arc-based border smoothing and concave-based border correction, the refined pulmonary parenchyma is obtained. The proposed scheme is evaluated on 45 volumes of chest scans, with volume difference (VD) 11.15 ± 69.63 cm 3 , volume overlap error (VOE) 3.5057 ± 1.3719%, average surface distance (ASD) 0.7917 ± 0.2741 mm, root mean square distance (RMSD) 1.6957 ± 0.6568 mm, maximum symmetric absolute surface distance (MSD) 21.3430 ± 8.1743 mm, and average time-cost 2 seconds per image. The preliminary results on accuracy and complexity prove that our scheme is a promising tool for lung segmentation with juxta-pleural nodules.

  5. Modeling emergent border-crossing behaviors during pandemics

    NASA Astrophysics Data System (ADS)

    Santos, Eunice E.; Santos, Eugene; Korah, John; Thompson, Jeremy E.; Gu, Qi; Kim, Keum Joo; Li, Deqing; Russell, Jacob; Subramanian, Suresh; Zhang, Yuxi; Zhao, Yan

    2013-06-01

    Modeling real-world scenarios is a challenge for traditional social science researchers, as it is often hard to capture the intricacies and dynamisms of real-world situations without making simplistic assumptions. This imposes severe limitations on the capabilities of such models and frameworks. Complex population dynamics during natural disasters such as pandemics is an area where computational social science can provide useful insights and explanations. In this paper, we employ a novel intent-driven modeling paradigm for such real-world scenarios by causally mapping beliefs, goals, and actions of individuals and groups to overall behavior using a probabilistic representation called Bayesian Knowledge Bases (BKBs). To validate our framework we examine emergent behavior occurring near a national border during pandemics, specifically the 2009 H1N1 pandemic in Mexico. The novelty of the work in this paper lies in representing the dynamism at multiple scales by including both coarse-grained (events at the national level) and finegrained (events at two separate border locations) information. This is especially useful for analysts in disaster management and first responder organizations who need to be able to understand both macro-level behavior and changes in the immediate vicinity, to help with planning, prevention, and mitigation. We demonstrate the capabilities of our framework in uncovering previously hidden connections and explanations by comparing independent models of the border locations with their fused model to identify emergent behaviors not found in either independent location models nor in a simple linear combination of those models.

  6. Segmentation of whole cells and cell nuclei from 3-D optical microscope images using dynamic programming.

    PubMed

    McCullough, D P; Gudla, P R; Harris, B S; Collins, J A; Meaburn, K J; Nakaya, M A; Yamaguchi, T P; Misteli, T; Lockett, S J

    2008-05-01

    Communications between cells in large part drive tissue development and function, as well as disease-related processes such as tumorigenesis. Understanding the mechanistic bases of these processes necessitates quantifying specific molecules in adjacent cells or cell nuclei of intact tissue. However, a major restriction on such analyses is the lack of an efficient method that correctly segments each object (cell or nucleus) from 3-D images of an intact tissue specimen. We report a highly reliable and accurate semi-automatic algorithmic method for segmenting fluorescence-labeled cells or nuclei from 3-D tissue images. Segmentation begins with semi-automatic, 2-D object delineation in a user-selected plane, using dynamic programming (DP) to locate the border with an accumulated intensity per unit length greater that any other possible border around the same object. Then the two surfaces of the object in planes above and below the selected plane are found using an algorithm that combines DP and combinatorial searching. Following segmentation, any perceived errors can be interactively corrected. Segmentation accuracy is not significantly affected by intermittent labeling of object surfaces, diffuse surfaces, or spurious signals away from surfaces. The unique strength of the segmentation method was demonstrated on a variety of biological tissue samples where all cells, including irregularly shaped cells, were accurately segmented based on visual inspection.

  7. SNW1 Is a Critical Regulator of Spatial BMP Activity, Neural Plate Border Formation, and Neural Crest Specification in Vertebrate Embryos

    PubMed Central

    Wu, Mary Y.; Ramel, Marie-Christine; Howell, Michael; Hill, Caroline S.

    2011-01-01

    Bone morphogenetic protein (BMP) gradients provide positional information to direct cell fate specification, such as patterning of the vertebrate ectoderm into neural, neural crest, and epidermal tissues, with precise borders segregating these domains. However, little is known about how BMP activity is regulated spatially and temporally during vertebrate development to contribute to embryonic patterning, and more specifically to neural crest formation. Through a large-scale in vivo functional screen in Xenopus for neural crest fate, we identified an essential regulator of BMP activity, SNW1. SNW1 is a nuclear protein known to regulate gene expression. Using antisense morpholinos to deplete SNW1 protein in both Xenopus and zebrafish embryos, we demonstrate that dorsally expressed SNW1 is required for neural crest specification, and this is independent of mesoderm formation and gastrulation morphogenetic movements. By exploiting a combination of immunostaining for phosphorylated Smad1 in Xenopus embryos and a BMP-dependent reporter transgenic zebrafish line, we show that SNW1 regulates a specific domain of BMP activity in the dorsal ectoderm at the neural plate border at post-gastrula stages. We use double in situ hybridizations and immunofluorescence to show how this domain of BMP activity is spatially positioned relative to the neural crest domain and that of SNW1 expression. Further in vivo and in vitro assays using cell culture and tissue explants allow us to conclude that SNW1 acts upstream of the BMP receptors. Finally, we show that the requirement of SNW1 for neural crest specification is through its ability to regulate BMP activity, as we demonstrate that targeted overexpression of BMP to the neural plate border is sufficient to restore neural crest formation in Xenopus SNW1 morphants. We conclude that through its ability to regulate a specific domain of BMP activity in the vertebrate embryo, SNW1 is a critical regulator of neural plate border formation and thus neural crest specification. PMID:21358802

  8. CLASSIFICATION AND QUANTITATIVE ANALYSIS OF GEOGRAPHIC ATROPHY JUNCTIONAL ZONE USING SPECTRAL DOMAIN OPTICAL COHERENCE TOMOGRAPHY.

    PubMed

    Qu, Jinfeng; Velaga, Swetha Bindu; Hariri, Amir H; Nittala, Muneeswar Gupta; Sadda, Srinivas

    2017-08-22

    The junctional zone at the border of areas of geographic atrophy (GA) in eyes with nonneovascular age-related macular degeneration is an important target region for future therapeutic strategies. The goal of this study was to perform a detailed classification and quantitative characterization of the junctional zone using spectral domain optical coherence tomography. Spectral domain optical coherence tomography volume cube scans (Spectralis OCT, 1024 × 37, Automatic Real Time > 9) were obtained from 15 eyes of 11 patients with GA because of nonneovascular age-related macular degeneration. Volume optical coherence tomography data were imported into previously described validated grading software (3D-OCTOR), and manual segmentation of the retinal pigment epithelium (RPE) and photoreceptor layers was performed on all B-scans (total of 555). Retinal pigment epithelium and photoreceptor defect maps were produced for each case. The borders of the photoreceptor defect area and RPE defect area were delineated individually on separate annotation layers. The two outlines were then superimposed to compare the areas of overlap and nonoverlap. The perimeter of the RPE defect area was calculated by the software in pixels. The superimposed outline of the photoreceptor defect area and the RPE defect area was scrutinized to classify the overlap configuration of the junctional zone into one of three categories: Type 0, exact correspondence between the edge of the RPE defect and photoreceptor defect; Type 1, loss of photoreceptors outside and beyond the edge of the RPE defect; Type 2, preservation of photoreceptors beyond the edge of the RPE defect. The relative proportion of the various border configurations was expressed as a percentage of the perimeter of the RPE defect. Each configuration was then classified into four subgroups according to irregularity of the RPE band and the presence of debris. Fifteen eyes of 11 patients (mean age: 79.3 ± 4.3 years; range: 79-94 years) were included in this study. Seventeen GA lesions were analyzed. Two hundred and thirty-two B-scans were found to pass through the GA lesions, yielding 612 individual GA borders which were separately analyzed and classified. The mean area of the RPE defect was 4.0 ± 4.4 mm, which was significantly smaller than that of the photoreceptor defect which measured 4.4 ± 4.1 mm (paired t test, P = 0.037). On average, 18.0 ± 9.6% (range, 2.3-36.6%) of the junctional zone was of the Type 0 configuration, 57.3 ± 19.0% (range, 21.3-96.8%) was Type 1, and 24.7 ± 18.0% (range, 0.9-64.4%) was Type 2. Type 1 was more prevalent than Type 0 and 2 (analysis of variance, P = 0.000). Debris was present at the margin of the defect in 24.3% (149 of 612) of all assessed junctional zones; 20.0% (14 of 70) of Type 0 junctions, 28.7% (120 of 418) of Type 1, and 12.1% (15 of 124) of Type 2. Debris was more common in Type 1 than Type 2 junctions (P < 0.001). Retinal pigment epithelial irregularity was present at the margin of the defect in 34.8% (213 of 612) of all assessed junctional zones; 52.9% (37 of 70) of Type 0 junctions, 38.0% (159 of 418) of Type 1, and 13.7% (17 of 124) of Type 2. Retinal pigment epithelial irregularity was present more often at Type 0 and Type 1 than at Type 2 junctions (P < 0.001 for both). The size of the optical coherence tomography-visible RPE and photoreceptor defect in GA lesions differ significantly. There were significant areas where the photoreceptor outer segments were preserved despite the absence of visible RPE cells, and also areas of photoreceptor outer segment loss despite apparent RPE preservation. These findings have implications for development of therapeutic strategies, particularly cell-replacement approaches.

  9. The late and dual origin of cerebrospinal fluid-contacting neurons in the mouse spinal cord.

    PubMed

    Petracca, Yanina L; Sartoretti, Maria Micaela; Di Bella, Daniela J; Marin-Burgin, Antonia; Carcagno, Abel L; Schinder, Alejandro F; Lanuza, Guillermo M

    2016-03-01

    Considerable progress has been made in understanding the mechanisms that control the production of specialized neuronal types. However, how the timing of differentiation contributes to neuronal diversity in the developing spinal cord is still a pending question. In this study, we show that cerebrospinal fluid-contacting neurons (CSF-cNs), an anatomically discrete cell type of the ependymal area, originate from surprisingly late neurogenic events in the ventral spinal cord. CSF-cNs are identified by the expression of the transcription factors Gata2 and Gata3, and the ionic channels Pkd2l1 and Pkd1l2. Contrasting with Gata2/3(+) V2b interneurons, differentiation of CSF-cNs is independent of Foxn4 and takes place during advanced developmental stages previously assumed to be exclusively gliogenic. CSF-cNs are produced from two distinct dorsoventral regions of the mouse spinal cord. Most CSF-cNs derive from progenitors circumscribed to the late-p2 and the oligodendrogenic (pOL) domains, whereas a second subset of CSF-cNs arises from cells bordering the floor plate. The development of these two subgroups of CSF-cNs is differentially controlled by Pax6, they adopt separate locations around the postnatal central canal and they display electrophysiological differences. Our results highlight that spatiotemporal mechanisms are instrumental in creating neural cell diversity in the ventral spinal cord to produce distinct classes of interneurons, motoneurons, CSF-cNs, glial cells and ependymal cells. © 2016. Published by The Company of Biologists Ltd.

  10. A cellular automata model for avascular solid tumor growth under the effect of therapy

    NASA Astrophysics Data System (ADS)

    Reis, E. A.; Santos, L. B. L.; Pinho, S. T. R.

    2009-04-01

    Tumor growth has long been a target of investigation within the context of mathematical and computer modeling. The objective of this study is to propose and analyze a two-dimensional stochastic cellular automata model to describe avascular solid tumor growth, taking into account both the competition between cancer cells and normal cells for nutrients and/or space and a time-dependent proliferation of cancer cells. Gompertzian growth, characteristic of some tumors, is described and some of the features of the time-spatial pattern of solid tumors, such as compact morphology with irregular borders, are captured. The parameter space is studied in order to analyze the occurrence of necrosis and the response to therapy. Our findings suggest that transitions exist between necrotic and non-necrotic phases (no-therapy cases), and between the states of cure and non-cure (therapy cases). To analyze cure, the control and order parameters are, respectively, the highest probability of cancer cell proliferation and the probability of the therapeutic effect on cancer cells. With respect to patterns, it is possible to observe the inner necrotic core and the effect of the therapy destroying the tumor from its outer borders inwards.

  11. Perichondrium phenotype and border function are regulated by Ext1 and heparan sulfate in developing long bones: a mechanism likely deranged in Hereditary Multiple Exostoses.

    PubMed

    Huegel, Julianne; Mundy, Christina; Sgariglia, Federica; Nygren, Patrik; Billings, Paul C; Yamaguchi, Yu; Koyama, Eiki; Pacifici, Maurizio

    2013-05-01

    During limb skeletogenesis the cartilaginous long bone anlagen and their growth plates become delimited by perichondrium with which they interact functionally. Yet, little is known about how, despite being so intimately associated with cartilage, perichondrium acquires and maintains its distinct phenotype and exerts its border function. Because perichondrium becomes deranged and interrupted by cartilaginous outgrowths in Hereditary Multiple Exostoses (HME), a pediatric disorder caused by EXT mutations and consequent heparan sulfate (HS) deficiency, we asked whether EXT genes and HS normally have roles in establishing its phenotype and function. Indeed, conditional Ext1 ablation in perichondrium and lateral chondrocytes flanking the epiphyseal region of mouse embryo long bone anlagen - a region encompassing the groove of Ranvier - caused ectopic cartilage formation. A similar response was observed when HS function was disrupted in long bone anlagen explants by genetic, pharmacological or enzymatic means, a response preceded by ectopic BMP signaling within perichondrium. These treatments also triggered excess chondrogenesis and cartilage nodule formation and overexpression of chondrogenic and matrix genes in limb bud mesenchymal cells in micromass culture. Interestingly, the treatments disrupted the peripheral definition and border of the cartilage nodules in such a way that many nodules overgrew and fused with each other into large amorphous cartilaginous masses. Interference with HS function reduced the physical association and interactions of BMP2 with HS and increased the cell responsiveness to endogenous and exogenous BMP proteins. In sum, Ext genes and HS are needed to establish and maintain perichondrium's phenotype and border function, restrain pro-chondrogenic signaling proteins including BMPs, and restrict chondrogenesis. Alterations in these mechanisms may contribute to exostosis formation in HME, particularly at the expense of regions rich in progenitor cells including the groove of Ranvier. Copyright © 2013 Elsevier Inc. All rights reserved.

  12. Visualization of structural organization of ventral membranes of sheared-open resorbing osteoclasts attached to apatite pellets.

    PubMed

    Akisaka, Toshitaka; Yoshida, Atsushi

    2015-05-01

    Osteoclasts are highly polarized cells from both morphological and functional points of view. Using quick-freeze, rotary-replication methods combined with cell-shearing, we clarified the variability of cytoplasmic surface of the polarized membranes of osteoclasts seeded on apatite. As to the organization of actin filaments and clathrin sheets, we confirmed almost the same ventral membrane specializations of osteoclasts on apatite as seen on glass plates. The organized actin filaments and membrane-associated particles supported the ruffled border membranes. Inside the actin sealing zone, membrane specializations were not always occupied with the ruffled border but also with other types of membranes. Some osteoclasts formed an actin ring but lacked the ruffled border projections. We report a unique and distinctive membrane modification of apatite-attached osteoclasts, i.e., the presence of dense aggregates of membrane-associated particles and related structures not found in the osteoclasts seeded on glass plates. Actin filament polarity in the podosomes was determined by decoration with myosin S1. The actin filament polarity within podosome appears to be oriented predominantly with its barbed ends toward the core, whereas the interconnecting F-actin appears to be mixed oriented. Two different types of clathrin plaques displayed different distributions: clathrin-dependent endocytosis was observed in the ruffled border regions, whereas flat clathrin sheets were found in the leading edge of lamellipodia and near podosomes. The clathrin sheets adhered to the apatite surface tightly on the ventral membranes overlaying the resorption lacunae. All these membrane specializations as mentioned above may indicate the functional variability of osteoclasts seeded on apatite.

  13. Regulation of protein expression and function of octn2 in forskolin-induced syncytialization in BeWo Cells.

    PubMed

    Huang, F-D; Kung, F-L; Tseng, Y-C; Chen, M-R; Chan, H-S; Lin, C-J

    2009-02-01

    Placental OCTN2 is a high-affinity carnitine transporter that can interact with a number of therapeutic agents. The process of syncytialization is associated with the expression of a variety of genes. However, the association between syncytialization and OCTN2 expression is not yet clear. Given that forskolin induces BeWo cells to undergo biochemical and morphological differentiation, the purpose of the present study was to investigate whether the function and expression of OCTN2 are influenced by forskolin treatment during syncytialization. The forskolin-induced differentiation of BeWo cells was validated by secretion of beta-human chorionic gonadotropin (beta-hCG) and syncytin expression. Cellular localization of OCTN2 was analyzed by confocal microscopy. Expression of OCTN2 and the modular proteins PDZK1, PDZK2, NHERF1 and NHERF2 was analyzed by Western blotting and carnitine uptake by BeWo cells was estimated and the kinetic properties of uptake measured. The results showed that forskolin treatment increased beta-hCG secretion and syncytin expression, suggesting induction of syncytialization. Confocal images of BeWo cells showed the localization of OCTN2 in the brush-border membrane. OCTN2 protein expression was upregulated in isolated brush-border membranes by long-term forskolin treatment, but the V(m) for carnitine uptake was unchanged, although the K(m) increased. PDZK1, NHERF1 and NHERF2 protein expression in the brush-border membrane was downregulated by forskolin treatment, whereas PDZK2 levels remained unchanged. In conclusion, protein expression and function of OCTN2 in BeWo cells can be regulated by forskolin treatment. While the presence of forskolin results in an increase in OCTN2 protein expression, the increase in uptake capacity may be compensated by the decreased expression of PDZK1, NHERF1 or NHERF2.

  14. Area 3a in the cat. I. A reevaluation of its location and architecture on the basis of Nissl, myelin, acetylcholinesterase, and cytochrome oxidase staining.

    PubMed

    Avendaño, C; Verdu, A

    1992-07-15

    Current knowledge on the anatomy of area 3a of the cat mainly derives from the cyto- and myeloarchitectonic study of Hassler and Muhs-Clement (J Hirnforsch 6:377, 1964). Previous investigations in the cat had failed to identify a cortical region comparable to monkey's area 3a. In the present study, Nissl, myelin, acetylcholinesterase, and cytochrome oxidase staining techniques were applied to coronal and sagittal serial sections of the cat brain. Area 3a appears as a slender band of cortex between areas 4 and 3b, and in Nissl-stained sections it is mainly characterized by an attenuated granular layer IV, overlying a thin layer V with pyramidal cells of various sizes, including a few large ones. These cytoarchitectonic features are sufficient to differentiate area 3a from neighboring areas, although the borders between them are not sharp in many cases. After the Nissl staining, the acetylcholinesterase staining proved to be the most helpful in defining the structure and borders of area 3a. Acetylcholinesterase staining was dense in layer I (in contrast with a lighter staining of outer layer I in area 4), and light in layers II and IIIa, changing to moderate in IIIc and IV (a pattern which is accentuated in area 3b). Myelin and cytochrome oxidase techniques also yielded differential staining patterns of area 3a and neighboring areas 4 and 3b, although the borders were not easily drawn with these techniques. Whereas our cyto- and myeloarchitectonic findings were comparable to those of Hassler and Muhs-Clement ('64) and applied well to area 3a in the convexity of the hemisphere, we found that most of the area 3a described by these authors in the medial face of the hemisphere had a number of distinguishing architectonic (as well as connectional and physiological) features which enabled us to define it as a separate area (7m). The techniques we used to delineate area 3a are compatible with most current procedures of histo- and immunohistochemical staining of the brain, and may also provide valuable supporting data for electrophysiological studies.

  15. Insecurity and Instability in the Sahel Region: The Case of Mali

    DTIC Science & Technology

    2012-03-19

    desertification . Great distances separate the three main regional capitals in the area. For example a nomad who knows the region can travel from Kidal to...which makes securing the border and providing government services very difficult. Also, desertification means that locals cannot necessarily rely on...historical Trans- Saharan commerce and its routes are the key factors for the trade.15 Difficult Socio-Political Environment. Desertification has put

  16. Geometric modeling of Plateau borders using the orthographic projection method for closed cell rigid polyurethane foam thermal conductivity prediction

    NASA Astrophysics Data System (ADS)

    Xu, Jie; Wu, Tao; Peng, Chuang; Adegbite, Stephen

    2017-09-01

    The geometric Plateau border model for closed cell polyurethane foam was developed based on volume integrations of approximated 3D four-cusp hypocycloid structure. The tetrahedral structure of convex struts was orthogonally projected into 2D three-cusp deltoid with three central cylinders. The idealized single unit strut was modeled by superposition. The volume of each component was calculated by geometric analyses. The strut solid fraction f s and foam porosity coefficient δ were calculated based on representative elementary volume of Kelvin and Weaire-Phelan structures. The specific surface area Sv derived respectively from packing structures and deltoid approximation model were put into contrast against strut dimensional ratio ɛ. The characteristic foam parameters obtained from this semi-empirical model were further employed to predict foam thermal conductivity.

  17. Cell cycle progression in irradiated endothelial cells cultured from bovine aorta

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

    Rubin, D.B.; Drab, E.A.; Ward, W.F.

    1988-11-01

    Logarithmically growing endothelial cells from bovine aortas were exposed to single doses of 0-10 Gy of 60Co gamma rays, and cell cycle phase distribution and progression were examined by flow cytometry and autoradiography. In some experiments, cells were synchronized in the cell cycle with hydroxyurea (1 mM). Cell number in sham-irradiated control cultures doubled in approximately 24 h. Estimated cycle stage times for control cells were 14.4 h for G1 phase, 7.2 h for S phase, and 2.4 h for G2 + M phase. Irradiated cells demonstrated a reduced distribution at the G1/S phase border at 4 h, and anmore » increased distribution in G2 + M phase at 24 h postirradiation. Autoradiographs of irradiated cells after continuous (3H)thymidine labeling indicated a block in G1 phase or at the G1/S-phase border. The duration of the block was dose dependent (2-3 min/cGy). Progression of the endothelial cells through S phase after removal of the hydroxyurea block also was retarded by irradiation, as demonstrated by increased distribution in early S phase and decreased distribution in late S phase. These results indicate that progression of asynchronous cultured bovine aortic endothelial cells through the DNA synthetic cycle is susceptible to radiation inhibition at specific sites in the cycle, resulting in redistribution and partial synchronization of the population. Thus aortic endothelial cells, diploid cells from a normal tissue, resemble many immortal cell types that have been examined in this regard in vitro.« less

  18. Bacillus thuringiensis pore-forming toxins trigger massive shedding of GPI-anchored aminopeptidase N from gypsy moth midgut epithelial cells

    Treesearch

    Algimantas P. Valaitis

    2008-01-01

    The insecticidal Cry proteins produced by Bacillus thuringiensis strains are pore-forming toxins (PFTs) that bind to the midgut brush border membrane and cause extensive damage to the midgut epithelial cells of susceptible insect larvae. Force-feeding B. thuringiensis PFTs to Lymantria dispar larvae elicited...

  19. The Foreign Body Giant Cell Cannot Resorb Bone, But Dissolves Hydroxyapatite Like Osteoclasts.

    PubMed

    ten Harkel, Bas; Schoenmaker, Ton; Picavet, Daisy I; Davison, Noel L; de Vries, Teun J; Everts, Vincent

    2015-01-01

    Foreign body multinucleated giant cells (FBGCs) and osteoclasts share several characteristics, like a common myeloid precursor cell, multinuclearity, expression of tartrate-resistant acid phosphatase (TRAcP) and dendritic cell-specific transmembrane protein (DC-STAMP). However, there is an important difference: osteoclasts form and reside in the vicinity of bone, while FBGCs form only under pathological conditions or at the surface of foreign materials, like medical implants. Despite similarities, an important distinction between these cell types is that osteoclasts can resorb bone, but it is unknown whether FBGCs are capable of such an activity. To investigate this, we differentiated FBGCs and osteoclasts in vitro from their common CD14+ monocyte precursor cells, using different sets of cytokines. Both cell types were cultured on bovine bone slices and analyzed for typical osteoclast features, such as bone resorption, presence of actin rings, formation of a ruffled border, and characteristic gene expression over time. Additionally, both cell types were cultured on a biomimetic hydroxyapatite coating to discriminate between bone resorption and mineral dissolution independent of organic matrix proteolysis. Both cell types differentiated into multinucleated cells on bone, but FBGCs were larger and had a higher number of nuclei compared to osteoclasts. FBGCs were not able to resorb bone, yet they were able to dissolve the mineral fraction of bone at the surface. Remarkably, FBGCs also expressed actin rings, podosome belts and sealing zones--cytoskeletal organization that is considered to be osteoclast-specific. However, they did not form a ruffled border. At the gene expression level, FBGCs and osteoclasts expressed similar levels of mRNAs that are associated with the dissolution of mineral (e.g., anion exchange protein 2 (AE2), carbonic anhydrase 2 (CAII), chloride channel 7 (CIC7), and vacuolar-type H+-ATPase (v-ATPase)), in contrast the matrix degrading enzyme cathepsin K, which was hardly expressed by FBGCs. Functionally, the latter cells were able to dissolve a biomimetic hydroxyapatite coating in vitro, which was blocked by inhibiting v-ATPase enzyme activity. These results show that FBGCs have the capacity to dissolve the mineral phase of bone, similar to osteoclasts. However, they are not able to digest the matrix fraction of bone, likely due to the lack of a ruffled border and cathepsin K.

  20. The Foreign Body Giant Cell Cannot Resorb Bone, But Dissolves Hydroxyapatite Like Osteoclasts

    PubMed Central

    ten Harkel, Bas; Schoenmaker, Ton; Picavet, Daisy I.; Davison, Noel L.; de Vries, Teun J.; Everts, Vincent

    2015-01-01

    Foreign body multinucleated giant cells (FBGCs) and osteoclasts share several characteristics, like a common myeloid precursor cell, multinuclearity, expression of tartrate-resistant acid phosphatase (TRAcP) and dendritic cell-specific transmembrane protein (DC-STAMP). However, there is an important difference: osteoclasts form and reside in the vicinity of bone, while FBGCs form only under pathological conditions or at the surface of foreign materials, like medical implants. Despite similarities, an important distinction between these cell types is that osteoclasts can resorb bone, but it is unknown whether FBGCs are capable of such an activity. To investigate this, we differentiated FBGCs and osteoclasts in vitro from their common CD14+ monocyte precursor cells, using different sets of cytokines. Both cell types were cultured on bovine bone slices and analyzed for typical osteoclast features, such as bone resorption, presence of actin rings, formation of a ruffled border, and characteristic gene expression over time. Additionally, both cell types were cultured on a biomimetic hydroxyapatite coating to discriminate between bone resorption and mineral dissolution independent of organic matrix proteolysis. Both cell types differentiated into multinucleated cells on bone, but FBGCs were larger and had a higher number of nuclei compared to osteoclasts. FBGCs were not able to resorb bone, yet they were able to dissolve the mineral fraction of bone at the surface. Remarkably, FBGCs also expressed actin rings, podosome belts and sealing zones—cytoskeletal organization that is considered to be osteoclast-specific. However, they did not form a ruffled border. At the gene expression level, FBGCs and osteoclasts expressed similar levels of mRNAs that are associated with the dissolution of mineral (e.g., anion exchange protein 2 (AE2), carbonic anhydrase 2 (CAII), chloride channel 7 (CIC7), and vacuolar-type H+-ATPase (v-ATPase)), in contrast the matrix degrading enzyme cathepsin K, which was hardly expressed by FBGCs. Functionally, the latter cells were able to dissolve a biomimetic hydroxyapatite coating in vitro, which was blocked by inhibiting v-ATPase enzyme activity. These results show that FBGCs have the capacity to dissolve the mineral phase of bone, similar to osteoclasts. However, they are not able to digest the matrix fraction of bone, likely due to the lack of a ruffled border and cathepsin K. PMID:26426806

  1. Topography and behavior of Sertoli cells in sparse culture during the transitional remodeling phase.

    PubMed

    Tung, P S; Choi, A H; Fritz, I B

    1988-01-01

    We report observations on the behavior of Sertoli cells in sparse culture during the period from the time of plating to the time of initial confluence (the transitional remodeling phase). Changes in shape, structure, and polarity of cells, as well as changes in migration patterns and cell-cell association patterns, have been followed during the transitional remodeling phase with the aid of topographical markers. These markers are based upon differences between ultrastructural features of the basolateral and apicolateral surfaces. The basolateral surface is characterized by plasmalemmal blebs, whereas the apicolateral surface is characterized by filopodial extensions. Structural differences observed in situ remain evident in Sertoli cells isolated by sequential enzymatic treatments that are described. Another marker is provided by laminin-binding sites, which are detected exclusively on the blebbed, basolateral surfaces of freshly prepared Sertoli cell aggregates. The orientation described is sustained during the initial radial migration of Sertoli cells explanted on uncoated glass coverslips. Under these conditions, blebs are detected only on the dorsal surfaces, and filopodial extensions are evident only on the ventral surfaces. In contrast, Sertoli cells sparsely plated on a reconstituted basement membrane (air-dried Matrigel) migrate rapidly, display an extraordinary capacity to form elaborate cytoplasmic extensions for cell-cell and cell-substratum contacts, and readily retract blebs and filopodial extensions. These cells do not form mosaic borders, whereas cells plated on uncoated glass do form a monolayer with mosaic-like borders. Cells sparsely seeded on gelated Matrigel migrate preferentially at gaps between adjacent cell explants, and develop a compact cell-cell association pattern. These cells display few, if any, cytoplasmic extensions. We compare the behavior of Sertoli cells sparsely plated on Matrigel with the behavior of Sertoli cells in situ during different stages of development.

  2. Intestinal epithelial cell surface glycosylation in mice. I. Effect of high-protein diet.

    PubMed

    Gupta, R; Jaswal, V M; Meenu Mahmood, A

    1992-01-01

    The effects of variation in dietary protein content have been investigated on brush border glycosylation and enzyme activities in mice small intestine. The comparison of different parameters was made between the mice fed 30% (high protein, HP) and 18% protein (pair-fed, PF, and ad libitum-fed) for 21 days. The activities of brush border sucrase, lactase, p-nitrophenyl (PNP)-beta-D-glucosidase and PNP-beta-D-galactosidase were reduced in the HP diet-fed mice compared to PF and ad libitum-fed controls. Alkaline phosphatase and leucine amino-peptidase activities were significantly enhanced while gamma-glutamyl transpeptidase activity was unaltered under these conditions. Total hexoses and sialic acid content in the brush borders were reduced significantly in the test group compared to the controls while hexosamine and fucose contents remained essentially similar in different groups. The results on the binding of wheat germ agglutinin and Ulex europaeus agglutininI to microvillus membranes corroborated the chemical analysis data on sialic acid and fucose contents of the membranes. Peanut agglutinin binding was enhanced in mice from the HP group. Incorporation of (14C)-mannose into membranes was significantly less in HP diet-fed mice. These results indicate that the feeding of HP diet to mice brings about marked alterations in small intestinal epithelial cell surface glycosylation and enzyme functions.

  3. Minimal Network Topologies for Signal Processing during Collective Cell Chemotaxis.

    PubMed

    Yue, Haicen; Camley, Brian A; Rappel, Wouter-Jan

    2018-06-19

    Cell-cell communication plays an important role in collective cell migration. However, it remains unclear how cells in a group cooperatively process external signals to determine the group's direction of motion. Although the topology of signaling pathways is vitally important in single-cell chemotaxis, the signaling topology for collective chemotaxis has not been systematically studied. Here, we combine mathematical analysis and simulations to find minimal network topologies for multicellular signal processing in collective chemotaxis. We focus on border cell cluster chemotaxis in the Drosophila egg chamber, in which responses to several experimental perturbations of the signaling network are known. Our minimal signaling network includes only four elements: a chemoattractant, the protein Rac (indicating cell activation), cell protrusion, and a hypothesized global factor responsible for cell-cell interaction. Experimental data on cell protrusion statistics allows us to systematically narrow the number of possible topologies from more than 40,000,000 to only six minimal topologies with six interactions between the four elements. This analysis does not require a specific functional form of the interactions, and only qualitative features are needed; it is thus robust to many modeling choices. Simulations of a stochastic biochemical model of border cell chemotaxis show that the qualitative selection procedure accurately determines which topologies are consistent with the experiment. We fit our model for all six proposed topologies; each produces results that are consistent with all experimentally available data. Finally, we suggest experiments to further discriminate possible pathway topologies. Copyright © 2018 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  4. Dbl3 drives Cdc42 signaling at the apical margin to regulate junction position and apical differentiation

    PubMed Central

    Zihni, Ceniz; Munro, Peter M.G.; Elbediwy, Ahmed; Keep, Nicholas H.; Terry, Stephen J.; Harris, John

    2014-01-01

    Epithelial cells develop morphologically characteristic apical domains that are bordered by tight junctions, the apical–lateral border. Cdc42 and its effector complex Par6–atypical protein kinase c (aPKC) regulate multiple steps during epithelial differentiation, but the mechanisms that mediate process-specific activation of Cdc42 to drive apical morphogenesis and activate the transition from junction formation to apical differentiation are poorly understood. Using a small interfering RNA screen, we identify Dbl3 as a guanine nucleotide exchange factor that is recruited by ezrin to the apical membrane, that is enriched at a marginal zone apical to tight junctions, and that drives spatially restricted Cdc42 activation, promoting apical differentiation. Dbl3 depletion did not affect junction formation but did affect epithelial morphogenesis and brush border formation. Conversely, expression of active Dbl3 drove process-specific activation of the Par6–aPKC pathway, stimulating the transition from junction formation to apical differentiation and domain expansion, as well as the positioning of tight junctions. Thus, Dbl3 drives Cdc42 signaling at the apical margin to regulate morphogenesis, apical–lateral border positioning, and apical differentiation. PMID:24379416

  5. Cross-border reprogenetic services.

    PubMed

    Couture, V; Drouin, R; Tan, S-L; Moutquin, J-M; Bouffard, C

    2015-01-01

    The purpose of this review is to synthesize the current knowledge on the international movement of patients and biopsied embryo cells for pre-implantation genetic diagnosis and its different applications. Thus far, few attempts have been made to identify the specific nature of this phenomenon called 'cross-border reprogenetic services'. There is scattered evidence, both empirical and speculative, suggesting that these services raise major issues in terms of service provision, risks for patients and the children-to-come, the legal liabilities of physicians, as well as social justice. To compile this evidence, this review uses the narrative overview protocol combined with thematic analysis. Five major themes have emerged from the literature at the conjunction of cross-border treatments and reprogenetics: 'scope', 'scale', 'motivations', 'concerns', and 'governance'. Similar themes have already been observed in the case of other medical tourism activities, but this review highlights their singularity with reprogenetic services. It emphasizes the diagnostic and autologous feature of reprogenetics, the constant risk of misdiagnosis, the restriction on certain tests for medically controversial conditions, and the uncertain accessibility of genetic counseling in cross-border settings. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  6. Object representation in the human auditory system

    PubMed Central

    Winkler, István; van Zuijen, Titia L.; Sussman, Elyse; Horváth, János; Näätänen, Risto

    2010-01-01

    One important principle of object processing is exclusive allocation. Any part of the sensory input, including the border between two objects, can only belong to one object at a time. We tested whether tones forming a spectro-temporal border between two sound patterns can belong to both patterns at the same time. Sequences were composed of low-, intermediate- and high-pitched tones. Tones were delivered with short onset-to-onset intervals causing the high and low tones to automatically form separate low and high sound streams. The intermediate-pitch tones could be perceived as part of either one or the other stream, but not both streams at the same time. Thus these tones formed a pitch ’border’ between the two streams. The tones were presented in a fixed, cyclically repeating order. Linking the intermediate-pitch tones with the high or the low tones resulted in the perception of two different repeating tonal patterns. Participants were instructed to maintain perception of one of the two tone patterns throughout the stimulus sequences. Occasional changes violated either the selected or the alternative tone pattern, but not both at the same time. We found that only violations of the selected pattern elicited the mismatch negativity event-related potential, indicating that only this pattern was represented in the auditory system. This result suggests that individual sounds are processed as part of only one auditory pattern at a time. Thus tones forming a spectro-temporal border are exclusively assigned to one sound object at any given time, as are spatio-temporal borders in vision. PMID:16836636

  7. Choroidal Haller's and Sattler's Layers Thickness in Normal Indian Eyes.

    PubMed

    Roy, Rupak; Saurabh, Kumar; Vyas, Chinmayi; Deshmukh, Kaustubh; Sharma, Preeti; Chandrasekharan, Dhileesh P; Bansal, Aditya

    2018-01-01

    This study aims to study normative choroidal thickness (CT) and Haller's and Sattler's layers thickness in normal Indian eyes. The choroidal imaging of 73 eyes of 43 healthy Indian individuals was done using enhanced depth imaging feature of spectralis optical coherence tomography. Rraster scan protocol centered at fovea was used for imaging separately by two observers. CT was defined as the length of the perpendicular line drown from the outer border of hypereflective RPE-Bruch's complex to inner margin of choroidoscleral junction. Choroidal vessel layer thickness was measured after defining a largest choroidal vessel lumen within 750 μ on either side of the subfoveal CT vector. A perpendicular line was drawn to the innermost border of this lumen, and the distance between the perpendicular line and innermost border of choroidoscleral junction gave large choroidal vessel layer thickness (LCVLT, Haller's layer). Medium choroidal vessel layer thickness (MCVLT, Sattler's layer) was measured as the distance between same perpendicular line and outer border of hypereflective RPE-Bruch's complex. The mean age of individuals was 28.23 ± 15.29 years (range 14-59 years). Overall, the mean subfoveal CT was 331.6 ± 63.9 μ. Mean LCVLT was 227.08 ± 51.24 μ and the mean MCVLT was 95.65 ± 23.62 μ. CT was maximum subfoveally with gradual reduction in the thickness as the distance from the fovea increased. This is the first study describing the choroidal sublayer thickness, i.e., Haller's and Sattler's layer thickness along with CT in healthy Indian population.

  8. Choroidal Haller's and Sattler's Layers Thickness in Normal Indian Eyes

    PubMed Central

    Roy, Rupak; Saurabh, Kumar; Vyas, Chinmayi; Deshmukh, Kaustubh; Sharma, Preeti; Chandrasekharan, Dhileesh P.; Bansal, Aditya

    2018-01-01

    AIM: This study aims to study normative choroidal thickness (CT) and Haller's and Sattler's layers thickness in normal Indian eyes. MATERIALS AND METHODS: The choroidal imaging of 73 eyes of 43 healthy Indian individuals was done using enhanced depth imaging feature of spectralis optical coherence tomography. Rraster scan protocol centered at fovea was used for imaging separately by two observers. CT was defined as the length of the perpendicular line drown from the outer border of hypereflective RPE-Bruch's complex to inner margin of choroidoscleral junction. Choroidal vessel layer thickness was measured after defining a largest choroidal vessel lumen within 750 μ on either side of the subfoveal CT vector. A perpendicular line was drawn to the innermost border of this lumen, and the distance between the perpendicular line and innermost border of choroidoscleral junction gave large choroidal vessel layer thickness (LCVLT, Haller's layer). Medium choroidal vessel layer thickness (MCVLT, Sattler's layer) was measured as the distance between same perpendicular line and outer border of hypereflective RPE-Bruch's complex. RESULTS: The mean age of individuals was 28.23 ± 15.29 years (range 14–59 years). Overall, the mean subfoveal CT was 331.6 ± 63.9 μ. Mean LCVLT was 227.08 ± 51.24 μ and the mean MCVLT was 95.65 ± 23.62 μ. CT was maximum subfoveally with gradual reduction in the thickness as the distance from the fovea increased. CONCLUSION: This is the first study describing the choroidal sublayer thickness, i.e., Haller's and Sattler's layer thickness along with CT in healthy Indian population. PMID:29899646

  9. Exploitable Vulnerabilities of Iran’s Islamic Revolutionary Guard Corps

    DTIC Science & Technology

    2011-10-12

    former IRGC Qods ( Jerusalem ) Force commander, He is also the subject of an Interpol Red Notice for his role in the 1994 attacks on Israeli and Jewish...maintain a capability to take the conflict outside its borders through unconventional tactics and asymmetric warfare. The Pasdaran’s Qods ( Jerusalem ...Force is its primary tool in that endeavor. The Jerusalem Force will be addressed separately in subsequent discussion of the IRGC’s influence

  10. [Common intralobular microcirculatory module peculliarities in cholestasis in white rats].

    PubMed

    Sulaberidze, G D; Kardzvia, D Dzh; Kikalishvili, L A; Khomeriki, Ts T

    2006-03-01

    The dynamics of intralobular microcirculatory module transformation in cholestasis was investigated. The liver tissues of 54 white Wistar rats were studied by Histology, TEM, TEM after injection of 1% Pb(NO(3))(3), vie common bile duct (CBD), SEM of corrosion casts prepared after methylmethacrylate injection vie CBD. It is demonstrated that liver is the basis of microcirculation of 4 different liquids: blood, bile, connective tissue liquid and lymph. Accordingly, in normal condition 4 different well formed compartments bordered from each-others by different types of cells exist. In the yearly stages of cholestasis (3-6 days) the function of bile drainage is partially overtaken by lymph pathways. This is accompanied by the beginning of the destruction of structures bordering above-mentioned microcirculatory beds. In the later stages of cholestasis (12-18 days) the final disorganization of microcirculatory compartments with mixture of all liquids is observed. It is accompanied by increased dystrophy of liver cells population.

  11. Analysis of high mass resolution PTR-TOF mass spectra from 1,3,5-trimethylbenzene (TMB) environmental chamber experiments

    NASA Astrophysics Data System (ADS)

    Müller, M.; Graus, M.; Wisthaler, A.; Hansel, A.; Metzger, A.; Dommen, J.; Baltensperger, U.

    2011-09-01

    A series of 1,3,5-trimethylbenzene (TMB) photo-oxidation experiments was performed in the 27-m3 Paul Scherrer Institute environmental chamber under various NOx conditions. A University of Innsbruck prototype high resolution Proton Transfer Reaction Time-of-Flight Mass Spectrometer (PTR-TOF) was used for measurements of gas and particulate phase organics. The gas phase mass spectrum displayed ~200 ion signals during the TMB photo-oxidation experiments. Molecular formulas CNmHnNoOp were determined and ion signals were separated and grouped according to their C, O and N numbers. This allowed to determine the time evolution of the O:C ratio and of the average carbon oxidation state OSC of the reaction mixture. Both quantities were compared with master chemical mechanism (MCMv3.1) simulations. The O:C ratio in the particle phase was about twice the O:C ratio in the gas phase. Average carbon oxidation states of secondary organic aerosol (SOA) samples OSCSOA were in the range of -0.34 to -0.31, in agreement with expected average carbon oxidation states of fresh SOA (OSC = -0.5 - 0).

  12. Analysis of high mass resolution PTR-TOF mass spectra from 1,3,5-trimethylbenzene (TMB) environmental chamber experiments

    NASA Astrophysics Data System (ADS)

    Müller, M.; Graus, M.; Wisthaler, A.; Hansel, A.; Metzger, A.; Dommen, J.; Baltensperger, U.

    2012-01-01

    A series of 1,3,5-trimethylbenzene (TMB) photo-oxidation experiments was performed in the 27-m3 Paul Scherrer Institute environmental chamber under various NOx conditions. A University of Innsbruck prototype high resolution Proton Transfer Reaction Time-of-Flight Mass Spectrometer (PTR-TOF) was used for measurements of gas and particulate phase organics. The gas phase mass spectrum displayed ~200 ion signals during the TMB photo-oxidation experiments. Molecular formulas CmHnNoOp were determined and ion signals were separated and grouped according to their C, O and N numbers. This allowed to determine the time evolution of the O:C ratio and of the average carbon oxidation state OSC of the reaction mixture. Both quantities were compared with master chemical mechanism (MCMv3.1) simulations. The O:C ratio in the particle phase was about twice the O:C ratio in the gas phase. Average carbon oxidation states of secondary organic aerosol (SOA) samples OSCSOA were in the range of -0.34 to -0.31, in agreement with expected average carbon oxidation states of fresh SOA (OSC = -0.5-0).

  13. Students Across Borders: A Summer Earth Science Workshop for Hispanic High School Students

    NASA Astrophysics Data System (ADS)

    Butler, R. F.; Kresan, P.; Baez, A.; Sheppard, P.; Forger, G.; Rendon-Coke, G.; Gray, F.

    2003-12-01

    Southern Arizona has a high school (HS) population that is 28% Hispanic. However this fast-growing minority group represents only 14% of undergraduate students at the University of Arizona and 11% of science and engineering majors. The Students Across Borders Program was designed to assist Hispanic HS students across borders that often separate them from higher education and careers in science. In June 2003, five person student-teacher teams from Tucson, Yuma, and northern Sonora, Mexico lived in dormitories and participated in a weeklong program based on the University of Arizona campus. Activities included: field trips featuring inquiry-based investigations of geology, water quality, and tree rings; tours of engineering and science laboratories; introduction to student support organizations such as the Society of Hispanic Professional Engineers; and counseling by Career Services and Admissions personnel. Technology training included instruction in web design, digital imaging and online communication tools. Web sites developed by the student teams were presented to participants and families at the conclusion of the on-campus program. Web site development is continuing during the academic year to foster continuing communication between the student teams and presentation of results of follow-on projects assisted by graduate and undergraduate CATTS fellows and university faculty.

  14. Synchrony and the binding problem in macaque visual cortex

    PubMed Central

    Dong, Yi; Mihalas, Stefan; Qiu, Fangtu; von der Heydt, Rüdiger; Niebur, Ernst

    2009-01-01

    We tested the binding-by-synchrony hypothesis which proposes that object representations are formed by synchronizing spike activity between neurons that code features of the same object. We studied responses of 32 pairs of neurons recorded with microelectrodes 3 mm apart in the visual cortex of macaques performing a fixation task. Upon mapping the receptive fields of the neurons, a quadrilateral was generated so that two of its sides were centered in the receptive fields at the optimal orientations. This one-figure condition was compared with a two-figure condition in which the neurons were stimulated by two separate figures, keeping the local edges in the receptive fields identical. For each neuron, we also determined its border ownership selectivity (H. Zhou, H. S. Friedman, & R. von der Heydt, 2000). We examined both synchronization and correlation at nonzero time lag. After correcting for effects of the firing rate, we found that synchrony did not depend on the binding condition. However, finding synchrony in a pair of neurons was correlated with finding border-ownership selectivity in both members of the pair. This suggests that the synchrony reflected the connectivity in the network that generates border ownership assignment. Thus, we have not found evidence to support the binding-by-synchrony hypothesis. PMID:19146262

  15. Localization of Bacillus thuringiensis Cry1A toxin-binding molecules in gypsy moth larval gut sections using fluorescence microscopy

    Treesearch

    Algimantas P. Valaitis

    2011-01-01

    The microbial insecticide Bacillus thuringiensis (Bt) produces Cry toxins, proteins that bind to the brush border membranes of gut epithelial cells of insects that ingest it, disrupting the integrity of the membranes, and leading to cell lysis and insect death. In gypsy moth, Lymantria dispar, two toxin-binding molecules for the...

  16. Actuation of chitosan-aptamer nanobrush borders for pathogen sensing.

    PubMed

    Hills, Katherine D; Oliveira, Daniela A; Cavallaro, Nicholas D; Gomes, Carmen L; McLamore, Eric S

    2018-03-26

    We demonstrate a sensing mechanism for rapid detection of Listeria monocytogenes in food samples using the actuation of chitosan-aptamer nanobrush borders. The bio-inspired soft material and sensing strategy mimic natural symbiotic systems, where low levels of bacteria are selectively captured from complex matrices. To engineer this biomimetic system, we first develop reduced graphene oxide/nanoplatinum (rGO-nPt) electrodes, and characterize the fundamental electrochemical behavior in the presence and absence of chitosan nanobrushes during actuation (pH-stimulated osmotic swelling). We then characterize the electrochemical behavior of the nanobrush when receptors (antibodies or DNA aptamers) are conjugated to the surface. Finally, we test various techniques to determine the most efficient capture strategy based on nanobrush actuation, and then apply the biosensors in a food product. Maximum cell capture occurs when aptamers conjugated to the nanobrush bind cells in the extended conformation (pH < 6), followed by impedance measurement in the collapsed nanobrush conformation (pH > 6). The aptamer-nanobrush hybrid material was more efficient than the antibody-nanobrush material, which was likely due to the relatively high adsorption capacity for aptamers. The biomimetic material was used to develop a rapid test (17 min) for selectively detecting L. monocytogenes at concentrations ranging from 9 to 107 CFU mL-1 with no pre-concentration, and in the presence of other Gram-positive cells (Listeria innocua and Staphylococcus aureus). Use of this bio-inspired material is among the most efficient for L. monocytogenes sensing to date, and does not require sample pretreatment, making nanobrush borders a promising new material for rapid pathogen detection in food.

  17. Exclusion of EDNRB and KIT as the basis for white spotting in Border Collies.

    PubMed

    Metallinos, D; Rine, J

    2000-01-01

    White spotting patterns in mammals can be caused by mutations in the genes for the endothelin B receptor and c-Kit, whose protein products are necessary for proper migration, differentiation or survival of the melanoblast population of cells. Although there are many different dog breeds that segregate white spotting patterns, no genes have been identified that are linked to these phenotypes. An intercross was generated from a female Newfoundland and a male Border Collie and the white spotting phenotypes of the intercross progeny were evaluated by measuring percentage surface area of white in the puppies. The Border Collie markings segregated as a simple autosomal recessive (7/25 intercross progeny had the phenotype). Two candidate genes, for the endothelin B receptor (EDNRB) and c-Kit (KIT), were evaluated for segregation with the white spotting pattern. Polymorphisms between the Border Collie and Newfoundland were identified for EDNRB using Southern analysis after a portion of the canine gene had been cloned. Polymorphisms for KIT were identified using a microsatellite developed from a bacterial artificial chromosome containing the canine gene. Both EDNRB and KIT were excluded as a cause of the white spotting pattern in at least two of the intercross progeny. Although these genes have been implicated in white spotting in other mammals, including horses, pigs, cows, mice and rats, they do not appear to be responsible for the white spotting pattern found in the Border Collie breed of dog.

  18. Exclusion of EDNRB and KIT as the basis for white spotting in Border Collies

    PubMed Central

    Metallinos, Danika; Rine, Jasper

    2000-01-01

    Background: White spotting patterns in mammals can be caused by mutations in the genes for the endothelin B receptor and c-Kit, whose protein products are necessary for proper migration, differentiation or survival of the melanoblast population of cells. Although there are many different dog breeds that segregate white spotting patterns, no genes have been identified that are linked to these phenotypes. Results: An intercross was generated from a female Newfoundland and a male Border Collie and the white spotting phenotypes of the intercross progeny were evaluated by measuring percentage surface area of white in the puppies. The Border Collie markings segregated as a simple autosomal recessive (7/25 intercross progeny had the phenotype). Two candidate genes, for the endothelin B receptor (EDNRB) and c-Kit (KIT), were evaluated for segregation with the white spotting pattern. Polymorphisms between the Border Collie and Newfoundland were identified for EDNRB using Southern analysis after a portion of the canine gene had been cloned. Polymorphisms for KIT were identified using a microsatellite developed from a bacterial artificial chromosome containing the canine gene. Conclusions: Both EDNRB and KIT were excluded as a cause of the white spotting pattern in at least two of the intercross progeny. Although these genes have been implicated in white spotting in other mammals, including horses, pigs, cows, mice and rats, they do not appear to be responsible for the white spotting pattern found in the Border Collie breed of dog. PMID:11178229

  19. Neuroglian stabilizes epithelial structure during Drosophila oogenesis.

    PubMed

    Wei, Jun; Hortsch, Michael; Goode, Scott

    2004-08-01

    The vertebrate L1 family of cell adhesion molecules (CAMs) and their fly homolog, Neuroglian, are members of the immunoglobulin (Ig) superfamily of CAMs. In general, Ig CAMs have been found to play critical roles in mediating axon guidance. One Ig CAM, NCAM, has also been implicated in maintaining epithelial integrity and suppressing metastatic dissemination of tumor cells. Other Ig CAMs, such as Nrg, are also expressed in epithelia. We thus tested the hypothesis that, like NCAM, Nrg might also be required for maintaining epithelial integrity and for inhibiting tumor invasion. We used the Drosophila follicular epithelium to determine the function of Nrg in vivo in maintaining epithelial structure, and in regulating the motility of migrating border cells and invasive tumorous follicle cells. Nrg(167) is expressed on the lateral membrane of follicle cells. Loss of Nrg(167) causes border cells to delay delamination and causes other follicle cells to delaminate inappropriately. The delaminated cells have aberrant epithelial polarity manifested as severe mislocalization of apical and basal membrane proteins, and uniform localization of lateral membrane proteins. Furthermore, loss of Nrg(167) dramatically enhances the invasive phenotype associated with loss of Discs Large, a neoplastic tumor suppressor. These results indicate that Nrg(167) stabilizes epithelial polarity by regulating junctional adhesion and function in normal and tumorous epithelia. Our data also suggest that Ig superfamily members have significant functional redundancy in maintaining epithelial polarity, with individual members playing subtle, unique roles during epithelial morphogenesis. Copyright 2004 Wiley-Liss, Inc.

  20. Identification of linker regions and domain borders of the transcription activator protein NtrC from Escherichia coli by limited proteolysis, in-gel digestion, and mass spectrometry.

    PubMed

    Bantscheff, M; Weiss, V; Glocker, M O

    1999-08-24

    We have developed a mass spectrometry based method for the identification of linker regions and domain borders in multidomain proteins. This approach combines limited proteolysis and in-gel proteolytic digestions and was applied to the determination of linkers in the transcription factor NtrC from Escherichia coli. Limited proteolysis of NtrC with thermolysin and papain revealed that initial digestion yielded two major bands in SDS-PAGE that were identified by mass spectrometry as the R-domain and the still covalently linked OC-domains. Subsequent steps in limited proteolysis afforded further cleavage of the OC-fragment into the O- and the C-domain at accessible amino acid residues. Mass spectrometric identification of the tryptic/thermolytic peptides obtained after in-gel total proteolysis of the SDS-PAGE-separated domains determined the domain borders and showed that the protease accessible linker between R- and O-domain comprised amino acids Val-131 and Gln-132 within the "Q-linker" in agreement with papain and subtilisin digestion. The region between amino acid residues Thr-389 and Gln-396 marked the hitherto unknown linker sequence that connects the O- with the C-domain. High abundances of proline-, alanine-, serine-, and glutamic acid residues were found in this linker structure (PASE-linker) of related NtrC response regulator proteins. While R- and C-domains remained stable under the applied limited proteolysis conditions, the O-domain was further truncated yielding a core fragment that comprised the sequence from Ile-140 to Arg-320. ATPase activity was lost after separation of the R-domain from the OC-fragment. However, binding of OC- and C- fragments to specific DNA was observed by characteristic band-shifts in migration retardation assays, indicating intact tertiary structures of the C-domain. The outlined strategy proved to be highly efficient and afforded lead information of tertiary structural features necessary for protein design and engineering and for structure-function studies.

  1. Avoiding Gluten Cross-Contamination

    MedlinePlus

    ... are a Key Part of Nutrition for Men's Health Nutrition for the Child with Sickle Cell Anemia Food ... Twitter Youtube True © 2018 eatright.org. Academy of Nutrition and Dietetics, All Rights ... style="display: none;" border="0" height="1" width="1" alt="Quantcast"/>

  2. Microimaging of Bacillus thuringiensis Toxin-binding proteins in gypsy moth larval gut using confocal fluorescence microscopy

    Treesearch

    Daniel J. Krofcheck; Algimantas P. Valaitis

    2010-01-01

    After ingestion by susceptible insect larvae, Bacillus thuringiensis (Bt) insecticidal proteins bind to the brush border membranes of gut epithelial cells and disrupt the integrity of the plasma membrane by forming...

  3. Organic particulate matter formation at varying relative humidity using surrogate secondary and primary organic compounds with activity corrections in the condensed phase obtained using a method based on the Wilson equation

    NASA Astrophysics Data System (ADS)

    Chang, E. I.; Pankow, J. F.

    2008-01-01

    Secondary organic aerosol (SOA) formation in the atmosphere is currently often modeled using a multiple lumped "two-product" (N·2p) approach. The N·2p approach neglects: 1) variation of activity coefficient (ζi) values and mean molecular weight MW in the particulate matter (PM) phase; 2) water uptake into the PM; and 3) the possibility of phase separation in the PM. This study considers these effects by adopting an (N·2p)ζ, MW ,θ approach (θ is a phase index). Specific chemical structures are assigned to 25 lumped SOA compounds and to 15 representative primary organic aerosol (POA) compounds to allow calculation of ζi and MW values. The SOA structure assignments are based on chamber-derived 2p gas/particle partition coefficient values coupled with known effects of structure on vapor pressure pL,i° (atm). To facilitate adoption of the (N·2p)ζ, MW, θ approach in large-scale models, this study also develops CP-Wilson.1, a group-contribution ζi-prediction method that is more computationally economical than the UNIFAC model of Fredenslund et al. (1975). Group parameter values required by CP-Wilson.1 are obtained by fitting ζi values to predictions from UNIFAC. The (N·2p)ζ,MW, θ approach is applied (using CP-Wilson.1) to several real α-pinene/O3 chamber cases for high reacted hydrocarbon levels (ΔHC≍400 to 1000 μg m-3) when relative humidity (RH) ≍50%. Good agreement between the chamber and predicted results is obtained using both the (N·2p)ζ, MW, θ and N·2p approaches, indicating relatively small water effects under these conditions. However, for a hypothetical α-pinene/O3 case at ΔHC=30 μg m-3 and RH=50%, the (N·2p)ζ, MW, θ approach predicts that water uptake will lead to an organic PM level that is more double that predicted by the N·2p approach. Adoption of the (N·2p)ζ, MW, θ approach using reasonable lumped structures for SOA and POA compounds is recommended for ambient PM modeling.

  4. Multiple scattering in chiral media: border effects, reduced depolarization, and sensitivity limit

    NASA Astrophysics Data System (ADS)

    Delplancke, Francoise; Badoz, Jacques P.; Boccara, A. Claude

    1997-10-01

    Suspensions of polystyrene latex beads in chiral solutions were investigated. The rotatory power, induced by solubilized sucrose, in near-forward scattering was measured via a method using polarization modulation by photo-elastic modulator. The sensitivity of the measurement was enhanced and optimized in order to measure sucrose concentrations as low as 5 mg/ml in a cell 5 mm thick only. Different concentrations and diameters of latex particles were used in combination with different sucrose concentrations going from 1 mg/ml up to saturation. The experiments showed that the apparent rotatory power is enhanced by multiple scattering, that depolarization effects are less important with highly concentrated sucrose solutions and that attention has to be paid to cell border effects in order to avoid important artifacts, in case of highly scattering suspensions. Qualitative and theoretical explanations of those observations are presented. One possible application of this method is to measure the sugar content in human blood, in vivo, non-invasively, through the skin. The concentration to be evaluated is at the sensitivity limit. So any artifact has to be removed carefully, e.g. skin cell birefringence or chirality.

  5. RhoB controls endothelial barrier recovery by inhibiting Rac1 trafficking to the cell border

    PubMed Central

    Marcos-Ramiro, Beatriz; García-Weber, Diego; Barroso, Susana; Feito, Jorge; Ortega, María C.; Cernuda-Morollón, Eva; Reglero-Real, Natalia; Fernández-Martín, Laura; Durán, Maria C.; Alonso, Miguel A.; Correas, Isabel; Cox, Susan; Ridley, Anne J.

    2016-01-01

    Endothelial barrier dysfunction underlies chronic inflammatory diseases. In searching for new proteins essential to the human endothelial inflammatory response, we have found that the endosomal GTPase RhoB is up-regulated in response to inflammatory cytokines and expressed in the endothelium of some chronically inflamed tissues. We show that although RhoB and the related RhoA and RhoC play additive and redundant roles in various aspects of endothelial barrier function, RhoB specifically inhibits barrier restoration after acute cell contraction by preventing plasma membrane extension. During barrier restoration, RhoB trafficking is induced between vesicles containing RhoB nanoclusters and plasma membrane protrusions. The Rho GTPase Rac1 controls membrane spreading and stabilizes endothelial barriers. We show that RhoB colocalizes with Rac1 in endosomes and inhibits Rac1 activity and trafficking to the cell border during barrier recovery. Inhibition of endosomal trafficking impairs barrier reformation, whereas induction of Rac1 translocation to the plasma membrane accelerates it. Therefore, RhoB-specific regulation of Rac1 trafficking controls endothelial barrier integrity during inflammation. PMID:27138256

  6. The Fate of the Red Cells: Insights from Two Models of Severe Malarial Anemia

    DTIC Science & Technology

    2011-03-07

    approximately 1%. The reticulocyte levels in these animals elevated to approximately 35% 2 days after the anemic crisis and then returned to basal...cells are destroyed for every parasitized red cell22. A prospective study in a Karen community on the western border of Thailand showed that in anemia...activation in severe Plasmodium falciparum malaria. Clin.Immunol.Immunopathol. 1997;85:166-171. 136. Facer CA, Bray RS, Brown J. Direct Coombs

  7. Id expression in amphioxus and lamprey highlights the role of gene cooption during neural crest evolution

    NASA Technical Reports Server (NTRS)

    Meulemans, Daniel; McCauley, David; Bronner-Fraser, Marianne

    2003-01-01

    Neural crest cells are unique to vertebrates and generate many of the adult structures that differentiate them from their closest invertebrate relatives, the cephalochordates. Id genes are robust markers of neural crest cells at all stages of development. We compared Id gene expression in amphioxus and lamprey to ask if cephalochordates deploy Id genes at the neural plate border and dorsal neural tube in a manner similar to vertebrates. Furthermore, we examined whether Id expression in these cells is a basal vertebrate trait or a derived feature of gnathostomes. We found that while expression of Id genes in the mesoderm and endoderm is conserved between amphioxus and vertebrates, expression in the lateral neural plate border and dorsal neural tube is a vertebrate novelty. Furthermore, expression of lamprey Id implies that recruitment of Id genes to these cells occurred very early in the vertebrate lineage. Based on expression in amphioxus we postulate that Id cooption conferred sensory cell progenitor-like properties upon the lateral neurectoderm, and pharyngeal mesoderm-like properties upon cranial neural crest. Amphioxus Id expression is also consistent with homology between the anterior neurectoderm of amphioxus and the presumptive placodal ectoderm of vertebrates. These observations support the idea that neural crest evolution was driven in large part by cooption of multipurpose transcriptional regulators from other tissues and cell types.

  8. JPRS Report, Near East and South Asia.

    DTIC Science & Technology

    1991-06-27

    Friday. The march started from the place of residence and moved to the border point ( La The Moroccan authorities intervened diplomatically. Firga...separating Gibraltar and Spanish La Linea. It was However, their efforts failed in the face of Spain’s followed by daily waves of human sit-ins on the...users of private cars is still pending the establishment of ities, and the Spanish and English media-strongly a land route between La Firga and Green

  9. Reality and the Physicist

    NASA Astrophysics Data System (ADS)

    D'Espagnat, Bernard; Whitehouse, Translated by J. C.

    1989-03-01

    Preface; Introduction; Part I. Instrumentalism and Science: 1. The positivism of the physicists; 2. Positivism and fallibilism: philosophical controversies; 3. Border areas of instrumentalism; Part II. Physical Realism and Contemporary Physics: 4. Physical realism and fallibilism; 5. Microrealism and non-separability; 6. Physical realism in trouble; Part III. Causality, Reality and Time: 7. Irreversibility; 8. Sensible reality; 9. Independent reality; 10. The dilemma of modern physics: reality or meaning?; 11. Questions and answers; 12. Summary and perspectives; Appendixes; Addendum; Notes; References; Index.

  10. Recent judicial developments in state income taxation of the oil and gas industry

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

    Hellerstein, W.

    1986-03-01

    The oil and gas industry has been at the center of activity involving state income taxation. The author reviews several recent judicial decisions whose implications extend beyond individual state borders. He describes how state interpretations of the Uniform Division of Income for Tax Purposes Act (UDITPA) property factor, the deductibility of the Windfall Profits Tax, and the constitutionality of Alaska's separate accounting for oil companies led to decisions which affect controversies in other states.

  11. Separation of abscission zone cells in detached Azolla roots depends on apoplastic pH.

    PubMed

    Fukuda, Kazuma; Yamada, Yoshiya; Miyamoto, Kensuke; Ueda, Junichi; Uheda, Eiji

    2013-01-01

    In studies on the mechanism of cell separation during abscission, little attention has been paid to the apoplastic environment. We found that the apoplastic pH surrounding abscission zone cells in detached roots of the water fern Azolla plays a major role in cell separation. Abscission zone cells of detached Azolla roots were separated rapidly in a buffer at neutral pH and slowly in a buffer at pH below 4.0. However, cell separation rarely occurred at pH 5.0-5.5. Light and electron microscopy revealed that cell separation was caused by a degradation of the middle lamella between abscission zone cells at both pH values, neutral and below 4.0. Low temperature and papain treatment inhibited cell separation. Enzyme(s) in the cell wall of the abscission zone cells might be involved in the degradation of the pectin of the middle lamella and the resultant, pH-dependent cell separation. By contrast, in Phaseolus leaf petioles, unlike Azolla roots, cell separation was slow and increased only at acidic pH. The rapid cell separation, as observed in Azolla roots at neutral pH, did not occur. Indirect immunofluorescence microscopy, using anti-pectin monoclonal antibodies, revealed that the cell wall pectins of the abscission zone cells of Azolla roots and Phaseolus leaf petioles looked similar and changed similarly during cell separation. Thus, the pH-related differences in cell separation mechanisms of Azolla and Phaseolus might not be due to differences in cell wall pectin, but to differences in cell wall-located enzymatic activities responsible for the degradation of pectic substances. A possible enzyme system is discussed. Copyright © 2012 Elsevier GmbH. All rights reserved.

  12. Fundamentals of affinity cell separations.

    PubMed

    Zhang, Ye; Lyons, Veronica; Pappas, Dimitri

    2018-03-01

    Cell separations using affinity methods continue to be an enabling science for a wide variety of applications. In this review, we discuss the fundamental aspects of affinity separation, including the competing forces for cell capture and elution, cell-surface interactions, and models for cell adhesion. Factors affecting separation performance such as bond affinity, contact area, and temperature are presented. We also discuss and demonstrate the effects of nonspecific binding on separation performance. Metrics for evaluating cell separations are presented, along with methods of comparing separation techniques for cell isolation using affinity capture. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Long life, rechargeable nickel-zinc battery

    NASA Technical Reports Server (NTRS)

    Luksha, E.

    1974-01-01

    A production version of the inorganic separator was evaluated for improving the life of the nickel-zinc system. Nickel-zinc cells (7-10 Ah capacities) of different electrode separator configurations were constructed and tested. The nickel-zinc cells using the inorganic separator encasing the zinc electrode, the nickel electrode, or both electrodes had shorter lives than cells using Visking and cellophane separation. Cells with the inorganic separation all fell below 70% of their theoretical capacity within 30 cycles, but the cells constructed with organic separation required 80 cycles. Failure of the cells using the ceramic separator was irreversible capacity degradation due to zinc loss through cracks developed in the inorganic separator. Zinc loss through the separator was minimized with the use of combinations of the inorganic separator with Visking and cellophane. Cells using the combined separation operated 130 duty cycles before degrading to 70% of their theoretical capacity.

  14. Visual area of the lateral suprasylvian gyrus (Clare—Bishop area) of the cat

    PubMed Central

    Hubel, David H.; Wiesel, Torsten N.

    1969-01-01

    On anatomical and physiological grounds a zone of cat cortex deep in the medial bank of the suprasylvian sulcus (the Clare—Bishop area) is known to receive strong visual projections both from the lateral geniculate body and area 17. We have mapped receptive fields of single cells in this area in eight cats. Active responses to visual stimuli were found over most of the medial bank of the suprasylvian sulcus extending to the depths and over to the lowest part of the lateral bank. The area is clearly topographically arranged. The first responsive cells, recorded over the lateral convexity and 2-3 mm down the medial bank, had receptive fields in the far periphery of the contralateral visual fields. The receptive fields tended to be large, but showed considerable variation in size and scatter in their positions. As the electrode advanced down the bank, fields of successively recorded cells gradually tended to move inwards, so that in the depths of the sulcus the inner borders of many of the fields reached the vertical mid line. Here the fields were smaller, though they still varied very much in size. Receptive fields were larger than in 17, 18, or 19, but otherwise were not obviously different from the complex and lower-order hypercomplex fields in those areas. No simple fields, or concentric fields of the retino-geniculate type, were seen. Cells with common receptive-field orientation were grouped together, but whether or not the grouping occurs in columns was not established. Most cells were driven independently by the two eyes. Fields in the two eyes seemed to be identical in organization. Cells dominated by the contralateral eye were much more common than ipsilaterally dominated ones, but when cells with parafoveal and peripheral fields were considered separately, the asymmetry was seen to apply mainly to cells with peripheral fields. PMID:5770897

  15. An Inversion Disrupting FAM134B Is Associated with Sensory Neuropathy in the Border Collie Dog Breed

    PubMed Central

    Forman, Oliver P.; Hitti, Rebekkah J.; Pettitt, Louise; Jenkins, Christopher A.; O’Brien, Dennis P.; Shelton, G. Diane; De Risio, Luisa; Quintana, Rodrigo Gutierrez; Beltran, Elsa; Mellersh, Cathryn

    2016-01-01

    Sensory neuropathy in the Border Collie is a severe neurological disorder caused by the degeneration of sensory and, to a lesser extent, motor nerve cells with clinical signs starting between 2 and 7 months of age. Using a genome-wide association study approach with three cases and 170 breed matched controls, a suggestive locus for sensory neuropathy was identified that was followed up using a genome sequencing approach. An inversion disrupting the candidate gene FAM134B was identified. Genotyping of additional cases and controls and RNAseq analysis provided strong evidence that the inversion is causal. Evidence of cryptic splicing resulting in novel exon transcription for FAM134B was identified by RNAseq experiments. This investigation demonstrates the identification of a novel sensory neuropathy associated mutation, by mapping using a minimal set of cases and subsequent genome sequencing. Through mutation screening, it should be possible to reduce the frequency of or completely eliminate this debilitating condition from the Border Collie breed population. PMID:27527794

  16. Arbovirus Surveillance near the Mexico-U.S. Border: Isolation and Sequence Analysis of Chikungunya Virus from Patients with Dengue-like Symptoms in Reynosa, Tamaulipas.

    PubMed

    Laredo-Tiscareño, S Viridiana; Machain-Williams, Carlos; Rodríguez-Pérez, Mario A; Garza-Hernandez, Javier A; Doria-Cobos, Gloria L; Cetina-Trejo, Rosa C; Bacab-Cab, Lucio A; Tangudu, Chandra S; Charles, Jermilia; De Luna-Santillana, Erick J; Garcia-Rejon, Julian E; Blitvich, Bradley J

    2018-05-14

    A total of 1,090 residents of the city of Reynosa, Tamaulipas, on the Mexico-U.S. border presented at hospitals and clinics of the Secretariat of Health, Mexico, in 2015 with symptoms characteristic of dengue. Dengue virus (DENV) antigen was detected by enzyme-linked immunosorbent assay in acute sera from 134 (12.3%) patients. Sera from select patients ( N = 34) were also tested for chikungunya virus (CHIKV) RNA by quantitative reverse transcription-polymerase chain reaction. Thirteen (38.2%) patients, including five DENV antigen-positive patients, were positive. Sera from three CHIKV RNA-positive patients were further assayed by virus isolation in cell culture and CHIKV was recovered on each occasion. The genome of one isolate and structural genes of the other two isolates were sequenced. In conclusion, we present evidence of CHIKV and DENV coinfections in patients who live near the Mexico-U.S. border and provide the first genome sequence of a CHIKV isolate from northern Mexico.

  17. Morocco and border of spain as seen from STS-59

    NASA Technical Reports Server (NTRS)

    1994-01-01

    The Atlantic Ocean, Straits of Gibraltar, and Alboran Sea (the westernmost portion of the Mediterranean Sea) separate Spain on the left from Morocco on the right. Algeciras Harbor is the prominent notch cut out of the eastern end of the north shore of the Strait; the Rock of Gibraltar is the tiny arrowhead that separates the notch from the Alboran Sea. The Sierra Nevada, farther away down the Spanish coast, lives up to its name in this April scene. The difference in elevation between the Sierra Morena and the Guadalquivir River valley is highlighted nicely by cumulus clouds. Tangier, Morocco can be seen as a light-toned spot on the southern shore of the Strait, near the entrance to the Atlantic Ocean.

  18. Environmental boundaries as a mechanism for correcting and anchoring spatial maps

    PubMed Central

    2016-01-01

    Abstract Ubiquitous throughout the animal kingdom, path integration‐based navigation allows an animal to take a circuitous route out from a home base and using only self‐motion cues, calculate a direct vector back. Despite variation in an animal's running speed and direction, medial entorhinal grid cells fire in repeating place‐specific locations, pointing to the medial entorhinal circuit as a potential neural substrate for path integration‐based spatial navigation. Supporting this idea, grid cells appear to provide an environment‐independent metric representation of the animal's location in space and preserve their periodic firing structure even in complete darkness. However, a series of recent experiments indicate that spatially responsive medial entorhinal neurons depend on environmental cues in a more complex manner than previously proposed. While multiple types of landmarks may influence entorhinal spatial codes, environmental boundaries have emerged as salient landmarks that both correct error in entorhinal grid cells and bind internal spatial representations to the geometry of the external spatial world. The influence of boundaries on error correction and grid symmetry points to medial entorhinal border cells, which fire at a high rate only near environmental boundaries, as a potential neural substrate for landmark‐driven control of spatial codes. The influence of border cells on other entorhinal cell populations, such as grid cells, could depend on plasticity, raising the possibility that experience plays a critical role in determining how external cues influence internal spatial representations. PMID:26563618

  19. Separation of cancer cells from white blood cells by pinched flow fractionation.

    PubMed

    Pødenphant, Marie; Ashley, Neil; Koprowska, Kamila; Mir, Kalim U; Zalkovskij, Maksim; Bilenberg, Brian; Bodmer, Walter; Kristensen, Anders; Marie, Rodolphe

    2015-12-21

    In this paper, the microfluidic size-separation technique pinched flow fractionation (PFF) is used to separate cancer cells from white blood cells (WBCs). The cells are separated at efficiencies above 90% for both cell types. Circulating tumor cells (CTCs) are found in the blood of cancer patients and can form new tumors. CTCs are rare cells in blood, but they are important for the understanding of metastasis. There is therefore a high interest in developing a method for the enrichment of CTCs from blood samples, which also enables further analysis of the separated cells. The separation is challenged by the size overlap between cancer cells and the 10(6) times more abundant WBCs. The size overlap prevents high efficiency separation, however we demonstrate that cell deformability can be exploited in PFF devices to gain higher efficiencies than expected from the size distribution of the cells.

  20. A simplified sheathless cell separation approach using combined gravitational-sedimentation-based prefocusing and dielectrophoretic separation.

    PubMed

    Luo, Tao; Fan, Lei; Zeng, Yixiao; Liu, Ya; Chen, Shuxun; Tan, Qiulin; Lam, Raymond H W; Sun, Dong

    2018-05-04

    Prefocusing of the cell mixture is necessary for achieving a high-efficiency and continuous dielectrophoretic (DEP) cell separation. However, prefocusing through sheath flow requires a complex and tedious peripheral system for multi-channel fluid control, hindering the integration of DEP separation systems with other microfluidic functionalities for comprehensive clinical and biological tasks. This paper presented a simplified sheathless cell separation approach that combines gravitational-sedimentation-based sheathless prefocusing and DEP separation methods. Through gravitational sedimentation in a tubing, which was inserted into the inlet of a microfluidic chip with an adjustable steering angle, the cells were focused into a stream at the upstream region of a microchannel prior to separation. Then, a DEP force was applied at the downstream region of the microchannel for the active separation of the cells. Through this combined strategy, the peripheral system for the sheath flow was no longer required, and thus the integration of cell separation system with additional microfluidic functionalities was facilitated. The proposed sheathless scheme focused the mixture of cells with different sizes and dielectric properties into a stream in a wide range of flow rates without changing the design of the microfluidic chip. The DEP method is a label-free approach that can continuously separate cells on the basis of the sizes or dielectric properties of the cells and thus capable of greatly flexible cell separation. The efficiency of the proposed approach was experimentally assessed according to its performance in the separation of human acute monocytic leukemia THP-1 cells from yeast cells with respect to different sizes and THP-1 cells from human acute myelomonocytic leukemia OCI-AML3 cells with respect to different dielectric properties. The experimental results revealed that the separation efficiency of the method can surpass 90% and thus effective in separating cells on the basis of either size or dielectric property.

  1. Federal Research and Development Funding: FY2010

    DTIC Science & Technology

    2009-10-22

    and represent the more research-oriented part of the RDT&E program. Budget activities 6.4 and 6.5 focus on the development of specific weapon ...Institute (HSI), which was funded as a separate item in FY2009. The House bill would provide $15 million more than the request for chemical and biological ...132 142 142 143 R&D, Acquisition, and Operations 692 800 826 825 844 Border and Maritime 25 33 40 40 40 Chemical and Biological 208 200 207 222

  2. Final Environmental Assessment: Lease with Omaha Public Power District (OPPD) to Support New United States Strategic Command (USSTRATCOM) Facility and Existing Base Load

    DTIC Science & Technology

    2013-02-01

    Biological Resources The area around and encompassing Offutt AFB is the western edge of the Eastern Deciduous Forest and borders on the ecotone...that separates the Eastern Deciduous Forest from the Tall and Mid Grass Prairies. Early photos of the Offutt AFB area indicate that it was grassland...regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden, to Washington

  3. Acetylated sialic acid residues and blood group antigens localise within the epithelium in microvillous atrophy indicating internal accumulation of the glycocalyx

    PubMed Central

    Phillips, A D; Brown, A; Hicks, S; Schüller, S; Murch, S H; Walker-Smith, J A; Swallow, D M

    2004-01-01

    Background: Microvillous atrophy, a disorder of intractable diarrhoea in infancy, is characterised by the intestinal epithelial cell abnormalities of abnormal accumulation of periodic acid-Schiff (PAS) positive secretory granules within the apical cytoplasm and the presence of microvillous inclusions. The identity of the PAS positive material is not known, and the aim of this paper was to further investigate its composition. Methods: Formaldehyde fixed sections were stained with alcian blue/PAS to identify the acidic or neutral nature of the material, phenylhydrazine blocking was employed to stain specifically for sialic acid, and saponification determined the presence of sialic acid acetylation. The specificity of sialic acid staining was tested by digestion with mild sulphuric acid. Expression of blood group related antigens was tested immunochemically. Results: Alcian blue/PAS staining identified a closely apposed layer of acidic material on the otherwise neutral (PAS positive) brush border in controls. In microvillous atrophy, a triple layer was seen with an outer acidic layer, an unstained brush border region, and accumulation within the epithelium of a neutral glycosubstance that contained acetylated sialic acid. Blood group antigens were detected on the brush border, in mucus, and within goblet cells in controls. In microvillous atrophy they were additionally expressed within the apical cytoplasm of epithelial cells mirroring the PAS abnormality. Immuno electron microscopy localised expression to secretory granules. Conclusions: A neutral, blood group antigen positive, glycosubstance that contains acetylated sialic acid accumulates in the epithelium in microvillous atrophy. Previous studies have demonstrated that the direct and indirect constitutive pathways are intact in this disorder and it is speculated that the abnormal staining pattern reflects accumulation of glycocalyx related material. PMID:15542511

  4. Identification of simple objects in image sequences

    NASA Astrophysics Data System (ADS)

    Geiselmann, Christoph; Hahn, Michael

    1994-08-01

    We present an investigation in the identification and location of simple objects in color image sequences. As an example the identification of traffic signs is discussed. Three aspects are of special interest. First regions have to be detected which may contain the object. The separation of those regions from the background can be based on color, motion, and contours. In the experiments all three possibilities are investigated. The second aspect focuses on the extraction of suitable features for the identification of the objects. For that purpose the border line of the region of interest is used. For planar objects a sufficient approximation of perspective projection is affine mapping. In consequence, it is near at hand to extract affine-invariant features from the border line. The investigation includes invariant features based on Fourier descriptors and moments. Finally, the object is identified by maximum likelihood classification. In the experiments all three basic object types are correctly identified. The probabilities for misclassification have been found to be below 1%

  5. HIV Risk Behaviors and Correlates of Inconsistent Condom Use Among Substance Using Migrants at the Mexico/Guatemala Border.

    PubMed

    Conners, Erin E; Swanson, Kate; Morales-Miranda, Sonia; Fernández Casanueva, Carmen; Mercer, Valerie J; Brouwer, Kimberly C

    2017-07-01

    This study assessed correlates of inconsistent condom use with casual partners and the prevalence of sexual risk behaviors and STIs in the Mexico/Guatemala border region using a sample of 392 migrants (303 men, 85 women) who reported current substance use or problem drinking. We ran separate univariate logistic regression models for men and women, and multivariate logistic regression models for men only. Prevalence of syphilis was 1.2% among women and 2.3% among men; HIV prevalence was 2.4% among women and 1.3% among men. Inconsistent condom use with casual partners was higher in women with greater education and lower among women who sold sex. In men, less access to free condoms, drug use with sexual partners, and drug use before sex were independently associated with inconsistent condom use with casual partners. Sexual and substance use risk behaviors were common, and HIV/STI prevention efforts should target both genders and expand beyond most-at risk populations.

  6. Thaumasioscolex didelphidis n. gen., n. sp. (Eucestoda: Proteocephalidae) from the black-eared opossum Didelphis marsupialis from Mexico, the first proteocephalidean tapeworm from a mammal.

    PubMed

    Cañeda-Guzmán, I C; de Chambrier, A; Scholz, T

    2001-06-01

    Thaumasioscolex didelphidis n. gen., n. sp. is described from the intestine of the black-eared opossum Didelphis marsupialis L. (Marsupialia: Didelphidae) from Los Tuxtlas, Veracruz, Mexico. The new genus differs from all proteocephalidean genera in the morphology of the scolex that is formed by 4 well separated lobes each containing 1 noncircular sucker opening laterally inside the exterolateral cavity, a large-sized body (length up to 1 m), a large number of testes, the shape of gravid proglottids that are inversely craspedote (the anterior border of a proglottid overlaps the posterior border of a preceding proglottid), eggs in groups mostly of 4-6 eggs each, and an embryophore bearing digitiform projections on its external surface. This is the first tapeworm of the Proteocephalidea, the members of which were previously reported exclusively from poikilotherm vertebrates (freshwater fishes, amphibians, and reptiles), found in a homoiotherm vertebrate.

  7. Antibody-immobilized column for quick cell separation based on cell rolling.

    PubMed

    Mahara, Atsushi; Yamaoka, Tetsuji

    2010-01-01

    Cell separation using methodological standards that ensure high purity is a very important step in cell transplantation for regenerative medicine and for stem cell research. A separation protocol using magnetic beads has been widely used for cell separation to isolate negative and positive cells. However, not only the surface marker pattern, e.g., negative or positive, but also the density of a cell depends on its developmental stage and differentiation ability. Rapid and label-free separation procedures based on surface marker density are the focus of our interest. In this study, we have successfully developed an antiCD34 antibody-immobilized cell-rolling column, that can separate cells depending on the CD34 density of the cell surfaces. Various conditions for the cell-rolling column were optimized including graft copolymerization, and adjustment of the column tilt angle, and medium flow rate. Using CD34-positive and -negative cell lines, the cell separation potential of the column was established. We observed a difference in the rolling velocities between CD34-positive and CD34-negative cells on antibody-immobilized microfluidic device. Cell separation was achieved by tilting the surface 20 degrees and the increasing medium flow. Surface marker characteristics of the isolated cells in each fraction were analyzed using a cell-sorting system, and it was found that populations containing high density of CD34 were eluted in the delayed fractions. These results demonstrate that cells with a given surface marker density can be continuously separated using the cell rolling column.

  8. Isolation of osteoprogenitors from human jaw periosteal cells: a comparison of two magnetic separation methods.

    PubMed

    Olbrich, Marcus; Rieger, Melanie; Reinert, Siegmar; Alexander, Dorothea

    2012-01-01

    Human jaw periosteum tissue contains osteoprogenitors that have potential for tissue engineering applications in oral and maxillofacial surgeries. To isolate osteoprogenitor cells from heterogeneous cell populations, we used the specific mesenchymal stem cell antigen-1 (MSCA-1) antibody and compared two magnetic separation methods. We analyzed the obtained MSCA-1(+) and MSCA-1(-) fractions in terms of purity, yield of positive/negative cells and proliferative and mineralization potentials. The analysis of cell viability after separation revealed that the EasySep method yielded higher viability rates, whereas the flow cytometry results showed a higher purity for the MACS-separated cell fractions. The mineralization capacity of the osteogenic induced MSCA-1(+) cells compared with the MSCA-1(-) controls using MACS was 5-fold higher, whereas the same comparison after EasySep showed no significant differences between both fractions. By analyzing cell proliferation, we detected a significant difference between the proliferative potential of the osteogenic cells versus untreated cells after the MACS and EasySep separations. The differentiated cells after MACS separation adjusted their proliferative capacity, whereas the EasySep-separated cells failed to do so. The protein expression analysis showed small differences between the two separation methods. Our findings suggest that MACS is a more suitable separation method to isolate osteoprogenitors from the entire jaw periosteal cell population.

  9. Elasticity of biomembranes studied by dynamic light scattering

    NASA Astrophysics Data System (ADS)

    Fujime, Satoru; Miyamoto, Shigeaki

    1991-05-01

    Combination of osmotic swelling and dynamic light scattering makes it possible to measure the elastic modulus of biomembranes. By this technique we have observed a drastic increase in membrane flexibility on activation of Na/glucose cotransporters in membrane vesicles prepared from brush-borders of rat small intestine and on activation by micromolar [Ca2] of exocytosis in secretory granules isolated from rat pancreatic acinar cells and bovine adrenal chromaffin cells. 1 .

  10. Rapid genotyping assays for the 4-base pair deletion of canine MDR1/ABCB1 gene and low frequency of the mutant allele in Border Collie dogs.

    PubMed

    Mizukami, Keijiro; Chang, Hye-Sook; Yabuki, Akira; Kawamichi, Takuji; Hossain, Mohammad A; Rahman, Mohammad M; Uddin, Mohammad M; Yamato, Osamu

    2012-01-01

    P-glycoprotein, encoded by the MDR1 or ABCB1 gene, is an integral component of the blood-brain barrier as an efflux pump for xenobiotics crucial in limiting drug uptake into the central nervous system. Dogs homozygous for a 4-base pair deletion of the canine MDR1 gene show altered expression or function of P-glycoprotein, resulting in neurotoxicosis after administration of the substrate drugs. In the present study, the usefulness of microchip electrophoresis for genotyping assays detecting this deletion mutation was evaluated. Mutagenically separated polymerase chain reaction (MS-PCR) and real-time PCR assays were newly developed and evaluated. Furthermore, a genotyping survey was carried out in a population of Border Collies dogs in Japan to determine the allele frequency in this breed. Microchip electrophoresis showed advantages in detection sensitivity and time saving over other modes of electrophoresis. The MS-PCR assay clearly discriminated all genotypes. Real-time PCR assay was most suitable for a large-scale survey due to its high throughput and rapidity. The genotyping survey demonstrated that the carrier and mutant allele frequencies were 0.49% and 0.25%, respectively, suggesting that the mutant allele frequency in Border Collies is markedly low compared to that in the susceptible dog breeds such as rough and smooth Collies.

  11. C-terminal polymorphism of Plasmodium falciparium merozoite surface protein-1 (MSP-1) from Tak Province, Thailand.

    PubMed

    Viputtigul, Kwanjai; Tungpukdee, Noppadon; Ruangareerate, Toon; Luplertlop, Natthanej; Wilairatana, Polrat; Gaywee, Jariyanart; Krudsood, Srivicha

    2013-01-01

    This study was undertaken to ascertain the extent of polymorphism in the C-terminal region of Plasmodium falciparum merozoite surface protein (MSP-1) from 119 malaria patients in Tak Province on the western border of Thailand, who were admitted to the Bangkok Hospital for Tropical Diseases, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. P. falciparum infection was confirmed by microscopic examination of peripheral blood smears. Clinical manifestations were categorized into 2 groups: uncomplicated (94 cases) and complicated/severe (25 cases). A 1,040 basepair fragment of P. falciparum MSP-1 gene was compared with MSP-1 of reference strains retrieved from GenBank. The consensus sequences of MSP-1 block 16 showed it belonged to MAD20 genotype, which is the major allele of falciparum malaria from the western border of Thailand. MSP-1 block 16 amino acid fragment could be separated into 2 groups: similar and dissimilar to reference sequence. Four variations in MSP-1 block 16 were -1494K, D1510G, D1556N, and K1696I. MSP-1 block 16 diversity is not significantly associated with clinical manifestation although MAD 20 genotype is the predominant genotype in this area. The genetic data of MSP1 gene of faciparum malaria isolated from western Thai border contribute to the existing genetic database of Thai P. falciparum strain.

  12. Culturally Specific Youth Substance Abuse Resistance Skills: Applicability across the U.S.-Mexico Border

    PubMed Central

    Marsiglia, Flavio F.; Kulis, Stephen; Rodriguez, Gregorio Martinez; Becerra, David; Castillo, Jason

    2009-01-01

    This study tests the applicability among adolescents in Mexico of the keepin' it REAL (refuse, explain, avoid, and leave) strategies that are common and effective ways that U.S. youth resist substance use. Following a social learning, communication competence and ecological theory integrated approach, the study draws on self-reported questionnaire data from a non-probability sample of 327 adolescents attending two public high schools in Monterrey, Nuevo León. Multivariate regressions were used to test whether the respondents' use of the REAL strategies by the participants could be predicted by key demographic variables. Separate models were estimated for the frequency of use of each strategy and for different substances. Findings indicate that most adolescents in this sample utilized each of the REAL strategies as well as other strategies to respond to offers of alcohol, cigarettes, or marijuana. Mexican and U.S. youth residing close to the US border appear to use similar drug resistance strategies. Use of the strategies varied considerably by the level of exposure to offers, but only minimally by gender and age. There were no notable differences by socioeconomic status or academic performance. Implications for prevention science, social work practice and social work research are discussed in the context of the bi-national border region and the applicability and prospect for dissemination of U.S. evidence based youth substance use prevention interventions. PMID:19924268

  13. Culturally Specific Youth Substance Abuse Resistance Skills: Applicability across the U.S.-Mexico Border.

    PubMed

    Marsiglia, Flavio F; Kulis, Stephen; Rodriguez, Gregorio Martinez; Becerra, David; Castillo, Jason

    2009-03-01

    This study tests the applicability among adolescents in Mexico of the keepin' it REAL (refuse, explain, avoid, and leave) strategies that are common and effective ways that U.S. youth resist substance use. Following a social learning, communication competence and ecological theory integrated approach, the study draws on self-reported questionnaire data from a non-probability sample of 327 adolescents attending two public high schools in Monterrey, Nuevo León. Multivariate regressions were used to test whether the respondents' use of the REAL strategies by the participants could be predicted by key demographic variables. Separate models were estimated for the frequency of use of each strategy and for different substances. Findings indicate that most adolescents in this sample utilized each of the REAL strategies as well as other strategies to respond to offers of alcohol, cigarettes, or marijuana. Mexican and U.S. youth residing close to the US border appear to use similar drug resistance strategies. Use of the strategies varied considerably by the level of exposure to offers, but only minimally by gender and age. There were no notable differences by socioeconomic status or academic performance. Implications for prevention science, social work practice and social work research are discussed in the context of the bi-national border region and the applicability and prospect for dissemination of U.S. evidence based youth substance use prevention interventions.

  14. Distribution of the cystine/glutamate antiporter system xc- in the brain, kidney, and duodenum.

    PubMed

    Burdo, Joseph; Dargusch, Richard; Schubert, David

    2006-05-01

    System x(c)(-), one of the main transporters responsible for central nervous system cystine transport, is comprised of two subunits, xCT and 4F2hc. The transport of cystine into cells is rate limiting for glutathione synthesis, the major antioxidant and redox cofactor in the brain. Alterations in glutathione status are prevalent in numerous neurodegenerative diseases, emphasizing the importance of proper cystine homeostasis. However, the distribution of xCT and 4F2hc within the brain and other areas has not been described. Using specific antibodies, both xCT and 4F2hc were localized predominantly to neurons in the mouse and human brain, but some glial cells were labeled as well. Border areas between the brain proper and periphery including the vascular endothelial cells, ependymal cells, choroid plexus, and leptomeninges were also highly positive for the system x(c)(-) components. xCT and 4F2hc are also present at the brush border membranes in the kidney and duodenum. These results indicate that system x(c)(-) is likely to play a role in cellular health throughout many areas of the brain as well as other organs by maintaining intracellular cystine levels, thereby resulting in low levels of oxidative stress.

  15. Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex

    NASA Astrophysics Data System (ADS)

    Ohki, Kenichi; Chung, Sooyoung; Ch'ng, Yeang H.; Kara, Prakash; Reid, R. Clay

    2005-02-01

    Neurons in the cerebral cortex are organized into anatomical columns, with ensembles of cells arranged from the surface to the white matter. Within a column, neurons often share functional properties, such as selectivity for stimulus orientation; columns with distinct properties, such as different preferred orientations, tile the cortical surface in orderly patterns. This functional architecture was discovered with the relatively sparse sampling of microelectrode recordings. Optical imaging of membrane voltage or metabolic activity elucidated the overall geometry of functional maps, but is averaged over many cells (resolution >100µm). Consequently, the purity of functional domains and the precision of the borders between them could not be resolved. Here, we labelled thousands of neurons of the visual cortex with a calcium-sensitive indicator in vivo. We then imaged the activity of neuronal populations at single-cell resolution with two-photon microscopy up to a depth of 400µm. In rat primary visual cortex, neurons had robust orientation selectivity but there was no discernible local structure; neighbouring neurons often responded to different orientations. In area 18 of cat visual cortex, functional maps were organized at a fine scale. Neurons with opposite preferences for stimulus direction were segregated with extraordinary spatial precision in three dimensions, with columnar borders one to two cells wide. These results indicate that cortical maps can be built with single-cell precision.

  16. 21 CFR 864.9245 - Automated blood cell separator.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated blood cell separator. 864.9245 Section... Blood and Blood Products § 864.9245 Automated blood cell separator. (a) Identification. An automated blood cell separator is a device that uses a centrifugal or filtration separation principle to...

  17. 21 CFR 864.9245 - Automated blood cell separator.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Automated blood cell separator. 864.9245 Section... Blood and Blood Products § 864.9245 Automated blood cell separator. (a) Identification. An automated blood cell separator is a device that uses a centrifugal or filtration separation principle to...

  18. 21 CFR 864.9245 - Automated blood cell separator.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Automated blood cell separator. 864.9245 Section... Blood and Blood Products § 864.9245 Automated blood cell separator. (a) Identification. An automated blood cell separator is a device that uses a centrifugal or filtration separation principle to...

  19. 21 CFR 864.9245 - Automated blood cell separator.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Automated blood cell separator. 864.9245 Section... Blood and Blood Products § 864.9245 Automated blood cell separator. (a) Identification. An automated blood cell separator is a device that uses a centrifugal or filtration separation principle to...

  20. 21 CFR 864.9245 - Automated blood cell separator.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Automated blood cell separator. 864.9245 Section... Blood and Blood Products § 864.9245 Automated blood cell separator. (a) Identification. An automated blood cell separator is a device that uses a centrifugal or filtration separation principle to...

  1. Patterns of globalized reproduction: Egg cells regulation in Israel and Austria

    PubMed Central

    2012-01-01

    Since the successful introduction of in vitro fertilization in 1978, medically assisted reproduction (MAR) has proliferated in multiple clinical innovations. Consequently, egg cells have become an object of demand for both infertility treatment and stem cell research, and this raises complex legal, ethical, social and economic issues. In this paper we compare how the procurement and use of human egg cells is regulated in two countries: Israel and Austria. Israel is known for its scientific leadership, generous public funding, high utilization and liberal regulation of assisted reproductive technology (ART). Austria lies at the other extreme of the regulatory spectrum in terms of restrictions on reproductive interventions. In both countries, however, there is a constant increase in the use of the technology, and recent legal developments make egg cells more accessible. Also, in both countries the scarcity of egg cells in concert with the rising demand for donations has led to the emergence of cross-border markets and global 'reproductive tourism' practices. In Israel, in particular, a scandal known as the 'eggs affair' was followed by regulation that allowed egg cell donations from outside the country under certain conditions. Cross-border markets are developed by medical entrepreneurs, driven by global economic gaps, made possible by trans-national regulatory lacunae and find expression as consumer demand. The transnational practice of egg cell donations indicates the emergence of a global public health issue, but there is a general lack of medical and epidemiological data on its efficacy and safety. We conclude that there is need for harmonisation of domestic laws and formulation of new instruments for international governance. PMID:22913734

  2. Boron Supply Enhances Aluminum Tolerance in Root Border Cells of Pea (Pisum sativum) by Interacting with Cell Wall Pectins

    PubMed Central

    Fang, Jing; Tao, Lin; Shen, Ren Fang; Li, Ya Lin; Xiao, Hong Dong; Feng, Ying Ming; Wen, Hai Xiang; Guan, Jia Hua; Wu, Li Shu; He, Yong Ming; Goldbach, Heiner E.; Yu, Min

    2017-01-01

    Aluminum (Al) toxicity is the primary factor limiting crop growth in acidic soils. Boron (B) alleviates Al toxicity in plants, which is mainly considered to be due to the formation of Rhamnogalacturonan II-B (RGII-B) complexes, which helps to stabilize the cytoskeleton. It is unclear yet whether this is due to the increasing of net negative charges and/or further mechanisms. Kinetics of Al accumulation and adsorption were investigated using entire cells, cell wall and pectin of root border cells (RBCs) of pea (Pisum sativum), to reveal the mechanism of B in interacting with alkali-soluble and chelator-soluble pectin for an increased Al tolerance in RBCs. The results show that B could rescue RBCs from Al-induced cell death by accumulating more Al in the cell wall, predominately in alkali-soluble pectin. Boron also promotes Al3+ adsorption and inhibits Al3+ desorption from alkali-soluble pectin. Thus, more Al3+ is immobilized within the alkali-soluble pectin fraction and less in the chelator-soluble pectin, rendering Al3+ less mobile. Boron induces an increase of RG-II (KDO,2-keto-3-deoxyoctonic acid) content for forming more borate-RGII complexes, and the decrease of pectin methyl-esterification, thus creates more negative charges to immobilize Al3+ in cell wall pectin. The study provides evidence that abundant B supply enhances the immobilization of Al in alkali-soluble pectin, thus most likely reducing the entry of Al3+ into the symplast from the surroundings. PMID:28533794

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

    PubMed Central

    Morin, M.; Morehouse, L. G.

    1974-01-01

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

  4. Patterns of globalized reproduction: Egg cells regulation in Israel and Austria.

    PubMed

    Shalev, Carmel; Werner-Felmayer, Gabriele

    2012-04-18

    Since the successful introduction of in vitro fertilization in 1978, medically assisted reproduction (MAR) has proliferated in multiple clinical innovations. Consequently, egg cells have become an object of demand for both infertility treatment and stem cell research, and this raises complex legal, ethical, social and economic issues.In this paper we compare how the procurement and use of human egg cells is regulated in two countries: Israel and Austria. Israel is known for its scientific leadership, generous public funding, high utilization and liberal regulation of assisted reproductive technology (ART). Austria lies at the other extreme of the regulatory spectrum in terms of restrictions on reproductive interventions.In both countries, however, there is a constant increase in the use of the technology, and recent legal developments make egg cells more accessible. Also, in both countries the scarcity of egg cells in concert with the rising demand for donations has led to the emergence of cross-border markets and global 'reproductive tourism' practices. In Israel, in particular, a scandal known as the 'eggs affair' was followed by regulation that allowed egg cell donations from outside the country under certain conditions.Cross-border markets are developed by medical entrepreneurs, driven by global economic gaps, made possible by trans-national regulatory lacunae and find expression as consumer demand. The transnational practice of egg cell donations indicates the emergence of a global public health issue, but there is a general lack of medical and epidemiological data on its efficacy and safety. We conclude that there is need for harmonisation of domestic laws and formulation of new instruments for international governance.

  5. The impact of Border policy effect on cross-border ethnic areas

    NASA Astrophysics Data System (ADS)

    Bie, Q. L.; Zhou, S. Y.; Li, C. S.

    2013-11-01

    Boundary effect analysis is related to border policy making in the cross-border ethnic area. The border effect literatures show that geographic boundaries have obvious impacts on economic, social and cultural relations in both sides of a nation border. Particularly in cross-border ethnic areas, each ethnic group has strong internal spatial structure relevance, and the boundary effect is more obvious. However, most of China's border areas are cross-border ethnic areas, each of border issues is unique. Under this perspective, we analyze the border effects of various boundaries can provide basis for formulating border management policies. For small scale of cross-border ethnic minority areas, how to formulate the boundary management policy is a good question to explore. This paper is demonstrated by a study of the impact of border management policies in Dehong Dai and Jingpo Autonomous Prefecture in Yunnan Province at the border area to Burma. The comparative method is used to analysis the border management policies in past 50 decades for the border area of Yunnan Province .This research aims to define trends within border policy and its influences to national security. This paper also examines Wendy Brown's liberal theory of border management policy. We found that it is not suitable for Sino-Burma border area. The conclusion is that the changes or instability of international economic and political situation has more influence to this cross-border ethnic area, and only innovative policy will be effective in cross-border ethnic area. So the border management policies should reflect the change of international context.

  6. Biocompatible and label-free separation of cancer cells from cell culture lines from white blood cells in ferrofluids.

    PubMed

    Zhao, Wujun; Cheng, Rui; Lim, So Hyun; Miller, Joshua R; Zhang, Weizhong; Tang, Wei; Xie, Jin; Mao, Leidong

    2017-06-27

    This paper reports a biocompatible and label-free cell separation method using ferrofluids that can separate a variety of low-concentration cancer cells from cell culture lines (∼100 cancer cells per mL) from undiluted white blood cells, with a throughput of 1.2 mL h -1 and an average separation efficiency of 82.2%. The separation is based on the size difference of the cancer cells and white blood cells, and is conducted in a custom-made biocompatible ferrofluid that retains not only excellent short-term viabilities but also normal proliferations of 7 commonly used cancer cell lines. A microfluidic device is designed and optimized specifically to shorten the time of live cells' exposure to ferrofluids from hours to seconds, by eliminating time-consuming off-chip sample preparation and extraction steps and integrating them on-chip to achieve a one-step process. As a proof-of-concept demonstration, a ferrofluid with 0.26% volume fraction was used in this microfluidic device to separate spiked cancer cells from cell lines at a concentration of ∼100 cells per mL from white blood cells with a throughput of 1.2 mL h -1 . The separation efficiencies were 80 ± 3%, 81 ± 5%, 82 ± 5%, 82 ± 4%, and 86 ± 6% for A549 lung cancer, H1299 lung cancer, MCF-7 breast cancer, MDA-MB-231 breast cancer, and PC-3 prostate cancer cell lines, respectively. The separated cancer cells' purity was between 25.3% and 28.8%. In addition, the separated cancer cells from this strategy showed an average short-term viability of 94.4 ± 1.3%, and these separated cells were cultured and demonstrated normal proliferation to confluence even after the separation process. Owing to its excellent biocompatibility and label-free operation and its ability to recover low concentrations of cancer cells from white blood cells, this method could lead to a promising tool for rare cell separation.

  7. Morphological Changes of Human Corneal Endothelial Cells after Rho-Associated Kinase Inhibitor Eye Drop (Ripasudil) Administration: A Prospective Open-Label Clinical Study

    PubMed Central

    Okumura, Naoki; Suganami, Hideki; Kinoshita, Shigeru

    2015-01-01

    Purpose To investigate the effect and safety of a selective Rho kinase inhibitor, ripasudil 0.4% eye drops, on corneal endothelial cells of healthy subjects. Design Prospective, interventional case series. Methods In this study, 6 healthy subjects were administered ripasudil 0.4% in the right eye twice daily for 1 week. Morphological changes and corneal endothelial cell density were examined by noncontact and contact specular microscopy. Central corneal thickness and corneal volume of 5 mm-diameter area of center cornea were analyzed by Pentacam Scheimpflug topography. All the above measurements were conducted in both eyes before administration, 1.5 and 6 hours after the initial administration on day 0; and in the same manner after the final administration on day 7. Results By noncontact specular microscopy, indistinct cell borders with pseudo guttae were observed, but by contact specular microscopy, morphological changes of corneal endothelial cells were mild and pseudo guttae was not observed after single and repeated administration of ripasudil in all subjects. These changes resolved prior to the next administration, and corneal endothelial cell density, central corneal thickness and corneal volume were not changed throughout the study period. Conclusion Transient morphological changes of corneal endothelial cells such as indistinct cell borders with pseudo guttae were observed by noncontact specular microscopy in healthy subjects after ripasudil administration. Corneal edema was not observed and corneal endothelial cell density did not decrease after 1 week repetitive administration. These morphological changes were reversible and corneal endothelial cell morphology returned to normal prior to the next administration. Trial Registration JAPIC Clinical Trials Information 142705 PMID:26367375

  8. A chemical screen in zebrafish embryonic cells establishes that Akt activation is required for neural crest development

    PubMed Central

    Ciarlo, Christie; Kaufman, Charles K; Kinikoglu, Beste; Michael, Jonathan; Yang, Song; D′Amato, Christopher; Blokzijl-Franke, Sasja; den Hertog, Jeroen; Schlaeger, Thorsten M; Zhou, Yi; Liao, Eric

    2017-01-01

    The neural crest is a dynamic progenitor cell population that arises at the border of neural and non-neural ectoderm. The inductive roles of FGF, Wnt, and BMP at the neural plate border are well established, but the signals required for subsequent neural crest development remain poorly characterized. Here, we conducted a screen in primary zebrafish embryo cultures for chemicals that disrupt neural crest development, as read out by crestin:EGFP expression. We found that the natural product caffeic acid phenethyl ester (CAPE) disrupts neural crest gene expression, migration, and melanocytic differentiation by reducing Sox10 activity. CAPE inhibits FGF-stimulated PI3K/Akt signaling, and neural crest defects in CAPE-treated embryos are suppressed by constitutively active Akt1. Inhibition of Akt activity by constitutively active PTEN similarly decreases crestin expression and Sox10 activity. Our study has identified Akt as a novel intracellular pathway required for neural crest differentiation. PMID:28832322

  9. Dynamic acoustic field activated cell separation (DAFACS).

    PubMed

    Skotis, G D; Cumming, D R S; Roberts, J N; Riehle, M O; Bernassau, A L

    2015-02-07

    Advances in diagnostics, cell and stem cell technologies drive the development of application-specific tools for cell and particle separation. Acoustic micro-particle separation offers a promising avenue for high-throughput, label-free, high recovery, cell and particle separation and isolation in regenerative medicine. Here, we demonstrate a novel approach utilizing a dynamic acoustic field that is capable of separating an arbitrary size range of cells. We first demonstrate the method for the separation of particles with different diameters between 6 and 45 μm and secondly particles of different densities in a heterogeneous medium. The dynamic acoustic field is then used to separate dorsal root ganglion cells. The shearless, label-free and low damage characteristics make this method of manipulation particularly suited for biological applications. Advantages of using a dynamic acoustic field for the separation of cells include its inherent safety and biocompatibility, the possibility to operate over large distances (centimetres), high purity (ratio of particle population, up to 100%), and high efficiency (ratio of separated particles over total number of particles to separate, up to 100%).

  10. Differential electrophoretic separation of cells and its effect on cell viability

    NASA Technical Reports Server (NTRS)

    Leise, E. M.; Lesane, F.

    1974-01-01

    An electrophoretic separation method was applied to the separation of cells. To determine the efficiency of the separation, it was necessary to apply existing methodology and develop new methods to assess the characteristics and functions of the separated subpopulations. Through appropriate application of the widely used isoelectric focusing procedure, a reproducible separation method was developed. Cells accumulated at defined pH and 70-80% remained viable. The cells were suitable for further biologic, biochemical and immunologic studies.

  11. Organic particulate matter formation at varying relative humidity using surrogate secondary and primary organic compounds with activity corrections in the condensed phase obtained using a method based on the Wilson equation

    NASA Astrophysics Data System (ADS)

    Chang, E. I.; Pankow, J. F.

    2010-06-01

    Secondary organic aerosol (SOA) formation in the atmosphere is currently often modeled using a multiple lumped "two-product" (N·2p) approach. The N·2p approach neglects: 1) variation of activity coefficient (ζi) values and mean molecular weight MW in the particulate matter (PM) phase; 2) water uptake into the PM; and 3) the possibility of phase separation in the PM. This study considers these effects by adopting an (N·2p)ζpMW,ζ approach (θ is a phase index). Specific chemical structures are assigned to 25 lumped SOA compounds and to 15 representative primary organic aerosol (POA) compounds to allow calculation of ζi and MW values. The SOA structure assignments are based on chamber-derived 2p gas/particle partition coefficient values coupled with known effects of structure on vapor pressure pL,io (atm). To facilitate adoption of the (N·2p)ζpMW,θ approach in large-scale models, this study also develops CP-Wilson.1 (Chang-Pankow-Wilson.1), a group-contribution ζi-prediction method that is more computationally economical than the UNIFAC model of Fredenslund et al. (1975). Group parameter values required by CP-Wilson.1 are obtained by fitting ζi values to predictions from UNIFAC. The (N·2p)ζpMW,θ approach is applied (using CP-Wilson.1) to several real α-pinene/O3 chamber cases for high reacted hydrocarbon levels (ΔHC≈400 to 1000 μg m-3) when relative humidity (RH) ≍50%. Good agreement between the chamber and predicted results is obtained using both the (N·2p)ζpMW,θ and N·2p approaches, indicating relatively small water effects under these conditions. However, for a hypothetical α-pinene/O3 case at ΔHC=30 μg m-3 and RH=50%, the (N·2p)ζpMW,θ approach predicts that water uptake will lead to an organic PM level that is more double that predicted by the N·2p approach. Adoption of the (N·2p)ζpMW,θ approach using reasonable lumped structures for SOA and POA compounds is recommended for ambient PM modeling.

  12. Analysis of Pvama1 genes from China-Myanmar border reveals little regional genetic differentiation of Plasmodium vivax populations.

    PubMed

    Zhu, Xiaotong; Zhao, Pan; Wang, Si; Liu, Fei; Liu, Jun; Wang, Jian; Yang, Zhaoqing; Yan, Guiyun; Fan, Qi; Cao, Yaming; Cui, Liwang

    2016-11-29

    With the premise of diminishing parasite genetic diversity following the reduction of malaria incidence, the analysis of polymorphic antigenic markers may provide important information about the impact of malaria control on local parasite populations. Here we evaluated the genetic diversity of Plasmodium vivax apical membrane antigen 1 (Pvama1) gene in a parasite population from the China-Myanmar border and compared it with global P. vivax populations. We performed evolutionary analysis to examine the genetic diversity, natural selection, and population differentiation of 73 Pvama1 sequences acquired from the China-Myanmar border as well as 615 publically available Pvama1 sequences from seven global P. vivax populations. A total of 308 Pvama1 haplotypes were identified among the global P. vivax isolates. The overall nucleotide diversity of Pvama1 gene among the 73 China-Myanmar border parasite isolates was 0.008 with 41 haplotypes being identified (Hd = 0.958). Domain I (DI) harbored the majority (26/33) of the polymorphic sites. The McDonald Kreitman test showed a significant positive selection across the ectodomain and the DI of Pvama1. The fixation index (F ST ) estimation between the China-Myanmar border, Thailand (0.01) and Myanmar (0.10) showed only slight geographical genetic differentiation. Notably, the Sal-I haplotype was not detected in any of the analyzed global isolates, whereas the Belem strain was restricted to the Thai population. The detected mutations are mapped outside the overlapped region of the predicted B-cell epitopes and intrinsically unstructured/disordered regions. This study revealed high levels of genetic diversity of Pvama1 in the P. vivax parasite population from the China-Myanmar border with DI displaying stronger diversifying selection than other domains. There were low levels of population subdivision among parasite populations from the Greater Mekong Subregion.

  13. Characterization of the camel skin cell line Dubca.

    PubMed

    Klopries, M; Wernery, U; Kaaden, O R

    1995-01-01

    A skin fibroblast cell culture was established from a 2-month-old dromedary foetus. The cells were transformed by infection with SV40 and cloned in soft agar. The established cell line is now designated Dubca cells (Dubai camel) and has been in permanent culture for 95 passages. The cell culture was examined morphologically, chromosome preparations made and DNA fingerprinting performed by hybridization with the oligonucleotide probe (GTG)5. SV40 large T antigen was detected by western blotting. The viral host range was determined by infection with viruses of different families. Camelpox virus (CaPV) bovine herpesvirus-1 (BHV-1), vesicular stomatitis virus (VSV) and border disease virus (BDV) could be propagated in these cells.

  14. The Cosmopolitanization of Science: Experience from Chinese Stem Cell Scientists.

    PubMed

    Zhang, Joy Yueyue

    2010-09-01

    It is commonly perceived that the 'globalization of science' may result in a 'Westernization of science'. In this paper, however, I use the case of stem cell science in China to demonstrate that developing countries are sometimes able to effectively shape the norms of global/local scientific exchange. Based on interviews with 38 stem cell scientists in six Chinese cities in early 2008, this paper elucidates Chinese scientists' outlook towards cross-border collaborations and the effects that the internationalization of science has had on everyday laboratory operations. Findings suggest that although there still exists an asymmetry of scientific influence, and in many aspects China is still 'catching-up' to the West, there is also a changing nature of communication beyond borders. One key aspect of recent international scientific development is the growing necessity for local stakeholders to acquire a global mindset and to compare, reflect and accommodate diverse interests. This is what I define as the 'cosmopolitanization of science'. The study empirically examines the sociological and methodological implications of the cosmopolitanization process and further develops Ulrich Beck's cosmopolitan theory by delineating four main features of the 'cosmopolitanization of science': shared future benefits, passive ethicization, reflexive negotiation, and continuous performance.

  15. A complete denture impression technique survey of postdoctoral prosthodontic programs in the United States.

    PubMed

    Mehra, Mamta; Vahidi, Farhad; Berg, Robert W

    2014-06-01

    The purpose of the study was to survey program directors of postdoctoral prosthodontic programs in the United States regarding their programs' complete denture impression techniques. The key objectives of the survey were to identify the current trends in complete denture impression making and to determine which techniques and materials are taught in US postdoctoral prosthodontic programs. An online survey was sent to all program directors of US postdoctoral prosthodontic programs. The survey comprised two sections: preliminary impressions and final impressions. The survey contained 22 questions that would take approximately 5 minutes to complete. All responses remained anonymous throughout the survey. The response rate for the survey was 87%. A majority of the programs did not separately border mold the tray prior to making the preliminary impressions (82%). The impression material of choice for the preliminary impression was irreversible hydrocolloid (88%). Selective pressure was the predominantly used impression philosophy (80%). All programs border molded the custom tray, and 95% recorded the borders in sections. The material of choice for border molding the custom tray was modeling plastic impression compound (71%). The most commonly used impression material for the final impressions was polyvinylsiloxane (PVS) (42%), and the second most commonly used impression material was polysulphide (32%). The most common technique for locating the posterior palatal seal was marking intraorally and transferring onto the final impression (65%). Most programs routinely advised their patients not to wear their existing dentures for at least 24 hours before the final impressions were made (83%). Based on the results of this study, the following conclusions can be drawn: (1) The most commonly used material for the preliminary impression was irreversible hydrocolloid and for the final impression was PVS. (2) Modeling plastic impression compound was used by most programs to border mold the custom trays. (3) Selective pressure was the predominantly used impression philosophy. (4) A majority of the programs made a special consideration for excessive movable (flabby) tissue. (5) Most programs routinely advised their patients to not wear their existing dentures for at least 24 hours before the final impressions were made. © 2013 by the American College of Prosthodontists.

  16. Cell separation using tilted-angle standing surface acoustic waves

    PubMed Central

    Ding, Xiaoyun; Peng, Zhangli; Lin, Sz-Chin Steven; Geri, Michela; Li, Sixing; Li, Peng; Chen, Yuchao; Dao, Ming; Suresh, Subra; Huang, Tony Jun

    2014-01-01

    Separation of cells is a critical process for studying cell properties, disease diagnostics, and therapeutics. Cell sorting by acoustic waves offers a means to separate cells on the basis of their size and physical properties in a label-free, contactless, and biocompatible manner. The separation sensitivity and efficiency of currently available acoustic-based approaches, however, are limited, thereby restricting their widespread application in research and health diagnostics. In this work, we introduce a unique configuration of tilted-angle standing surface acoustic waves (taSSAW), which are oriented at an optimally designed inclination to the flow direction in the microfluidic channel. We demonstrate that this design significantly improves the efficiency and sensitivity of acoustic separation techniques. To optimize our device design, we carried out systematic simulations of cell trajectories, matching closely with experimental results. Using numerically optimized design of taSSAW, we successfully separated 2- and 10-µm-diameter polystyrene beads with a separation efficiency of ∼99%, and separated 7.3- and 9.9-µm-polystyrene beads with an efficiency of ∼97%. We illustrate that taSSAW is capable of effectively separating particles–cells of approximately the same size and density but different compressibility. Finally, we demonstrate the effectiveness of the present technique for biological–biomedical applications by sorting MCF-7 human breast cancer cells from nonmalignant leukocytes, while preserving the integrity of the separated cells. The method introduced here thus offers a unique route for separating circulating tumor cells, and for label-free cell separation with potential applications in biological research, disease diagnostics, and clinical practice. PMID:25157150

  17. Cell separation using tilted-angle standing surface acoustic waves.

    PubMed

    Ding, Xiaoyun; Peng, Zhangli; Lin, Sz-Chin Steven; Geri, Michela; Li, Sixing; Li, Peng; Chen, Yuchao; Dao, Ming; Suresh, Subra; Huang, Tony Jun

    2014-09-09

    Separation of cells is a critical process for studying cell properties, disease diagnostics, and therapeutics. Cell sorting by acoustic waves offers a means to separate cells on the basis of their size and physical properties in a label-free, contactless, and biocompatible manner. The separation sensitivity and efficiency of currently available acoustic-based approaches, however, are limited, thereby restricting their widespread application in research and health diagnostics. In this work, we introduce a unique configuration of tilted-angle standing surface acoustic waves (taSSAW), which are oriented at an optimally designed inclination to the flow direction in the microfluidic channel. We demonstrate that this design significantly improves the efficiency and sensitivity of acoustic separation techniques. To optimize our device design, we carried out systematic simulations of cell trajectories, matching closely with experimental results. Using numerically optimized design of taSSAW, we successfully separated 2- and 10-µm-diameter polystyrene beads with a separation efficiency of ∼ 99%, and separated 7.3- and 9.9-µm-polystyrene beads with an efficiency of ∼ 97%. We illustrate that taSSAW is capable of effectively separating particles-cells of approximately the same size and density but different compressibility. Finally, we demonstrate the effectiveness of the present technique for biological-biomedical applications by sorting MCF-7 human breast cancer cells from nonmalignant leukocytes, while preserving the integrity of the separated cells. The method introduced here thus offers a unique route for separating circulating tumor cells, and for label-free cell separation with potential applications in biological research, disease diagnostics, and clinical practice.

  18. Continuous separation of breast cancer cells from blood samples using multi-orifice flow fractionation (MOFF) and dielectrophoresis (DEP).

    PubMed

    Moon, Hui-Sung; Kwon, Kiho; Kim, Seung-Il; Han, Hyunju; Sohn, Joohyuk; Lee, Soohyeon; Jung, Hyo-Il

    2011-03-21

    Circulating tumor cells (CTCs) are highly correlated with the invasive behavior of cancer, so their isolations and quantifications are important for biomedical applications such as cancer prognosis and measuring the responses to drug treatments. In this paper, we present the development of a microfluidic device for the separation of CTCs from blood cells based on the physical properties of cells. For use as a CTC model, we successfully separated human breast cancer cells (MCF-7) from a spiked blood cell sample by combining multi-orifice flow fractionation (MOFF) and dielectrophoretic (DEP) cell separation technique. Hydrodynamic separation takes advantage of the massive and high-throughput filtration of blood cells as it can accommodate a very high flow rate. DEP separation plays a role in precise post-processing to enhance the efficiency of the separation. The serial combination of these two different sorting techniques enabled high-speed continuous flow-through separation without labeling. We observed up to a 162-fold increase in MCF-7 cells at a 126 µL min(-1) flow rate. Red and white blood cells were efficiently removed with separation efficiencies of 99.24% and 94.23% respectively. Therefore, we suggest that our system could be used for separation and detection of CTCs from blood cells for biomedical applications. This journal is © The Royal Society of Chemistry 2011

  19. The functional and structural borders between the CSF- and blood-dominated milieus in the choroid plexuses and the area postrema of the rat.

    PubMed

    Krisch, B

    1986-01-01

    In the borderline area between the hemal milieu of the choroid plexuses (PC) and the interstitial cerebrospinal-fluid (CSF) compartment, ground substances displaying increased amounts of basal lamina-like material and containing negatively charged sulfated glycosaminoglycans appear to be endowed with selective properties. They may function as a sieve or filtration barrier gradually controlling the passage of substances between the two milieus, depending on their charge and molecular weight. Special structural features and functional properties of ependymal cells are associated with such bordering structures. These ependymal cells are transitional elements between choroid epithelium and ciliated ependymal cells. As judged from experiments with horseradish peroxidase and conventional electron microscopy, occluding junctions at the basal pole of these cells prevent a rapid alteration in the milieu conditions, enabling gradual change from hemal to CSF composition near the bases of these transitional ependymal cells. The borderline structures between the hemal milieu of the PC and the area postrema are established by leptomeningeal cells which face a hemal milieu, are endowed with conspicuous tight junctions, and produce a flocculent substance, the light-microscopic equivalent of which is PAS positive. These structures probably establish an effective barrier between the two milieus of different composition. The functional characteristics and the morphology of the meningeal cells facing the hemal milieu of neurohemal regions resemble closely the neurothelial cells, which are interposed between the CSF milieu and the hemal milieu in the dura mater. The present results suggest that the location between the hemal and the CSF milieu is decisive for the transformation of leptomeningeal cells into "neurothelial" elements.

  20. V1 mechanisms and some figure-ground and border effects.

    PubMed

    Li, Zhaoping

    2003-01-01

    V1 neurons have been observed to respond more strongly to figure than background regions. Within a figure region, the responses are usually stronger near figure boundaries (the border effect), than further inside the boundaries. Sometimes the medial axes of the figures (e.g., the vertical midline of a vertical figure strip) induce secondary, intermediate, response peaks (the medial axis effect). Related is the physiologically elusive "cross-orientation facilitation", the observation that a cell's response to a grating patch can be facilitated by an orthogonally oriented grating in the surround. Higher center feedbacks have been suggested to cause these figure-ground effects. It has been shown, using a V1 model, that the causes could be intra-cortical interactions within V1 that serve pre-attentive visual segmentation, particularly, object boundary detection. Furthermore, whereas the border effect is robust, the figure-ground effects in the interior of a figure, in particular, the medial axis effect, are by-products of the border effect and are predicted to diminish to zero for larger figures. This model prediction (of the figure size dependence) was subsequently confirmed physiologically, and supported by findings that the response modulations by texture surround do not depend on feedbacks from V2. In addition, the model explains the "cross-orientation facilitation" as caused by a dis-inhibition, to the cell responding to the center of the central grating, by the background grating. Furthermore, the elusiveness of this phenomena was accounted for by the insight that it depends critically on the size of the figure grating. The model is applied to understand some figure-ground effects and segmentation in psychophysics: in particular, that contrast discrimination threshold is lower within and at the center of a closed contour than that in the background, and that a very briefly presented vernier target can perceptually shine through a subsequently presented large grating centered at the same location.

  1. Birc7: A Late Fiber Gene of the Crystalline Lens.

    PubMed

    De Maria, Alicia; Bassnett, Steven

    2015-07-01

    A distinct subset of genes, so-called "late fiber genes," is expressed in cells bordering the central, organelle-free zone (OFZ) of the lens. The purpose of this study was to identify additional members of this group. Fiber cells were harvested from various layers of the lens by laser micro-dissection and subjected to microarray, in situ hybridization, and Western blot analysis. Expression of Livin, a member of the inhibitor of apoptosis protein (IAP) family encoded by Birc7, was strongly upregulated in deep cortical fiber cells. The depth-dependent distribution of Livin mRNA was confirmed by quantitative PCR and in situ hybridization. The onset of Livin expression coincided with loss of organelles from primary fiber cells. Livin expression peaked at 1 month but was sustained even in aged lenses. Antibodies raised against mouse Livin labeled multiple bands on immunoblots, reflecting progressive proteolysis of the parent molecule during differentiation. Mice harboring a floxed Birc7 allele were generated and used to conditionally delete Birc7 in lens. Lenses from knockout mice grew normally and retained their transparency, suggesting that Livin does not have an indispensable role in fiber cell differentiation. Birc7 is a late fiber gene of the mouse lens. In tumor cells, Livin acts as an antiapoptotic protein, but its function in the lens is enigmatic. Livin is a RING domain protein with putative E3 ubiquitin ligase activity. Its expression in cells bordering the OFZ is consistent with a role in organelle degradation, a process in which the ubiquitin proteasome pathway has been implicated previously.

  2. Birc7: A Late Fiber Gene of the Crystalline Lens

    PubMed Central

    De Maria, Alicia; Bassnett, Steven

    2015-01-01

    Purpose A distinct subset of genes, so-called “late fiber genes,” is expressed in cells bordering the central, organelle-free zone (OFZ) of the lens. The purpose of this study was to identify additional members of this group. Methods Fiber cells were harvested from various layers of the lens by laser micro-dissection and subjected to microarray, in situ hybridization, and Western blot analysis. Results Expression of Livin, a member of the inhibitor of apoptosis protein (IAP) family encoded by Birc7, was strongly upregulated in deep cortical fiber cells. The depth-dependent distribution of Livin mRNA was confirmed by quantitative PCR and in situ hybridization. The onset of Livin expression coincided with loss of organelles from primary fiber cells. Livin expression peaked at 1 month but was sustained even in aged lenses. Antibodies raised against mouse Livin labeled multiple bands on immunoblots, reflecting progressive proteolysis of the parent molecule during differentiation. Mice harboring a floxed Birc7 allele were generated and used to conditionally delete Birc7 in lens. Lenses from knockout mice grew normally and retained their transparency, suggesting that Livin does not have an indispensable role in fiber cell differentiation. Conclusions Birc7 is a late fiber gene of the mouse lens. In tumor cells, Livin acts as an antiapoptotic protein, but its function in the lens is enigmatic. Livin is a RING domain protein with putative E3 ubiquitin ligase activity. Its expression in cells bordering the OFZ is consistent with a role in organelle degradation, a process in which the ubiquitin proteasome pathway has been implicated previously. PMID:26218911

  3. Novel platform for minimizing cell loss on separation process: Droplet-based magnetically activated cell separator

    NASA Astrophysics Data System (ADS)

    Kim, Youngho; Hong, Su; Lee, Sang Ho; Lee, Kangsun; Yun, Seok; Kang, Yuri; Paek, Kyeong-Kap; Ju, Byeong-Kwon; Kim, Byungkyu

    2007-07-01

    To reduce the problem of cell loss due to adhesion, one of the basic phenomena in microchannel, we proposed the droplet-based magnetically activated cell separator (DMACS). Based on the platform of the DMACS—which consists of permanent magnets, a coverslip with a circle-shaped boundary, and an injection tube—we could collect magnetically (CD45)-labeled (positive) cells with high purity and minimize cell loss due to adhesion. To compare separation efficiency between the MACS and the DMACS, the total number of cells before and after separation with both the separators was counted by flow cytometry. We could find that the number (3241/59940) of cells lost in the DMACS is much less than that (22360/59940) in the MACS while the efficiency of cell separation in the DMACS (96.07%) is almost the same as that in the MACS (96.72%). Practically, with fluorescent images, it was visually confirmed that the statistical data are reliable. From the viability test by using Hoechst 33 342, it was also demonstrated that there was no cell damage on a gas-liquid interface. Conclusively, DMACS will be a powerful tool to separate rare cells and applicable as a separator, key component of lab-on-a-chip.

  4. Separation of cells from the rat anterior pituitary gland

    NASA Technical Reports Server (NTRS)

    Hymer, Wesley C.; Hatfield, J. Michael

    1983-01-01

    Various techniques for separating the hormone-producing cell types from the rat anterior pituitary gland are examined. The purity, viability, and responsiveness of the separated cells depend on the physiological state of the donor, the tissue dissociation procedures, the staining technique used for identification of cell type, and the cell separation technique. The chamber-gradient setup and operation, the characteristics of the gradient materials, and the separated cell analysis of velocity sedimentation techniques (in particular Staput and Celsep) are described. Consideration is given to the various types of materials used in density gradient centrifugation and the operation of a gradient generating device. The use of electrophoresis to separate rat pituitary cells is discussed.

  5. Imaging cell picker: A morphology-based automated cell separation system on a photodegradable hydrogel culture platform.

    PubMed

    Shibuta, Mayu; Tamura, Masato; Kanie, Kei; Yanagisawa, Masumi; Matsui, Hirofumi; Satoh, Taku; Takagi, Toshiyuki; Kanamori, Toshiyuki; Sugiura, Shinji; Kato, Ryuji

    2018-06-09

    Cellular morphology on and in a scaffold composed of extracellular matrix generally represents the cellular phenotype. Therefore, morphology-based cell separation should be interesting method that is applicable to cell separation without staining surface markers in contrast to conventional cell separation methods (e.g., fluorescence activated cell sorting and magnetic activated cell sorting). In our previous study, we have proposed a cloning technology using a photodegradable gelatin hydrogel to separate the individual cells on and in hydrogels. To further expand the applicability of this photodegradable hydrogel culture platform, we here report an image-based cell separation system imaging cell picker for the morphology-based cell separation on a photodegradable hydrogel. We have developed the platform which enables the automated workflow of image acquisition, image processing and morphology analysis, and collection of a target cells. We have shown the performance of the morphology-based cell separation through the optimization of the critical parameters that determine the system's performance, such as (i) culture conditions, (ii) imaging conditions, and (iii) the image analysis scheme, to actually clone the cells of interest. Furthermore, we demonstrated the morphology-based cloning performance of cancer cells in the mixture of cells by automated hydrogel degradation by light irradiation and pipetting. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  6. Automatic detection of blood versus non-blood regions on intravascular ultrasound (IVUS) images using wavelet packet signatures

    NASA Astrophysics Data System (ADS)

    Katouzian, Amin; Baseri, Babak; Konofagou, Elisa E.; Laine, Andrew F.

    2008-03-01

    Intravascular ultrasound (IVUS) has been proven a reliable imaging modality that is widely employed in cardiac interventional procedures. It can provide morphologic as well as pathologic information on the occluded plaques in the coronary arteries. In this paper, we present a new technique using wavelet packet analysis that differentiates between blood and non-blood regions on the IVUS images. We utilized the multi-channel texture segmentation algorithm based on the discrete wavelet packet frames (DWPF). A k-mean clustering algorithm was deployed to partition the extracted textural features into blood and non-blood in an unsupervised fashion. Finally, the geometric and statistical information of the segmented regions was used to estimate the closest set of pixels to the lumen border and a spline curve was fitted to the set. The presented algorithm may be helpful in delineating the lumen border automatically and more reliably prior to the process of plaque characterization, especially with 40 MHz transducers, where appearance of the red blood cells renders the border detection more challenging, even manually. Experimental results are shown and they are quantitatively compared with manually traced borders by an expert. It is concluded that our two dimensional (2-D) algorithm, which is independent of the cardiac and catheter motions performs well in both in-vivo and in-vitro cases.

  7. Novel microfluidic device for the continuous separation of cancer cells using dielectrophoresis.

    PubMed

    Alazzam, Anas; Mathew, Bobby; Alhammadi, Falah

    2017-03-01

    We describe the design, microfabrication, and testing of a microfluidic device for the separation of cancer cells based on dielectrophoresis. Cancer cells, specifically green fluorescent protein-labeled MDA-MB-231, are successfully separated from a heterogeneous mixture of the same and normal blood cells. MDA-MB-231 cancer cells are separated with an accuracy that enables precise detection and counting of circulating tumor cells present among normal blood cells. The separation is performed using a set of planar interdigitated transducer electrodes that are deposited on the surface of a glass wafer and slightly protrude into the separation microchannel at one side. The device includes two parts, namely, a glass wafer and polydimethylsiloxane element. The device is fabricated using standard microfabrication techniques. All experiments are conducted with low conductivity sucrose-dextrose isotonic medium. The variation in response between MDA-MB-231 cancer cells and normal cells to a certain band of alternating-current frequencies is used for continuous separation of cells. The fabrication of the microfluidic device, preparation of cells and medium, and flow conditions are detailed. The proposed microdevice can be used to detect and separate malignant cells from heterogeneous mixture of cells for the purpose of early screening for cancer. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Remodeling of nuclear architecture by the thiodioxoxpiperazine metabolite chaetocin

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

    Illner, Doris; Zinner, Roman; Handtke, Violet

    2010-06-10

    Extensive changes of higher order chromatin arrangements can be observed during prometaphase, terminal cell differentiation and cellular senescence. Experimental systems where major reorganization of nuclear architecture can be induced under defined conditions, may help to better understand the functional implications of such changes. Here, we report on profound chromatin reorganization in fibroblast nuclei by chaetocin, a thiodioxopiperazine metabolite. Chaetocin induces strong condensation of chromosome territories separated by a wide interchromatin space largely void of DNA. Cell viability is maintained irrespective of this peculiar chromatin phenotype. Cell cycle markers, histone signatures, and tests for cellular senescence and for oxidative stress indicatemore » that chaetocin induced chromatin condensation/clustering (CICC) represents a distinct entity among nuclear phenotypes associated with condensed chromatin. The territorial organization of entire chromosomes is maintained in CICC nuclei; however, the conventional nuclear architecture harboring gene-dense chromatin in the nuclear interior and gene-poor chromatin at the nuclear periphery is lost. Instead gene-dense and transcriptionally active chromatin is shifted to the periphery of individual condensed chromosome territories where nascent RNA becomes highly enriched around their outer surface. This chromatin reorganization makes CICC nuclei an attractive model system to study this border zone as a distinct compartment for transcription. Induction of CICC is fully inhibited by thiol-dependent antioxidants, but is not related to the production of reactive oxygen species. Our results suggest that chaetocin functionally impairs the thioredoxin (Trx) system, which is essential for deoxynucleotide synthesis, but in addition involved in a wide range of cellular functions. The mechanisms involved in CICC formation remain to be fully explored.« less

  9. Collective cell migration in development

    PubMed Central

    Scarpa, Elena

    2016-01-01

    During embryonic development, tissues undergo major rearrangements that lead to germ layer positioning, patterning, and organ morphogenesis. Often these morphogenetic movements are accomplished by the coordinated and cooperative migration of the constituent cells, referred to as collective cell migration. The molecular and biomechanical mechanisms underlying collective migration of developing tissues have been investigated in a variety of models, including border cell migration, tracheal branching, blood vessel sprouting, and the migration of the lateral line primordium, neural crest cells, or head mesendoderm. Here we review recent advances in understanding collective migration in these developmental models, focusing on the interaction between cells and guidance cues presented by the microenvironment and on the role of cell–cell adhesion in mechanical and behavioral coupling of cells within the collective. PMID:26783298

  10. The minimal local-asperity hypothesis of early retinal lateral inhibition.

    PubMed

    Balboa, R M; Grzywacz, N M

    2000-07-01

    Recently we found that the theories related to information theory existent in the literature cannot explain the behavior of the extent of the lateral inhibition mediated by retinal horizontal cells as a function of background light intensity. These theories can explain the fall of the extent from intermediate to high intensities, but not its rise from dim to intermediate intensities. We propose an alternate hypothesis that accounts for the extent's bell-shape behavior. This hypothesis proposes that the lateral-inhibition adaptation in the early retina is part of a system to extract several image attributes, such as occlusion borders and contrast. To do so, this system would use prior probabilistic knowledge about the biological processing and relevant statistics in natural images. A key novel statistic used here is the probability of the presence of an occlusion border as a function of local contrast. Using this probabilistic knowledge, the retina would optimize the spatial profile of lateral inhibition to minimize attribute-extraction error. The two significant errors that this minimization process must reduce are due to the quantal noise in photoreceptors and the straddling of occlusion borders by lateral inhibition.

  11. An Inversion Disrupting FAM134B Is Associated with Sensory Neuropathy in the Border Collie Dog Breed.

    PubMed

    Forman, Oliver P; Hitti, Rebekkah J; Pettitt, Louise; Jenkins, Christopher A; O'Brien, Dennis P; Shelton, G Diane; De Risio, Luisa; Quintana, Rodrigo Gutierrez; Beltran, Elsa; Mellersh, Cathryn

    2016-09-08

    Sensory neuropathy in the Border Collie is a severe neurological disorder caused by the degeneration of sensory and, to a lesser extent, motor nerve cells with clinical signs starting between 2 and 7 months of age. Using a genome-wide association study approach with three cases and 170 breed matched controls, a suggestive locus for sensory neuropathy was identified that was followed up using a genome sequencing approach. An inversion disrupting the candidate gene FAM134B was identified. Genotyping of additional cases and controls and RNAseq analysis provided strong evidence that the inversion is causal. Evidence of cryptic splicing resulting in novel exon transcription for FAM134B was identified by RNAseq experiments. This investigation demonstrates the identification of a novel sensory neuropathy associated mutation, by mapping using a minimal set of cases and subsequent genome sequencing. Through mutation screening, it should be possible to reduce the frequency of or completely eliminate this debilitating condition from the Border Collie breed population. Copyright © 2016 Forman et al.

  12. Numerical simulation of dielectrophoretic separation of live/dead cells using a three-dimensional nonuniform AC electric field in micro-fabricated devices.

    PubMed

    Tada, Shigeru

    2015-01-01

    The analysis of cell separation has many important biological and medical applications. Dielectrophoresis (DEP) is one of the most effective and widely used techniques for separating and identifying biological species. In the present study, a DEP flow channel, a device that exploits the differences in the dielectric properties of cells in cell separation, was numerically simulated and its cell-separation performance examined. The samples of cells used in the simulation were modeled as human leukocyte (B cell) live and dead cells. The cell-separation analysis was carried out for a flow channel equipped with a planar electrode on the channel's top face and a pair of interdigitated counter electrodes on the bottom. This yielded a three-dimensional (3D) nonuniform AC electric field in the entire space of the flow channel. To investigate the optimal separation conditions for mixtures of live and dead cells, the strength of the applied electric field was varied. With appropriately selected conditions, the device was predicted to be very effective at separating dead cells from live cells. The major advantage of the proposed method is that a large volume of sample can be processed rapidly because of a large spacing of the channel height.

  13. CHARACTERIZATION OF A BRUSH BORDER FERRIREDUCTASE IN LOBSTER HEPATOPANCREATIC EPITHELIAL CELLS AND ITS POSSIBLE ROLE IN IRON TRANSPORT. (R823068)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  14. High-resolution Identification and Separation of Living Cell Types by Multiple microRNA-responsive Synthetic mRNAs.

    PubMed

    Endo, Kei; Hayashi, Karin; Saito, Hirohide

    2016-02-23

    The precise identification and separation of living cell types is critical to both study cell function and prepare cells for medical applications. However, intracellular information to distinguish live cells remains largely inaccessible. Here, we develop a method for high-resolution identification and separation of cell types by quantifying multiple microRNA (miRNA) activities in live cell populations. We found that a set of miRNA-responsive, in vitro synthesized mRNAs identify a specific cell population as a sharp peak and clearly separate different cell types based on less than two-fold differences in miRNA activities. Increasing the number of miRNA-responsive mRNAs enhanced the capability for cell identification and separation, as we precisely and simultaneously distinguished different cell types with similar miRNA profiles. In addition, the set of synthetic mRNAs separated HeLa cells into subgroups, uncovering heterogeneity of the cells and the level of resolution achievable. Our method could identify target live cells and improve the efficiency of cell purification from heterogeneous populations.

  15. Cosine Directional Tuning of Theta Cell Burst Frequencies: Evidence for Spatial Coding by Oscillatory Interference

    PubMed Central

    Welday, Adam C.; Shlifer, I. Gary; Bloom, Matthew L.; Zhang, Kechen

    2011-01-01

    The rodent septohippocampal system contains “theta cells,” which burst rhythmically at 4–12 Hz, but the functional significance of this rhythm remains poorly understood (Buzsáki, 2006). Theta rhythm commonly modulates the spike trains of spatially tuned neurons such as place (O'Keefe and Dostrovsky, 1971), head direction (Tsanov et al., 2011a), grid (Hafting et al., 2005), and border cells (Savelli et al., 2008; Solstad et al., 2008). An “oscillatory interference” theory has hypothesized that some of these spatially tuned neurons may derive their positional firing from phase interference among theta oscillations with frequencies that are modulated by the speed and direction of translational movements (Burgess et al., 2005, 2007). This theory is supported by studies reporting modulation of theta frequency by movement speed (Rivas et al., 1996; Geisler et al., 2007; Jeewajee et al., 2008a), but modulation of theta frequency by movement direction has never been observed. Here we recorded theta cells from hippocampus, medial septum, and anterior thalamus of freely behaving rats. Theta cell burst frequencies varied as the cosine of the rat's movement direction, and this directional tuning was influenced by landmark cues, in agreement with predictions of the oscillatory interference theory. Computer simulations and mathematical analysis demonstrated how a postsynaptic neuron can detect location-dependent synchrony among inputs from such theta cells, and thereby mimic the spatial tuning properties of place, grid, or border cells. These results suggest that theta cells may serve a high-level computational function by encoding a basis set of oscillatory signals that interfere with one another to synthesize spatial memory representations. PMID:22072668

  16. High Mobility Group Box 1 Promotes Angiogenesis from Bone Marrow-derived Endothelial Progenitor Cells after Myocardial Infarction.

    PubMed

    Nakamura, Yuichi; Suzuki, Satoshi; Shimizu, Takeshi; Miyata, Makiko; Shishido, Tetsuro; Ikeda, Kazuhiko; Saitoh, Shu-Ichi; Kubota, Isao; Takeishi, Yasuchika

    2015-01-01

    High mobility group box 1 (HMGB1) is a DNA-binding protein secreted into the extracellular space from necrotic cells that acts as a cytokine. We examined the role of HMGB1 in angiogenesis from bone marrow-derived cells in the heart using transgenic mice exhibiting the cardiac-specific overexpression of HMGB1 (HMGB1-TG). HMGB1-TG mice and wild-type littermate (WT) mice were lethally irradiated and injected with bone marrow cells from green fluorescent protein mice through the tail vein. After bone marrow transplantation, the left anterior descending artery was ligated to induce myocardial infarction (MI). Flow cytometry revealed that the levels of circulating endothelial progenitor cells (EPCs) mobilized from the bone marrow increased after MI in the HMGB-TG mice versus the WT mice. In addition, the size of MI was smaller in the HMGB1-TG mice than in the WT mice, and immunofluorescence staining demonstrated that the number of engrafted vascular endothelial cells derived from bone marrow in the border zones of the MI areas was increased in the HMGB1-TG mice compared to that observed in the WT mice. Moreover, the levels of cardiac vascular endothelial growth factor after MI were higher in the HMGB1-TG mice than in the WT mice. The present study demonstrated that HMGB1 promotes angiogenesis and reduces the MI size by enhancing the mobilization and differentiation of bone marrow cells to EPCs as well as their migration to the border zones of the MI areas and engraftment as vascular endothelial cells in new capillaries or arterioles in the infarcted heart.

  17. Neurons identified by NeuN/Fox-3 immunoreactivity have a novel distribution in the hamster and mouse suprachiasmatic nucleus.

    PubMed

    Morin, Lawrence P; Hefton, Sara; Studholme, Keith M

    2011-11-03

    The suprachiasmatic nucleus (SCN) has several structural characteristics and cell phenotypes shared across species. Here, we describe a novel feature of SCN anatomy that is seen in both hamster and mouse. Frozen sections through the SCN were obtained from fixed brains and stained for the presence of immunoreactivity to neuronal nuclear protein (NeuN-IR) using a mouse monoclonal antibody which is known to exclusively identify neurons. NeuN-IR did not identify all SCN neurons as medial NeuN-IR neurons were generally not present. In the hamster, NeuN-IR cells are present rostrally, scattered in the dorsal half of the nucleus. More caudally, the NeuN-IR cells are largely, but not exclusively, scattered inside the lateral and dorsolateral border. At mid- to mid-caudal SCN levels, a dense group of NeuN-IR cells extends from the dorsolateral border ventromedially to encompass the central subnucleus of the SCN (SCNce). The pattern is similar in the mouse SCN. NeuN-IR does not co-localize with either cholecystokinin- or vasoactive intestinal polypeptide, but does with vasopressin-IR in the caudal SCN. In the hamster SCNce, numerous cells contain both calbindin- and NeuN-IR. The distribution of NeuN-IR cells in the SCN is unique, especially with regard to its generally lateral location through the length of the nucleus. The distribution of NeuN-IR cells is not consistent with most schemas representing SCN organization or with terminology referring to its widely accepted subdivisions. NeuN has recently been identified as Fox-3 protein. Its function in the SCN is not known, nor is it known why a large proportion of SCN cells do not contain NeuN-IR. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Modeling the Time-Course of Responses for the Border Ownership Selectivity Based on the Integration of Feedforward Signals and Visual Cortical Interactions

    PubMed Central

    Wagatsuma, Nobuhiko; Sakai, Ko

    2017-01-01

    Border ownership (BO) indicates which side of a contour owns a border, and it plays a fundamental role in figure-ground segregation. The majority of neurons in V2 and V4 areas of monkeys exhibit BO selectivity. A physiological work reported that the responses of BO-selective cells show a rapid transition when a presented square is flipped along its classical receptive field (CRF) so that the opposite BO is presented, whereas the transition is significantly slower when a square with a clear BO is replaced by an ambiguous edge, e.g., when the square is enlarged greatly. The rapid transition seemed to reflect the influence of feedforward processing on BO selectivity. Herein, we investigated the role of feedforward signals and cortical interactions for time-courses in BO-selective cells by modeling a visual cortical network comprising V1, V2, and posterior parietal (PP) modules. In our computational model, the recurrent pathways among these modules gradually established the visual progress and the BO assignments. Feedforward inputs mainly determined the activities of these modules. Surrounding suppression/facilitation of early-level areas modulates the activities of V2 cells to provide BO signals. Weak feedback signals from the PP module enhanced the contrast gain extracted in V1, which underlies the attentional modulation of BO signals. Model simulations exhibited time-courses depending on the BO ambiguity, which were caused by the integration delay of V1 and V2 cells and the local inhibition therein given the difference in input stimulus. However, our model did not fully explain the characteristics of crucially slow transition: the responses of BO-selective physiological cells indicated the persistent activation several times longer than that of our model after the replacement with the ambiguous edge. Furthermore, the time-course of BO-selective model cells replicated the attentional modulation of response time in human psychophysical experiments. These attentional modulations for time-courses were induced by selective enhancement of early-level features due to interactions between V1 and PP. Our proposed model suggests fundamental roles of surrounding suppression/facilitation based on feedforward inputs as well as the interactions between early and parietal visual areas with respect to the ambiguity dependence of the neural dynamics in intermediate-level vision. PMID:28163688

  19. Modeling the Time-Course of Responses for the Border Ownership Selectivity Based on the Integration of Feedforward Signals and Visual Cortical Interactions.

    PubMed

    Wagatsuma, Nobuhiko; Sakai, Ko

    2016-01-01

    Border ownership (BO) indicates which side of a contour owns a border, and it plays a fundamental role in figure-ground segregation. The majority of neurons in V2 and V4 areas of monkeys exhibit BO selectivity. A physiological work reported that the responses of BO-selective cells show a rapid transition when a presented square is flipped along its classical receptive field (CRF) so that the opposite BO is presented, whereas the transition is significantly slower when a square with a clear BO is replaced by an ambiguous edge, e.g., when the square is enlarged greatly. The rapid transition seemed to reflect the influence of feedforward processing on BO selectivity. Herein, we investigated the role of feedforward signals and cortical interactions for time-courses in BO-selective cells by modeling a visual cortical network comprising V1, V2, and posterior parietal (PP) modules. In our computational model, the recurrent pathways among these modules gradually established the visual progress and the BO assignments. Feedforward inputs mainly determined the activities of these modules. Surrounding suppression/facilitation of early-level areas modulates the activities of V2 cells to provide BO signals. Weak feedback signals from the PP module enhanced the contrast gain extracted in V1, which underlies the attentional modulation of BO signals. Model simulations exhibited time-courses depending on the BO ambiguity, which were caused by the integration delay of V1 and V2 cells and the local inhibition therein given the difference in input stimulus. However, our model did not fully explain the characteristics of crucially slow transition: the responses of BO-selective physiological cells indicated the persistent activation several times longer than that of our model after the replacement with the ambiguous edge. Furthermore, the time-course of BO-selective model cells replicated the attentional modulation of response time in human psychophysical experiments. These attentional modulations for time-courses were induced by selective enhancement of early-level features due to interactions between V1 and PP. Our proposed model suggests fundamental roles of surrounding suppression/facilitation based on feedforward inputs as well as the interactions between early and parietal visual areas with respect to the ambiguity dependence of the neural dynamics in intermediate-level vision.

  20. Imagining and Imaging Borders: Understanding Borderlands for Global Sustainability

    NASA Astrophysics Data System (ADS)

    Konrad, V.

    2013-11-01

    Borders are increasingly complex human responses and social constructions in a world where globalizing forces confront basic human concerns for security and certainty. In an effort to provide a background to assess research directions for imaging borders, this paper explores what we know about borders, and what we do not know well about borders. Borders in globalization are the meeting points of globalizing forces of security, trade and migration flows with emerging technologies, self determination and regionalization around the world. We need to know more about how: self determination fuels secessions and new borders; borders result from complex rather than simple policy and governance issues; borders depend on the political clout of borderland communities; market and migration flows impact borders; and borders are always in motion. The paper shows how these organizing principles underlie the basic themes of border governance, flows, culture, history, security and sustainability. Finally, the paper offers two brief illustrations of border imaging to link this presentation to the following discussion of the workshop.

  1. Electrophoretic cell separation using microspheres. [purification of lymphocytes

    NASA Technical Reports Server (NTRS)

    Smolka, A.; Sachs, G.

    1980-01-01

    Methods of cell separation based on the electrokinetic properties of the cell membrane offer a degree of discrimination among cell populations which is not available with methods based on cell size or density alone. Studies aimed at extending red cell separations using microspheres to purification of lymphocytes.

  2. Evaluation of Inorganic/Organic Separators

    NASA Technical Reports Server (NTRS)

    Donnel, C. P., III

    1976-01-01

    Thirty-six (36) experimental 40AH sealed silver-zinc cells were constructed during phase I of this two (2) phase program. These cells were divided into six (6) groups of six (6) cells each. Each group of six (6) cells was evenly divided into two batches of three (3) cells each. Groups 1 through 4 each featured a different inorganic filler material in the slurry used to coat the separator substrate. Groups 5 and 6 featured an alternate method of separator bag construction. With the exception of the various separator materials, the parts and processes used to produce these thirty-six (36) cells were the same as those used to make the HR40-7 cell. The two (2) batches of cells in each cell group differed only in the lots of solutions and other separator slurry components used. Each cell was given two formation charge/discharge cycles prior to being shipped to NASA Lewis Research Center. Phase II of the program consisted of constructing another thirty-six (36) 40AH experimental cells in six (6) groups of six (6) cells each. Each group was distinguished by the type of precoated separator material used to fabricate separator bags. A new method of separator bag construction was used in this phase of the program. These cells were given two (2) formation cycles and shipped to NASA Lewis Research Center.

  3. Label-free cancer cell separation from human whole blood using inertial microfluidics at low shear stress.

    PubMed

    Lee, Myung Gwon; Shin, Joong Ho; Bae, Chae Yun; Choi, Sungyoung; Park, Je-Kyun

    2013-07-02

    We report a contraction-expansion array (CEA) microchannel device that performs label-free high-throughput separation of cancer cells from whole blood at low Reynolds number (Re). The CEA microfluidic device utilizes hydrodynamic field effect for cancer cell separation, two kinds of inertial effects: (1) inertial lift force and (2) Dean flow, which results in label-free size-based separation with high throughput. To avoid cell damages potentially caused by high shear stress in conventional inertial separation techniques, the CEA microfluidic device isolates the cells with low operational Re, maintaining high-throughput separation, using nondiluted whole blood samples (hematocrit ~45%). We characterized inertial particle migration and investigated the migration of blood cells and various cancer cells (MCF-7, SK-BR-3, and HCC70) in the CEA microchannel. The separation of cancer cells from whole blood was demonstrated with a cancer cell recovery rate of 99.1%, a blood cell rejection ratio of 88.9%, and a throughput of 1.1 × 10(8) cells/min. In addition, the blood cell rejection ratio was further improved to 97.3% by a two-step filtration process with two devices connected in series.

  4. Intestinal surfactant permeation enhancers and their interaction with enterocyte cell membranes in a mucosal explant system.

    PubMed

    Danielsen, E Michael; Hansen, Gert H

    2017-07-03

    Intestinal permeation enhancers (PEs) are agents aimed to improve oral delivery of therapeutic drugs with poor bioavailability. The main permeability barrier for oral delivery is the intestinal epithelium, and PEs act to increase the paracellular and/or transcellular passage of drugs. Transcellular passage can be achieved by cell membrane permeabilization and/or by endocytic uptake and subsequent transcytosis. One broad class of PEs is surfactants which act by inserting into the cell membrane, thereby perturbing its integrity, but little is known about how the dynamics of the membrane are affected. In the present work, the interaction of the surfactants lauroyl-L-carnitine, 1-decanoyl-rac-glycerol, and nonaethylene glycol monododecyl ether with the intestinal epithelium was studied in organ cultured pig jejunal mucosal explants. As expected, at 2 mM, these agents rapidly permeabilized the enterocytes for the fluorescent polar tracer lucifer yellow, but surprisingly, they all also blocked both constitutive -and receptor-mediated pathways of endocytosis from the brush border, indicating a complete arrest of apical membrane trafficking. At the ultrastructural level, the PEs caused longitudinal fusion of brush border microvilli. Such a membrane fusogenic activity could also explain the observed formation of vesicle-like structures and large vacuoles along the lateral cell membranes of the enterocytes induced by the PEs. We conclude that the surfactant action of the PEs selected in this study not only permeabilized the enterocytes, but profoundly changed the dynamic properties of their constituent cell membranes.

  5. Further analyses of human kidney cell populations separated on the Space Shuttle

    NASA Technical Reports Server (NTRS)

    Stewart, Robin M.; Todd, Paul; Cole, Kenneth D.; Morrison, Dennis R.

    1992-01-01

    Cultured human embryonic kidney cells were separated into electrophoretic subpopulations in laboratory experiments and in two separation experiments on the STS-8 (Challenger) Space Shuttle flight using the mid-deck Continuous Flow Electrophoretic Separator (CFES). Populations of cells from each fraction were cultured for the lifetime of the cells, and supernatant medium was withdrawn and replaced at 4-day intervals. Withdrawn medium was frozen at -120 C for subsequent analysis. Enzyme assays, antibodies and gel electrophoresis were used as analytical tools for the detection and quantization of plasminogen activators in these samples. These assays of frozen-culture supernatant fluids confirmed the electrophoretic separation of plasminogen-activator-producing cells from nonproducing cells, the isolation of cells capable of sustained production, and the separation of cells that produce different plasminogen activators from one other.

  6. Quality testing of an innovative cascade separation system for multiple cell separation

    NASA Astrophysics Data System (ADS)

    Pierzchalski, Arkadiusz; Moszczynska, Aleksandra; Albrecht, Bernd; Heinrich, Jan-Michael; Tarnok, Attila

    2012-03-01

    Isolation of different cell types from mixed samples in one separation step by FACS is feasible but expensive and slow. It is cheaper and faster but still challenging by magnetic separation. An innovative bead-based cascade-system (pluriSelect GmbH, Leipzig, Germany) relies on simultaneous physical separation of different cell types. It is based on antibody-mediated binding of cells to beads of different size and isolation with sieves of different mesh-size. We validated pluriSelect system for single parameter (CD3) and simultaneous separation of CD3 and CD15 cells from EDTA blood-samples. Results were compared with those obtained by MACS (Miltenyi-Biotech) magnetic separation (CD3 separation). pluriSelect separation was done in whole blood, MACS on Ficoll gradient isolated leukocytes, according to the manufacturer's protocols. Isolated and residual cells were immunophenotyped (7-color 8-antibody panel (CD3; CD16/56; CD4; CD8; CD14; CD19; CD45; HLADR) on a CyFlowML flow cytometer (Partec GmbH). Cell count (Coulter), purity, yield and viability (7-AAD exclusion) were determined. There were no significant differences between both systems regarding purity (92-98%), yield (50-60%) and viability (92-98%) of isolated cells. PluriSelect separation was slightly faster than MACS (1.15 h versus 1.5h). Moreover, no preenrichment steps were necessary. In conclusion, pluriSelect is a fast, simple and gentle system for efficient simultaneous separation of two cell subpopulation directly from whole blood and can provide a simple alternative to FACS. The isolated cells can be used for further research applications.

  7. Solitary intramuscular nasal Juvenile Xanthogranuloma: a case report with review of literature.

    PubMed

    Aggarwal, Kiran; Pahuja, Sangeeta

    2002-07-01

    Juvenile Xanthogranuloma is a non-langerhans cell histiocytosis characterized by yellowish cutaneous nodules that usually appear in early infancy and childhood. Intramuscular variant is a rare form, with only eight reported cases, and none reported in ala of nose. Sheets of histiocytes, few touton giant cells and infiltrative borders makes it susceptible to misdiagnosis as childhood sarcomas or lymphoproliferative disorders. Awareness of the lesion aided by immunohistochemistry helps in reaching the proper diagnosis.

  8. Fresenius AS.TEC204 blood cell separator.

    PubMed

    Sugai, Mikiya

    2003-02-01

    Fresenius AS.TEC204 is a third-generation blood cell separator that incorporates the continuous centrifugal separation method and automatic control of the cell separation process. Continuous centrifugation separates cell components according to their specific gravity, and different cell components are either harvested or eliminated as needed. The interface between the red blood cell and plasma is optically detected, and the Interface Control (IFC) cooperates with different pumps, monitors and detectors to harvest required components automatically. The system is composed of three major sections; the Front Panel Unit; the Pump Unit, and the Centrifuge Unit. This unit can be used for a wide variety of clinical applications including collection of platelets, peripheral blood stem cells, bone marrow stem cells, granulocytes, mononuclear cells, and exchange of plasma or red cells, and for plasma treatment.

  9. Immunolocalization of matrix metalloproteinase-13 on bone surface under osteoclasts in rat tibia.

    PubMed

    Nakamura, Hiroaki; Sato, Ginga; Hirata, Azumi; Yamamoto, Toshio

    2004-01-01

    Matrix metalloproteinase (MMP)-13 (an interstitial collagenase also called collagenase 3) is involved in degradation of extracellular matrix in various tissues. Using immunohistochemistry and Western blotting, we investigated localization of MMP-13 in rat tibia, to clarify the role of MMP-13 in bone resorption. MMP-13 reactivity was mainly seen on bone surfaces under osteoclasts, and in some osteocytes and their lacunae near osteoclasts. However, immunoreactivity was not seen in chondrocytes or osteoclasts. MMP-13 was also localized on cement lines in the epiphysis. In the growth plate erosion zone, perivascular cells showed MMP-13 reactivity. Immunoelectron microscopy revealed that MMP-13 was localized on the bone surfaces, under the ruffled borders and some clear zones of osteoclasts. Gold-labeled MMP-13 was closely associated with collagen fibrils. Gold labeling was also detected in Golgi apparatus of osteocytes adjacent to osteoclasts and bone lining cells. Western blotting showed that MMP-13 was mainly associated with mineralized bone matrix. These findings suggest that MMP-13 synthesized and secreted by osteoblast-lineage cells is localized under the ruffled borders of osteoclasts. MMP-13 may play an important role in degradation of type I collagen in bone matrix, acting in concert with cathepsin K and MMP-9 produced by osteoclasts. MMP-13 in perivascular cells may be involved in removal of cartilage matrix proteins such as type II collagen and aggrecan.

  10. The characterization of small hypoattenuating renal masses on contrast-enhanced CT☆

    PubMed Central

    Patel, Neesha S.; Poder, Liina; Wang, Zhen J.; Yeh, Benjamin M.; Qayyum, Aliya; Jin, Hua; Coakley, Fergus V.

    2011-01-01

    Purpose To determine if small hypoattenuating renal masses can be characterized as simple cysts or renal cell carcinomas on contrast-enhanced computed tomography (CT). Materials and methods We retrospectively identified 20 small (≤1.5 cm) hypoattenuating renal masses seen on contrast enhanced CT, consisting of 14 simple cysts and six renal cell carcinomas. Three independent readers recorded subjective visual impression (five-point scale from 1=definitely fluid to 5=definitely solid), CT attenuation, border (well circumscribed or ill defined), and shape (ovoid or irregular) for each lesion. Results The overall area under the receiver operator characteristic curves for subjective visual impression, CT attenuation, border, and shape were 0.97, 0.82, 0.59, and 0.55, respectively. Using dichotomized ratings (1–2=cyst and 3–5=carcinoma), subjective impression had a sensitivity and specificity of 100% and 79–100%, respectively, for the diagnosis of renal cell carcinoma. Using a threshold of 50 Hounsfield Units (HU) or more, CT attenuation had a sensitivity and specificity of 100% and 43–64%, respectively. Conclusion Small hypoattenuating renal masses can be characterized with reasonable accuracy by subjective impression and CT attenuation; lesions that appear solid on visual inspection or have an attenuation value of 50 HU or more are likely to be renal cell carcinoma. © 2009 Elsevier Inc. All rights reserved. PMID:19559352

  11. Reconnaissance geologic map of the Muwassam Quadrangle, sheet 16/42 D, Kingdom of Saudi Arabia

    USGS Publications Warehouse

    Blank, H. Richard

    1985-01-01

    All of the mapped area is occupied by coastal plain bordering the Red Sea. The flat coastal plain is covered by Quaternary surficial deposits overlying a sequence of Tertiary rocks as much as 5 km thick. The coastal plain is separated from the Red Sea by zone of supratidal sabkha deposits, offshore bars, islands, tidal md flats, and shallow lagoons. The sea is shallow, less than 200 m deep, and forms part of the shelf marginal to the main axial trough of the Red Sea.

  12. Back to the future: sweatshop conditions on the Mexico-U.S. border. I. Community health impact of maquiladora industrial activity.

    PubMed

    Moure-Eraso, R; Wilcox, M; Punnett, L; Copeland, L; Levenstein, C

    1994-03-01

    Present working conditions in one of the most active areas of the maquiladora system along the Mexico-U.S. border are reminiscent of nineteenth-century U.S. sweatshops. This conclusion was reached after evaluating two separate but interrelated surveys among Mexican nationals living near the Mexico-U.S. border, one of community leaders (Paper I), and one of workers in maquiladora enterprises in the towns of Matamoros and Reynosa, Mexico (Paper II). Paper I evaluates the results of the community leaders' survey. Criteria for selection of the leaders were: level of responsibility in the community; knowledge of the industry in the region, and length of residence in the area (more than 3 years). Representatives from government, maquiladora industry management, labor union leadership, labor union activists, and community improvement activists were interviewed. Structured questionnaires with opportunities for open-ended answers were used by trained Spanish speaking interviewers. The questions covered community demographics, health care structures, governance of the region, knowledge of working conditions, and knowledge of environmental impact on the region and the community. Community leaders were ambivalent on the purported benefits of the development of these types of industries in their communities. A substantial majority (21 of 25) thought that the maquiladoras brought few positive developments, other than creating jobs. Serious concerns about overextending weak social infrastructures and about environmental deterioration were voiced. Immediate (preventive) measures appear necessary to develop community infrastructures and to protect environmental health.

  13. Mexican Pharmacies and Antibiotic Consumption at the US-Mexico Border.

    PubMed

    Homedes, Núria; Ugalde, Antonio

    2012-12-01

    To study antibiotic dispensing to US and Mexican residents, at Mexican pharmacies at the US-Mexico border, and the pharmacy clerks' capability to promote appropriate use. The site selected was Ciudad Juarez, Chihuahua (pop. 1.2 million) separated from El Paso, Texas (pop. 800,000) by the Rio Grande River. A convenience sample of 32 pharmacies located near the international bridges, major shopping centers, and interior neighborhoods was selected. Pharmacy clients were interviewed (n=230) and 152 interactions between clients and pharmacy clerks were observed. Information was obtained about education and pharmaceutical training of 113 clerks working in 25 pharmacies. A senior pharmacy clerk in each of the 25 pharmacies was interviewed and asked for their recommendations to clients presenting two clinical scenarios and seven diagnoses. Professionally trained pharmacists only spend a few hours a week in some pharmacies. Clerks' education levels are very low; some have only completed primary education. There is no required pharmaceutical training and their knowledge about pharmaceuticals comes mostly from representatives of the pharmaceutical industry. Clerks' knowledge of antibiotics, the most frequently sold class of medicines (65% without prescription), is very limited. Clients trust pharmacy clerks and tend to follow their advice. The findings raise concerns about dispensing of antibiotics at Mexican border pharmacies and antibiotic overuse due to lack of control. Because inappropriate antibiotic use contributes to increased resistance, pharmacy clerks should receive independent training to dispense antibiotics and promote their appropriate use.

  14. Strong Paleoearthquakes along the Aksuu Border Fault according to the Results of Dating the Offset Terrace Complex of the Chon-Aksuu River, Northern Tien Shan

    NASA Astrophysics Data System (ADS)

    Korzhenkov, A. M.; Arrowsmith, J. R.; Crosby, C. J.; Guralnik, B.; Rogozhin, E. A.; Sorokin, A. A.; Abdieva, S. V.; Fortuna, A. B.; Yudakhin, A. S.; Agatova, A. R.; Deev, E. V.; Mazeika, J. V.; Rodkin, M. V.; Shen, J.

    2018-03-01

    The study and radiocarbon dating of the low alluvial terraces of the Chon-Aksuu River, in the Northern Issyk-Kul region, which were broken by the Kebin (Kemin) earthquake of 1911 (Ms = 8.2, Io = 10 to 11), are carried out. The obtained radiocarbon dated ages refer to the second half of the Holocene. Since that time, at least eight strong earthquakes took place along this (Chon-Aksuu) segment of the Aksuu border fault. Three seismic events, including the earthquake of 1911 occurred in the second millennium A.D. This outburst of seismic energy was preceded by two millennia of seismic quiescence, which set in after another pulse of seismic activation. The latter lasted for 1.5 millennia and included five strong earthquakes. The recurrence period of seismic events during the activations is 300-600 years. Hence, the seismic regime along the Chon-Aksuu segment of the Aksuu border fault in the second half of the Holocene was a succession of two seismic activations, each with a duration of 1.0-1.5 ka, which were separated by a 2-ka interval of seismic quiescence. Therefore, the absolute datings of the river terraces of different ages which have been broken by a seismogenic rupture can serve as a reliable source of information about the age of the strong earthquakes that occurred along the seismogenic fault.

  15. Mexican Pharmacies and Antibiotic Consumption at the US-Mexico Border

    PubMed Central

    Homedes, Núria; Ugalde, Antonio

    2012-01-01

    Objective: To study antibiotic dispensing to US and Mexican residents, at Mexican pharmacies at the US-Mexico border, and the pharmacy clerks’ capability to promote appropriate use. Methods: The site selected was Ciudad Juarez, Chihuahua (pop. 1.2 million) separated from El Paso, Texas (pop. 800,000) by the Rio Grande River. A convenience sample of 32 pharmacies located near the international bridges, major shopping centers, and interior neighborhoods was selected. Pharmacy clients were interviewed (n=230) and 152 interactions between clients and pharmacy clerks were observed. Information was obtained about education and pharmaceutical training of 113 clerks working in 25 pharmacies. A senior pharmacy clerk in each of the 25 pharmacies was interviewed and asked for their recommendations to clients presenting two clinical scenarios and seven diagnoses. Findings: Professionally trained pharmacists only spend a few hours a week in some pharmacies. Clerks’ education levels are very low; some have only completed primary education. There is no required pharmaceutical training and their knowledge about pharmaceuticals comes mostly from representatives of the pharmaceutical industry. Clerks’ knowledge of antibiotics, the most frequently sold class of medicines (65% without prescription), is very limited. Clients trust pharmacy clerks and tend to follow their advice. Conclusions: The findings raise concerns about dispensing of antibiotics at Mexican border pharmacies and antibiotic overuse due to lack of control. Because inappropriate antibiotic use contributes to increased resistance, pharmacy clerks should receive independent training to dispense antibiotics and promote their appropriate use. PMID:23532456

  16. Evolution of Spiral and Scroll Waves of Excitation in a Mathematical Model of Ischaemic Border Zone

    PubMed Central

    Biktashev, Vadim N.; Biktasheva, Irina V.; Sarvazyan, Narine A.

    2011-01-01

    Abnormal electrical activity from the boundaries of ischemic cardiac tissue is recognized as one of the major causes in generation of ischemia-reperfusion arrhythmias. Here we present theoretical analysis of the waves of electrical activity that can rise on the boundary of cardiac cell network upon its recovery from ischaemia-like conditions. The main factors included in our analysis are macroscopic gradients of the cell-to-cell coupling and cell excitability and microscopic heterogeneity of individual cells. The interplay between these factors allows one to explain how spirals form, drift together with the moving boundary, get transiently pinned to local inhomogeneities, and finally penetrate into the bulk of the well-coupled tissue where they reach macroscopic scale. The asymptotic theory of the drift of spiral and scroll waves based on response functions provides explanation of the drifts involved in this mechanism, with the exception of effects due to the discreteness of cardiac tissue. In particular, this asymptotic theory allows an extrapolation of 2D events into 3D, which has shown that cells within the border zone can give rise to 3D analogues of spirals, the scroll waves. When and if such scroll waves escape into a better coupled tissue, they are likely to collapse due to the positive filament tension. However, our simulations have shown that such collapse of newly generated scrolls is not inevitable and that under certain conditions filament tension becomes negative, leading to scroll filaments to expand and multiply leading to a fibrillation-like state within small areas of cardiac tissue. PMID:21935402

  17. Simulating spinal border cells and cerebellar granule cells under locomotion--a case study of spinocerebellar information processing.

    PubMed

    Spanne, Anton; Geborek, Pontus; Bengtsson, Fredrik; Jörntell, Henrik

    2014-01-01

    The spinocerebellar systems are essential for the brain in the performance of coordinated movements, but our knowledge about the spinocerebellar interactions is very limited. Recently, several crucial pieces of information have been acquired for the spinal border cell (SBC) component of the ventral spinocerebellar tract (VSCT), as well as the effects of SBC mossy fiber activation in granule cells of the cerebellar cortex. SBCs receive monosynaptic input from the reticulospinal tract (RST), which is an important driving system under locomotion, and disynaptic inhibition from Ib muscle afferents. The patterns of activity of RST neurons and Ib afferents under locomotion are known. The activity of VSCT neurons under fictive locomotion, i.e. without sensory feedback, is also known, but there is little information on how these neurons behave under actual locomotion and for cerebellar granule cells receiving SBC input this is completely unknown. But the available information makes it possible to simulate the interactions between the spinal and cerebellar neuronal circuitries with a relatively large set of biological constraints. Using a model of the various neuronal elements and the network they compose, we simulated the modulation of the SBCs and their target granule cells under locomotion and hence generated testable predictions of their general pattern of modulation under this condition. This particular system offers a unique opportunity to simulate these interactions with a limited number of assumptions, which helps making the model biologically plausible. Similar principles of information processing may be expected to apply to all spinocerebellar systems.

  18. Enhancing Centrifugal Separation With Electrophoresis

    NASA Technical Reports Server (NTRS)

    Herrmann, F. T.

    1986-01-01

    Separation of biological cells by coil-planet centrifuge enhanced by electrophoresis. By itself, coil-planet centrifuge offers relatively gentle method of separating cells under low centrifugal force in physiological medium that keeps cells alive. With addition of voltage gradient to separation column of centrifuge, separation still gentle but faster and more complete. Since separation apparatus contains no rotary seal, probability of leakage, contamination, corrosion, and short circuits reduced.

  19. High speed flow cytometric separation of viable cells

    DOEpatents

    Sasaki, D.T.; Van den Engh, G.J.; Buckie, A.M.

    1995-11-14

    Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

  20. High speed flow cytometric separation of viable cells

    DOEpatents

    Sasaki, Dennis T.; Van den Engh, Gerrit J.; Buckie, Anne-Marie

    1995-01-01

    Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

  1. Paramagnetic capture mode magnetophoretic microseparator for high efficiency blood cell separations.

    PubMed

    Han, Ki-Ho; Frazier, A Bruno

    2006-02-01

    This paper presents the characterization of continuous single-stage and three-stage cascade paramagnetic capture (PMC) mode magnetophoretic microseparators for high efficiency separation of red and white blood cells from diluted whole blood based on their native magnetic properties. The separation mechanism for both PMC microseparators is based on a high gradient magnetic separation (HGMS) method. This approach enables separation of blood cells without the use of additives such as magnetic beads. Experimental results for the single-stage PMC microseparator show that 91.1% of red blood cells were continuously separated from the sample at a volumetric flow rate of 5 microl h-1. In addition, the three-stage cascade PMC microseparator continuously separated 93.5% of red blood cells and 97.4% of white blood cells from whole blood at a volumetric flow rate of 5 microl h-1.

  2. Deterministic Migration-Based Separation of White Blood Cells.

    PubMed

    Kim, Byeongyeon; Choi, Young Joon; Seo, Hyekyung; Shin, Eui-Cheol; Choi, Sungyoung

    2016-10-01

    Functional and phenotypic analyses of peripheral white blood cells provide useful clinical information. However, separation of white blood cells from peripheral blood requires a time-consuming, inconvenient process and thus analyses of separated white blood cells are limited in clinical settings. To overcome this limitation, a microfluidic separation platform is developed to enable deterministic migration of white blood cells, directing the cells into designated positions according to a ridge pattern. The platform uses slant ridge structures on the channel top to induce the deterministic migration, which allows efficient and high-throughput separation of white blood cells from unprocessed whole blood. The extent of the deterministic migration under various rheological conditions is explored, enabling highly efficient migration of white blood cells in whole blood and achieving high-throughput separation of the cells (processing 1 mL of whole blood less than 7 min). In the separated cell population, the composition of lymphocyte subpopulations is well preserved, and T cells secrete cytokines without any functional impairment. On the basis of the results, this microfluidic platform is a promising tool for the rapid enrichment of white blood cells, and it is useful for functional and phenotypic analyses of peripheral white blood cells. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. A miniaturized multipurpose platform for rapid, label-free, and simultaneous separation, patterning, and in vitro culture of primary and rare cells.

    PubMed

    Didar, Tohid Fatanat; Bowey, Kristen; Almazan, Guillermina; Tabrizian, Maryam

    2014-02-01

    Given that current cell isolation techniques are expensive, time consuming, yield low isolation purities, and/or alter target cell properties, a versatile, cost effective, and easy-to-operate microchip with the capability to simultaneously separate, capture, pattern, and culture rare and primary cells in vitro is developed. The platform is based on target cell adhesion onto the micro-fabricated interfaces produced by microcontact printing of cell-specific antibodies. Results show over 95% separation efficiency in less than 10 min for the separation of oligodendrocyte progenitor cells (OPCs) and cardiomyocytes from rat brain and heart mixtures, respectively. Target cell attachment and single cell spreading can be precisely controlled on the basis of the designed patterns. Both cell types can maintain their biofunctionality. Indeed, isolated OPCs can proliferate and differentiate into mature oligodendrocytes, while isolated cardiomyocytes retain their contractile properties on the separation platform. Successful separation of two dissimilar cell types present in varying concentrations in their respective cell mixtures and the demonstration of their integrity after separation open new avenues for time and cost-effective sorting of various cell types using the developed miniaturized platform. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. A micropatterning approach for imaging Cx43 dynamic trafficking to cell-cell borders

    PubMed Central

    Zhang, Shan-Shan; Hong, SoonGweon; Kléber, André G.; Lee, Luke P.; Shaw, Robin M.

    2014-01-01

    The precise expression and timely delivery of connexin 43 (Cx43) proteins to form gap junctions are essential for electrical coupling of cardiomyocytes. Growing evidence supports a cytoskeletal-based trafficking paradigm for Cx43 delivery directly to adherens junctions at the intercalated disc. A limitation of Cx43 localization assays in cultured cells, in which cell-cell contacts are essential, is the inability to control for cell geometry or reproducibly generate contact points. Here we present a micropatterned cell pairing system well suited for live microscopy to examine how the microtubule and actin cytoskeleton confer specificity to Cx43 trafficking to precisely defined cell-cell junctions. This system can also be adapted for other cell types and used to study dynamic intracellular movements of other proteins important for cell-cell communication‥ PMID:24444605

  5. The Role of Chronic Mucosal Trauma in Oral Cancer: A Review of Literature

    PubMed Central

    Singhvi, Hitesh Rajendra; Malik, Akshat; Chaturvedi, Pankaj

    2017-01-01

    Chronic mucosal trauma resulting from sharp teeth, dentures, faulty restoration, or implants has frequently been associated with the development of oral cancer. The definitive evidence for the same is lacking. We undertook a search using the terms – dental trauma, mucosal trauma, oral cancer, squamous cell carcinoma, risk factor, potentially malignant lesion, dental factor, mechanical irritation, dental irritation, and cancer in the following electronic databases: MEDLINE, PubMed, ScienceDirect, Cochrane Database of Systematic Reviews, and Wiley InterScience. The search yielded 788 articles. Of these articles, only 22 articles described chronic mucosal trauma as risk factors for oral cancers and were considered in this review. The review shows that chronic mucosal irritation resulting from ill-fitting dentures may be considered a risk factor for the development of oral cancer, such cancers occur commonly over the lateral border of the tongue. However, no association has been proven between the duration of denture use and cancer formation. In patients without any addiction, such cancers occur more frequently in females. These cancers may present with an early nodal disease but their prognosis and outcomes have not been studied separately till now. PMID:28469336

  6. Molten carbonate fuel cell separator

    DOEpatents

    Nickols, Richard C.

    1986-09-02

    In a stacked array of molten carbonate fuel cells, a fuel cell separator is positioned between adjacent fuel cells to provide isolation as well as a conductive path therebetween. The center portion of the fuel cell separator includes a generally rectangular, flat, electrical conductor. Around the periphery of the flat portion of the separator are positioned a plurality of elongated resilient flanges which form a gas-tight seal around the edges of the fuel cell. With one elongated flange resiliently engaging a respective edge of the center portion of the separator, the sealing flanges, which are preferably comprised of a noncorrosive material such as an alloy of yttrium, iron, aluminum or chromium, form a tight-fitting wet seal for confining the corrosive elements of the fuel cell therein. This arrangement permits a good conductive material which may be highly subject to corrosion and dissolution to be used in combination with a corrosion-resistant material in the fuel cell separator of a molten carbonate fuel cell for improved fuel cell conductivity and a gas-tight wet seal.

  7. Molten carbonate fuel cell separator

    DOEpatents

    Nickols, R.C.

    1984-10-17

    In a stacked array of molten carbonate fuel cells, a fuel cell separator is positioned between adjacent fuel cells to provide isolation as well as a conductive path therebetween. The center portion of the fuel cell separator includes a generally rectangular, flat, electrical conductor. Around the periphery of the flat portion of the separator are positioned a plurality of elongated resilient flanges which form a gas-tight seal around the edges of the fuel cell. With one elongated flange resiliently engaging a respective edge of the center portion of the separator, the sealing flanges, which are preferably comprised of a noncorrosive material such as an alloy of yttrium, iron, aluminum or chromium, form a tight-fitting wet seal for confining the corrosive elements of the fuel cell therein. This arrangement permits a good conductive material which may be highly subject to corrosion and dissolution to be used in combination with a corrosion-resistant material in the fuel cell separator of a molten carbonate fuel cell for improved fuel cell conductivity and a gas-tight wet seal.

  8. Separation of CHO cells using hydrocyclones.

    PubMed

    Pinto, Rodrigo C V; Medronho, Ricardo A; Castilho, Leda R

    2008-01-01

    Hydrocyclones are simple and robust separation devices with no moving parts. In the past few years, their use in animal cell separation has been proposed. In this work, the use of different hydrocyclone configurations for Chinese hamster ovary (CHO) cell separation was investigated following an experimental design. It was shown that cell separation efficiencies for cultures of the wild-type CHO.K1 cell line and of a recombinant CHO cell line producing granulocyte-macrophage colony stimulating factor (GM-CSF) were kept above 97%. Low viability losses were observed, as measured by trypan blue exclusion and by determination of intracellular lactate dehydrogenase (LDH) released to the culture medium. Mathematical models were proposed to predict the flow rate, flow ratio and separation efficiency as a function of hydrocyclone geometry and pressure drop. When cells were monitored for any induction of apoptosis upon passage through the hydrocyclones, no increase in apoptotic cell concentration was observed within 48 h of hydrocycloning. Thus, based on the high separation efficiencies, the robustness of the equipment, and the absence of apoptosis induction, hydrocyclones seem to be specially suited for use as cell retention devices in long-term perfusion runs.

  9. Cell separator for use in bipolar-stack energy storage devices

    DOEpatents

    Mayer, Steven T.; Feikert, John H.; Kachmitter, James L.; Pekala, Richard W.

    1995-01-01

    An improved multi-cell electrochemical energy storage device, such as a battery, fuel cell, or double layer capacitor using a cell separator which allows cells to be stacked and interconnected with low electrical resistance and high reliability while maximizing packaging efficiency. By adding repeating cells, higher voltages can be obtained. The cell separator is formed by applying an organic adhesive on opposing surfaces of adjacent carbon electrodes or surfaces of aerogel electrodes of a pair of adjacent cells prior to or after pyrolysis thereof to form carbon aerogel electrodes. The cell separator is electronically conductive, but ionically isolating, preventing an electrolytic conduction path between adjacent cells in the stack.

  10. Separation and sorting of cells in microsystems using physical principles

    NASA Astrophysics Data System (ADS)

    Lee, Gi-Hun; Kim, Sung-Hwan; Ahn, Kihoon; Lee, Sang-Hoon; Park, Joong Yull

    2016-01-01

    In the last decade, microfabrication techniques have been combined with microfluidics and applied to cell biology. Utilizing such new techniques, various cell studies have been performed for the research of stem cells, immune cells, cancer, neurons, etc. Among the various biological applications of microtechnology-based platforms, cell separation technology has been highly regarded in biological and clinical fields for sorting different types of cells, finding circulating tumor cells (CTCs), and blood cell separation, amongst other things. Many cell separation methods have been created using various physical principles. Representatively, these include hydrodynamic, acoustic, dielectrophoretic, magnetic, optical, and filtering methods. In this review, each of these methods will be introduced, and their physical principles and sample applications described. Each physical principle has its own advantages and disadvantages. The engineers who design the systems and the biologists who use them should understand the pros and cons of each method or principle, to broaden the use of microsystems for cell separation. Continuous development of microsystems for cell separation will lead to new opportunities for diagnosing CTCs and cancer metastasis, as well as other elements in the bloodstream.

  11. Deformability and size-based cancer cell separation using an integrated microfluidic device.

    PubMed

    Pang, Long; Shen, Shaofei; Ma, Chao; Ma, Tongtong; Zhang, Rui; Tian, Chang; Zhao, Lei; Liu, Wenming; Wang, Jinyi

    2015-11-07

    Cell sorting by filtration techniques offers a label-free approach for cell separation on the basis of size and deformability. However, filtration is always limited by the unpredictable variation of the filter hydrodynamic resistance due to cell accumulation and clogging in the microstructures. In this study, we present a new integrated microfluidic device for cell separation based on the cell size and deformability by combining the microstructure-constricted filtration and pneumatic microvalves. Using this device, the cell populations sorted by the microstructures can be easily released in real time for subsequent analysis. Moreover, the periodical sort and release of cells greatly avoided cell accumulation and clogging and improved the selectivity. Separation of cancer cells (MCF-7, MDA-MB-231 and MDA231-LM2) with different deformability showed that the mixture of the less flexible cells (MCF-7) and the flexible cells (MDA-MB-231 and MDA231-LM2) can be well separated with more than 75% purity. Moreover, the device can be used to separate cancer cells from the blood samples with more than 90% cell recovery and more than 80% purity. Compared with the current filtration methods, the device provides a new approach for cancer cell separation with high collection recovery and purity, and also, possesses practical potential to be applied as a sample preparation platform for fundamental studies and clinical applications.

  12. Design and simulation of a microfluidic device for acoustic cell separation.

    PubMed

    Shamloo, Amir; Boodaghi, Miad

    2018-03-01

    Experimental acoustic cell separation methods have been widely used to perform separation for different types of blood cells. However, numerical simulation of acoustic cell separation has not gained enough attention and needs further investigation since by using numerical methods, it is possible to optimize different parameters involved in the design of an acoustic device and calculate particle trajectories in a simple and low cost manner before spending time and effort for fabricating these devices. In this study, we present a comprehensive finite element-based simulation of acoustic separation of platelets, red blood cells and white blood cells, using standing surface acoustic waves (SSAWs). A microfluidic channel with three inlets, including the middle inlet for sheath flow and two symmetrical tilted angle inlets for the cells were used to drive the cells through the channel. Two interdigital transducers were also considered in this device and by implementing an alternating voltage to the transducers, an acoustic field was created which can exert the acoustic radiation force to the cells. Since this force is dependent to the size of the cells, the cells are pushed towards the midline of the channel with different path lines. Particle trajectories for different cells were obtained and compared with a theoretical equation. Two types of separations were observed as a result of varying the amplitude of the acoustic field. In the first mode of separation, white blood cells were sorted out through the middle outlet and in the second mode of separation, platelets were sorted out through the side outlets. Depending on the clinical needs and by using the studied microfluidic device, each of these modes can be applied to separate the desired cells. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Cell partition in two phase polymer systems

    NASA Technical Reports Server (NTRS)

    Brooks, D. E.

    1979-01-01

    Aqueous phase-separated polymer solutions can be used as support media for the partition of biological macromolecules, organelles and cells. Cell separations using the technique have proven to be extremely sensitive to cell surface properties but application of the systems are limited to cells or aggregates which do not significantly while the phases are settling. Partition in zero g in principle removes this limitation but an external driving force must be applied to induce the phases to separate since their density difference disappears. We have recently shown that an applied electric field can supply the necessary driving force. We are proposing to utilize the NASA FES to study field-driven phase separation and cell partition on the ground and in zero g to help define the separation/partition process, with the ultimate goal being to develop partition as a zero g cell separation technique.

  14. The role of the bi-compartmental stem cell niche in delaying cancer

    NASA Astrophysics Data System (ADS)

    Shahriyari, Leili; Komarova, Natalia L.

    2015-10-01

    In recent years, by using modern imaging techniques, scientists have found evidence of collaboration between different types of stem cells (SCs), and proposed a bi-compartmental organization of the SC niche. Here we create a class of stochastic models to simulate the dynamics of such a heterogeneous SC niche. We consider two SC groups: the border compartment, S1, is in direct contact with transit-amplifying (TA) cells, and the central compartment, S2, is hierarchically upstream from S1. The S1 SCs differentiate or divide asymmetrically when the tissue needs TA cells. Both groups proliferate when the tissue requires SCs (thus maintaining homeostasis). There is an influx of S2 cells into the border compartment, either by migration, or by proliferation. We examine this model in the context of double-hit mutant generation, which is a rate-limiting step in the development of many cancers. We discover that this type of a cooperative pattern in the stem niche with two compartments leads to a significantly smaller rate of double-hit mutant production compared with a homogeneous, one-compartmental SC niche. Furthermore, the minimum probability of double-hit mutant generation corresponds to purely symmetric division of SCs, consistent with the literature. Finally, the optimal architecture (which minimizes the rate of double-hit mutant production) requires a large proliferation rate of S1 cells along with a small, but non-zero, proliferation rate of S2 cells. This result is remarkably similar to the niche structure described recently by several authors, where one of the two SC compartments was found more actively engaged in tissue homeostasis and turnover, while the other was characterized by higher levels of quiescence (but contributed strongly to injury recovery). Both numerical and analytical results are presented.

  15. Validity of endothelial cell analysis methods and recommendations for calibration in Topcon SP-2000P specular microscopy.

    PubMed

    van Schaick, Willem; van Dooren, Bart T H; Mulder, Paul G H; Völker-Dieben, Hennie J M

    2005-07-01

    To report on the calibration of the Topcon SP-2000P specular microscope and the Endothelial Cell Analysis Module of the IMAGEnet 2000 software, and to establish the validity of the different endothelial cell density (ECD) assessment methods available in these instruments. Using an external microgrid, we calibrated the magnification of the SP-2000P and the IMAGEnet software. In both eyes of 36 volunteers, we validated 4 ECD assessment methods by comparing these methods to the gold standard manual ECD, manual counting of cells on a video print. These methods were: the estimated ECD, estimation of ECD with a reference grid on the camera screen; the SP-2000P ECD, pointing out whole contiguous cells on the camera screen; the uncorrected IMAGEnet ECD, using automatically drawn cell borders, and the corrected IMAGEnet ECD, with manual correction of incorrectly drawn cell borders in the automated analysis. Validity of each method was evaluated by calculating both the mean difference with the manual ECD and the limits of agreement as described by Bland and Altman. Preset factory values of magnification were incorrect, resulting in errors in ECD of up to 9%. All assessments except 1 of the estimated ECDs differed significantly from manual ECDs, with most differences being similar (< or =6.5%), except for uncorrected IMAGEnet ECD (30.2%). Corrected IMAGEnet ECD showed the narrowest limits of agreement (-4.9 to +19.3%). We advise checking the calibration of magnification in any specular microscope or endothelial analysis software as it may be erroneous. Corrected IMAGEnet ECD is the most valid of the investigated methods in the Topcon SP-2000P/IMAGEnet 2000 combination.

  16. Slow cycling cells in the continuous dental lamina of Scyliorhinus canicula: new evidence for stem cells in sharks.

    PubMed

    Vandenplas, Sam; Vandeghinste, Robbe; Boutet, Agnes; Mazan, Sylvie; Huysseune, Ann

    2016-05-01

    In the lesser spotted catshark (Scyliorhinus canicula), as in most non-mammalian vertebrates, the dentition renews throughout life. To contribute to our understanding of how continuous tooth replacement is achieved, we searched for evidence for the presence of stem cells in this species. Three-dimensional reconstructions of juvenile (2-3 weeks post-hatch) specimens showed that tooth families merge imperceptibly with so-called interdental zones within a continuous and permanent dental lamina. Interdental regions are composed of three layers, continuous with cervical loop, middle, and outer dental epithelium of the tooth families, respectively. A BrdU pulse-chase experiment revealed that cell proliferation is initiated in the lingual part of the dental lamina and the resulting population shifts one tooth position towards the oral epithelium in around four to five weeks. In the longest chase time (114 days) label-retaining and arguably non-differentiated cells were present at the lingual border of the dental lamina. These were found in the outer and middle dental epithelium, both within and between tooth families. This area of the dental lamina did not show expression or distribution of Sox2. Our data support the hypothesis that stem cells reside at the lingual border of the continuous dental lamina, more specifically in the middle dental epithelium at the level of the tooth families, and in its extension between the tooth families. To demonstrate their true stemness and their role in continuous tooth replacement, it remains to be shown that these cells have the potential to give rise to a complete new successor. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Information Analysis Methodology for Border Security Deployment Prioritization and Post Deployment Evaluation

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

    Booker, Paul M.; Maple, Scott A.

    2010-06-08

    Due to international commerce, cross-border conflicts, and corruption, a holistic, information driven, approach to border security is required to best understand how resources should be applied to affect sustainable improvements in border security. The ability to transport goods and people by land, sea, and air across international borders with relative ease for legitimate commercial purposes creates a challenging environment to detect illicit smuggling activities that destabilize national level border security. Smuggling activities operated for profit or smuggling operations driven by cross border conflicts where militant or terrorist organizations facilitate the transport of materials and or extremists to advance a causemore » add complexity to smuggling interdiction efforts. Border security efforts are further hampered when corruption thwarts interdiction efforts or reduces the effectiveness of technology deployed to enhance border security. These issues necessitate the implementation of a holistic approach to border security that leverages all available data. Large amounts of information found in hundreds of thousands of documents can be compiled to assess national or regional borders to identify variables that influence border security. Location data associated with border topics of interest may be extracted and plotted to better characterize the current border security environment for a given country or region. This baseline assessment enables further analysis, but also documents the initial state of border security that can be used to evaluate progress after border security improvements are made. Then, border security threats are prioritized via a systems analysis approach. Mitigation factors to address risks can be developed and evaluated against inhibiting factor such as corruption. This holistic approach to border security helps address the dynamic smuggling interdiction environment where illicit activities divert to a new location that provides less resistance to smuggling activities after training or technology is deployed at a given location. This paper will present an approach to holistic border security information analysis.« less

  18. Vulnerability of Xylem Vessels to Cavitation in Sugar Maple. Scaling from Individual Vessels to Whole Branches1

    PubMed Central

    Melcher, Peter J.; Zwieniecki, Maciej A.; Holbrook, N. Michele

    2003-01-01

    The relation between xylem vessel age and vulnerability to cavitation of sugar maple (Acer saccharum Marsh.) was quantified by measuring the pressure required to force air across bordered pit membranes separating individual xylem vessels. We found that the bordered pit membranes of vessels located in current year xylem could withstand greater applied gas pressures (3.8 MPa) compared with bordered pit membranes in vessels located in older annular rings (2.0 MPa). A longitudinal transect along 6-year-old branches indicated that the pressure required to push gas across bordered pit membranes of current year xylem did not vary with distance from the growing tip. To understand the contribution of age-related changes in vulnerability to the overall resistance to cavitation, we combined data on the pressure thresholds of individual xylem vessels with measurements of the relative flow rate through each annual ring. The annual ring of the current year contributed only 16% of the total flow measured on 10-cm-long segments cut from 6-year-old branches, but it contributed more than 70% of the total flow when measured through 6-year-old branches to the point of leaf attachment. The vulnerability curve calculated using relative flow rates measured on branch segments were similar to vulnerability curves measured on 6-year-old branches (pressure that reduces hydraulic conductance by 50% = 1.6–2.4 MPa), whereas the vulnerability curve calculated using relative flow rates measured on 6-year-old branches were similar to ones measured on the extension growth of the current year (pressure that reduces hydraulic conductance by 50% = 3.8 MPa). These data suggest that, in sugar maple, the xylem of the current year can withstand larger xylem tensions than older wood and dominates water delivery to leaves. PMID:12692336

  19. Suicidal behavior by burns among women in two bordering provinces in Iran

    PubMed Central

    Alaghehbandan, R.; Dinn, N.A.; Rastegar Lari, E.; Rastegar Lari, A.

    2015-01-01

    Summary The objective of the study was to describe epidemiologic features of suicidal behavior by burns among women in two bordering provinces in Iran. A prospective hospital-based study of all suicidal behaviors by burns among women was carried out in the Iranian provinces of Khorasan (which borders Afghanistan) and Ilam (which borders Iraq) in 2006. These two provinces suffered from war and economic instability with high rates of poverty and unemployment. Data were obtained from patients, family members, and/or significant others through interviews during the course of hospitalization. Of 181 hospitalized patients, 130 (71.8%) were female, representing a hospital separation rate of 5.4 per 100,000 person-year (P-Y). The rate of suicidal behavior involving burns among the rural population was higher than that in the urban population (6.3 vs. 4.7 per 100,000 P-Y, P = 0.09). The median age of the patients was 23 years old, with a mean (±SD) of 27.9 (±14.3) years old. Approximately 68.5% of women were married. Quarrels with “a family member, spouse, a relative, or a friend” (marital plus other conflicts) was identified as the most common precipitating factor (88.5%) in attempting suicide. More than one third of women were abused (physically and/or verbally) by family members (i.e. spouse, father-in-law). Case fatality rate in this study was 52.3% (68/130). Suicide by burns is still a socio-epidemiological problem in Iran, particularly among young, married women in rural areas. Social and economic conditions, as well as violence against women, play a major role in the high rate of suicide by burns. The findings of this study highlight the need for the implementation of a well-organized approach to reduce the rate of suicide by burns among the most vulnerable populations in Iran. PMID:27252614

  20. Evaluation of results of cell electrophoresis experiments on space shuttle STS-3 including pre-flight and post-flight laboratory experiments

    NASA Technical Reports Server (NTRS)

    Todd, P. W.

    1985-01-01

    The objectives of the red blood cell experiments were to provide a visual check on the electrophoretic process and especially electroosmotic flow in space as well as to provide test separations of non-degradable standard particles for comparison with the separations of the three viable cell types studied on the Apollo-Soyuz Test Project. Determination of the maximum concentrations of cells that can be separated in column electrophore was a significant goal. Two of the eight columns were available for red cell experiments, so two concentrations of human and rabbit RBC mixtures were used. The objectives of another experiment were to evaluate the reproducibility of microgravity electrophoretic separation of living kidney cells, to separate cells with highly viability despite two freeze-thaw cycles, and to optimize the physical conditions of cell separation. Owing to the uncertain heterogeneity of the starting material, the experimental design does not assess resolution in microgravity, but improved separability was sought in comparison to density-gradient electrophoresis or continuous-flow electrophoresis. Efforts were made to increase cell yield and cell viability and to assess reproducibility directly.

  1. 9 CFR 130.6 - User fees for inspection of live animals at land border ports along the United States-Mexico border.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... animals at land border ports along the United States-Mexico border. 130.6 Section 130.6 Animals and Animal... User fees for inspection of live animals at land border ports along the United States-Mexico border. (a... importation into or entry into the United States through a land border port along the United States-Mexico...

  2. 9 CFR 130.6 - User fees for inspection of live animals at land border ports along the United States-Mexico border.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... animals at land border ports along the United States-Mexico border. 130.6 Section 130.6 Animals and Animal... User fees for inspection of live animals at land border ports along the United States-Mexico border. (a... importation into or entry into the United States through a land border port along the United States-Mexico...

  3. 9 CFR 130.6 - User fees for inspection of live animals at land border ports along the United States-Mexico border.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... animals at land border ports along the United States-Mexico border. 130.6 Section 130.6 Animals and Animal... User fees for inspection of live animals at land border ports along the United States-Mexico border. (a... importation into or entry into the United States through a land border port along the United States-Mexico...

  4. 9 CFR 130.6 - User fees for inspection of live animals at land border ports along the United States-Mexico border.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... animals at land border ports along the United States-Mexico border. 130.6 Section 130.6 Animals and Animal... User fees for inspection of live animals at land border ports along the United States-Mexico border. (a... importation into or entry into the United States through a land border port along the United States-Mexico...

  5. 9 CFR 130.6 - User fees for inspection of live animals at land border ports along the United States-Mexico border.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... animals at land border ports along the United States-Mexico border. 130.6 Section 130.6 Animals and Animal... User fees for inspection of live animals at land border ports along the United States-Mexico border. (a... importation into or entry into the United States through a land border port along the United States-Mexico...

  6. A New Cell Separation Method Based on Antibody-Immobilized Nanoneedle Arrays for the Detection of Intracellular Markers.

    PubMed

    Kawamura, Ryuzo; Miyazaki, Minami; Shimizu, Keita; Matsumoto, Yuta; Silberberg, Yaron R; Sathuluri, Ramachandra Rao; Iijima, Masumi; Kuroda, Shun'ichi; Iwata, Futoshi; Kobayashi, Takeshi; Nakamura, Chikashi

    2017-11-08

    Focusing on intracellular targets, we propose a new cell separation technique based on a nanoneedle array (NNA) device, which allows simultaneous insertion of multiple needles into multiple cells. The device is designed to target and lift ("fish") individual cells from a mixed population of cells on a substrate using an antibody-functionalized NNA. The mechanics underlying this approach were validated by force analysis using an atomic force microscope. Accurate high-throughput separation was achieved using one-to-one contacts between the nanoneedles and the cells by preparing a single-cell array in which the positions of the cells were aligned with 10,000 nanoneedles in the NNA. Cell-type-specific separation was realized by controlling the adhesion force so that the cells could be detached in cell-type-independent manner. Separation of nestin-expressing neural stem cells (NSCs) derived from human induced pluripotent stem cells (hiPSCs) was demonstrated using the proposed technology, and successful differentiation to neuronal cells was confirmed.

  7. Stable isotope ratios of marijuana. I. Carbon and nitrogen stable isotopes describe growth conditions.

    PubMed

    West, Jason B; Hurley, Janet M; Ehleringer, James R

    2009-01-01

    There remains significant uncertainty in illicit marijuana cultivation. We analyzed the delta(13)C and delta(15)N of 508 domestic samples from known U.S.A. counties, 31 seized from a single location, 5 samples grown in Mexico and Colombia, and 10 northwest border seizures. For a subset, inflorescences and leaves were analyzed separately. These data revealed a strong correspondence, with inflorescences having slightly higher delta(13)C and delta(15)N values than leaves. A framework for interpreting these results is introduced and evaluated. Samples identified as outdoor-grown by delta(13)C were generally recorded as such by the Drug Enforcement Administration (DEA). DEA-classified indoor-grown samples had the most negative delta(13)C values, consistent with indoor cultivation, although many were also in the outdoor-grown domain. Delta(15)N indicated a wide range of fertilizers across the dataset. Samples seized at the single location suggested multiple sources. Northwest border delta(13)C values suggested indoor growth, whereas for the Mexican and Colombian samples they indicated outdoor growth.

  8. Direct in situ measurement of specific capacitance, monolayer tension, and bilayer tension in a droplet interface bilayer

    DOE PAGES

    Taylor, Graham J.; Venkatesan, Guru A.; Collier, C. Patrick; ...

    2015-08-05

    In this study, thickness and tension are important physical parameters of model cell membranes. However, traditional methods to measure these quantities require multiple experiments using separate equipment. This work introduces a new multi-step procedure for directly accessing in situ multiple physical properties of droplet interface bilayers (DIB), including specific capacitance (related to thickness), lipid monolayer tension in the Plateau-Gibbs border, and bilayer tension. The procedure employs a combination of mechanical manipulation of bilayer area followed by electrowetting of the capacitive interface to examine the sensitivities of bilayer capacitance to area and contact angle to voltage, respectively. These data allow formore » determining the specific capacitance of the membrane and surface tension of the lipid monolayer, which are then used to compute bilayer thickness and tension, respectively. The use of DIBs affords accurate optical imaging of the connected droplets in addition to electrical measurements of bilayer capacitance, and it allows for reversibly varying bilayer area. After validating the accuracy of the technique with diphytanoyl phosphatidylcholine (DPhPC) DIBs in hexadecane, the method is applied herein to quantify separately the effects on membrane thickness and tension caused by varying the solvent in which the DIB is formed and introducing cholesterol into the bilayer. Because the technique relies only on capacitance measurements and optical images to determine both thickness and tension, this approach is specifically well-suited for studying the effects of peptides, biomolecules, natural and synthetic nanoparticles, and other species that accumulate within membranes without altering bilayer conductance.« less

  9. Retrospective cohort study of prognostic factors in patients with oral cavity and oropharyngeal squamous cell carcinoma.

    PubMed

    Carrillo, José F; Carrillo, Liliana C; Cano, Ana; Ramirez-Ortega, Margarita C; Chanona, Jorge G; Avilés, Alejandro; Herrera-Goepfert, Roberto; Corona-Rivera, Jaime; Ochoa-Carrillo, Francisco J; Oñate-Ocaña, Luis F

    2016-04-01

    Prognostic factors in oral cavity and oropharyngeal squamous cell carcinoma (SCC) are debated. The purpose of this study was to investigate the association of prognostic factors with oncologic outcomes. Patients with oral cavity and oropharyngeal SCC treated from 1997 to 2012 were included in this retrospective cohort study. Associations of prognostic factors with locoregional recurrence (LRR) or overall survival (OS) were analyzed using the logistic regression and the Cox models. Six hundred thirty-four patients were included in this study; tumor size, surgical margins, and N classification were associated with LRR (p < .0001); considering histopathology: perineural invasion, lymphocytic infiltration, infiltrative borders, and N classification were significant determinants of LRR. Tumor size, N classification, alcoholism, and surgical margins were associated with OS (p < .0001); considering pathologic prognostic factors, perivascular invasion, islands borders, and surgical margins were independently associated with OS (p < .0001). Surgical margins, perineural and perivascular invasion, lymphocytic infiltration, and infiltrative patterns of tumor invasion are significant prognostic factors in oral cavity and oropharyngeal SCC. © 2015 Wiley Periodicals, Inc.

  10. Haematology and serum chemistry of Pyrenean chamois (Rupicapra pyrenaica) naturally infected with a border disease virus.

    PubMed

    Fernández-Sirera, L; Mentaberre, G; López-Olvera, J R; Cuenca, R; Lavín, S; Marco, I

    2011-06-01

    In 2005 and 2006 an outbreak of disease associated with border disease virus (BDV) infection caused high mortality in the Pyrenean chamois (Rupicapra pyrenaica) in the Catalan Pyrenees (NE Spain). The aim of this study was to determine values for different haematological and serum biochemical analytes in 32 free-ranging Pyrenean chamois affected by the disease and to compare them with those obtained from healthy chamois. In the affected chamois red blood cell counts, haemoglobin concentrations, packed cell volumes, mean corpuscular volumes and lymphocyte counts were all lower, while the neutrophil and platelet counts were higher. Glucose, lactate, triglycerides, creatinine, total protein concentrations and alkaline phosphatase activity were also lower, in contrast to the concentrations of total bilirubin, urea and aspartate aminotransferase activity, which were higher. Most of the observed changes could be associated with cachexia and inflammation in the affected chamois. Lymphopenia could be directly related to the BDV, which would lead to immunosuppression and explain the high rate of secondary infection observed in these animals. Copyright © 2010 Elsevier Ltd. All rights reserved.

  11. Fundamentals and Application of Magnetic Particles in Cell Isolation and Enrichment

    PubMed Central

    Plouffe, Brian D.; Murthy, Shashi K.; Lewis, Laura H.

    2014-01-01

    Magnetic sorting using magnetic beads has become a routine methodology for the separation of key cell populations from biological suspensions. Due to the inherent ability of magnets to provide forces at a distance, magnetic cell manipulation is now a standardized process step in numerous processes in tissue engineering, medicine, and in fundamental biological research. Herein we review the current status of magnetic particles to enable isolation and separation of cells, with a strong focus on the fundamental governing physical phenomena, properties and syntheses of magnetic particles and on current applications of magnet-based cell separation in laboratory and clinical settings. We highlight the contribution of cell separation to biomedical research and medicine and detail modern cell separation methods (both magnetic and non-magnetic). In addition to a review of the current state-of-the-art in magnet-based cell sorting, we discuss current challenges and available opportunities for further research, development and commercialization of magnetic particle-based cell separation systems. PMID:25471081

  12. In Vivo, Villin Is Required for Ca2+-Dependent F-Actin Disruption in Intestinal Brush Borders

    PubMed Central

    Ferrary, Evelyne; Cohen-Tannoudji, Michel; Pehau-Arnaudet, Gérard; Lapillonne, Alexandre; Athman, Rafika; Ruiz, Tereza; Boulouha, Lilia; El Marjou, Fatima; Doye, Anne; Fontaine, Jean-Jacques; Antony, Claude; Babinet, Charles; Louvard, Daniel; Jaisser, Frédéric; Robine, Sylvie

    1999-01-01

    Villin is an actin-binding protein localized in intestinal and kidney brush borders. In vitro, villin has been demonstrated to bundle and sever F-actin in a Ca2+-dependent manner. We generated knockout mice to study the role of villin in vivo. In villin-null mice, no noticeable changes were observed in the ultrastructure of the microvilli or in the localization and expression of the actin-binding and membrane proteins of the intestine. Interestingly, the response to elevated intracellular Ca2+ differed significantly between mutant and normal mice. In wild-type animals, isolated brush borders were disrupted by the addition of Ca2+, whereas Ca2+ had no effect in villin-null isolates. Moreover, increase in intracellular Ca2+ by serosal carbachol or mucosal Ca2+ ionophore A23187 application abolished the F-actin labeling only in the brush border of wild-type animals. This F-actin disruption was also observed in physiological fasting/refeeding experiments. Oral administration of dextran sulfate sodium, an agent that causes colonic epithelial injury, induced large mucosal lesions resulting in a higher death probability in mice lacking villin, 36 ± 9.6%, compared with wild-type mice, 70 ± 8.8%, at day 13. These results suggest that in vivo, villin is not necessary for the bundling of F-actin microfilaments, whereas it is necessary for the reorganization elicited by various signals. We postulate that this property might be involved in cellular plasticity related to cell injury. PMID:10459016

  13. Similarity in replication timing between polytene and diploid cells is associated with the organization of the Drosophila genome

    PubMed Central

    Goncharov, Fedor P.; Zhimulev, Igor F.

    2018-01-01

    Morphologically, polytene chromosomes of Drosophila melanogaster consist of compact “black” bands alternating with less compact “grey” bands and interbands. We developed a comprehensive approach that combines cytological mapping data of FlyBase-annotated genes and novel tools for predicting cytogenetic features of chromosomes on the basis of their protein composition and determined the genomic coordinates for all black bands of polytene chromosome 2R. By a PCNA immunostaining assay, we obtained the replication timetable for all the bands mapped. The results allowed us to compare replication timing between polytene chromosomes in salivary glands and chromosomes from cultured diploid cell lines and to observe a substantial similarity in the global replication patterns at the band resolution level. In both kinds of chromosomes, the intervals between black bands correspond to early replication initiation zones. Black bands are depleted of replication initiation events and are characterized by a gradient of replication timing; therefore, the time of replication completion correlates with the band length. The bands are characterized by low gene density, contain predominantly tissue-specific genes, and are represented by silent chromatin types in various tissues. The borders of black bands correspond well to the borders of topological domains as well as to the borders of the zones showing H3K27me3, SUUR, and LAMIN enrichment. In conclusion, the characteristic pattern of polytene chromosomes reflects partitioning of the Drosophila genome into two global types of domains with contrasting properties. This partitioning is conserved in different tissues and determines replication timing in Drosophila. PMID:29659604

  14. Alcohol-related social problems among Mexican Americans living in U.S.-Mexico border and non-border areas.

    PubMed

    Vaeth, Patrice A C; Caetano, Raul; Mills, Britain A; Rodriguez, Lori A

    2012-08-01

    This paper examines alcohol-related social problems among Mexican Americans living along the U.S.-Mexico border and in non-border areas. Interviews were conducted among Mexican Americans in the border regions of California, Arizona, New Mexico, and Texas (N=1307). Non-border respondents were interviewed primarily in Houston and Los Angeles (N=1288) as part of the Hispanic Americans Baseline Alcohol Survey (HABLAS). Both the border and HABLAS surveys employed multistage cluster sample designs (response rates were 67% and 76%, respectively). In the bivariate analysis, there were no significant differences between border and non-border areas in the proportion of those with one or more social problem. In non-border areas, the prevalence of alcohol problems did not differ significantly by age. However, along the border the prevalence of alcohol problems was significantly different across age groups, with 18 to 29year old men and women having the highest prevalence. The final models showed no residence effect on problem likelihood. Drinking was strongly associated with problems. Although young border residents had higher problem prevalence rates than older residents, the logistic regression models showed no effect of border residence on the likelihood of problems, indicating that problems are due to alcohol consumption, not the border environment. The border, however, did appear to influence more drinking among young people. Regardless of residence, alcohol treatment and preventive interventions tailored to Mexican Americans are essential and special attention should be focused on younger individuals near the border. Published by Elsevier Ltd.

  15. Alcohol-related Social Problems among Mexican Americans Living in U.S.-Mexico Border and Non-border Areas

    PubMed Central

    Vaeth, Patrice A.C.; Caetano, Raul; Mills, Britain A.; Rodriguez, Lori A.

    2012-01-01

    This paper examines alcohol-related social problems among Mexican Americans living along the U.S.-Mexico border and in non-border areas. Interviews were conducted among Mexican Americans in the border regions of California, Arizona, New Mexico, and Texas (N=1,307). Non-border respondents were interviewed primarily in Houston and Los Angeles (N=1,288) as part of the Hispanic Americans Baseline Alcohol Survey (HABLAS). Both the border and HABLAS surveys employed multistage cluster sample designs (response rates were 67% and 76%, respectively). In the bivariate analysis, there were no significant differences between border and non-border areas in the proportion of those with one or more social problem. In non-border areas, the prevalence of alcohol problems did not differ significantly by age. However, along the border the prevalence of alcohol problems was significantly different across age groups, with 18 to 29 year old men and women having the highest prevalence. The final models showed no residence effect on problem likelihood. Drinking was strongly associated with problems. Although young border residents had higher problem prevalence rates than older residents, the logistic regression models showed no effect of border residence on the likelihood of problems, indicating that problems are due to alcohol consumption, not the border environment. The border, however, did appear to influence more drinking among young people. Regardless of residence, alcohol treatment and preventive interventions tailored to Mexican Americans are essential and special attention should be focused on younger individuals near the border. PMID:22564755

  16. Repair of Traumatic Skeletal Muscle Injury with Bone-Marrow-Derived Mesenchymal Stem Cells Seeded on Extracellular Matrix

    DTIC Science & Technology

    2010-06-02

    fully restored muscle fibers and blood vessels is not known; however, FIG. 4. von Willebrand factor (vWF). Masson’s Trichrome stain of sections of...dystrophic skeletal muscle is able to partially restore expression of dystrophin within the fibers .30,35,49,50 Conflict exists as to whether or not the...significantly higher number of fibers expressed in regions closer to the border with native muscle tissue indicate that engraftment of cells was not the main

  17. Evolution of vertebrates: a view from the crest

    PubMed Central

    Bronner, Marianne E.

    2016-01-01

    The origin of vertebrates was accompanied by the advent of a novel cell type: the neural crest. Emerging from the central nervous system, these cells migrate to diverse locations and differentiate into numerous derivatives. By coupling morphological and gene regulatory information from vertebrates and other chordates, we describe how addition of the neural crest specification program may have enabled cells at the neural plate border to acquire multipotency and migratory ability. Analyzing the topology of the neural crest gene regulatory network can serve as a useful template for understanding vertebrate evolution, including elaboration of neural crest derivatives. PMID:25903629

  18. Continuous flow microfluidic separation and processing of rare cells and bioparticles found in blood - A review.

    PubMed

    Antfolk, Maria; Laurell, Thomas

    2017-05-01

    Rare cells in blood, such as circulating tumor cells or fetal cells in the maternal circulation, posses a great prognostic or diagnostic value, or for the development of personalized medicine, where the study of rare cells could provide information to more specifically targeted treatments. When conventional cell separation methods, such as flow cytometry or magnetic activated cell sorting, have fallen short other methods are desperately sought for. Microfluidics have been extensively used towards isolating and processing rare cells as it offers possibilities not present in the conventional systems. Furthermore, microfluidic methods offer new possibilities for cell separation as they often rely on non-traditional biomarkers and intrinsic cell properties. This offers the possibility to isolate cell populations that would otherwise not be targeted using conventional methods. Here, we provide an extensive review of the latest advances in continuous flow microfluidic rare cell separation and processing with each cell's specific characteristics and separation challenges as a point of view. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. High-throughput, low-loss, low-cost, and label-free cell separation using electrophysiology-activated cell enrichment.

    PubMed

    Faraghat, Shabnam A; Hoettges, Kai F; Steinbach, Max K; van der Veen, Daan R; Brackenbury, William J; Henslee, Erin A; Labeed, Fatima H; Hughes, Michael P

    2017-05-02

    Currently, cell separation occurs almost exclusively by density gradient methods and by fluorescence- and magnetic-activated cell sorting (FACS/MACS). These variously suffer from lack of specificity, high cell loss, use of labels, and high capital/operating cost. We present a dielectrophoresis (DEP)-based cell-separation method, using 3D electrodes on a low-cost disposable chip; one cell type is allowed to pass through the chip whereas the other is retained and subsequently recovered. The method advances usability and throughput of DEP separation by orders of magnitude in throughput, efficiency, purity, recovery (cells arriving in the correct output fraction), cell losses (those which are unaccounted for at the end of the separation), and cost. The system was evaluated using three example separations: live and dead yeast; human cancer cells/red blood cells; and rodent fibroblasts/red blood cells. A single-pass protocol can enrich cells with cell recovery of up to 91.3% at over 300,000 cells per second with >3% cell loss. A two-pass protocol can process 300,000,000 cells in under 30 min, with cell recovery of up to 96.4% and cell losses below 5%, an effective processing rate >160,000 cells per second. A three-step protocol is shown to be effective for removal of 99.1% of RBCs spiked with 1% cancer cells while maintaining a processing rate of ∼170,000 cells per second. Furthermore, the self-contained and low-cost nature of the separator device means that it has potential application in low-contamination applications such as cell therapies, where good manufacturing practice compatibility is of paramount importance.

  20. Separator development and testing of nickel-hydrogen cells

    NASA Technical Reports Server (NTRS)

    Gonzalez-Sanabria, O. D.; Manzo, M. A.

    1984-01-01

    The components, design, and operating characteristics of Ni-H2 cells batteries were improved. A separator development program was designed to develop a separator that is resistant to penetration by oxygen and loose active material from then nickel electrode, while retraining the required chemical and thermal stability, reservoir capability, and high ionic conductivity. The performance of the separators in terms of cell operating voltage was to at least match that of state-of-the-art separators while eliminating the separator problems. The separators were submitted to initial screening tests and those which successfully completed the tests were built into Ni-H2 cells for short term testing. The separators with the best performance are tested for long term performance and life.

  1. Method for forming a cell separator for use in bipolar-stack energy storage devices

    DOEpatents

    Mayer, Steven T.; Feikert, John H.; Kaschmitter, James L.; Pekala, Richard W.

    1994-01-01

    An improved multi-cell electrochemical energy storage device, such as a battery, fuel cell, or double layer capacitor using a cell separator which allows cells to be stacked and interconnected with low electrical resistance and high reliability while maximizing packaging efficiency. By adding repeating cells, higher voltages can be obtained. The cell separator is formed by applying an organic adhesive on opposing surfaces of adjacent carbon electrodes or surfaces of aerogel electrodes of a pair of adjacent cells prior to or after pyrolysis thereof to form carbon aerogel electrodes. The cell separator is electronically conductive, but ionically isolating, preventing an electrolytic conduction path between adjacent cells in the stack.

  2. Cell separator for use in bipolar-stack energy storage devices

    DOEpatents

    Mayer, S.T.; Feikert, J.H.; Kachmitter, J.L.; Pekala, R.W.

    1995-02-28

    An improved multi-cell electrochemical energy storage device is described, such as a battery, fuel cell, or double layer capacitor using a cell separator which allows cells to be stacked and interconnected with low electrical resistance and high reliability while maximizing packaging efficiency. By adding repeating cells, higher voltages can be obtained. The cell separator is formed by applying an organic adhesive on opposing surfaces of adjacent carbon electrodes or surfaces of aerogel electrodes of a pair of adjacent cells prior to or after pyrolysis thereof to form carbon aerogel electrodes. The cell separator is electronically conductive, but ionically isolating, preventing an electrolytic conduction path between adjacent cells in the stack. 2 figs.

  3. Method for forming a cell separator for use in bipolar-stack energy storage devices

    DOEpatents

    Mayer, S.T.; Feikert, J.H.; Kaschmitter, J.L.; Pekala, R.W.

    1994-08-09

    An improved multi-cell electrochemical energy storage device, such as a battery, fuel cell, or double layer capacitor using a cell separator which allows cells to be stacked and interconnected with low electrical resistance and high reliability while maximizing packaging efficiency. By adding repeating cells, higher voltages can be obtained. The cell separator is formed by applying an organic adhesive on opposing surfaces of adjacent carbon electrodes or surfaces of aerogel electrodes of a pair of adjacent cells prior to or after pyrolysis thereof to form carbon aerogel electrodes. The cell separator is electronically conductive, but ionically isolating, preventing an electrolytic conduction path between adjacent cells in the stack. 2 figs.

  4. Separation of rare oligodendrocyte progenitor cells from brain using a high-throughput multilayer thermoplastic-based microfluidic device.

    PubMed

    Didar, Tohid Fatanat; Li, Kebin; Veres, Teodor; Tabrizian, Maryam

    2013-07-01

    Despite the advances made in the field of regenerative medicine, the progress in cutting-edge technologies for separating target therapeutic cells are still at early stage of development. These cells are often rare, such as stem cells or progenitor cells that their overall properties should be maintained during the separation process for their subsequent application in regenerative medicine. This work, presents separation of oligodendrocyte progenitor cells (OPCs) from rat brain primary cultures using an integrated thermoplastic elastomeric (TPE)- based multilayer microfluidic device fabricated using hot-embossing technology. OPCs are frequently used in recovery, repair and regeneration of central nervous system after injuries. Indeed, their ability to differentiate in vitro into myelinating oligodendrocytes, are extremely important for myelin repair. OPCs form 5-10% of the glial cells population. The traditional macroscale techniques for OPCs separation require pre-processing of cells and/or multiple time consuming steps with low efficiency leading very often to alteration of their properties. The proposed methodology implies to separate OPCs based on their smaller size compared to other cells from the brain tissue mixture. Using aforementioned microfluidic chip embedded with a 5 μm membrane pore size and micropumping system, a separation efficiency more than 99% was achieved. This microchip was able to operate at flow rates up to 100 μl/min, capable of separating OPCs from a confluent 75 cm(2) cell culture flask in less than 10 min, which provides us with a high-throughput and highly efficient separation expected from any cell sorting techniques. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Rapid cell separation with minimal manipulation for autologous cell therapies

    NASA Astrophysics Data System (ADS)

    Smith, Alban J.; O'Rorke, Richard D.; Kale, Akshay; Rimsa, Roberts; Tomlinson, Matthew J.; Kirkham, Jennifer; Davies, A. Giles; Wälti, Christoph; Wood, Christopher D.

    2017-02-01

    The ability to isolate specific, viable cell populations from mixed ensembles with minimal manipulation and within intra-operative time would provide significant advantages for autologous, cell-based therapies in regenerative medicine. Current cell-enrichment technologies are either slow, lack specificity and/or require labelling. Thus a rapid, label-free separation technology that does not affect cell functionality, viability or phenotype is highly desirable. Here, we demonstrate separation of viable from non-viable human stromal cells using remote dielectrophoresis, in which an electric field is coupled into a microfluidic channel using shear-horizontal surface acoustic waves, producing an array of virtual electrodes within the channel. This allows high-throughput dielectrophoretic cell separation in high conductivity, physiological-like fluids, overcoming the limitations of conventional dielectrophoresis. We demonstrate viable/non-viable separation efficacy of >98% in pre-purified mesenchymal stromal cells, extracted from human dental pulp, with no adverse effects on cell viability, or on their subsequent osteogenic capabilities.

  6. The Implications for Yugoslav Borders of the Dissolution of the Yugoslav State

    DTIC Science & Technology

    1992-03-31

    region: While Romance speaking groups, closely related to the Romanians , live across the border, Romania has to deal with the discontent of ill- 20...closely related to the Romanians , live across the border, Romania has to deal with the discontent of ill- treated minorities at home. Serbia is... Romanian Border 19 V. The Bulakrian Bordei- 21 1,. Tho P,,vlgarian - Serbian Border 22 2. The ulgarian - Macedonian Border 23 VI,: The Greek Border 26

  7. HLA-targeted flow cytometric sorting of blood cells allows separation of pure and viable microchimeric cell populations.

    PubMed

    Drabbels, Jos J M; van de Keur, Carin; Kemps, Berit M; Mulder, Arend; Scherjon, Sicco A; Claas, Frans H J; Eikmans, Michael

    2011-11-10

    Microchimerism is defined by the presence of low levels of nonhost cells in a person. We developed a reliable method for separating viable microchimeric cells from the host environment. For flow cytometric cell sorting, HLA antigens were targeted with human monoclonal HLA antibodies (mAbs). Optimal separation of microchimeric cells (present at a proportion as low as 0.01% in artificial mixtures) was obtained with 2 different HLA mAbs, one targeting the chimeric cells and the other the background cells. To verify purity of separated cell populations, flow-sorted fractions of 1000 cells were processed for DNA analysis by HLA-allele-specific and Y-chromosome-directed real-time quantitative PCR assays. After sorting, PCR signals of chimeric DNA markers in the positive fractions were significantly enhanced compared with those in the presort samples, and they were similar to those in 100% chimeric control samples. Next, we demonstrate applicability of HLA-targeted FACS sorting after pregnancy by separating chimeric maternal cells from child umbilical cord mononuclear cells. Targeting allelic differences with anti-HLA mAbs with FACS sorting allows maximal enrichment of viable microchimeric cells from a background cell population. The current methodology enables reliable microchimeric cell detection and separation in clinical specimens.

  8. Effects of protease, phytase and a Bacillus sp. direct-fed microbial on nutrient and energy digestibility, ileal brush border digestive enzyme activity and cecal short-chain fatty acid concentration in broiler chickens.

    PubMed

    Murugesan, Ganapathi R; Romero, Luis F; Persia, Michael E

    2014-01-01

    Two experiments were conducted to determine the effects of protease and phytase (PP) and a Bacillus sp. direct-fed microbial (DFM) on dietary energy and nutrient utilization in broiler chickens. In the first experiment, Ross 308 broiler chicks were fed diets supplemented with PP and DFM in a 2×2 factorial arrangement. The 4 diets (control (CON), CON + PP, CON + DFM, and CON + PP + DFM) were fed from 15-21 days of age. In Experiment 1, significant interaction (P≤0.01) between PP and DFM on the apparent ileal digestibility coefficient for starch, crude protein, and amino acid indicated that both additives increased the digestibility. Both additives increased the nitrogen retention coefficient with a significant interaction (P≤0.01). Although no interaction was observed, significant main effects (P≤0.01) for nitrogen-corrected apparent ME (AMEn) for PP or DFM indicated an additive response. In a follow-up experiment, Ross 308 broiler chicks were fed the same experimental diets from 1-21 days of age. Activities of ileal brush border maltase, sucrase, and L-alanine aminopeptidase were increased (P≤0.01) by PP addition, while a trend (P = 0.07) for increased sucrase activity was observed in chickens fed DFM, in Experiment 2. The proportion of cecal butyrate was increased (P≤0.01) by DFM addition. Increased nutrient utilization and nitrogen retention appear to involve separate but complementary mechanisms for PP and DFM, however AMEn responses appear to have separate and additive mechanisms.

  9. Automated Microfluidic Instrument for Label-Free and High-Throughput Cell Separation.

    PubMed

    Zhang, Xinjie; Zhu, Zhixian; Xiang, Nan; Long, Feifei; Ni, Zhonghua

    2018-03-20

    Microfluidic technologies for cell separation were reported frequently in recent years. However, a compact microfluidic instrument enabling thoroughly automated cell separation is still rarely reported until today due to the difficult hybrid between the macrosized fluidic control system and the microsized microfluidic device. In this work, we propose a novel and automated microfluidic instrument to realize size-based separation of cancer cells in a label-free and high-throughput manner. Briefly, the instrument is equipped with a fully integrated microfluidic device and a set of robust fluid-driven and control units, and the instrument functions of precise fluid infusion and high-throughput cell separation are guaranteed by a flow regulatory chip and two cell separation chips which are the key components of the microfluidic device. With optimized control programs, the instrument is successfully applied to automatically sort human breast adenocarcinoma cell line MCF-7 from 5 mL of diluted human blood with a high recovery ratio of ∼85% within a rapid processing time of ∼23 min. We envision that our microfluidic instrument will be potentially useful in many biomedical applications, especially cell separation, enrichment, and concentration for the purpose of cell culture and analysis.

  10. Thinner, More-Efficient Oxygen-Separation Cells

    NASA Technical Reports Server (NTRS)

    Clark, Douglas J.; Galica, Leo M.; Losey, Robert W.

    1992-01-01

    Better gas-distribution plates fabricated more easily. Oxygen-separation cell redesigned to make it more efficient, smaller, lighter, and easier to manufacture. Potential applications include use as gas separators, filters, and fuel cells.

  11. Effects of Vegetated Field Borders on Arthropods in Cotton Fields in Eastern North Carolina

    PubMed Central

    Outward, Randy; Sorenson, Clyde E.; Bradley, J. R.

    2008-01-01

    The influence, if any, of 5m wide, feral, herbaceous field borders on pest and beneficial arthropods in commercial cotton, Gossypium hirsutum (L.) (Malvales: Malvaceae), fields was measured through a variety of sampling techniques over three years. In each year, 5 fields with managed, feral vegetation borders and five fields without such borders were examined. Sampling was stratified from the field border or edge in each field in an attempt to elucidate any edge effects that might have occurred. Early season thrips populations appeared to be unaffected by the presence of a border. Pitfall sampling disclosed no differences in ground-dwelling predaceous arthropods but did detect increased populations of crickets around fields with borders. Cotton aphid (Aphis gossypii Glover) (Hemiptera: Aphididae) populations were too low during the study to adequately assess border effects. Heliothines, Heliothis virescens (F.) and Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), egg numbers and damage rates were largely unaffected by the presence or absence of a border, although in one instance egg numbers were significantly lower in fields with borders. Overall, foliage-dwelling predaceous arthropods were somewhat more abundant in fields with borders than in fields without borders. Tarnished plant bugs, Lygus lineolaris (Palisot de Beauvois) (Heteroptera: Miridae) were significantly more abundant in fields with borders, but stink bugs, Acrosternum hilare (Say), and Euschistus servus (Say) (Hemiptera: Pentatomidae) numbers appeared to be largely unaffected by border treatment. Few taxa clearly exhibited distributional edge effects relative to the presence or absence of border vegetation. Field borders like those examined in this study likely will have little impact on insect pest management in cotton under current insect management regimens. PMID:20345293

  12. A 3D cell culture system: separation distance between INS-1 cell and endothelial cell monolayers co-cultured in fibrin influences INS-1 cells insulin secretion.

    PubMed

    Sabra, Georges; Vermette, Patrick

    2013-02-01

    The aim of this study was to develop an in vitro cell culture system allowing studying the effect of separation distance between monolayers of rat insulinoma cells (INS-1) and human umbilical vein endothelial cells (HUVEC) co-cultured in fibrin over INS-1 cell insulin secretion. For this purpose, a three-dimensional (3D) cell culture chamber was designed, built using micro-fabrication techniques and validated. The co-culture was successfully carried out and the effect on INS-1 cell insulin secretion was investigated. After 48 and 72 h, INS-1 cells co-cultured with HUVEC separated by a distance of 100 µm revealed enhanced insulin secretion compared to INS-1 cells cultured alone or co-cultured with HUVEC monolayers separated by a distance of 200 µm. These results illustrate the importance of the separation distance between two cell niches for cell culture design and the possibility to further enhance the endocrine function of beta cells when this factor is considered. Copyright © 2012 Wiley Periodicals, Inc.

  13. Label-free density difference amplification-based cell sorting.

    PubMed

    Song, Jihwan; Song, Minsun; Kang, Taewook; Kim, Dongchoul; Lee, Luke P

    2014-11-01

    The selective cell separation is a critical step in fundamental life sciences, translational medicine, biotechnology, and energy harvesting. Conventional cell separation methods are fluorescent activated cell sorting and magnetic-activated cell sorting based on fluorescent probes and magnetic particles on cell surfaces. Label-free cell separation methods such as Raman-activated cell sorting, electro-physiologically activated cell sorting, dielectric-activated cell sorting, or inertial microfluidic cell sorting are, however, limited when separating cells of the same kind or cells with similar sizes and dielectric properties, as well as similar electrophysiological phenotypes. Here we report a label-free density difference amplification-based cell sorting (dDACS) without using any external optical, magnetic, electrical forces, or fluidic activations. The conceptual microfluidic design consists of an inlet, hydraulic jump cavity, and multiple outlets. Incoming particles experience gravity, buoyancy, and drag forces in the separation chamber. The height and distance that each particle can reach in the chamber are different and depend on its density, thus allowing for the separation of particles into multiple outlets. The separation behavior of the particles, based on the ratio of the channel heights of the inlet and chamber and Reynolds number has been systematically studied. Numerical simulation reveals that the difference between the heights of only lighter particles with densities close to that of water increases with increasing the ratio of the channel heights, while decreasing Reynolds number can amplify the difference in the heights between the particles considered irrespective of their densities.

  14. Separation of malignant human breast cancer epithelial cells from healthy epithelial cells using an advanced dielectrophoresis-activated cell sorter (DACS).

    PubMed

    An, Jaemin; Lee, Jangwon; Lee, Sang Ho; Park, Jungyul; Kim, Byungkyu

    2009-06-01

    In this paper, we successfully separated malignant human breast cancer epithelial cells (MCF 7) from healthy breast cells (MCF 10A) and analyzed the main parameters that influence the separation efficiency with an advanced dielectrophoresis (DEP)-activated cell sorter (DACS). Using the efficient DACS, the malignant cancer cells (MCF 7) were isolated successfully by noninvasive methods from normal cells with similar cell size distributions (MCF 10A), depending on differences between their material properties such as conductivity and permittivity, because our system was able to discern the subtle differences in the properties by generating continuously changed electrical field gradients. In order to evaluate the separation performance without considering size variations, the cells collected from each outlet were divided into size-dependent groups and counted statistically. Following that, the quantitative relative ratio of numbers between MCF 7 and MCF 10A cells in each size-dependent group separated by the DEP were compared according to applied frequencies in the range 48, 51, and 53 MHz with an applied amplitude of 8 V(pp). Finally, under the applied voltage of 48 MHz-8 V(pp) and a flow rate of 290 microm/s, MCF 7 and MCF 10A cells were separated with a maximum efficiency of 86.67% and 98.73% respectively. Therefore, our suggested system shows it can be used for detection and separation of cancerous epithelial cells from noncancerous cells in clinical applications.

  15. The evaluation of layered separators for nickel-hydrogen cells

    NASA Technical Reports Server (NTRS)

    Gahn, Randall F.

    1991-01-01

    The concept of using layered separators to achieve the required electrolyte retention and bubble pressure fo nickel-hydrogen cells was evaluated in a boilerplate cell test. Zircar cloth, polyethylene paper and polypropylene felt were combined with a layer of radiation-grafted polyethylene film to achieve the required properties. Three cells of each layered separator were built and tested by characterization cycling and by low earth orbit cycling for 5000 cycles at 80 percent DOD. Three cells containing asbestos separators were used as the reference.

  16. Cell separation: Terminology and practical considerations

    PubMed Central

    Tomlinson, Sophie; Yang, Xuebin B; Kirkham, Jennifer

    2013-01-01

    Cell separation is a powerful tool in biological research. Increasing usage, particularly within the tissue engineering and regenerative medicine communities, means that researchers from a diverse range of backgrounds are utilising cell separation technologies. This review aims to offer potential solutions to cell sorting problems and to clarify common ambiguities in terminology and experimental design. The frequently used cell separation terms of ‘purity’, ‘recovery’ and ‘viability’ are discussed, and attempts are made to reach a consensus view of their sometimes ambiguous meanings. The importance of appropriate experimental design is considered, with aspects such as marker expression, tissue isolation and original cell population analysis discussed. Finally, specific technical issues such as cell clustering, dead cell removal and non-specific antibody binding are considered and potential solutions offered. The solutions offered may provide a starting point to improve the quality of cell separations achieved by both the novice and experienced researcher alike. PMID:23440031

  17. Neural crest cells: from developmental biology to clinical interventions.

    PubMed

    Noisa, Parinya; Raivio, Taneli

    2014-09-01

    Neural crest cells are multipotent cells, which are specified in embryonic ectoderm in the border of neural plate and epiderm during early development by interconnection of extrinsic stimuli and intrinsic factors. Neural crest cells are capable of differentiating into various somatic cell types, including melanocytes, craniofacial cartilage and bone, smooth muscle, and peripheral nervous cells, which supports their promise for cell therapy. In this work, we provide a comprehensive review of wide aspects of neural crest cells from their developmental biology to applicability in medical research. We provide a simplified model of neural crest cell development and highlight the key external stimuli and intrinsic regulators that determine the neural crest cell fate. Defects of neural crest cell development leading to several human disorders are also mentioned, with the emphasis of using human induced pluripotent stem cells to model neurocristopathic syndromes. © 2014 Wiley Periodicals, Inc.

  18. Neural Computation of Surface Border Ownership and Relative Surface Depth from Ambiguous Contrast Inputs.

    PubMed

    Dresp-Langley, Birgitta; Grossberg, Stephen

    2016-01-01

    The segregation of image parts into foreground and background is an important aspect of the neural computation of 3D scene perception. To achieve such segregation, the brain needs information about border ownership; that is, the belongingness of a contour to a specific surface represented in the image. This article presents psychophysical data derived from 3D percepts of figure and ground that were generated by presenting 2D images composed of spatially disjoint shapes that pointed inward or outward relative to the continuous boundaries that they induced along their collinear edges. The shapes in some images had the same contrast (black or white) with respect to the background gray. Other images included opposite contrasts along each induced continuous boundary. Psychophysical results demonstrate conditions under which figure-ground judgment probabilities in response to these ambiguous displays are determined by the orientation of contrasts only, not by their relative contrasts, despite the fact that many border ownership cells in cortical area V2 respond to a preferred relative contrast. Studies are also reviewed in which both polarity-specific and polarity-invariant properties obtain. The FACADE and 3D LAMINART models are used to explain these data.

  19. Neural Computation of Surface Border Ownership and Relative Surface Depth from Ambiguous Contrast Inputs

    PubMed Central

    Dresp-Langley, Birgitta; Grossberg, Stephen

    2016-01-01

    The segregation of image parts into foreground and background is an important aspect of the neural computation of 3D scene perception. To achieve such segregation, the brain needs information about border ownership; that is, the belongingness of a contour to a specific surface represented in the image. This article presents psychophysical data derived from 3D percepts of figure and ground that were generated by presenting 2D images composed of spatially disjoint shapes that pointed inward or outward relative to the continuous boundaries that they induced along their collinear edges. The shapes in some images had the same contrast (black or white) with respect to the background gray. Other images included opposite contrasts along each induced continuous boundary. Psychophysical results demonstrate conditions under which figure-ground judgment probabilities in response to these ambiguous displays are determined by the orientation of contrasts only, not by their relative contrasts, despite the fact that many border ownership cells in cortical area V2 respond to a preferred relative contrast. Studies are also reviewed in which both polarity-specific and polarity-invariant properties obtain. The FACADE and 3D LAMINART models are used to explain these data. PMID:27516746

  20. Application of smart phone in "Better Border Healthcare Program": a module for mother and child care.

    PubMed

    Kaewkungwal, Jaranit; Singhasivanon, Pratap; Khamsiriwatchara, Amnat; Sawang, Surasak; Meankaew, Pongthep; Wechsart, Apisit

    2010-11-03

    To assess the application of cell phone integrating into the healthcare system to improve antenatal care (ANC) and expanded programme on immunization (EPI) services for the under-served population in border area. A module combining web-based and mobile technology was developed to generate ANC/EPI visit schedule dates in which the healthcare personnel can cross-check, identify and update the mother's ANC and child's EPI status at the healthcare facility or at the household location when performing home visit; with additional feature of sending appointment reminder directly to the scheduled mother in the community. The module improved ANC/EPI coverage in the study area along the country border including for both Thai and non-Thai mothers and children who were either permanent resident or migrants; numbers of ANC and EPI visit on-time as per schedule significantly increased; there was less delay of antenatal visits and immunizations. The module integrated and functioned successfully as part of the healthcare system; it is proved for its feasibility and the extent to which community healthcare personnel in the low resource setting could efficiently utilize it to perform their duties.

  1. Integrated fuel cell stack shunt current prevention arrangement

    DOEpatents

    Roche, Robert P.; Nowak, Michael P.

    1992-01-01

    A fuel cell stack includes a plurality of fuel cells juxtaposed with one another in the stack and each including a pair of plate-shaped anode and cathode electrodes that face one another, and a quantity of liquid electrolyte present at least between the electrodes. A separator plate is interposed between each two successive electrodes of adjacent ones of the fuel cells and is unified therewith into an integral separator plate. Each integral separator plate is provided with a circumferentially complete barrier that prevents flow of shunt currents onto and on an outer peripheral surface of the separator plate. This barrier consists of electrolyte-nonwettable barrier members that are accommodated, prior to the formation of the integral separator plate, in corresponding edge recesses situated at the interfaces between the electrodes and the separator plate proper. Each barrier member extends over the entire length of the associated marginal portion and is flush with the outer periphery of the integral separator plate. This barrier also prevents cell-to-cell migration of any electrolyte that may be present at the outer periphery of the integral separator plate while the latter is incorporated in the fuel cell stack.

  2. Quantitative Magnetic Separation of Particles and Cells using Gradient Magnetic Ratcheting

    PubMed Central

    Murray, Coleman; Pao, Edward; Tseng, Peter; Aftab, Shayan; Kulkarni, Rajan; Rettig, Matthew; Di Carlo, Dino

    2016-01-01

    Extraction of rare target cells from biosamples is enabling for life science research. Traditional rare cell separation techniques, such as magnetic activated cell sorting (MACS), are robust but perform coarse, qualitative separations based on surface antigen expression. We report a quantitative magnetic separation technology using high-force magnetic ratcheting over arrays of magnetically soft micro-pillars with gradient spacing, and use the system to separate and concentrate magnetic beads based on iron oxide content (IOC) and cells based on surface expression. The system consists of a microchip of permalloy micro-pillar arrays with increasing lateral pitch and a mechatronic device to generate a cycling magnetic-field. Particles with higher IOC separate and equilibrate along the miro-pillar array at larger pitches. We develop a semi-analytical model that predicts behavior for particles and cells. Using the system, LNCaP cells were separated based on the bound quantity of 1μm anti-EpCAM particles as a metric for expression. The ratcheting cytometry system was able to resolve a ±13 bound particle differential, successfully distinguishing LNCaP from PC3 populations based on EpCAM expression, correlating with flow cytometry analysis. As a proof of concept, EpCAM-labeled cells from patient blood were isolated with 74% purity, demonstrating potential towards a quantitative magnetic separation instrument. PMID:26890496

  3. Large silver-cadmium technology program

    NASA Technical Reports Server (NTRS)

    Charlip, S.; Lerner, S.

    1971-01-01

    The effects of varying cell design on operation factors on the electrochemical performance of sealed, silver-cadmium cells were determined. A factorial experiment was conducted for all test cells constructed with organic separators. Three operating factors were evaluated: temperature, depth of discharge, and charge rate. The six construction factors considered were separator, absorber, electrolyte quantity, cadmium electrode type, cadmium-to-silver ratio, and auxiliary electrode. Test cells of 4 ampere-hour capacity were fabricated and cycled. The best performing cells, on a 94 minute orbit, at 40% depth of discharge, were those containing silver-treated fibrous sausage casings as the separator, and Teflon-ated, pressed cadmium electrodes. Cycling data of cells with inorganic separators (Astroset) are given. Best performance was shown by cells with nonwoven nylon absorbers. Rigid inorganic separators provided the best barrier to silver migration.

  4. Improved Separators For Rechargeable Lithium Cells

    NASA Technical Reports Server (NTRS)

    Shen, David; Surampudi, Subbarao; Huang, Chen-Kuo; Halpert, Gerald

    1994-01-01

    Improved pairs of separators proposed for use in rechargeable lithium cells operating at ambient temperature. Block growth of lithium dendrites and help prevent short circuits. Each cell contains one separator made of microporous polypropylene placed next to anode, and one separator made of microporous polytetrafluoroethylene (PTFE) next to cathode. Separators increase cycle lives of secondary lithium cells. Cells to which concept applicable those of Li/TiS(2), Li/NbSe(3), Li/CoO(2), Li/MoS(2), Li/VO(x), and Li/MnO(2) chemical systems. Advantageous in spacecraft, military, communications, automotive, and other applications in which high energy density and rechargeability needed.

  5. Intracellular trafficking pathways of Cx43 gap junction channels.

    PubMed

    Epifantseva, Irina; Shaw, Robin M

    2018-01-01

    Gap Junction (GJ) channels, including the most common Connexin 43 (Cx43), have fundamental roles in excitable tissues by facilitating rapid transmission of action potentials between adjacent cells. For instance, synchronization during each heartbeat is regulated by these ion channels at the cardiomyocyte cell-cell border. Cx43 protein has a short half-life, and rapid synthesis and timely delivery of those proteins to particular subdomains are crucial for the cellular organization of gap junctions and maintenance of intracellular coupling. Impairment in gap junction trafficking contributes to dangerous complications in diseased hearts such as the arrhythmias of sudden cardiac death. Of recent interest are the protein-protein interactions with the Cx43 carboxy-terminus. These interactions have significant impact on the full length Cx43 lifecycle and also contribute to trafficking of Cx43 as well as possibly other functions. We are learning that many of the known non-canonical roles of Cx43 can be attributed to the recently identified six endogenous Cx43 truncated isoforms which are produced by internal translation. In general, alternative translation is a new leading edge for proteome expansion and therapeutic drug development. This review highlights recent mechanisms identified in the trafficking of gap junction channels, involvement of other proteins contributing to the delivery of channels to the cell-cell border, and understanding of possible roles of the newly discovered alternatively translated isoforms in Cx43 biology. This article is part of a Special Issue entitled: Gap Junction Proteins edited by Jean Claude Herve. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Fuel-Cell Water Separator

    NASA Technical Reports Server (NTRS)

    Burke, Kenneth Alan; Fisher, Caleb; Newman, Paul

    2010-01-01

    The main product of a typical fuel cell is water, and many fuel-cell configurations use the flow of excess gases (i.e., gases not consumed by the reaction) to drive the resultant water out of the cell. This two-phase mixture then exits through an exhaust port where the two fluids must again be separated to prevent the fuel cell from flooding and to facilitate the reutilization of both fluids. The Glenn Research Center (GRC) has designed, built, and tested an innovative fuel-cell water separator that not only removes liquid water from a fuel cell s exhaust ports, but does so with no moving parts or other power-consuming components. Instead it employs the potential and kinetic energies already present in the moving exhaust flow. In addition, the geometry of the separator is explicitly intended to be integrated into a fuel-cell stack, providing a direct mate with the fuel cell s existing flow ports. The separator is also fully scalable, allowing it to accommodate a wide range of water removal requirements. Multiple separators can simply be "stacked" in series or parallel to adapt to the water production/removal rate. GRC s separator accomplishes the task of water removal by coupling a high aspect- ratio flow chamber with a highly hydrophilic, polyethersulfone membrane. The hydrophilic membrane readily absorbs and transports the liquid water away from the mixture while simultaneously resisting gas penetration. The expansive flow path maximizes the interaction of the water particles with the membrane while minimizing the overall gas flow restriction. In essence, each fluid takes its corresponding path of least resistance, and the two fluids are effectively separated. The GRC fuel-cell water separator has a broad range of applications, including commercial hydrogen-air fuel cells currently being considered for power generation in automobiles.

  7. Quantitative Magnetic Separation of Particles and Cells Using Gradient Magnetic Ratcheting.

    PubMed

    Murray, Coleman; Pao, Edward; Tseng, Peter; Aftab, Shayan; Kulkarni, Rajan; Rettig, Matthew; Di Carlo, Dino

    2016-04-13

    Extraction of rare target cells from biosamples is enabling for life science research. Traditional rare cell separation techniques, such as magnetic activated cell sorting, are robust but perform coarse, qualitative separations based on surface antigen expression. A quantitative magnetic separation technology is reported using high-force magnetic ratcheting over arrays of magnetically soft micropillars with gradient spacing, and the system is used to separate and concentrate magnetic beads based on iron oxide content (IOC) and cells based on surface expression. The system consists of a microchip of permalloy micropillar arrays with increasing lateral pitch and a mechatronic device to generate a cycling magnetic field. Particles with higher IOC separate and equilibrate along the miropillar array at larger pitches. A semi-analytical model is developed that predicts behavior for particles and cells. Using the system, LNCaP cells are separated based on the bound quantity of 1 μm anti-epithelial cell adhesion molecule (EpCAM) particles as a metric for expression. The ratcheting cytometry system is able to resolve a ±13 bound particle differential, successfully distinguishing LNCaP from PC3 populations based on EpCAM expression, correlating with flow cytometry analysis. As a proof-of-concept, EpCAM-labeled cells from patient blood are isolated with 74% purity, demonstrating potential toward a quantitative magnetic separation instrument. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Effects of ozone and peroxone on algal separation via dispersed air flotation.

    PubMed

    Nguyen, Truc Linh; Lee, D J; Chang, J S; Liu, J C

    2013-05-01

    Effects of pre-oxidation on algal separation by dispersed air flotation were examined. Ozone (O3) and peroxone (O3 and H2O2) could induce cell lysis, release of intracellular organic matter (IOM), and mineralization of organic substances. Separation efficiency of algal cells improved when pre-oxidized. Total of 76.4% algal cells was separated at 40 mg/L of N-cetyl-N-N-N-trimethylammonium bromide (CTAB), while 95% were separated after 30-min ozonation. Pre-oxidation by ozone and peroxone also enhanced flotation separation efficiency of dissolved organic carbon (DOC), polysaccharide, and protein, in which peroxone process exerted more significantly than O3. Two main mechanisms were involved in flotation separation of unoxidized algal suspension, namely hydrophobic cell surface and cell flocculation resulting from CTAB adsorption. However, flocculation by CTAB was hindered for pre-oxidized algal suspensions. It implied that the compositional changes in extracellular organic matter (EOM) by pre-oxidation were more determined for flotation separation of pre-oxidized cells. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Fuel cell system with separating structure bonded to electrolyte

    DOEpatents

    Bourgeois, Richard Scott; Gudlavalleti, Sauri; Quek, Shu Ching; Hasz, Wayne Charles; Powers, James Daniel

    2010-09-28

    A fuel cell assembly comprises a separating structure configured for separating a first reactant and a second reactant wherein the separating structure has an opening therein. The fuel cell assembly further comprises a fuel cell comprising a first electrode, a second electrode, and an electrolyte interposed between the first and second electrodes, and a passage configured to introduce the second reactant to the second electrode. The electrolyte is bonded to the separating structure with the first electrode being situated within the opening, and the second electrode being situated within the passage.

  10. Outer brain barriers in rat and human development

    PubMed Central

    Brøchner, Christian B.; Holst, Camilla B.; Møllgård, Kjeld

    2015-01-01

    Complex barriers at the brain's surface, particularly in development, are poorly defined. In the adult, arachnoid blood-cerebrospinal fluid (CSF) barrier separates the fenestrated dural vessels from the CSF by means of a cell layer joined by tight junctions. Outer CSF-brain barrier provides diffusion restriction between brain and subarachnoid CSF through an initial radial glial end feet layer covered with a pial surface layer. To further characterize these interfaces we examined embryonic rat brains from E10 to P0 and forebrains from human embryos and fetuses (6–21st weeks post-conception) and adults using immunohistochemistry and confocal microscopy. Antibodies against claudin-11, BLBP, collagen 1, SSEA-4, MAP2, YKL-40, and its receptor IL-13Rα2 and EAAT1 were used to describe morphological characteristics and functional aspects of the outer brain barriers. Claudin-11 was a reliable marker of the arachnoid blood-CSF barrier. Collagen 1 delineated the subarachnoid space and stained pial surface layer. BLBP defined radial glial end feet layer and SSEA-4 and YKL-40 were present in both leptomeningeal cells and end feet layer, which transformed into glial limitans. IL-13Rα2 and EAAT1 were present in the end feet layer illustrating transporter/receptor presence in the outer CSF-brain barrier. MAP2 immunostaining in adult brain outlined the lower border of glia limitans; remnants of end feet were YKL-40 positive in some areas. We propose that outer brain barriers are composed of at least 3 interfaces: blood-CSF barrier across arachnoid barrier cell layer, blood-CSF barrier across pial microvessels, and outer CSF-brain barrier comprising glial end feet layer/pial surface layer. PMID:25852456

  11. Outer brain barriers in rat and human development.

    PubMed

    Brøchner, Christian B; Holst, Camilla B; Møllgård, Kjeld

    2015-01-01

    Complex barriers at the brain's surface, particularly in development, are poorly defined. In the adult, arachnoid blood-cerebrospinal fluid (CSF) barrier separates the fenestrated dural vessels from the CSF by means of a cell layer joined by tight junctions. Outer CSF-brain barrier provides diffusion restriction between brain and subarachnoid CSF through an initial radial glial end feet layer covered with a pial surface layer. To further characterize these interfaces we examined embryonic rat brains from E10 to P0 and forebrains from human embryos and fetuses (6-21st weeks post-conception) and adults using immunohistochemistry and confocal microscopy. Antibodies against claudin-11, BLBP, collagen 1, SSEA-4, MAP2, YKL-40, and its receptor IL-13Rα2 and EAAT1 were used to describe morphological characteristics and functional aspects of the outer brain barriers. Claudin-11 was a reliable marker of the arachnoid blood-CSF barrier. Collagen 1 delineated the subarachnoid space and stained pial surface layer. BLBP defined radial glial end feet layer and SSEA-4 and YKL-40 were present in both leptomeningeal cells and end feet layer, which transformed into glial limitans. IL-13Rα2 and EAAT1 were present in the end feet layer illustrating transporter/receptor presence in the outer CSF-brain barrier. MAP2 immunostaining in adult brain outlined the lower border of glia limitans; remnants of end feet were YKL-40 positive in some areas. We propose that outer brain barriers are composed of at least 3 interfaces: blood-CSF barrier across arachnoid barrier cell layer, blood-CSF barrier across pial microvessels, and outer CSF-brain barrier comprising glial end feet layer/pial surface layer.

  12. Dynamics of the Glycophorin A Dimer in Membranes of Native-Like Composition Uncovered by Coarse-Grained Molecular Dynamics Simulations.

    PubMed

    Flinner, Nadine; Schleiff, Enrico

    2015-01-01

    Membranes are central for cells as borders to the environment or intracellular organelle definition. They are composed of and harbor different molecules like various lipid species and sterols, and they are generally crowded with proteins. The membrane system is very dynamic and components show lateral, rotational and translational diffusion. The consequence of the latter is that phase separation can occur in membranes in vivo and in vitro. It was documented that molecular dynamics simulations of an idealized plasma membrane model result in formation of membrane areas where either saturated lipids and cholesterol (liquid-ordered character, Lo) or unsaturated lipids (liquid-disordered character, Ld) were enriched. Furthermore, current discussions favor the idea that proteins are sorted into the liquid-disordered phase of model membranes, but experimental support for the behavior of isolated proteins in native membranes is sparse. To gain insight into the protein behavior we built a model of the red blood cell membrane with integrated glycophorin A dimer. The sorting and the dynamics of the dimer were subsequently explored by coarse-grained molecular dynamics simulations. In addition, we inspected the impact of lipid head groups and the presence of cholesterol within the membrane on the dynamics of the dimer within the membrane. We observed that cholesterol is important for the formation of membrane areas with Lo and Ld character. Moreover, it is an important factor for the reproduction of the dynamic behavior of the protein found in its native environment. The protein dimer was exclusively sorted into the domain of Ld character in the model red blood cell plasma membrane. Therefore, we present structural information on the glycophorin A dimer distribution in the plasma membrane in the absence of other factors like e.g. lipid anchors in a coarse grain resolution.

  13. Stripe-patterned thermo-responsive cell culture dish for cell separation without cell labeling.

    PubMed

    Kumashiro, Yoshikazu; Ishihara, Jun; Umemoto, Terumasa; Itoga, Kazuyoshi; Kobayashi, Jun; Shimizu, Tatsuya; Yamato, Masayuki; Okano, Teruo

    2015-02-11

    A stripe-patterned thermo-responsive surface is prepared to enable cell separation without labeling. The thermo-responsive surface containing a 3 μm striped pattern exhibits various cell adhesion and detachment properties. A mixture of three cell types is separated on the patterned surface based on their distinct cell-adhesion properties, and the composition of the cells is analyzed by flow cytometry. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. High-throughput separation of cells by dielectrophoresis enhanced with 3D gradient AC electric field.

    PubMed

    Tada, Shigeru; Hayashi, Masako; Eguchi, Masanori; Tsukamoto, Akira

    2017-11-01

    We propose a novel, high-performance dielectrophoretic (DEP) cell-separation flow chamber with a parallel-plate channel geometry. The flow chamber, consisting of a planar electrode on the top and an interdigitated-pair electrode array at the bottom, was developed to facilitate the separation of cells by creating a nonuniform AC electric field throughout the volume of the flow chamber. The operation and performance of the device were evaluated using live and dead human epithermal breast (MCF10A) cells. The separation dynamics of the cell suspension in the flow chamber was also investigated by numerically simulating the trajectories of individual cells. A theoretical model to describe the dynamic cell behavior under the action of DEP, including dipole-dipole interparticle, viscous, and gravitational forces, was developed. The results demonstrated that the live cells traveling through the flow chamber congregated into sites where the electric field gradient was minimal, in the middle of the flow stream slightly above the centerlines of the grounded electrodes at the bottom. Meanwhile, the dead cells were trapped on the edges of the high-voltage electrodes at the bottom. Cells were thus successfully separated with a remarkably high separation ratio (∼98%) at the appropriately tuned field frequency and applied voltage. The numerically predicted behavior and spatial distribution of the cells during separation also showed good agreement with those observed experimentally.

  15. The lid wiper and muco-cutaneous junction anatomy of the human eyelid margins: an in vivo confocal and histological study

    PubMed Central

    Knop, Erich; Knop, Nadja; Zhivov, Andrey; Kraak, Robert; Korb, Donald R; Blackie, Caroline; Greiner, Jack V; Guthoff, Rudolf

    2011-01-01

    The inner border of the eyelid margin is critically important for ocular surface integrity because it guarantees the thin spread of the tear film. Its exact morphology in the human is still insufficiently known. The histology in serial sections of upper and lower lid margins in whole-mount specimens from 10 human body donors was compared to in vivo confocal microscopy of eight eyes with a Heidelberg retina-tomograph (HRT II) and attached Rostock cornea module. Behind the posterior margin of the Meibomian orifices, the cornified epidermis stopped abruptly and was replaced by a continuous layer of para-keratinized (pk) cells followed by discontinuous pk cells. The pk cells covered the muco-cutaneous junction (MCJ), the surface of which corresponded to the line of Marx (0.2–0.3 mm wide). Then a stratified epithelium with a conjunctival structure of cuboidal cells, some pk cells, and goblet cells formed an epithelial elevation of typically about 100 μm initial thickness (lid wiper). This continued for 0.3–1.5 mm and formed a slope. The MCJ and lid wiper extended all along the lid margin from nasal to temporal positions in the upper and lower lids. Details of the epithelium and connective tissue were also detectable using the Rostock cornea module. The human inner lid border has distinct zones. Due to its location and morphology, the epithelial lip of the lid wiper appears a suitable structure to spread the tear film and is distinct from the MCJ/line of Marx. Better knowledge of the lid margin appears important for understanding dry eye disease and its morphology can be analysed clinically by in vivo confocal microscopy. PMID:21413985

  16. Induction of a reversible cardiac lipidosis by a dietary long-chain fatty acid (erucic acid). Relationship to lipid accumulation in border zones of myocardial infarcts.

    PubMed Central

    Chien, K. R.; Bellary, A.; Nicar, M.; Mukherjee, A.; Buja, L. M.

    1983-01-01

    Previous studies have demonstrated that cardiac myocytes in the border zone of acute myocardial infarction become markedly overloaded with neutral lipid during the transition from reversible to irreversible injury. To examine directly the role of these changes in neutral lipid metabolism in the development of irreversible cellular injury and associated increases in tissue Ca2+ content, the authors fed rats large amounts of a fatty acid (erucic acid) that is poorly oxidized by the heart and that subsequently accumulates as neutral lipid. Rats fed a high erucic acid (C22:1) diet in the form of 20% rapeseed oil for 3-5 days had a fourfold increase in triglyceride (49.5 +/- 3.8 SEM mg/g wet wt versus 13.6 +/- 13, n = 4) and a 60% increase in long-chain acyl CoA content (166.0 +/- 21.9 versus 91.5 +/- 9.0 nM/g wet wt, n = 4), compared with controls. However, there was no change in long-chain acyl carnitine or total phospholipid content. Histochemical studies showed accumulation of numerous lipid droplets in the myocytes, and electron microscopy revealed localization of lipid vesicles in direct contact with mitochondria, thus mimicking the lipid-laden cells in the border zone regions of acute myocardial infarcts. The acute lipidosis was reversible with either continued feeding of erucic acid for several weeks or conversion to a normal diet. It was not associated with an increased tissue Ca2+ content, nor with cell necrosis. However, continued erucic acid intake for 3 months was associated with focal myocardial degeneration and loss of myocytes. These results suggest that acute increases in neutral lipids, as found in the border zone of acute myocardial infarction, may not be the cause of progression to irreversible damage during acute myocardial injury, but that the persistent presence of similar lipid material over months may result in focal myocardial degeneration. Images Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 PMID:6859230

  17. --No Title--

    Science.gov Websites

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  18. Histological and Lectin Histochemical Studies on the Olfactory and Respiratory Mucosae of the Sheep

    PubMed Central

    IBRAHIM, Dalia; NAKAMUTA, Nobuaki; TANIGUCHI, Kazumi; YAMAMOTO, Yoshio; TANIGUCHI, Kazuyuki

    2013-01-01

    ABSTRACT The olfactory and respiratory mucosae of the Corriedale sheep were examined using lectin histochemistry in order to clarify the histochemical and glycohistochemical differences between these two tissues. The olfactory epithelium was stained with 13 lectins out of 21 lectins examined, while the respiratory epithelium was positive to 16 lectins. The free border of both of the olfactory and respiratory epithelia was stained with 12 lectins: Wheat germ agglutinin (WGA), succinylated-wheat germ agglutinin (s-WGA), Lycopersicon esculentum lectin (LEL), Solanum tuberosum lectin (STL), Datura stramonium lectin (DSL), Soybean agglutinin (SBA), Bandeiraea simplicifolia lectin-I (BSL-I), Ricinus communis agglutinin-I (RCA-120), Erythrina cristagalli lectin (ECL), Concanavalin A (Con A), Phaseolus vulgaris agglutinin-E (PHA-E) and Phaseolus vulgaris agglutinin-L (PHA-L). The associated glands of the olfactory mucosa, Bowman’s glands, were stained with 13 lectins. While both the goblet cells and mucous nasal glands were stained with 8 lectins; five of them (WGA, s-WGA, STL, Vicia villosa agglutinin (VVA) and ECL) were mutually positive among the Bowman’s glands, mucous nasal glands and the goblet cells. These findings indicate that the glycohistochemical characteristics of the free borders of both olfactory and respiratory epithelia are similar to each other, suggesting that secretions from the Bowman’s glands and those of the goblet cells and mucous nasal glands are partially exchanged between the surface of two epithelia to contribute the functions of the respiratory epithelium and the olfactory receptor cells, respectively. PMID:24200894

  19. Electrophoretic separation and analysis of living cells from solid tissues by several methods - Human embryonic kidney cell cultures as a model

    NASA Technical Reports Server (NTRS)

    Todd, Paul; Plank, Lindsay D.; Kunze, M. Elaine; Lewis, Marian L.; Morrison, Dennis R.

    1986-01-01

    The use of free-fluid electrophoresis methods to separate tissue cells having a specific function is discussed. It is shown that cells suspended by trypsinization from cultures of human embryonic kidney are electrophoretically heterogeneous and tolerate a wide range of electrophoresis buffers and conditions without significant attenuation of function. Moreover, these cells do not separate electrophoretically on the basis of size or cell position alone and can be separated according to their ability to give rise to progeny that produce specific plasminogen activators.

  20. Separation of cells from the rat anterior pituitary gland

    NASA Technical Reports Server (NTRS)

    Hymer, W. C.; Hatfield, J. Michael

    1984-01-01

    Data concerned with analyzing the cellular organization of the rat anterior pituitary gland are examined. The preparation of the cell suspensions and the methods used to separate pituitary cell types are described. Particular emphasis is given to velocity sedimentation at unit gravity, density gradient centrifugation, affinity methods, fluorescence activated cell sorting, and density gradient and continuous-flow electrophoresis. The difficulties encountered when attempting to compare data from different pituitary cell separation studies are discussed, and results from various experiments are presented. The functional capabilities of the separated cell populations can be tested in various culture systems.

  1. A feed-forward spiking model of shape-coding by IT cells

    PubMed Central

    Romeo, August; Supèr, Hans

    2014-01-01

    The ability to recognize a shape is linked to figure-ground (FG) organization. Cell preferences appear to be correlated across contrast-polarity reversals and mirror reversals of polygon displays, but not so much across FG reversals. Here we present a network structure which explains both shape-coding by simulated IT cells and suppression of responses to FG reversed stimuli. In our model FG segregation is achieved before shape discrimination, which is itself evidenced by the difference in spiking onsets of a pair of output cells. The studied example also includes feature extraction and illustrates a classification of binary images depending on the dominance of vertical or horizontal borders. PMID:24904494

  2. A Binational Overview of Reproductive Health Outcomes Among US Hispanic and Mexican Women in the Border Region

    PubMed Central

    Mojarro, Octavio; Sutton, Paul D.; Ventura, Stephanie J.

    2013-01-01

    Introduction The US–Mexico border region has 15 million residents and 300,000 births annually. Reproductive health concerns have been identified on both sides of the border, but comparable information about reproductive health is not available. The objective of this study was to compare reproductive health indicators among populations in this region. Methods We used 2009 US Hispanic and Mexican birth certificate data to compare births inside the border region, elsewhere within the border states, and in the United States and Mexico overall. We examined trends in total fertility and birth rates using birth data from 2000 through 2009 and intercensal population estimates. Results Among women in the border region, US women had more lifetime births than Mexican women in 2009 (2.69 births vs 2.15 births) and throughout the decade. Birth rates in the group aged 15 to 19 years were high in both the US (73.8/1,000) and Mexican (86.7/1,000) border regions. Late or no prenatal care was nearly twice as prevalent in the border regions as in the nonborder regions of border states. Low birth weight and preterm and early-term birth were more prevalent in the US border than in the Mexican border region; US border rates were higher and Mexican rates were lower than their corresponding nonborder and national rates. We found some variations within border states. Conclusion These findings constitute the first population-based information on the reproductive health of the entire Hispanic US–Mexico border population. Evidence of disparities warrants exploration at state and local levels. Teen pregnancy and inadequate prenatal care are shared problems in US–Mexico border communities and suggest an area for binational cooperation. PMID:23948338

  3. A binational overview of reproductive health outcomes among US Hispanic and Mexican women in the border region.

    PubMed

    McDonald, Jill A; Mojarro, Octavio; Sutton, Paul D; Ventura, Stephanie J

    2013-08-15

    The US-Mexico border region has 15 million residents and 300,000 births annually. Reproductive health concerns have been identified on both sides of the border, but comparable information about reproductive health is not available. The objective of this study was to compare reproductive health indicators among populations in this region. We used 2009 US Hispanic and Mexican birth certificate data to compare births inside the border region, elsewhere within the border states, and in the United States and Mexico overall. We examined trends in total fertility and birth rates using birth data from 2000 through 2009 and intercensal population estimates. Among women in the border region, US women had more lifetime births than Mexican women in 2009 (2.69 births vs 2.15 births) and throughout the decade. Birth rates in the group aged 15 to 19 years were high in both the US (73.8/1,000) and Mexican (86.7/1,000) border regions. Late or no prenatal care was nearly twice as prevalent in the border regions as in the nonborder regions of border states. Low birth weight and preterm and early-term birth were more prevalent in the US border than in the Mexican border region; US border rates were higher and Mexican rates were lower than their corresponding nonborder and national rates. We found some variations within border states. These findings constitute the first population-based information on the reproductive health of the entire Hispanic US-Mexico border population. Evidence of disparities warrants exploration at state and local levels. Teen pregnancy and inadequate prenatal care are shared problems in US-Mexico border communities and suggest an area for binational cooperation.

  4. Incremental evolution of the neural crest, neural crest cells and neural crest-derived skeletal tissues

    PubMed Central

    Hall, Brian K; Gillis, J Andrew

    2013-01-01

    Urochordates (ascidians) have recently supplanted cephalochordates (amphioxus) as the extant sister taxon of vertebrates. Given that urochordates possess migratory cells that have been classified as ‘neural crest-like’– and that cephalochordates lack such cells – this phylogenetic hypothesis may have significant implications with respect to the origin of the neural crest and neural crest-derived skeletal tissues in vertebrates. We present an overview of the genes and gene regulatory network associated with specification of the neural crest in vertebrates. We then use these molecular data – alongside cell behaviour, cell fate and embryonic context – to assess putative antecedents (latent homologues) of the neural crest or neural crest cells in ascidians and cephalochordates. Ascidian migratory mesenchymal cells – non-pigment-forming trunk lateral line cells and pigment-forming ‘neural crest-like cells’ (NCLC) – are unlikely latent neural crest cell homologues. Rather, Snail-expressing cells at the neural plate of border of urochordates and cephalochordates likely represent the extent of neural crest elaboration in non-vertebrate chordates. We also review evidence for the evolutionary origin of two neural crest-derived skeletal tissues – cartilage and dentine. Dentine is a bona fide vertebrate novelty, and dentine-secreting odontoblasts represent a cell type that is exclusively derived from the neural crest. Cartilage, on the other hand, likely has a much deeper origin within the Metazoa. The mesodermally derived cellular cartilages of some protostome invertebrates are much more similar to vertebrate cartilage than is the acellular ‘cartilage-like’ tissue in cephalochordate pharyngeal arches. Cartilage, therefore, is not a vertebrate novelty, and a well-developed chondrogenic program was most likely co-opted from mesoderm to the neural crest along the vertebrate stem. We conclude that the neural crest is a vertebrate novelty, but that neural crest cells and their derivatives evolved and diversified in a step-wise fashion – first by elaboration of neural plate border cells, then by the innovation or co-option of new or ancient metazoan cell fates. PMID:22414251

  5. Optimization of yield in magnetic cell separations using nickel nanowires of different lengths.

    PubMed

    Hultgren, Anne; Tanase, Monica; Felton, Edward J; Bhadriraju, Kiran; Salem, Aliasger K; Chen, Christopher S; Reich, Daniel H

    2005-01-01

    Ferromagnetic nanowires are shown to perform both high yield and high purity single-step cell separations on cultures of NIH-3T3 mouse fibroblast cells. The nanowires are made by electrochemical deposition in nanoporous templates, permitting detailed control of their chemical and physical properties. When added to fibroblast cell cultures, the nanowires are internalized by the cells via the integrin-mediated adhesion pathway. The effectiveness of magnetic cell separations using Ni nanowires 350 nm in diameter and 5-35 micrometers long in field gradients of 40 T/m was compared to commercially available superparamagnetic beads. The percent yield of the separated populations is found to be optimized when the length of the nanowire is matched to the diameter of the cells in the culture. Magnetic cell separations performed under these conditions achieve 80% purity and 85% yield, a 4-fold increase over the beads. This effect is shown to be robust when the diameter of the cell is changed within the same cell line using mitomycin-C.

  6. Free flow electrophoresis in space shuttle program (biotex)

    NASA Astrophysics Data System (ADS)

    Hannig, Kurt; Bauer, Johann

    In the space shuttle program free flow electrophoresis will be applied for separation of proteins, biopolymers and cells. Proteins are to be separated according to the ``Feldsprung-Gradienten'' procedure by Prof. H. Wagner, University of Saarbruecken, biopolymers are to be separated by the isotachophoresis technique by Prof. Schmitz, University of Muenster and we intend to separate cells in order to increase the efficiency of recovery of hybrid cells after electrofusion performed under microgravity in collaboration with Prof. U. Zimmermann, University of Wuerzburg. There are supposed two ways for reaching this goal: Enrichment of cells before electrofusion may enhance the probability that the cells of interest are immortalized. Separation of cells after electrofusion may help to clone the hybrid cells of interest. Under microgravity, the combination of improved electrophoresis with higher electrofusion rates may provide new possibilities for immortalization of cells. This may be a new way to obtain cellular products, which are physiologically glycosylated.

  7. Identification of column edges of DNA fragments by using K-means clustering and mean algorithm on lane histograms of DNA agarose gel electrophoresis images

    NASA Astrophysics Data System (ADS)

    Turan, Muhammed K.; Sehirli, Eftal; Elen, Abdullah; Karas, Ismail R.

    2015-07-01

    Gel electrophoresis (GE) is one of the most used method to separate DNA, RNA, protein molecules according to size, weight and quantity parameters in many areas such as genetics, molecular biology, biochemistry, microbiology. The main way to separate each molecule is to find borders of each molecule fragment. This paper presents a software application that show columns edges of DNA fragments in 3 steps. In the first step the application obtains lane histograms of agarose gel electrophoresis images by doing projection based on x-axis. In the second step, it utilizes k-means clustering algorithm to classify point values of lane histogram such as left side values, right side values and undesired values. In the third step, column edges of DNA fragments is shown by using mean algorithm and mathematical processes to separate DNA fragments from the background in a fully automated way. In addition to this, the application presents locations of DNA fragments and how many DNA fragments exist on images captured by a scientific camera.

  8. Toward a United States Arctic research policy

    NASA Astrophysics Data System (ADS)

    Roederer, Juan G.

    Of all countries bordering on the Arctic, the United States is the only one without a national institute, laboratory, or any other organization devoted to the sustained planning and support of Arctic research. Up to now, the responsibility for planning, implementing, and funding Arctic research has been divided between several federal agencies, the state of Alaska, and private groups whose mandates or objectives are often unconnected.The result of this pluralistic approach to U.S. science in the Arctic is that basic research has been conducted in piecemeal fashion. Individual studies are proposed and supported separately, and their costly logistic requirements must be funded in competition with research carried out under less-demanding environmental conditions in the rest of the country. Fundamental data-gathering and interpretation of information has been the responsibility of public agencies whose missions are separate and whose budgets may not reflect the priorities of Arctic issues.

  9. Continuous cell introduction and rapid dynamic lysis for high-throughput single-cell analysis on microfludic chips with hydrodynamic focusing.

    PubMed

    Xu, Chun-Xiu; Yin, Xue-Feng

    2011-02-04

    A chip-based microfluidic system for high-throughput single-cell analysis is described. The system was integrated with continuous introduction of individual cells, rapid dynamic lysis, capillary electrophoretic (CE) separation and laser induced fluorescence (LIF) detection. A cross microfluidic chip with one sheath-flow channel located on each side of the sampling channel was designed. The labeled cells were hydrodynamically focused by sheath-flow streams and sequentially introduced into the cross section of the microchip under hydrostatic pressure generated by adjusting liquid levels in the reservoirs. Combined with the electric field applied on the separation channel, the aligned cells were driven into the separation channel and rapidly lysed within 33ms at the entry of the separation channel by Triton X-100 added in the sheath-flow solution. The maximum rate for introducing individual cells into the separation channel was about 150cells/min. The introduction of sheath-flow streams also significantly reduced the concentration of phosphate-buffered saline (PBS) injected into the separation channel along with single cells, thus reducing Joule heating during electrophoretic separation. The performance of this microfluidic system was evaluated by analysis of reduced glutathione (GSH) and reactive oxygen species (ROS) in single erythrocytes. A throughput of 38cells/min was obtained. The proposed method is simple and robust for high-throughput single-cell analysis, allowing for analysis of cell population with considerable size to generate results with statistical significance. Copyright © 2010 Elsevier B.V. All rights reserved.

  10. Separation technologies for stem cell bioprocessing.

    PubMed

    Diogo, Maria Margarida; da Silva, Cláudia Lobato; Cabral, Joaquim M S

    2012-11-01

    Stem cells have been the focus of an intense research due to their potential in Regenerative Medicine, drug discovery, toxicology studies, as well as for fundamental studies on developmental biology and human disease mechanisms. To fully accomplish this potential, the successful application of separation processes for the isolation and purification of stem cells and stem cell-derived cells is a crucial issue. Although separation methods have been used over the past decades for the isolation and enrichment of hematopoietic stem/progenitor cells for transplantation in hemato-oncological settings, recent achievements in the stem cell field have created new challenges including the need for novel scalable separation processes with a higher resolution and more cost-effective. Important examples are the need for high-resolution methods for the separation of heterogeneous populations of multipotent adult stem cells to study their differential biological features and clinical utility, as well as for the depletion of tumorigenic cells after pluripotent stem cell differentiation. Focusing on these challenges, this review presents a critical assessment of separation processes that have been used in the stem cell field, as well as their current and potential applications. The techniques are grouped according to the fundamental principles that govern cell separation, which are defined by the main physical, biophysical, and affinity properties of cells. A special emphasis is given to novel and promising approaches such as affinity-based methods that take advantage of the use of new ligands (e.g., aptamers, lectins), as well as to novel biophysical-based methods requiring no cell labeling and integrated with microscale technologies. Copyright © 2012 Wiley Periodicals, Inc.

  11. Cross-border policy effects on alcohol outcomes: drinking without thinking on the u.s.-Mexico border?

    PubMed

    Mills, Britain A; Caetano, Raul; Vaeth, Patrice

    2014-11-01

    Rates of alcohol-related outcomes are sensitive to policy differences in politically distinct, adjacent territories. Factors that shape these cross-border effects, particularly when the policy differences are longstanding, remain poorly understood. We compared the ability of 2 classes of variables with theoretical relevance to the U.S.-Mexico border context-bar attendance and alcohol-related social-cognitive variables-to explain elevated drinking on the U.S. side of the border relative to other areas of the United States. Data were collected from multistage cluster samples of adult Mexican Americans on and off the U.S.-Mexico border (current drinker N = 1,351). Structural equation models were used to test drinking context (frequency of bar attendance) and 6 different social-cognitive variables (including alcohol-related attitudes, norms, motives, and beliefs) as mediators of border effects on a composite drinking index. The border effect on drinking varied by age (with younger adults showing a stronger effect), consistent with previous findings and known risk factors in the region. Contrary to theoretical expectations, 6 different social-cognitive variables-despite relating strongly with drinking-were comparable in border and nonborder areas (within and across age) and played no role in elevated drinking on the border. Conversely, elevated drinking among border youth was mediated by bar attendance. This mediated moderation effect held after adjusting for potential sociodemographic and neighborhood-level confounders. Increased drinking among U.S.-Mexico border youth is explained by patterns of bar attendance, but not by more permissive alcohol-related social-cognitive variables in border areas: Border youth attend bars and drink more than their nonborder counterparts, despite having comparable alcohol-related beliefs, attitudes, norms, and motives for use. Alcohol's heightened availability and visibility on both sides of the border may create opportunities for border youth to drink that otherwise would not be considered. Copyright © 2014 by the Research Society on Alcoholism.

  12. Reprint of: Dream vs. reality: seven case-studies on the desirability and feasibility of cross-border hospital collaboration in Europe.

    PubMed

    Glinos, Irene A; Baeten, Rita

    2015-01-01

    Despite being a niche phenomenon, cross-border health care collaboration receives a lot of attention in the EU and figures visibly on the policy agenda, in particular since the policy process which eventually led to the adoption of Directive 2011/24/EU. One of the underlying assumptions is that cross-border collaboration is desirable, providing justification to both the European Commission and to border-region stakeholders for promoting it. The purpose of this paper is to question this assumption and to examine the role of actors in pushing (or not) for cross-border collaboration. The analysis takes place in two parts. First, the EU policies to promote cross-border collaboration and the tools employed are examined, namely (a) use of European funds to sponsor concrete border-region collaboration projects, (b) use of European funds to sponsor research which gives visibility to cross-border collaboration, and (c) use of the European Commission's newly acquired legal mandate to encourage "Member States to cooperate in cross-border health care provision in border-regions" (Art. 10) and support "Member States in the development of European reference networks between health care providers and centres of expertise" (Art. 12). Second, evidence gathered in 2011-2013 from seven European border-regions on hospital cross-border collaboration is systematically reviewed to assess the reality of cross-border collaboration - can it work and when, and why do actors engage in cross-border collaboration? The preliminary findings suggest that while the EU plays a prominent role in some border-region initiatives, cross-border collaboration needs such a specific set of circumstances to work that it is questionable whether it can effectively be promoted. Moreover, local actors make use of the EU (as a source of funding, legislation or legitimisation) to serve their needs. Copyright © 2014. Published by Elsevier Ltd.

  13. Dream vs. reality: seven case-studies on the desirability and feasibility of cross-border hospital collaboration in Europe.

    PubMed

    Glinos, Irene A; Baeten, Rita

    2014-09-01

    Despite being a niche phenomenon, cross-border health care collaboration receives a lot of attention in the EU and figures visibly on the policy agenda, in particular since the policy process which eventually led to the adoption of Directive 2011/24/EU. One of the underlying assumptions is that cross-border collaboration is desirable, providing justification to both the European Commission and to border-region stakeholders for promoting it. The purpose of this paper is to question this assumption and to examine the role of actors in pushing (or not) for cross-border collaboration. The analysis takes place in two parts. First, the EU policies to promote cross-border collaboration and the tools employed are examined, namely (a) use of European funds to sponsor concrete border-region collaboration projects, (b) use of European funds to sponsor research which gives visibility to cross-border collaboration, and (c) use of the European Commission's newly acquired legal mandate to encourage "Member States to cooperate in cross-border health care provision in border-regions" (Art. 10) and support "Member States in the development of European reference networks between health care providers and centres of expertise" (Art. 12). Second, evidence gathered in 2011-2013 from seven European border-regions on hospital cross-border collaboration is systematically reviewed to assess the reality of cross-border collaboration - can it work and when, and why do actors engage in cross-border collaboration? The preliminary findings suggest that while the EU plays a prominent role in some border-region initiatives, cross-border collaboration needs such a specific set of circumstances to work that it is questionable whether it can effectively be promoted. Moreover, local actors make use of the EU (as a source of funding, legislation or legitimisation) to serve their needs. Copyright © 2014. Published by Elsevier Ltd.

  14. "Border Sexualities, Border Families in Schools": Queering Education

    ERIC Educational Resources Information Center

    Ryan, John

    2012-01-01

    This essay reviews Maria Pallotta-Chiarolli's (2010) Lambda Award-winning monograph "Border Sexualities, Border Families in Schools", in which queer and mestizage pedagogies frame a groundbreaking and highly accessible exploration of the issues that sexual border dwellers experience. Her particular focus areas are bisexual "sexually fluid"…

  15. Exposure to Engineered Nanomaterial Results in Disruption of Brush Borders in Epithelia Models in vitro

    NASA Astrophysics Data System (ADS)

    Faust, James J.

    Engineered nanoparticles (NP; 10-9 m) have found use in a variety of consumer goods and medical devices because of the unique changes in material properties that occur when synthesized on the nanoscale. Although many definitions for nanoparticle exist, from the perspective of size, nanoparticle is defined as particles with diameters less than 100 nm in any external dimension. Examples of their use include titanium dioxide added as a pigment in products intended to be ingested by humans, silicon dioxide NPs are used in foods as an anticaking agent, and gold or iron oxide NPs can be used as vectors for drug delivery or contrast agents for specialized medical imaging. Although the intended use of these NPs is often to improve human health, it has come to the attention of investigators that NPs can have unintended or even detrimental effects on the organism. This work describes one such unintended effect of NP exposure from the perspective of exposure via the oral route. First, this Dissertation will explain an event referred to as brush border disruption that occurred after nanoparticles interacted with an in vitro model of the human intestinal epithelium. Second, this Dissertation will identify and characterize several consumer goods that were shown to contain titanium dioxide that are intended to be ingested. Third, this Dissertation shows that sedimentation due to gravity does not artifactually result in disruption of brush borders as a consequence of exposure to food grade titanium dioxide in vitro. Finally, this Dissertation will demonstrate that iron oxide nanoparticles elicited similar effects after exposure to an in vitro brush border expressing model of the human placenta. Together, these data suggest that brush border disruption is not an artifact of the material/cell culture model, but instead represents a bona fide biological response as a result of exposure to nanomaterial.

  16. Gravity separation of fat, somatic cells, and bacteria in raw and pasteurized milks.

    PubMed

    Caplan, Z; Melilli, C; Barbano, D M

    2013-04-01

    The objective of experiment 1 was to determine if the extent of gravity separation of milk fat, bacteria, and somatic cells is influenced by the time and temperature of gravity separation or the level of contaminating bacteria present in the raw milk. The objective of experiment 2 was to determine if different temperatures of milk heat treatment affected the gravity separation of milk fat, bacteria, and somatic cells. In raw milk, fat, bacteria, and somatic cells rose to the top of columns during gravity separation. About 50 to 80% of the fat and bacteria were present in the top 8% of the milk after gravity separation of raw milk. Gravity separation for 7h at 12°C or for 22h at 4°C produced equivalent separation of fat, bacteria, and somatic cells. The completeness of gravity separation of fat was influenced by the level of bacteria in the milk before separation. Milk with a high bacterial count had less (about 50 to 55%) gravity separation of fat than milk with low bacteria count (about 80%) in 22h at 4°C. Gravity separation caused fat, bacteria, and somatic cells to rise to the top of columns for raw whole milk and high temperature, short-time pasteurized (72.6°C, 25s) whole milk. Pasteurization at ≥76.9°C for 25s prevented all 3 components from rising, possibly due to denaturation of native bovine immunoglobulins that normally associate with fat, bacteria, and somatic cells during gravity separation. Gravity separation can be used to produce reduced-fat milk with decreased bacterial and somatic cell counts, and may be a critical factor in the history of safe and unique traditional Italian hard cheeses produced from gravity-separated raw milk. A better understanding of the mechanism of this natural process could lead to the development of new nonthermal thermal technology (that does not involve heating the milk to high temperatures) to remove bacteria and spores from milk or other liquids. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  17. Stacking Oxygen-Separation Cells

    NASA Technical Reports Server (NTRS)

    Schroeder, James E.

    1991-01-01

    Simplified configuration and procedure developed for assembly of stacks of solid-electrolyte cells separating oxygen from air electrochemically. Reduces number of components and thus reduces probability of such failures as gas leaks, breakdown of sensitive parts, and electrical open or short circuits. Previous, more complicated version of cell described in "Improved Zirconia Oxygen-Separation Cell" (NPO-16161).

  18. Antibody enhancement of free-flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Cohly, H. H. P.; Morrison, Dennis R.; Atassi, M. Zouhair

    1988-01-01

    Specific T cell clones and antibodies (ABs) were developed to study the efficiency of purifying closely associated T cells using Continuous Flow Electrophoresis System. Enhanced separation is accomplished by tagging cells first with ABs directed against the antigenic determinants on the cell surface and then with ABs against the Fc portion of the first AB. This second AB protrudes sufficiently beyond the cell membrane and glycocalyx to become the major overall cell surface potential determinant and thus causes a reduction of electrophoretic mobility. This project was divided into three phases. Phase one included development of specific T cell clones and separation of these specific clones. Phase two extends these principles to the separation of T cells from spleen cells and immunized lymph node cells. Phase three applies this double antibody technique to the separation of T cytotoxic cells from bone marrow.

  19. Separation of mouse testis cells on a Celsep (TM) apparatus and their usefulness as a source of high molecular weight DNA or RNA

    NASA Technical Reports Server (NTRS)

    Wolgemuth, D. J.; Gizang-Ginsberg, E.; Engelmyer, E.; Gavin, B. J.; Ponzetto, C.

    1985-01-01

    The use of a self-contained unit-gravity cell separation apparatus for separation of populations of mouse testicular cells is described. The apparatus, a Celsep (TM), maximizes the unit area over which sedimentation occurs, reduces the amount of separation medium employed, and is quite reproducible. Cells thus isolated have been good sources for isolation of DNA, and notably, high molecular weight RNA.

  20. 78 FR 73875 - Agency Information Collection Activities: Canadian Border Boat Landing Permit (CBP Form I-68)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-09

    ... Activities: Canadian Border Boat Landing Permit (CBP Form I-68) AGENCY: U.S. Customs and Border Protection... information collection requirement concerning the Canadian Border Boat Landing Permit (Form I-68). This... soliciting comments concerning the following information collection: Title: Canadian Border Boat Landing...

  1. Border Malaria Associated with Multidrug Resistance on Thailand-Myanmar and Thailand-Cambodia Borders: Transmission Dynamic, Vulnerability, and Surveillance

    PubMed Central

    Bhumiratana, Adisak; Intarapuk, Apiradee; Sorosjinda-Nunthawarasilp, Prapa; Maneekan, Pannamas; Koyadun, Surachart

    2013-01-01

    This systematic review elaborates the concepts and impacts of border malaria, particularly on the emergence and spread of Plasmodium falciparum and Plasmodium vivax multidrug resistance (MDR) malaria on Thailand-Myanmar and Thailand-Cambodia borders. Border malaria encompasses any complex epidemiological settings of forest-related and forest fringe-related malaria, both regularly occurring in certain transmission areas and manifesting a trend of increased incidence in transmission prone areas along these borders, as the result of interconnections of human settlements and movement activities, cross-border population migrations, ecological changes, vector population dynamics, and multidrug resistance. For regional and global perspectives, this review analyzes and synthesizes the rationales pertaining to transmission dynamics and the vulnerabilities of border malaria that constrain surveillance and control of the world's most MDR falciparum and vivax malaria on these chaotic borders. PMID:23865048

  2. CO-OCCURRENCE OF ALCOHOL, DRUG USE, DSM-5 ALCOHOL USE DISORDER AND SYMPTOMS OF DRUG USE DISORDER ON BOTH SIDES OF THE U.S.-MEXICO BORDER

    PubMed Central

    Borges, Guilherme; Zemore, Sarah; Orozco, Ricardo; Cherpitel, Cheryl J.; Ye, Yu; Bond, Jason; Maxwell, Jane Carlisle; Wallisch, Lynn

    2015-01-01

    Background The U.S.-Mexico border displays elevated rates of hazardous alcohol and drug use. Whether the co-occurrence of alcohol and drug use and disorders is also high in the border area is unknown. Methods Data are from the U.S.-Mexico Study on Alcohol and Related Conditions, a cross-sectional survey of randomly selected respondents interviewed from 2011–2013. Participants included 1,690 Mexican Americans from Texas (572 in an off-border city and 1,118 from 3 border cities) and 1,293 Mexicans from Nuevo Leon and Tamaulipas (415 in an off-border city and 878 from 3 Mexican cities bordering Texas) who reported drinking in the last 12 months. Participants were interviewed regarding the prevalence of and risk factors for: a) co-occurring hazardous alcohol use (5+/4+ at least monthly) and drug use (medical and illicit), and b) co-occurring presence of a DSM-5 alcohol use disorder (AUD) and 2 symptoms (hazardous use and quit/control) of drug use disorders (DUD symptoms). Results Co-occurring hazardous alcohol and drug use was more common in the U.S. border cities (14.7%) than off-border (7.2%), but similar for Mexican border (1.2%) and off-border (1.4%) cities. Co-occurrence of AUD and DUD symptoms was likewise more common at the U.S. border (6.8%) than off-border (3.3%), as well as at the Mexican border (1.3%), compared to off-border (0.6%), but not statistically significant for Mexico. In models adjusting for demographics, mobility factors and exposure to the U.S. culture, border residence in both countries related to a nearly two-fold increase in prevalence ratios (PR) of co-occurring AUD and DUD symptoms (PR=1.97, 95%CI=1.36–2.85). Conclusions Increased rates of co-occurring alcohol and drug use disorders suggest an added negative impact on already difficult conditions of the border population. PMID:25833029

  3. Optical cell separation from three-dimensional environment in photodegradable hydrogels for pure culture techniques.

    PubMed

    Tamura, Masato; Yanagawa, Fumiki; Sugiura, Shinji; Takagi, Toshiyuki; Sumaru, Kimio; Matsui, Hirofumi; Kanamori, Toshiyuki

    2014-05-07

    Cell sorting is an essential and efficient experimental tool for the isolation and characterization of target cells. A three-dimensional environment is crucial in determining cell behavior and cell fate in biological analysis. Herein, we have applied photodegradable hydrogels to optical cell separation from a 3D environment using a computer-controlled light irradiation system. The hydrogel is composed of photocleavable tetra-arm polyethylene glycol and gelatin, which optimized cytocompatibility to adjust a composition of crosslinker and gelatin. Local light irradiation could degrade the hydrogel corresponding to the micropattern image designed on a laptop; minimum resolution of photodegradation was estimated at 20 µm. Light irradiation separated an encapsulated fluorescent microbead without any contamination of neighbor beads, even at multiple targets. Upon selective separation of target cells in the hydrogels, the separated cells have grown on another dish, resulting in pure culture. Cell encapsulation, light irradiation and degradation products exhibited negligible cytotoxicity in overall process.

  4. Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells.

    PubMed

    Sanchez-Antequera, Yolanda; Mykhaylyk, Olga; van Til, Niek P; Cengizeroglu, Arzu; de Jong, J Henk; Huston, Marshall W; Anton, Martina; Johnston, Ian C D; Pojda, Zygmunt; Wagemaker, Gerard; Plank, Christian

    2011-04-21

    Research applications and cell therapies involving genetically modified cells require reliable, standardized, and cost-effective methods for cell manipulation. We report a novel nanomagnetic method for integrated cell separation and gene delivery. Gene vectors associated with magnetic nanoparticles are used to transfect/transduce target cells while being passaged and separated through a high gradient magnetic field cell separation column. The integrated method yields excellent target cell purity and recovery. Nonviral and lentiviral magselectofection is efficient and highly specific for the target cell population as demonstrated with a K562/Jurkat T-cell mixture. Both mouse and human enriched hematopoietic stem cell pools were effectively transduced by lentiviral magselectofection, which did not affect the hematopoietic progenitor cell number determined by in vitro colony assays. Highly effective reconstitution of T and B lymphocytes was achieved by magselectofected murine wild-type lineage-negative Sca-1(+) cells transplanted into Il2rg(-/-) mice, stably expressing GFP in erythroid, myeloid, T-, and B-cell lineages. Furthermore, nonviral, lentiviral, and adenoviral magselectofection yielded high transfection/transduction efficiency in human umbilical cord mesenchymal stem cells and was fully compatible with their differentiation potential. Upscaling to a clinically approved automated cell separation device was feasible. Hence, once optimized, validated, and approved, the method may greatly facilitate the generation of genetically engineered cells for cell therapies.

  5. Evaluation program for secondary spacecraft cells: Initial evaluation tests of Eagle-Picher Industries, Incorporated 6.0 ampere-hour, nickel-cadmium spacecraft cells for separator material evaluation

    NASA Technical Reports Server (NTRS)

    Harkness, J. D.

    1975-01-01

    Several groups of nickel cadmium cells were tested for the durability of their separator materials. The cells were rated at 6.0 ampere-hours, and contained double ceramic seals. Two cells in each group were fitted with pressure gauge assemblies. Results are presented for various brands of separator materials.

  6. Method for separating biological cells. [suspended in aqueous polymer systems

    NASA Technical Reports Server (NTRS)

    Brooks, D. E. (Inventor)

    1980-01-01

    A method for separating biological cells by suspending a mixed cell population in a two-phase polymer system is described. The polymer system consists of droplet phases with different surface potentials for which the cell populations exhibit different affinities. The system is subjected to an electrostatic field of sufficient intensity to cause migration of the droplets with an attendant separation of cells.

  7. Anatomical Evidence for Classical and Extra-classical Receptive Field Completion Across the Discontinuous Horizontal Meridian Representation of Primate Area V2

    PubMed Central

    Jeffs, Janelle; Ichida, Jennifer M.; Federer, Frederick

    2009-01-01

    In primates, a split of the horizontal meridian (HM) representation at the V2 rostral border divides this area into dorsal (V2d) and ventral (V2v) halves (representing lower and upper visual quadrants, respectively), causing retinotopically neighboring loci across the HM to be distant within V2. How is perceptual continuity maintained across this discontinuous HM representation? Injections of neuroanatomical tracers in marmoset V2d demonstrated that cells near the V2d rostral border can maintain retinotopic continuity within their classical and extra-classical receptive field (RF), by making both local and long-range intra- and interareal connections with ventral cortex representing the upper visual quadrant. V2d neurons located <0.9–1.3 mm from the V2d rostral border, whose RFs presumably do not cross the HM, make nonretinotopic horizontal connections with V2v neurons in the supra- and infragranular layers. V2d neurons located <0.6–0.9 mm from the border, whose RFs presumably cross the HM, in addition make retinotopic local connections with V2v neurons in layer 4. V2d neurons also make interareal connections with upper visual field regions of extrastriate cortex, but not of MT or MTc outside the foveal representation. Labeled connections in ventral cortex appear to represent the “missing” portion of the connectional fields in V2d across the HM. We conclude that connections between dorsal and ventral cortex can create visual field continuity within a second-order discontinuous visual topography. PMID:18755777

  8. Pore size engineering applied to the design of separators for nickel-hydrogen cells and batteries

    NASA Technical Reports Server (NTRS)

    Abbey, K. M.; Britton, D. L.

    1983-01-01

    Pore size engineering in starved alkaline multiplate cells involves adopting techniques to widen the volume tolerance of individual cells. Separators with appropriate pore size distributions and wettability characteristics (capillary pressure considerations) to have wider volume tolerances and an ability to resist dimensional changes in the electrodes were designed. The separators studied for potential use in nickel-hydrogen cells consist of polymeric membranes as well as inorganic microporous mats. In addition to standard measurements, the resistance and distribution of electrolyte as a function of total cell electrolyte content were determined. New composite separators consisting of fibers, particles and/or binders deposited on Zircar cloth were developed in order to engineer the proper capillary pressure characteristics in the separator. These asymmetric separators were prepared from a variety of fibers, particles and binders.

  9. [The perichromatin compartment of the cell nucleus].

    PubMed

    Bogoliubov, D S

    2014-01-01

    In this review, the data on the structure and composition of the perichromatin compartment, a special border area between the condensed chromatin and the interchromatin space of the cell nucleus, are discussed in the light of the concept of nuclear functions in complex nuclear architectonics. Morphological features, molecular composition and functions of main extrachromosomal structures of the perichromatin compartment, perichromatin fibrils (PFs) and perichromatin granules (PGs) including nuclear stress-bodies (nSBs) that are derivates of the PGs under heat shock, are presented. A special attention was paid to the features of the molecular compositions of PFs and PGs in different cell types and at different physiological conditions.

  10. In vitro motility of cells from human epidermoid carcinomas. A study by phase-contrast and reflection-contrast cinematography.

    PubMed

    Haemmerli, G; Sträuli, P

    1981-05-15

    The motile behavior of six cell lines derived from human squamous carcinomas (two from the larynx, four from the tongue) was studied by cinematography under phase- and reflection-contrast illumination. The recorded cell activities consist in spreading, stationary and translocation motility, and aggregate formation. Within this common pattern, quantitative modifications ("sub-pattern") are stable properties of the individual cells lines. Such modifications are particularly evident with regard to the dynamic texture of the aggregates which ranges from loose, netlike structures to compact islands with smooth borders. Accordingly, the intensity of cell traffic within and around the aggregates varies considerably. It is discussed to what extent the in vitro motility of the carcinoma cell populations reflects their behavior in the organism and thus the significance of cell movements for invasion.

  11. Separating biological cells

    NASA Technical Reports Server (NTRS)

    Brooks, D. E.

    1979-01-01

    Technique utilizing electric field to promote biological cell separation from suspending medium in zero gravity increases speed, reduces sedimentation, and improves efficiency of separation in normal gravity.

  12. Type of monocyte immunomagnetic separation affects the morphology of monocyte-derived dendritic cells, as investigated by scanning electron microscopy.

    PubMed

    Kowalewicz-Kulbat, M; Ograczyk, E; Krawczyk, K; Rudnicka, W; Fol, M

    2016-12-01

    Dendritic cells (DCs) are increasingly being used for multiple applications and are useful tools for many immunotherapeutic strategies. The understanding of the possible impact of the DCs-generation methods on the biological capacities of these cells is therefore essential. Although the immunomagnetic separation is regarded as a fast and accurate method yielding cells with the high purity and efficiency, still little is known about its impact on the properties of the generated DCs. The aim of this study was to compare the morphology of the monocyte derived dendritic cells (MoDCs), generated from monocytes selected with anti-CD14 mAbs (positive separation) and treated with anti-CD3, -CD7, -CD16, -CD19, -CD56, -CD123, glycophorin A (negative separation), using laser scanning microscopy. We found that the type of the immunomagnetic separation method used strongly influences the shape and cell dimension of the MoDCs. We observed that the height of both immature and LPS-matured DCs generated from monocytes isolated by negative separation was significantly higher compared to the cells obtained by positive separation. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Prognostic factors in breast phyllodes tumors: a nomogram based on a retrospective cohort study of 404 patients.

    PubMed

    Zhou, Zhi-Rui; Wang, Chen-Chen; Sun, Xiang-Jie; Yang, Zhao-Zhi; Chen, Xing-Xing; Shao, Zhi-Ming; Yu, Xiao-Li; Guo, Xiao-Mao

    2018-04-01

    The aim of this study was to explore the independent prognostic factors related to postoperative recurrence-free survival (RFS) in patients with breast phyllodes tumors (PTBs). A retrospective analysis was conducted in Fudan University Shanghai Cancer Center. According to histological type, patients with benign PTBs were classified as a low-risk group, while borderline and malignant PTBs were classified as a high-risk group. The Cox regression model was adopted to identify factors affecting postoperative RFS in the two groups, and a nomogram was generated to predict recurrence-free survival at 1, 3, and 5 years. Among the 404 patients, 168 (41.6%) patients had benign PTB, 184 (45.5%) had borderline PTB, and 52 (12.9%) had malignant PTB. Fifty-five patients experienced postoperative local recurrence, including six benign cases, 26 borderline cases, and 22 malignant cases; the three histological types of PTB had local recurrence rates of 3.6%, 14.1%, and 42.3%, respectively. Stromal cell atypia was an independent prognostic factor for RFS in the low-risk group, while the surgical approach and tumor border were independent prognostic factors for RFS in the high-risk group, and patients receiving simple excision with an infiltrative tumor border had a higher recurrence rate. A nomogram developed based on clinicopathologic features and surgical approaches could predict recurrence-free survival at 1, 3, and 5 years. For high-risk patients, this predictive nomogram based on tumor border, tumor residue, mitotic activity, degree of stromal cell hyperplasia, and atypia can be applied for patient counseling and clinical management. The efficacy of adjuvant radiotherapy remains uncertain. © 2018 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

  14. Recharge of valley-fill aquifers in the glaciated northeast from upland runoff

    USGS Publications Warehouse

    Williams, J.H.; Morrissey, D.J.

    1996-01-01

    Channeled and unchanneled runoff from till-covered bedrock uplands is a major source of recharge to valley-fill aquifers in the glaciated northeastern United States. Streamflow measurements and model simulation of average steady-state conditions indicate that upland runoff accounted for more recharge to two valley-fill aquifers in moderately high topographic-relief settings than did direct infiltration of precipitation. Recharge from upland runoff to a modeled valley-fill aquifer in an area of lower relief was significant but less than that from direct infiltration of precipitation. The amount of upland runoff available for recharging valley-fill aquifers in the glaciated Northeast ranges from about 1.5 to 2.5 cubic feet per second per square mile of drainage area that borders the aquifer. Stream losses from tributaries that drain the uplands commonly range from 0.3 to 1.5 cubic feet per second per 1,000 feet of wetted channel where the tributaries cross alluvial fans in the main valleys. Recharge of valley-fill aquifers from channeled runoff was estimated from measured losses and average runoff rates and was represented in aquifer models as specified fluxes or simulated by head-dependent fluxes with streamflow routing in the model cells that represent the tributary streams. Unchanneled upland runoff, which includes overland and subsurface flow, recharges the valley-fill aquifers at the contact between the aquifer and uplands near the base of the bordering till-covered hillslopes. Recharge from unchanneled runoff was estimated from average runoff rates and the hillslope area that borders the aquifer and was represented as specified fluxes to model-boundary cells along the valley walls.

  15. Isolation of a virulent Newcastle disease virus from confiscated LaSota vaccine.

    PubMed

    Pedersen, Janice C; Hines, Nichole L; Killian, Mary Lea; Predgen, Ann S; Schmitt, Beverly J

    2013-06-01

    Vials of Newcastle disease vaccine labeled as LaSota were confiscated by the Arizona Division of Customs and Border Protection officials. Two different avian type 1 paramyxoviruses were isolated from all three vials of vaccine submitted to the National Veterinary Services Laboratories. The LaSota strain of avian paramyxovirus type 1 virus was isolated from all three vials and analyzed by nucleotide sequence analysis. A virulent Newcastle disease virus was also present in all three vials, but in low concentration. The virulence of the Newcastle disease virus was characterized by the intracerebral chicken pathogenicity index chicken inoculation assay but could not be determined by nucleotide sequence analysis from the virus isolated from embryonating chicken eggs. The intracerebral chicken pathogenicity index value for the isolated Newcastle disease virus was 1.55. Strains of Newcastle disease virus with intracerebral pathogenicity indexes significantly above 1.0 have been found to selectively kill many types of cancer cells while not affecting normal nonneoplastic cells and are considered to be a viable option for cancer treatment in humans by alternative medical researchers; however, the treatment is not approved for use in the United States by the Food and Drug Administration. Customs and Border Protection officials have been notified of an increased risk of Newcastle disease virus entering the United States for use as a nonapproved cancer treatment. Illegal importation of Newcastle disease vaccine for vaccination of backyard poultry is also a threat. This case report emphasizes the importance of conducting chicken inoculation for complete virus pathotyping and demonstrates the need for stringent security procedures at U.S. borders to detect known livestock pathogens that may be smuggled in for use in animal agriculture and reasons unrelated to animal agriculture.

  16. Regional Joint Border Commands: A Pathway to Improving Collaboration and Effectiveness for Border Control

    DTIC Science & Technology

    2009-12-01

    Police xii SBI Secure Border Initiative SES Senior Executive Service SSP Secretaria de Seguridad Publica TSA Transportation Security...also been involvement from the Mexican law enforcement agency, Secretaria de Seguridad Publica (SSP) on the southwest border (CBP, 2009; ICE, 2009, p. 5...The U.S. border is comprised of nearly 20,000 miles of a mix of coastline and rugged land borders with Mexico and Canada; this includes parts of

  17. Modeling the U.S. Border Patrol Tucson Sector for the Deployment and Operations of Border Security Forces

    DTIC Science & Technology

    2006-03-01

    Crime_Trends_2005.pdf (accessed August 3, 2005). Consejo de Seguridad Publica , “Programa de Mediano Plazo 2004 - 2009: Seguridad Publica ,” http...international Southwest border. The issue of illegal human smuggling is not new to the United States- Mexico border or to law enforcement agencies...Operation Gatekeeper; Operation Hold the Line; Operation Stonegarden; Tucson Sector; US Border Patrol; US- Mexico border. 16. PRICE CODE 17. SECURITY

  18. Two-dimensional numerical modeling for separation of deformable cells using dielectrophoresis.

    PubMed

    Ye, Ting; Li, Hua; Lam, K Y

    2015-02-01

    In this paper, we numerically explore the possibility of separating two groups of deformable cells, by a very small dielectrophoretic (DEP) microchip with the characteristic length of several cell diameters. A 2D two-fluid model is developed to describe the separation process, where three types of forces are considered, the aggregation force for cell-cell interaction, the deformation force for cell deformation, and the DEP force for cell dielectrophoresis. As a model validation, we calculate the levitation height of a cell subject to DEP force, and compare it with the experimental data. After that, we simulate the separation of two groups of cells with different dielectric properties at high and low frequencies, respectively. The simulation results show that the deformable cells can be separated successfully by a very small DEP microchip, according to not only their different permittivities at the high frequency, but also their different conductivities at the low frequency. In addition, both two groups of cells have a shape deformation from an original shape to a lopsided slipper shape during the separation process. It is found that the cell motion is mainly determined by the DEP force arising from the electric field, causing the cells to deviate from the centerline of microchannel. However, the cell deformation is mainly determined by the deformation force arising from the fluid flow, causing the deviated cells to undergo an asymmetric motion with the deformation of slipper shape. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Wall effects in continuous microfluidic magneto-affinity cell separation.

    PubMed

    Wu, Liqun; Zhang, Yong; Palaniapan, Moorthi; Roy, Partha

    2010-05-01

    Continuous microfluidic magneto-affinity cell separator combines unique microscale flow phenomenon with advantageous nanobead properties, to isolate cells with high specificity. Owing to the comparable size of the cell-bead complexes and the microchannels, the walls of the microchannel exert a strong influence on the separation of cells by this method. We present a theoretical and experimental study that provides a quantitative description of hydrodynamic wall interactions and wall rolling velocity of cells. A transient convection model describes the transport of cells in two-phase microfluidic flow under the influence of an external magnetic field. Transport of cells along the microchannel walls is also considered via an additional equation. Results show the variation of cell flux in the fluid phases and the wall as a function of a dimensionless parameter arising in the equations. Our results suggest that conditions may be optimized to maximize cell separation while minimizing contact with the wall surfaces. Experimentally measured cell rolling velocities on the wall indicate the presence of other near-wall forces in addition to fluid shear forces. Separation of a human colon carcinoma cell line from a mixture of red blood cells, with folic acid conjugated 1 microm and 200 nm beads, is reported.

  20. On Borders: From Ancient to Postmodern Times

    NASA Astrophysics Data System (ADS)

    Bellezza, G.

    2013-11-01

    The article deals with the evolution of the concept of borders between human groups and with its slow evolution from the initial no men's land zones to the ideal single-dimension linear borders. In ancient times the first borders were natural, such as mountain ranges or large rivers until, with the development of Geodesy, astronomical borders based on meridians and parallels became a favourite natural base. Actually, Modern States adopted these to fix limits in unknown conquered territories. The postmodern thought led give more importance to cultural borders until, in the most recent times, is becoming rather impossible to fix borders in the virtual cyberspace.

  1. EPA blocks TNF-α-induced inhibition of sugar uptake in Caco-2 cells via GPR120 and AMPK.

    PubMed

    Castilla-Madrigal, Rosa; Barrenetxe, Jaione; Moreno-Aliaga, María J; Lostao, María Pilar

    2018-03-01

    The aim of the present work was to investigate in Caco-2 cells whether eicosapentaenoic acid (EPA), an omega-3 polyunsaturated fatty acid, could block the inhibitory effect of tumor necrosis factor-α (TNF-α) on sugar transport, and identify the intracellular signaling pathways involved. After pre-incubation of the Caco-2 cells with TNF-α and EPA for 1 hr, EPA prevented the inhibitory effect of the cytokine on α-methyl-d-glucose (αMG) uptake (15 min) and on SGLT1 expression at the brush border membrane, measured by Western blot. The ERK1/2 inhibitor PD98059 and the AMPK activator AICAR also prevented the inhibitory effect of TNF-α on both αMG uptake and SGLT1 expression. Interestingly, the AMPK inhibitor, Compound C, abolished the ability of EPA to prevent TNF-α-induced reduction of sugar uptake and transporter expression. The GPR120 antagonist, AH7614, also blocked the preventive effect of EPA on TNF-α-induced decrease of αMG uptake and AMPK phosphorylation. In summary, TNF-α inhibits αMG uptake by decreasing SGLT1 expression in the brush border membrane through the activation of ERK1/2 pathway. EPA prevents the inhibitory effect of TNF-α through the involvement of GPR120 and AMPK activation. © 2017 Wiley Periodicals, Inc.

  2. Beneficial Insect Borders Provide Northern Bobwhite Brood Habitat

    PubMed Central

    Moorman, Christopher E.; Plush, Charles J.; Orr, David B.; Reberg-Horton, Chris

    2013-01-01

    Strips of fallow vegetation along cropland borders are an effective strategy for providing brood habitat for declining populations of upland game birds (Order: Galliformes), including northern bobwhite (Colinus virginianus), but fallow borders lack nectar-producing vegetation needed to sustain many beneficial insect populations (e.g., crop pest predators, parasitoids, and pollinator species). Planted borders that contain mixes of prairie flowers and grasses are designed to harbor more diverse arthropod communities, but the relative value of these borders as brood habitat is unknown. We used groups of six human-imprinted northern bobwhite chicks as a bioassay for comparing four different border treatments (planted native grass and prairie flowers, planted prairie flowers only, fallow vegetation, or mowed vegetation) as northern bobwhite brood habitat from June-August 2009 and 2010. All field border treatments were established around nine organic crop fields. Groups of chicks were led through borders for 30-min foraging trials and immediately euthanized, and eaten arthropods in crops and gizzards were measured to calculate a foraging rate for each border treatment. We estimated arthropod prey availability within each border treatment using a modified blower-vac to sample arthropods at the vegetation strata where chicks foraged. Foraging rate did not differ among border treatments in 2009 or 2010. Total arthropod prey densities calculated from blower-vac samples did not differ among border treatments in 2009 or 2010. Our results showed plant communities established to attract beneficial insects should maximize the biodiversity potential of field border establishment by providing habitat for beneficial insects and young upland game birds. PMID:24376759

  3. Patriot War and the Fenian Raids: Case Studies in Border Security on the U.S. Canada Border in the Nineteenth Century

    DTIC Science & Technology

    2017-06-09

    THE PATRIOT WAR AND THE FENIAN RAIDS: CASE STUDIES IN BORDER SECURITY ON THE U.S.-CANADA BORDER IN THE NINETEENTH CENTURY...3. DATES COVERED (From - To) AUG 2016 – JUN 2017 4. TITLE AND SUBTITLE The Patriot War and the Fenian Raids: Case Studies in Border Security on ...nineteenth century. The Patriot War and Fenian Raids are examined as case studies on the same border occurring roughly thirty years apart. The

  4. Efficiency and Impact of Positive and Negative Magnetic Separation on Monocyte Derived Dendritic Cell Generation.

    PubMed

    Kowalewicz-Kulbat, Magdalena; Ograczyk, Elżbieta; Włodarczyk, Marcin; Krawczyk, Krzysztof; Fol, Marek

    2016-06-01

    The immunomagnetic separation technique is the basis of monocyte isolation and further generation of monocyte-derived dendritic cells. To compare the efficiency of monocyte positive and negative separation, concentration of beads, and their impact on generated dendritic cells. Monocytes were obtained using monoclonal antibody-coated magnetic beads followed the Ficoll-Paque gradient separation of mononuclear cell fraction from the peripheral blood of 6 healthy volunteers. CD14 expression was analyzed by flow cytometry. Both types of magnetic separation including recommended and reduced concentrations of beads did not affect the yield and the purity of monocytes and their surface CD14 expression. However, DCs originated from the "positively" separated monocytes had noticeable higher expression of CD80.

  5. Quantification of deep percolation from two flood-irrigated alfalfa field, Roswell Basin, New Mexico

    USGS Publications Warehouse

    Roark, D. Michael; Healy, D.F.

    1998-01-01

    For many years water management in the Roswell ground-water basin (Roswell Basin) and other declared basins in New Mexico has been the responsibility of the State of New Mexico. One of the water management issues requiring better quantification is the amount of deep percolation from applied irrigation water. Two adjacent fields, planted in alfalfa, were studied to determine deep percolation by the water-budget, volumetric-moisture, and chloride mass-balance methods. Components of the water-budget method were measured, in study plots called borders, for both fields during the 1996 irrigation season. The amount of irrigation water applied in the west border was 95.8 centimeters and in the east border was 169.8 centimeters. The total amount of precipitation that fell during the irrigation season was 21.9 centimeters. The increase in soil-moisture storage from the beginning to the end of the irrigation season was 3.2 centimeters in the west border and 8.8 centimeters in the east border. Evapotranspiration, as estimated by the Bowen ratio energy balance technique, in the west border was 97.8 centimeters and in the east border was 101.0 centimeters. Deep percolation determined using the water-budget method was 16.4 centimeters in the west border and 81.6 centimeters in the east border. An average deep percolation of 22.3 centimeters in the west border and 31.6 centimeters in the east border was determined using the volumetric-moisture method. The chloride mass-balance method determined the multiyear deep percolation to be 15.0 centimeters in the west border and 38.0 centimeters in the east border. Large differences in the amount of deep percolation between the two borders calculated by the water-budget method are due to differences in the amount of water that was applied to each border. More water was required to flood the east border because of the greater permeability of the soils in that field and the smaller rate at which water could be applied.

  6. Cell design concepts for aqueous lithium-oxygen batteries: A model-based assessment

    NASA Astrophysics Data System (ADS)

    Grübl, Daniel; Bessler, Wolfgang G.

    2015-11-01

    Seven cell design concepts for aqueous (alkaline) lithium-oxygen batteries are investigated using a multi-physics continuum model for predicting cell behavior and performance in terms of the specific energy and specific power. Two different silver-based cathode designs (a gas diffusion electrode and a flooded cathode) and three different separator designs (a porous separator, a stirred separator chamber, and a redox-flow separator) are compared. Cathode and separator thicknesses are varied over a wide range (50 μm-20 mm) in order to identify optimum configurations. All designs show a considerable capacity-rate effect due to spatiotemporally inhomogeneous precipitation of solid discharge product LiOH·H2O. In addition, a cell design with flooded cathode and redox-flow separator including oxygen uptake within the external tank is suggested. For this design, the model predicts specific power up to 33 W/kg and specific energy up to 570 Wh/kg (gravimetric values of discharged cell including all cell components and catholyte except housing and piping).

  7. U.S. border patrol potential applications of internetted unattended ground sensors

    NASA Astrophysics Data System (ADS)

    Eaton, Wilbur W., Jr.; Schatzmann, Larry A.

    1997-07-01

    The U.S. Border Patrol monitors the traffic on the Mexican/U.S. Border, the Canadian/U.S. Border and along some coastal areas. Measures have been taken to reduce or eliminate illegal immigration and smuggling. An automated border surveillance sub-system based on the DARPA Internetted Unattended Ground Sensors Program is discussed.

  8. International Issues, High-Stakes Testing, and Border Pedagogy: Social Studies at Border High School

    ERIC Educational Resources Information Center

    Cashman, Timothy G.; McDermott, Benjamin R.

    2013-01-01

    A recently constructed border wall stands within walking distance of Border High School (BHS) and was created to impede the flow of people, goods, fauna, and contraband from Mexico into the United States (U.S.). The reality, however, is that this geopolitical border is fluid, allowing connections between sociopolitical zones. The researchers…

  9. The Intersectionality of Border Pedagogy and Latino/a Youth: Enacting Border Pedagogy in Multiple Spaces

    ERIC Educational Resources Information Center

    Ramirez, Pablo C.; Ross, Lydia; Jimenez-Silva, Margarita

    2016-01-01

    In this one-year qualitative study, the authors examined how border pedagogy is enacted by two Latino/a high school teachers in a border community in Southern California. Through classroom observations, the authors documented powerful student discussions that named complex borders (Giroux, 1992) that existed in their daily lives. We drew from…

  10. 75 FR 61508 - Agency Information Collection Activities: Canadian Border Boat Landing Permit (CBP Form I-68)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-05

    ... Activities: Canadian Border Boat Landing Permit (CBP Form I-68) AGENCY: U.S. Customs and Border Protection... information collection requirement concerning the Canadian Border Boat Landing Permit (Form I- [[Page 61509... Boat Landing Permit. OMB Number: 1651-0108. Form Number: CBP Form I-68. Abstract: The Canadian Border...

  11. --No Title--

    Science.gov Websites

    {background-color:#0079C2;text-align:center}.feature-border{border-top:3px solid #0079C2}.news{border-top:11px }.banner-continuum .issue-info,.feature .number,.multimedia-container{background-color:#5E6A71}.analysis -issue .banner-continuum{background-color:#a5acaf}.analysis-issue .feature-border{border-top:3px solid

  12. Rapid and label-free separation of Burkitt's lymphoma cells from red blood cells by optically-induced electrokinetics.

    PubMed

    Liang, Wenfeng; Zhao, Yuliang; Liu, Lianqing; Wang, Yuechao; Dong, Zaili; Li, Wen Jung; Lee, Gwo-Bin; Xiao, Xiubin; Zhang, Weijing

    2014-01-01

    Early stage detection of lymphoma cells is invaluable for providing reliable prognosis to patients. However, the purity of lymphoma cells in extracted samples from human patients' marrow is typically low. To address this issue, we report here our work on using optically-induced dielectrophoresis (ODEP) force to rapidly purify Raji cells' (a type of Burkitt's lymphoma cell) sample from red blood cells (RBCs) with a label-free process. This method utilizes dynamically moving virtual electrodes to induce negative ODEP force of varying magnitudes on the Raji cells and RBCs in an optically-induced electrokinetics (OEK) chip. Polarization models for the two types of cells that reflect their discriminate electrical properties were established. Then, the cells' differential velocities caused by a specific ODEP force field were obtained by a finite element simulation model, thereby established the theoretical basis that the two types of cells could be separated using an ODEP force field. To ensure that the ODEP force dominated the separation process, a comparison of the ODEP force with other significant electrokinetics forces was conducted using numerical results. Furthermore, the performance of the ODEP-based approach for separating Raji cells from RBCs was experimentally investigated. The results showed that these two types of cells, with different concentration ratios, could be separated rapidly using externally-applied electrical field at a driven frequency of 50 kHz at 20 Vpp. In addition, we have found that in order to facilitate ODEP-based cell separation, Raji cells' adhesion to the OEK chip's substrate should be minimized. This paper also presents our experimental results of finding the appropriate bovine serum albumin concentration in an isotonic solution to reduce cell adhesion, while maintaining suitable medium conductivity for electrokinetics-based cell separation. In short, we have demonstrated that OEK technology could be a promising tool for efficient and effective purification of Raji cells from RBCs.

  13. The Cosmopolitanization of Science1

    PubMed Central

    Zhang, Joy Yueyue

    2013-01-01

    It is commonly perceived that the ‘globalization of science’ may result in a ‘Westernization of science’. In this paper, however, I use the case of stem cell science in China to demonstrate that developing countries are sometimes able to effectively shape the norms of global/local scientific exchange. Based on interviews with 38 stem cell scientists in six Chinese cities in early 2008, this paper elucidates Chinese scientists’ outlook towards cross-border collaborations and the effects that the internationalization of science has had on everyday laboratory operations. Findings suggest that although there still exists an asymmetry of scientific influence, and in many aspects China is still ‘catching-up’ to the West, there is also a changing nature of communication beyond borders. One key aspect of recent international scientific development is the growing necessity for local stakeholders to acquire a global mindset and to compare, reflect and accommodate diverse interests. This is what I define as the ‘cosmopolitanization of science’. The study empirically examines the sociological and methodological implications of the cosmopolitanization process and further develops Ulrich Beck’s cosmopolitan theory by delineating four main features of the ‘cosmopolitanization of science’: shared future benefits, passive ethicization, reflexive negotiation, and continuous performance. PMID:24409002

  14. Border screening vs. community level disease control for infectious diseases: Timing and effectiveness

    NASA Astrophysics Data System (ADS)

    Kim, Sehjeong; Chang, Dong Eui

    2017-06-01

    There have been many studies of the border screening using a simple math model or a statistical analysis to investigate the ineffectiveness of border screening during 2003 and 2009 pandemics. However, the use of border screening is still a controversial issue. It is due to focusing only on the functionality of border screening without considering the timing to use. In this paper, we attempt to qualitatively answer whether the use of border screening is a desirable action during a disease pandemic. Thus, a novel mathematical model with a transition probability of status change during flight and border screening is developed. A condition to check a timing of the border screening is established in terms of a lower bound of the basic reproduction number. If the lower bound is greater than one, which indicates a pandemic, then the border screening may not be effective and the disease persists. In this case, a community level control strategy should be conducted.

  15. Lateral separation of colloids or cells by dielectrophoresis augmented by AC electroosmosis.

    PubMed

    Zhou, Hao; White, Lee R; Tilton, Robert D

    2005-05-01

    Colloidal particles and biological cells are patterned and separated laterally adjacent to a micropatterned electrode array by applying AC electric fields that are principally oriented normally to the electrode array. This is demonstrated for yeast cells, red blood cells, and colloidal polystyrene particles of different sizes and zeta-potentials. The separation mechanism is observed experimentally to depend on the applied field frequency and voltage. At high frequencies, particles position themselves in a manner that is consistent with dielectrophoresis, while at low frequencies, the positioning is explained in terms of a strong coupling between gravity, the vertical component of the dielectrophoretic force, and the Stokes drag on particles induced by AC electroosmotic flow. Compared to high frequency dielectrophoretic separations, the low frequency separations are faster and require lower applied voltages. Furthermore, the AC electroosmosis coupling with dielectrophoresis may enable cell separations that are not feasible based on dielectrophoresis alone.

  16. EGF Induced Centrosome Separation Promotes Mitotic Progression and Cell Survival

    PubMed Central

    Mardin, Balca R.; Isokane, Mayumi; Cosenza, Marco R.; Krämer, Alwin; Ellenberg, Jan; Fry, Andrew M.; Schiebel, Elmar

    2014-01-01

    Summary Timely and accurate assembly of the mitotic spindle is critical for the faithful segregation of chromosomes and centrosome separation is a key step in this process. The timing of centrosome separation varies dramatically between cell types; however, the mechanisms responsible for these differences and its significance are unclear. Here, we show that activation of epidermal growth factor receptor (EGFR) signaling determines the timing of centrosome separation. Premature separation of centrosomes decreases the requirement for the major mitotic kinesin Eg5 for spindle assembly, accelerates mitosis and decreases the rate of chromosome missegregation. Importantly, EGF stimulation impacts upon centrosome separation and mitotic progression to different degrees in different cell lines. Cells with high EGFR levels fail to arrest in mitosis upon Eg5 inhibition. This has important implications for cancer therapy since cells with high centrosomal response to EGF are more susceptible to combinatorial inhibition of EGFR and Eg5. PMID:23643362

  17. Pore size engineering applied to the design of separators for nickel-hydrogen cells and batteries

    NASA Technical Reports Server (NTRS)

    Abbey, K. M.; Britton, D. L.

    1983-01-01

    Pore size engineering in starved alkaline multiplate cells involves adopting techniques to widen the volume tolerance of individual cells. Separators with appropriate pore size distributions and wettability characteristics (capillary pressure considerations) to have wider volume tolerances and an ability to resist dimensional changes in the electrodes were designed. The separators studied for potential use in nickel-hydrogen cells consist of polymeric membranes as well as inorganic microporous mats. In addition to standard measurements, the resistance and distribution of electrolyte as a function of total cell electrolyte content were determined. New composite separators consisting of fibers, particles and/or binders deposited on Zircar cloth were developed in order to engineer the proper capillary pressure characteristics in the separator. These asymmetric separators were prepared from a variety of fibers, particles and binders. Previously announced in STAR as N83-24571

  18. Water outlet control mechanism for fuel cell system operation in variable gravity environments

    NASA Technical Reports Server (NTRS)

    Vasquez, Arturo (Inventor); McCurdy, Kerri L. (Inventor); Bradley, Karla F. (Inventor)

    2007-01-01

    A self-regulated water separator provides centrifugal separation of fuel cell product water from oxidant gas. The system uses the flow energy of the fuel cell's two-phase water and oxidant flow stream and a regulated ejector or other reactant circulation pump providing the two-phase fluid flow. The system further uses a means of controlling the water outlet flow rate away from the water separator that uses both the ejector's or reactant pump's supply pressure and a compressibility sensor to provide overall control of separated water flow either back to the separator or away from the separator.

  19. New separators for nickel-zinc batteries

    NASA Technical Reports Server (NTRS)

    Sheibley, D. W.

    1976-01-01

    Flexible separators consisting of a substrate coated with a mixture of a polymer and organic and inorganic additives were cycle tested in nickel-zinc cells. By substituting a rubber-based resin for polyphenylene oxide in the standard inorganic-organic separator, major improvements in both cell life and flexibility were made. Substituting newsprint for asbestos as the substrate shows promise for use on the zinc electrode and reduces separator cost. The importance of ample electrolyte in the cells was noted. Cycle lives and the characteristics of these flexible, low-cost separators were compared with those of a standard microporous polypropylene separator.

  20. An Artificial Intelligence Approach to Automatic Left Ventricular Border Detection in 2-D Echocardiography

    PubMed Central

    Buchan, Iris; Covvey, H. Dominic; Rakowski, Harry

    1985-01-01

    A program has been developed for left ventricular (LV) border tracking on ultrasound images. For each frame, forty border points at equally-spaced angles around the LV center are found gradually during three passes. Pass 1 uses adaptive thresholding to find the most obvious border points. Pass 2 then uses an artificial intelligence technique of finding possible border path segments, associating a score with each, and, from paths with superior scores, obtaining more of the border points. Pass 3 closes any remaining gaps by interpolation. The program tracks the LV border quite well in spite of dropout and interference from intracardiac structures, except during end-systole. Multi-level passes provide a very useful structure for border tracking, with increasingly slow but more sophisticated algorithms possible at higher levels for use when earlier passes recognise failure.

  1. The Structure of Borders in a Small World

    PubMed Central

    Thiemann, Christian; Theis, Fabian; Grady, Daniel; Brune, Rafael; Brockmann, Dirk

    2010-01-01

    Territorial subdivisions and geographic borders are essential for understanding phenomena in sociology, political science, history, and economics. They influence the interregional flow of information and cross-border trade and affect the diffusion of innovation and technology. However, it is unclear if existing administrative subdivisions that typically evolved decades ago still reflect the most plausible organizational structure of today. The complexity of modern human communication, the ease of long-distance movement, and increased interaction across political borders complicate the operational definition and assessment of geographic borders that optimally reflect the multi-scale nature of today's human connectivity patterns. What border structures emerge directly from the interplay of scales in human interactions is an open question. Based on a massive proxy dataset, we analyze a multi-scale human mobility network and compute effective geographic borders inherent to human mobility patterns in the United States. We propose two computational techniques for extracting these borders and for quantifying their strength. We find that effective borders only partially overlap with existing administrative borders, and show that some of the strongest mobility borders exist in unexpected regions. We show that the observed structures cannot be generated by gravity models for human traffic. Finally, we introduce the concept of link significance that clarifies the observed structure of effective borders. Our approach represents a novel type of quantitative, comparative analysis framework for spatially embedded multi-scale interaction networks in general and may yield important insight into a multitude of spatiotemporal phenomena generated by human activity. PMID:21124970

  2. The structure of borders in a small world.

    PubMed

    Thiemann, Christian; Theis, Fabian; Grady, Daniel; Brune, Rafael; Brockmann, Dirk

    2010-11-18

    Territorial subdivisions and geographic borders are essential for understanding phenomena in sociology, political science, history, and economics. They influence the interregional flow of information and cross-border trade and affect the diffusion of innovation and technology. However, it is unclear if existing administrative subdivisions that typically evolved decades ago still reflect the most plausible organizational structure of today. The complexity of modern human communication, the ease of long-distance movement, and increased interaction across political borders complicate the operational definition and assessment of geographic borders that optimally reflect the multi-scale nature of today's human connectivity patterns. What border structures emerge directly from the interplay of scales in human interactions is an open question. Based on a massive proxy dataset, we analyze a multi-scale human mobility network and compute effective geographic borders inherent to human mobility patterns in the United States. We propose two computational techniques for extracting these borders and for quantifying their strength. We find that effective borders only partially overlap with existing administrative borders, and show that some of the strongest mobility borders exist in unexpected regions. We show that the observed structures cannot be generated by gravity models for human traffic. Finally, we introduce the concept of link significance that clarifies the observed structure of effective borders. Our approach represents a novel type of quantitative, comparative analysis framework for spatially embedded multi-scale interaction networks in general and may yield important insight into a multitude of spatiotemporal phenomena generated by human activity.

  3. Separation of active and inactive fractions from starved culture of Vibrio parahaemolyticus by density dependent cell sorting.

    PubMed

    Nayak, Binaya Bhusan; Kamiya, Eriko; Nishino, Tomohiko; Wada, Minoru; Nishimura, Masahiko; Kogure, Kazuhiro

    2005-01-01

    The co-existence of physiologically different cells in bacterial cultures is a general phenomenon. We have examined the applicability of the density dependent cell sorting (DDCS) method to separate subpopulations from a long-term starvation culture of Vibrio parahaemolyticus. The cells were subjected to Percoll density gradient and separated into 12 fractions of different buoyant densities, followed by measuring the cell numbers, culturability, respiratory activity and leucine incorporation activity. While more than 78% of cells were in lighter fractions, about 95% of culturable cells were present in heavier fractions. The high-density subpopulations also had high proportion of cells capable of forming formazan granules. Although this was accompanied by the cell specific INT-reduction rate, both leucine incorporation rates and INT-reduction rates per cell had a peak at mid-density fraction. The present results indicated that DDCS could be used to separate subpopulations of different physiological conditions.

  4. Microfluidic immunomagnetic cell separation from whole blood.

    PubMed

    Bhuvanendran Nair Gourikutty, Sajay; Chang, Chia-Pin; Puiu, Poenar Daniel

    2016-02-01

    Immunomagnetic-based separation has become a viable technique for the separation of cells and biomolecules. Here we report on the design and analysis of a simple and efficient microfluidic device for high throughput and high efficiency capture of cells tagged with magnetic particles. This is made possible by using a microfluidic chip integrated with customized arrays of permanent magnets capable of creating large magnetic field gradients, which determine the effective capturing of the tagged cells. This method is based on manipulating the cells which are under the influence of a combination of magnetic and fluid dynamic forces in a fluid under laminar flow through a microfluidic chip. A finite element analysis (FEA) model is developed to analyze the cell separation process and predict its behavior, which is validated subsequently by the experimental results. The magnetic field gradients created by various arrangements of magnetic arrays have been simulated using FEA and the influence of these field gradients on cell separation has been studied with the design of our microfluidic chip. The proof-of-concept for the proposed technique is demonstrated by capturing white blood cells (WBCs) from whole human blood. CD45-conjugated magnetic particles were added into whole blood samples to label WBCs and the mixture was flown through our microfluidic device to separate the labeled cells. After the separation process, the remaining WBCs in the elute were counted to determine the capture efficiency, and it was found that more than 99.9% WBCs have been successfully separated from whole blood. The proposed design can be used for positive selection as well as for negative enrichment of rare cells. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Correlation of simulation/finite element analysis to the separation of intrinsically magnetic spores and red blood cells using a microfluidic magnetic deposition system.

    PubMed

    Sun, Jianxin; Moore, Lee; Xue, Wei; Kim, James; Zborowski, Maciej; Chalmers, Jeffrey J

    2018-05-01

    Magnetic separation of cells has been, and continues to be, widely used in a variety of applications, ranging from healthcare diagnostics to detection of food contamination. Typically, these technologies require cells labeled with antibody magnetic particle conjugate and a high magnetic energy gradient created in the flow containing the labeled cells (i.e., a column packed with magnetically inducible material), or dense packing of magnetic particles next to the flow cell. Such designs, while creating high magnetic energy gradients, are not amenable to easy, highly detailed, mathematic characterization. Our laboratories have been characterizing and developing analysis and separation technology that can be used on intrinsically magnetic cells or spores which are typically orders of magnitude weaker than typically immunomagnetically labeled cells. One such separation system is magnetic deposition microscopy (MDM) which not only separates cells, but deposits them in specific locations on slides for further microscopic analysis. In this study, the MDM system has been further characterized, using finite element and computational fluid mechanics software, and separation performance predicted, using a model which combines: 1) the distribution of the intrinsic magnetophoretic mobility of the cells (spores); 2) the fluid flow within the separation device; and 3) accurate maps of the values of the magnetic field (max 2.27 T), and magnetic energy gradient (max of 4.41 T 2 /mm) within the system. Guided by this model, experimental studies indicated that greater than 95% of the intrinsically magnetic Bacillus spores can be separated with the MDM system. Further, this model allows analysis of cell trajectories which can assist in the design of higher throughput systems. © 2018 Wiley Periodicals, Inc.

  6. Electrically Conductive Porous Membrane

    NASA Technical Reports Server (NTRS)

    Burke, Kenneth Alan (Inventor)

    2014-01-01

    The present invention relates to an electrically conductive membrane that can be configured to be used in fuel cell systems to act as a hydrophilic water separator internal to the fuel cell, or as a water separator used with water vapor fed electrolysis cells, or as a water separator used with water vapor fed electrolysis cells, or as a capillary structure in a thin head pipe evaporator, or as a hydrophobic gas diffusion layer covering the fuel cell electrode surface in a fuel cell.

  7. Effects of Protease, Phytase and a Bacillus sp. Direct-Fed Microbial on Nutrient and Energy Digestibility, Ileal Brush Border Digestive Enzyme Activity and Cecal Short-Chain Fatty Acid Concentration in Broiler Chickens

    PubMed Central

    Murugesan, Ganapathi R.; Romero, Luis F.; Persia, Michael E.

    2014-01-01

    Two experiments were conducted to determine the effects of protease and phytase (PP) and a Bacillus sp. direct-fed microbial (DFM) on dietary energy and nutrient utilization in broiler chickens. In the first experiment, Ross 308 broiler chicks were fed diets supplemented with PP and DFM in a 2×2 factorial arrangement. The 4 diets (control (CON), CON + PP, CON + DFM, and CON + PP + DFM) were fed from 15–21 days of age. In Experiment 1, significant interaction (P≤0.01) between PP and DFM on the apparent ileal digestibility coefficient for starch, crude protein, and amino acid indicated that both additives increased the digestibility. Both additives increased the nitrogen retention coefficient with a significant interaction (P≤0.01). Although no interaction was observed, significant main effects (P≤0.01) for nitrogen-corrected apparent ME (AMEn) for PP or DFM indicated an additive response. In a follow-up experiment, Ross 308 broiler chicks were fed the same experimental diets from 1–21 days of age. Activities of ileal brush border maltase, sucrase, and L-alanine aminopeptidase were increased (P≤0.01) by PP addition, while a trend (P = 0.07) for increased sucrase activity was observed in chickens fed DFM, in Experiment 2. The proportion of cecal butyrate was increased (P≤0.01) by DFM addition. Increased nutrient utilization and nitrogen retention appear to involve separate but complementary mechanisms for PP and DFM, however AMEn responses appear to have separate and additive mechanisms. PMID:25013936

  8. Thin-skinned tectonics of the Upper Ojai Valley and Sulphur Mountain area, Ventura basin, California

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

    Huftile, G.J.

    1991-08-01

    By integrating surface mapping with subsurface well data and drawing cross sections and subsurface maps, the geometry of shallow structures and their geologic history of the Upper Ojai Valley of California can be reconstructed. The geometry of shallow structures, the geologic history, and the location of earthquake foci then offer constraints on the deep structure of this complex area. The Upper Ojai Valley is a tectonic depression between opposing reverse faults. Its northern border is formed by the active, north-dipping San Cayetano fault, which has 6.0 km of stratigraphic separation in the Silverthread area of the Ojai oil field andmore » 2.6 km of stratigraphic separation west of Sisar Creek. The fault dies out farther west in Ojai Valley, where the south-vergent shortening is transferred to a blind thrust. The southern border of the Upper Ojai Valley is formed by the Quaternary Lion fault set, which dips south and merges into the Sisar decollement within the south-dipping, ductile, lower Miocene Rincon formation. By the middle Pleistocene, the Sulphur Mountain anticlinorium and the Big Canyon syncline began forming as a fault-propagation fold; the fault-propagation fold is rooted in the Sisar decollement, a passive backthrust rising from a blind thrust at depth. The formation of the Sulphur Mountain anticlinorium was followed closely by the ramping of the south-dipping Lion fault set to the surface over the nonmarine upper Pleistocene Saugus Formation. To the east, the San Cayetano fault overrides and folds the Lion Fault set near the surface. Area-balancing of the deformation shows shortening of 15.5 km, and suggests a 17 km depth to the brittle-ductile transition.« less

  9. Delineation of the Species Haemophilus influenzae by Phenotype, Multilocus Sequence Phylogeny, and Detection of Marker Genes▿ †

    PubMed Central

    Nørskov-Lauritsen, Niels; Overballe, Merete D.; Kilian, Mogens

    2009-01-01

    To obtain more information on the much-debated definition of prokaryotic species, we investigated the borders of Haemophilus influenzae by comparative analysis of H. influenzae reference strains with closely related bacteria including strains assigned to Haemophilus haemolyticus, cryptic genospecies biotype IV, and the never formally validated species “Haemophilus intermedius”. Multilocus sequence phylogeny based on six housekeeping genes separated a cluster encompassing the type and the reference strains of H. influenzae from 31 more distantly related strains. Comparison of 16S rRNA gene sequences supported this delineation but was obscured by a conspicuously high number of polymorphic sites in many of the strains that did not belong to the core group of H. influenzae strains. The division was corroborated by the differential presence of genes encoding H. influenzae adhesion and penetration protein, fuculokinase, and Cu,Zn-superoxide dismutase, whereas immunoglobulin A1 protease activity or the presence of the iga gene was of limited discriminatory value. The existence of porphyrin-synthesizing strains (“H. intermedius”) closely related to H. influenzae was confirmed. Several chromosomally encoded hemin biosynthesis genes were identified, and sequence analysis showed these genes to represent an ancestral genotype rather than recent transfers from, e.g., Haemophilus parainfluenzae. Strains previously assigned to H. haemolyticus formed several separate lineages within a distinct but deeply branching cluster, intermingled with strains of “H. intermedius” and cryptic genospecies biotype IV. Although H. influenzae is phenotypically more homogenous than some other Haemophilus species, the genetic diversity and multicluster structure of strains traditionally associated with H. influenzae make it difficult to define the natural borders of that species. PMID:19060144

  10. Importing the Poor: Welfare Magnetism and Cross-Border Welfare Migration

    PubMed Central

    McKinnish, Terra

    2011-01-01

    I test for welfare-induced migration by comparing AFDC participation in border counties to interior counties in the same state. If migration costs are lower for border county residents, border counties on the high-benefit side of a state border should have higher welfare participation relative to the state’s interior counties. Border counties on the low-benefit side should have lower welfare participation relative to the state’s interior counties. The results obtained using county-level data from 1970–90 indicate that having a neighbor with benefits that are $100 lower increases AFDC expenditures in border counties by 4.0–6.8 percent relative to interior counties. PMID:21949446

  11. Embryonic stem cell grafting in normal and infarcted myocardium: serial assessment with MR imaging and PET dual detection.

    PubMed

    Qiao, Hui; Zhang, Hualei; Zheng, Yuanjie; Ponde, Datta E; Shen, Dinggang; Gao, Fabao; Bakken, Ashley B; Schmitz, Alexander; Kung, Hank F; Ferrari, Victor A; Zhou, Rong

    2009-03-01

    To use magnetic resonance (MR) imaging and positron emission tomography (PET) dual detection of cardiac-grafted embryonic stem cells (ESCs) to examine (a) survival and proliferation of ESCs in normal and infarcted myocardium, (b) host macrophage versus grafted ESC contribution to serial MR imaging signal over time, and (c) cardiac function associated with the formation of grafts and whether improvement in cardiac function is related to cardiac differentiation of ESCs. All animal procedures were approved by the institutional animal care and use committee. Murine ESCs were stably transfected with a mutant version of herpes simplex virus type 1 thymidine kinase, HSV1-sr39tk, and also were labeled with superparamagnetic iron oxide (SPIO) particles. Cells were injected directly in the border zone of the infarcted heart or in corresponding regions of normal hearts in athymic rats. PET and MR imaging were performed longitudinally for 4 weeks in the same animals. ESCs survived and underwent proliferation in the infarcted and normal hearts, as demonstrated by serial increases in 9-(4-[(18)F]fluoro-3-hydroxymethylbutyl) guanine PET signals. In parallel, the hypointense areas on MR images at the injection sites decreased over time. Double staining for host macrophages and SPIO particles revealed that the majority of SPIO-containing cells were macrophages at week 4 after injection. Left ventricular ejection fraction increased in the ESC-treated rats but decreased in culture media-treated rats, and border-zone function was preserved in ESC-treated animals; however, cardiac differentiation of ESCs was less than 0.5%. Dual-modality imaging permits complementary information in regard to cell survival and proliferation, graft formation, and effects on cardiac function. http://radiology.rsnajnls.org/cgi/content/full/250/3/821/DC1. RSNA, 2009

  12. A Comparative Study of Intratesticular Ductules in the Spermatogenically Active Testes of Shortfin Mako and Thresher Sharks.

    PubMed

    McClusky, Leon Mendel; Sulikowski, James

    2016-10-01

    This comparative study of the radial testes of sexually mature thresher sharks (Alopias vulpinus) and shortfin mako sharks (Isurus oxyrinchus) describes the histology of the three-tiered network of sperm-carrying ductules in the testis and the lymphomyeloid tissue associated with it, namely the epigonal organ. In both species, a testis → epigonal gradient was evident regarding the thickness of the ductule epithelial lining and subepithelial investment of connective tissue. Ductules straddling the testis-epigonal border often displayed luminal leukocytes and various signs of regression, including the progressive thickening of the ductule epithelial lining, dissolution of the cytoplasm, and loss of normal histoarchitecture. In Isurus, large amorphous areas formed due to the fusion of neighboring regressing ductules. The epigonal organ of Alopias additionally revealed circular degenerative sperm-containing, Hassall-like bodies with either a degenerate or cellular appearance, the latter the result of cell proliferative activity (as shown by proliferating cell nuclear antigen (PCNA) immunohistochemistry) in an expanding outer border comprising cells with intensely PCNA immunoreactive slender and oblong nuclei. The latter cells exhibited a periphery-to-center transformation of their nuclei, at which stage they were PCNA-negative and most likely in a terminally differentiated state as they phagocytized the cell debris in the degenerate core. Intermediate stages of these circular bodies were a rarity. The relationship between these degenerate bodies, and the common occurrence of blind pockets in the epithelial linings and non-apoptosis-related degenerate patches in the apical cytoplasmic regions of the irregular shaped ductules in Alopias is unclear, and needs further elucidation. Anat Rec, 299:1435-1448, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  13. Overexpression of protein kinase C ɛ improves retention and survival of transplanted mesenchymal stem cells in rat acute myocardial infarction.

    PubMed

    He, H; Zhao, Z-H; Han, F-S; Liu, X-H; Wang, R; Zeng, Y-J

    2016-01-21

    We assessed the effects of protein kinase C ɛ (PKCɛ) for improving stem cell therapy for acute myocardial infarction (AMI). Primary mesenchymal stem cells (MSCs) were harvested from rat bone marrow. PKCɛ-overexpressed MSCs and control MSCs were transplanted into infarct border zones in a rat AMI model. MSCs and PKCɛ distribution and expression of principal proteins involved in PKCɛ signaling through the stromal cell-derived factor 1 (SDF-1)/CXC chemokine receptor type 4 (CXCR4) axis and the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT) pathway were analyzed by immunofluorescence and western blot 1 day after transplantation. Echocardiographic measurements and histologic studies were performed at 4 weeks after transplantation, and MSC survival, expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), transforming growth factor β (TGFβ), cardiac troponin I (cTnI), von Willebrand factor (vWF), smooth muscle actin (SMA) and factor VIII and apoptosis in infarct border zones were assessed. Rat heart muscles retained more MSCs and SDF-1, CXCR4, PI3K and phosphorylated AKT increased with PKCɛ overexpression 1 day after transplantation. MSC survival and VEGF, bFGF, TGFβ, cTnI, vWF, SMA and factor VIII expression increased in animals with PKCɛ-overexpressed MSCs at 4 weeks after transplantation and cardiac dysfunction and remodeling improved. Infarct size and apoptosis decreased as well. Inhibitory actions of CXCR4 or PI3K partly attenuated the effects of PKCɛ. Activation of PKCɛ may improve retention, survival and differentiation of transplanted MSCs in myocardia. Augmentation of PKCɛ expression may enhance the therapeutic effects of stem cell therapy for AMI.

  14. BMP, Wnt and FGF signals are integrated through evolutionarily conserved enhancers to achieve robust expression of Pax3 and Zic genes at the zebrafish neural plate border

    PubMed Central

    Garnett, Aaron T.; Square, Tyler A.; Medeiros, Daniel M.

    2012-01-01

    Neural crest cells generate a range of cells and tissues in the vertebrate head and trunk, including peripheral neurons, pigment cells, and cartilage. Neural crest cells arise from the edges of the nascent central nervous system, a domain called the neural plate border (NPB). NPB induction is known to involve the BMP, Wnt and FGF signaling pathways. However, little is known about how these signals are integrated to achieve temporally and spatially specific expression of genes in NPB cells. Furthermore, the timing and relative importance of these signals in NPB formation appears to differ between vertebrate species. Here, we use heat-shock overexpression and chemical inhibitors to determine whether, and when, BMP, Wnt and FGF signaling are needed for expression of the NPB specifiers pax3a and zic3 in zebrafish. We then identify four evolutionarily conserved enhancers from the pax3a and zic3 loci and test their response to BMP, Wnt and FGF perturbations. We find that all three signaling pathways are required during gastrulation for the proper expression of pax3a and zic3 in the zebrafish NPB. We also find that, although the expression patterns driven by the pax3a and zic3 enhancers largely overlap, they respond to different combinations of BMP, Wnt and FGF signals. Finally, we show that the combination of the two pax3a enhancers is less susceptible to signaling perturbations than either enhancer alone. Taken together, our results reveal how BMPs, FGFs and Wnts act cooperatively and redundantly through partially redundant enhancers to achieve robust, specific gene expression in the zebrafish NPB. PMID:23034628

  15. Rapid Turnover of Stereocilia Membrane Proteins: Evidence from the Trafficking and Mobility of Plasma Membrane Ca2+-ATPase 2

    PubMed Central

    Grati, M'hamed; Schneider, Mark E.; Lipkow, Karen; Strehler, Emanuel E.; Wenthold, Robert J.; Kachar, Bechara

    2007-01-01

    We studied the spatial distribution, mobility, and trafficking of plasma membrane Ca2+ATPase-2 (PMCA2), a protein enriched in the hair cell apical membrane and essential for hair cell function. Using immunofluorescence, we determined that PMCA2 is enriched in the stereocilia and present at a relatively low concentration in the kinocilium and in the remaining apical membrane. Using an antibody to the extracellular domain of PMCA2 as a probe, we observed that PMCA2 diffuses laterally from the stereocilia membrane and is internalized at the apical cell border maintaining an estimated half-life of residency in the stereocilia of ∼5–7 h. A computer simulation of our data indicates that PMCA2 has an estimated global diffusion coefficient of 0.01– 0.005 μm2/s. Using a green fluorescent protein tag, we observed that PMCA2 is rapidly delivered to the apical cell border from where it diffuses to the entire stereocilia surface. Fluorescence recovery after photobleaching experiments show that ∼60% of PMCA2 in the stereocilia exhibit high mobility with a diffusion coefficient of 0.1– 0.2 μm2/s, whereas the remaining pool represents a relatively immobile fraction. These results suggest that PMCA2 molecules maintain transient interactions with other components of the stereocilia, and the mobile pool of PMCA2 mediates the exchange between the stereocilia and the removal and delivery sites at the periphery of the apical cell surface. This rapid turnover of a major stereocilia membrane protein matches the previously described rapid turnover of proteins of the stereocilia actin core, further demonstrating that these organelles undergo rapid continuous renewal. PMID:16763047

  16. Mapping and assessing the environmental impacts of border tactical infrastructure in the Sky Island Region

    Treesearch

    Caroline Patrick-Birdwell; Sergio Avila-Villegas; Jenny Neeley; Louise Misztal

    2013-01-01

    In this project we mapped the different types of border barriers, identified impacts of border infrastructure on public and private lands and conducted spatial analyses within the approximately 200 miles of international border in the Sky Island region. The Sky Island region, bisected by the U.S.-Mexico border, is critically important for its biodiversity and...

  17. Decision Model for U.S.- Mexico Border Security Measures

    DTIC Science & Technology

    2017-09-01

    and money assigned to border security investments. 14. SUBJECT TERMS Department of Homeland Security (DHS), border security, U.S.–Mexico border...and money assigned to border security investments. vi THIS PAGE INTENTIONALLY LEFT BLANK vii TABLE OF CONTENTS I. INTRODUCTION...FAA Federal Aviation Administration FAMS Federal Air Marshals Service FAST Free and Secure Trade GSA General Services Administration HIR Human

  18. Gross anatomy of pancreatic surgery related fascia and fascial spaces.

    PubMed

    Zhou, Xiaobo; Ding, Zihai

    2014-01-01

    The study aims to provide anatomical basis and identify surgical planes or safe routes for pancreatic surgery by studying adult or fresh cadaver specimens. Thirty formalin-fixed adult cadavers (16 males and 14 females), provided by the Department of Anatomy at Southern Medical University, China, were perfused with red latex through the abdominal aorta, treated with antiseptic and antibacterial agents and then refrigerated. Fifteen abdomen specimens from fresh adult cadavers (Southern Medical body donation reception centers) were also perfused with red latex through the femoral artery and placed in -20 C freezer for 1 week before surgery and anatomic observation. The renal fascia surrounding pancreas and duodenum were mostly filled with loose connective tissues and adipose tissues. They were mutually connected with clear fascial borders but easily to be separated, suitable for surgical operations. Also, the integrating spaces were the connecting borders between different tissues without nerves or blood vessels inside. They may serve as ideal surgical planes for pancreatic surgery. A better understanding of the anatomy of the renal fascia and fascial spaces may provide guidance for identifying surgical landmarks and planes, and help to reduce bleeding and unnecessary side injuries in pancreatic surgery.

  19. Parafermionic zero modes in gapless edge states

    NASA Astrophysics Data System (ADS)

    Clarke, David

    It has been recently demonstrated1 that Majorana zero modes may occur in the gapless edge of Abelian quantum Hall states at a boundary between different edge phases bordering the same bulk. Such a zero mode is guaranteed to occur when an edge phase that supports fermionic excitations borders one that does not. Here we generalize to the non-charge conserving case such as may occur when a superconductor abuts the quantum Hall edge. We find that not only Majorana zero modes, but their ℤN generalizations (known as parafermionic zero modes) may occur at boundaries between edge phases in a fractional quantum Hall state. In particular, we find thst the ν = 1 / 3 fractional quantum Hall state supports topologically distinct edge phases separated by ℤ3 parafermionic zero modes when charge conservation is broken. Paradoxically, an arrangement of phases can be made such that only an odd number of localized parafermionic zero modes occur around the edge of a quantum Hall droplet. Such an arrangement is not allowed in a gapped system, but here the paradox is resolved due to an extended zero mode in the edge spectrum. LPS-MPO-CMTC, JQI-NSF-PFC, Microsoft Station Q.

  20. Chiral pathways in DNA dinucleotides using gradient optimized refinement along metastable borders

    NASA Astrophysics Data System (ADS)

    Romano, Pablo; Guenza, Marina

    We present a study of DNA breathing fluctuations using Markov state models (MSM) with our novel refinement procedure. MSM have become a favored method of building kinetic models, however their accuracy has always depended on using a significant number of microstates, making the method costly. We present a method which optimizes macrostates by refining borders with respect to the gradient along the free energy surface. As the separation between macrostates contains highest discretization errors, this method corrects for any errors produced by limited microstate sampling. Using our refined MSM methods, we investigate DNA breathing fluctuations, thermally induced conformational changes in native B-form DNA. Running several microsecond MD simulations of DNA dinucleotides of varying sequences, to include sequence and polarity effects, we've analyzed using our refined MSM to investigate conformational pathways inherent in the unstacking of DNA bases. Our kinetic analysis has shown preferential chirality in unstacking pathways that may be critical in how proteins interact with single stranded regions of DNA. These breathing dynamics can help elucidate the connection between conformational changes and key mechanisms within protein-DNA recognition. NSF Chemistry Division (Theoretical Chemistry), the Division of Physics (Condensed Matter: Material Theory), XSEDE.

  1. Development of Indonesia-Papua New Guinea border, Muara Tami District, Jayapura City through agropolitan concept

    NASA Astrophysics Data System (ADS)

    Subagiyo, A.; Dwiproborini, F.; Sari, N.

    2017-06-01

    The border of RI-PNG Muara Tami district is located on the eastern part of Jayapura city, which has agricultural potential. The past paradigm put the border as the backyard caused underdevelopment in border RI-PNG Muara Tami district, so that needed acceleration development through agropolitan concept. The purpose of the research is to define the aspect of physical, social, economic and border security to support agropolitan concept in border RI-PNG Muara Tami district. The analytical research method are border interactionan analysis, border security analysis, land capability analysis, land availability analysis, schallogram analysis, institutional analysis, leading comodity analysis (LQ and Growth Share), agribusiness linkage system analysis, accessibility analysis and A’WOT analysis. The result shown that mobilization from PNG to Muara Tami district could increase the economic opportunities with agricultural based. Border security of RI-PNG Muara Tami district is vulnerable, yet still condusive to mobilization. There is 12.977,94 Ha potensial land for agricultural (20,93%). There are six leading commodities to developed are rice, watermelon, banana, coconut, areca nut and cocoa. The border of RI-PNG Muara Tami district is ready enough to support agropolitan concept, but still have problems in social and economy aspect.

  2. Myofilament dysfunction contributes to impaired myocardial contraction in the infarct border zone

    PubMed Central

    Shimkunas, Rafael; Makwana, Om; Spaulding, Kimberly; Bazargan, Mona; Khazalpour, Michael; Takaba, Kiyoaki; Soleimani, Mehrdad; Myagmar, Bat-Erdene; Lovett, David H.; Simpson, Paul C.; Ratcliffe, Mark B.

    2014-01-01

    After myocardial infarction, a poorly contracting nonischemic border zone forms adjacent to the infarct. The cause of border zone dysfunction is unclear. The goal of this study was to determine the myofilament mechanisms involved in postinfarction border zone dysfunction. Two weeks after anteroapical infarction of sheep hearts, we studied in vitro isometric and isotonic contractions of demembranated myocardium from the infarct border zone and a zone remote from the infarct. Maximal force development (Fmax) of the border zone myocardium was reduced by 31 ± 2% versus the remote zone myocardium (n = 6/group, P < 0.0001). Decreased border zone Fmax was not due to a reduced content of contractile material, as assessed histologically, and from myosin content. Furthermore, decreased border zone Fmax did not involve altered cross-bridge kinetics, as assessed by muscle shortening velocity and force development kinetics. Decreased border zone Fmax was associated with decreased cross-bridge formation, as assessed from muscle stiffness in the absence of ATP where cross-bridge formation should be maximized (rigor stiffness was reduced 34 ± 6%, n = 5, P = 0.011 vs. the remote zone). Furthermore, the border zone myocardium had significantly reduced phosphorylation of myosin essential light chain (ELC; 41 ± 10%, n = 4, P < 0.05). However, for animals treated with doxycycline, an inhibitor of matrix metalloproteinases, rigor stiffness and ELC phosphorylation were not reduced in the border zone myocardium, suggesting that doxycycline had a protective effect. In conclusion, myofilament dysfunction contributes to postinfarction border zone dysfunction, myofilament dysfunction involves impaired cross-bridge formation and decreased ELC phosphorylation, and matrix metalloproteinase inhibition may be beneficial for limiting postinfarct border zone dysfunction. PMID:25128171

  3. Alcohol consumption and binge drinking among U.S.-Mexico border and non-border Mexican Americans

    PubMed Central

    Caetano, Raul; Mills, Britain; Vaeth, Patrice A. C.

    2011-01-01

    Background This paper examines differences in drinking and binge drinking between Mexican Americans living along the U.S.-Mexico border and those living in two metropolitan areas away from the border (Houston, Texas, and Los Angeles, California). Methods Respondents in the non-border area (Houston and Los Angeles) constitute a multistage probability sample (N=1,288) who were interviewed as part of the 2006 Hispanic Americans Baseline Alcohol Survey (HABLAS). Respondents in the border area (N=1,307) constitute a household probability sample of Mexican Americans living on the U.S.-Mexico border. In both surveys, data were collected during computer assisted interviews conducted in respondents’ homes. The HABLAS and the border sample response rates were 76% and 67%, respectively. Results There were no differences between border and non-border Mexican American men in the proportion of drinkers, the proportion who binge drink at least once a year and volume of alcohol consumption. However, within each location, there were significant differences in drinking by age, indicating that younger men drank more than men who were older. Border women showed significant differences across age groups in the proportion of drinkers, in binge drinking and volume of alcohol consumption, which were not seen among non-border women. Conclusions Women’s drinking seems to be more affected than men’s by their residence on or off the U.S.-Mexico border. This is seen most clearly among young women 18–29 years old and it is associated with an increased proportion of drinkers, a higher volume of drinking and an increased proportion of women who report binge drinking. Increased drinking in this group of younger women seems to be associated with drinking in Mexico. PMID:22017228

  4. Coutts/Sweetgrass automated border crossing : phase I

    DOT National Transportation Integrated Search

    1999-03-01

    The Coutts/Sweetgrass Automated Border Crossing Project was intended to improve operational efficiency of this rural border crossing facility using ITS applications. Phase I of the Coutts/Sweetgrass Automated Border Crossing Project was intended to r...

  5. MRI Findings of Suprasellar Germ Cell Tumors in Two Dogs.

    PubMed

    Cook, Laurie; Tensley, Michelle; Drost, Wm Tod; Koivisto, Christopher; Oglesbee, Michael

    A 4 yr old border collie presenting for mydriasis and decreased mentation and a 7 yr old Boston terrier presenting for obtundation, head tilt, and paraparesis were both evaluated using MRI. Findings in both included mass lesions of the thalamus and brainstem that were hypo- to isointense on T1-weighted images and hyperintense on T2-weighted images with regions of hypointensity, and robust contrast enhancement and displacement of adjacent structures. Postmortem histopathology findings, tumor location, and a mixed pattern of epithelial cell differentiation were consistent with germ cell tumor in both cases. Germ cell tumor of the suprasellar region is an infrequently reported neoplasm of dogs and imaging findings in this species have not been well described in the prior literature.

  6. Continuous high throughput molecular adhesion based cell sorting using ridged microchannels

    NASA Astrophysics Data System (ADS)

    Tasadduq, Bushra; Wang, Gonghao; Alexeev, Alexander; Sarioglu, Ali Fatih; Sulchek, Todd

    2016-11-01

    Cell molecular interactions govern important physiological processes such as stem cell homing, inflammation and cancer metastasis. But due to a lack of effective separation technologies selective to these interactions it is challenging to specifically sort cells. Other label free separation techniques based on size, stiffness and shape do not provide enough specificity to cell type, and correlation to clinical condition. We propose a novel microfluidic device capable of high throughput molecule dependent separation of cells by flowing them through a microchannel decorated with molecule specific coated ridges. The unique aspect of this sorting design is the use of optimized gap size which is small enough to lightly squeeze the cells while flowing under the ridged part of the channel to increase the surface area for interaction between the ligand on cell surface and coated receptor molecule but large enough so that biomechanical markers, stiffness and viscoelasticity, do not dominate the cell separation mechanism. We are able to separate Jurkat cells based on its expression of PSGL-1ligand using ridged channel coated with P selectin at a flow rate of 0.045ml/min and achieve 2-fold and 5-fold enrichment of PSGL-1 positive and negative Jurkat cells respectively.

  7. Electrophoretic cell separation by means of immunomicrospheres

    NASA Technical Reports Server (NTRS)

    Rembaum, A.; Smolka, A. J. K.

    1980-01-01

    The electrophoretic mobility of fixed human red blood cells immunologically labeled with polymeric (4-vinyl)pyridine or polyglutaraldehyde microspheres was altered to a considerable extent. This observation was utilized in the preparative scale electrophoretic separation of human and turkey fixed red blood cells, whose mobilities under normal physiological conditions do not differ sufficiently to allow their separation by continuous flow electrophoresis. It is suggested that resolution in the electrophoretic separation of cell subpopulations, currently limited by finite and often overlapping mobility distributions, may be significantly enhanced by immuno-specific labeling of target populations using microspheres.

  8. Immunomagnetic separation can enrich fixed solid tumors for epithelial cells.

    PubMed

    Yaremko, M L; Kelemen, P R; Kutza, C; Barker, D; Westbrook, C A

    1996-01-01

    Immunomagnetic separation is a highly specific technique for the enrichment or isolation of cells from a variety of fresh tissues and microorganisms or molecules from suspensions. Because new techniques for molecular analysis of solid tumors are now applicable to fixed tissue but sometimes require or benefit from enrichment for tumor cells, we tested the efficacy of immunomagnetic separation for enriching fixed solid tumors for malignant epithelial cells. We applied it to two different tumors and fixation methods to separate neoplastic from non-neoplastic cells in primary colorectal cancers and metastatic breast cancers, and were able to enrich to a high degree of purity. Immunomagnetic separation was effective in unembedded fixed tissue as well as fixed paraffin-embedded tissue. The magnetically separated cells were amenable to fluorescence in situ hybridization and polymerase chain reaction amplification of their DNA with minimal additional manipulation. The high degree of enrichment achieved before amplification contributed to interpretation of loss of heterozygosity in metastatic breast cancers, and simplified fluorescence in situ hybridization analysis because only neoplastic cells were hybridized and counted. Immunomagnetic separation is effective for the enrichment of fixed solid tumors, can be performed with widely available commercial antibodies, and requires little specialized instrumentation. It can contribute to interpretation of results in situations where enrichment by other methods is difficult or not possible.

  9. Material review of Li ion battery separators

    NASA Astrophysics Data System (ADS)

    Weber, Christoph J.; Geiger, Sigrid; Falusi, Sandra; Roth, Michael

    2014-06-01

    Separators for Li Ion batteries have a strong impact on cell production, cell performance, life, as well as reliability and safety. The separator market volume is about 500 million m2 mainly based on consumer applications. It is expected to grow strongly over the next decade for mobile and stationary applications using large cells. At present, the market is essentially served by polyolefine membranes. Such membranes have some technological limitations, such as wettability, porosity, penetration resistance, shrinkage and meltdown. The development of a cell failure due to internal short circuit is potentially closely related to separator material properties. Consequently, advanced separators became an intense area of worldwide research and development activity in academia and industry. New separator technologies are being developed especially to address safety and reliability related property improvements.

  10. Can Plant Viruses Cross the Kingdom Border and Be Pathogenic to Humans?

    PubMed Central

    Balique, Fanny; Lecoq, Hervé; Raoult, Didier; Colson, Philippe

    2015-01-01

    Phytoviruses are highly prevalent in plants worldwide, including vegetables and fruits. Humans, and more generally animals, are exposed daily to these viruses, among which several are extremely stable. It is currently accepted that a strict separation exists between plant and vertebrate viruses regarding their host range and pathogenicity, and plant viruses are believed to infect only plants. Accordingly, plant viruses are not considered to present potential pathogenicity to humans and other vertebrates. Notwithstanding these beliefs, there are many examples where phytoviruses circulate and propagate in insect vectors. Several issues are raised here that question if plant viruses might further cross the kingdom barrier to cause diseases in humans. Indeed, there is close relatedness between some plant and animal viruses, and almost identical gene repertoires. Moreover, plant viruses can be detected in non-human mammals and humans samples, and there are evidence of immune responses to plant viruses in invertebrates, non-human vertebrates and humans, and of the entry of plant viruses or their genomes into non-human mammal cells and bodies after experimental exposure. Overall, the question raised here is unresolved, and several data prompt the additional extensive study of the interactions between phytoviruses and non-human mammals and humans, and the potential of these viruses to cause diseases in humans. PMID:25903834

  11. Ion transport restriction in mechanically strained separator membranes

    NASA Astrophysics Data System (ADS)

    Cannarella, John; Arnold, Craig B.

    2013-03-01

    We use AC impedance methods to investigate the effect of mechanical deformation on ion transport in commercial separator membranes and lithium-ion cells as a whole. A Bruggeman type power law relationship is found to provide an accurate correlation between porosity and tortuosity of deformed separators, which allows the impedance of a separator membrane to be predicted as a function of deformation. By using mechanical compression to vary the porosity of the separator membranes during impedance measurements it is possible to determine both the α and γ parameters from the modified Bruggeman relation for individual separator membranes. From impedance testing of compressed pouch cells it is found that separator deformation accounts for the majority of the transport restrictions arising from compressive stress in a lithium-ion cell. Finally, a charge state dependent increase in the impedance associated with charge transfer is observed with increasing cell compression.

  12. Role of Sphingolipids in Infant Gut Health and Immunity.

    PubMed

    Nilsson, Åke

    2016-06-01

    Sphingomyelin (SM), glycosphingolipids, and gangliosides are important polar lipids in the milk fat globule membrane but are not found in standard milk replacement formulas. Because digestion and absorption of SM and glycosphingolipids generate the bioactive metabolites ceramide, sphingosine, and sphingosine-1-phosphate (S1P), and because intact gangliosides may have beneficial effects in the gut, this may be important for gut integrity and immune maturation in the neonate. The brush border enzymes that hydrolyze milk SM, alkaline sphingomyelinase (nucleotide phosphodiesterase pyrophosphatase 7), and neutral ceramidase are expressed at birth in both term and preterm infants. Released sphingosine is absorbed, phosphorylated to S1P, and converted to palmitic acid via S1P-lyase in the gut mucosa. Hypothetically, S1P also may be released from absorptive cells and exert important paracrine actions favoring epithelial integrity and renewal, as well as immune function, including secretory IgA production and migration of T lymphocyte subpopulations. Gluco-, galacto-, and lactosylceramide are hydrolyzed to ceramide by lactase-phlorizin hydrolase, which also hydrolyzes lactose. Gangliosides may adhere to the brush border and is internalized, modified, and possibly transported into blood, and may exert protective functions by their interactions with bacteria, bacterial toxins, and the brush border. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Isolation of the Lateral Border Recycling Compartment using a diaminobenzidine-induced density shift

    PubMed Central

    Sullivan, David P.; Rüffer, Claas; Muller, William A.

    2014-01-01

    The migration of leukocytes across the endothelium and into tissue is critical to mounting an inflammatory response. The Lateral Border Recycling Compartment (LBRC), a complex vesicular-tubule invagination of the plasma membrane found at endothelial cell borders, plays an important role in the this process. Although a few proteins have been shown to be present in the LBRC, no unique marker is known. Here we detail methods that can be used to characterize a subcellular compartment that lacks an identifying marker. Initial characterization of the LBRC was performed using standard subcellular fractionation with sucrose gradients and took advantage of the observation that the compartment migrated at a lower density than other membrane compartments. To isolate larger quantities of the compartment, we modified a classic technique known as a diaminobenzidine (DAB)-induced density shift. The DAB-induced density shift allowed for specific isolation of membranes labeled with HRP conjugated antibody. Because the LBRC could be differentially labeled at 4°C and 37°C, we were able to identify proteins that are enriched in the compartment, despite lacking a unique marker. These methods serve as a model to others studying poorly characterized compartments and organelles and are applicable to a wide variety of biological systems. PMID:24915828

  14. Characterization and Separation of Cancer Cells with a Wicking Fiber Device.

    PubMed

    Tabbaa, Suzanne M; Sharp, Julia L; Burg, Karen J L

    2017-12-01

    Current cancer diagnostic methods lack the ability to quickly, simply, efficiently, and inexpensively screen cancer cells from a mixed population of cancer and normal cells. Methods based on biomarkers are unreliable due to complexity of cancer cells, plasticity of markers, and lack of common tumorigenic markers. Diagnostics are time intensive, require multiple tests, and provide limited information. In this study, we developed a novel wicking fiber device that separates cancer and normal cell types. To the best of our knowledge, no previous work has used vertical wicking of cells through fibers to identify and isolate cancer cells. The device separated mouse mammary tumor cells from a cellular mixture containing normal mouse mammary cells. Further investigation showed the device separated and isolated human cancer cells from a heterogeneous mixture of normal and cancerous human cells. We report a simple, inexpensive, and rapid technique that has potential to identify and isolate cancer cells from large volumes of liquid samples that can be translated to on-site clinic diagnosis.

  15. Separation of human bone marrow by counterflow centrifugation monitored by DNA-flowcytometry.

    PubMed

    de Witte, T; Plas, A; Koekman, E; Blankenborg, G; Salden, M; Wessels, J; Haanen, C

    1984-10-01

    Human bone marrow was fractionated by counterflow centrifugation into 16 fractions with increasing cell size. Three distinct subpopulations could be recognized: small lymphocytic cells, medium-sized nucleated erythroid cells and large myeloid elements. DNA-flowcytometry and 3H-thymidine uptake showed that within the erythroid and myeloid cell populations counterflow centrifugation separates each population according to the cell cycle phase. Hypotonic treatment of bone marrow for removal of the erythroid nucleated cells resulted in a complete abrogation of the proliferating erythroid cell population. Counterflow centrifugation also separates the small non-proliferating myeloid and erythroid committed stem cells from the larger proliferating stem cells. It appeared feasible to separate the small lymphocytic cells from the majority of BFU-E and CFU-GM, due to the larger size of the proliferating normoblasts and the committed progenitor cells. Elimination of the mature lymphocytes from the haematopoietic stem cells by counterflow centrifugation may offer an alternative approach to the prevention of graft versus host disease (GvHD).

  16. High temperature 1 MHz capacitance-voltage method for evaluation of border traps in 4H-SiC MOS system

    NASA Astrophysics Data System (ADS)

    Peng, Zhao-Yang; Wang, Sheng-Kai; Bai, Yun; Tang, Yi-Dan; Chen, Xi-Ming; Li, Cheng-Zhan; Liu, Ke-An; Liu, Xin-Yu

    2018-04-01

    In this work, border traps located in SiO2 at different depths in 4H-SiC MOS system are evaluated by a simple and effective method based on capacitance-voltage (C-V) measurements. This method estimates the border traps between two adjacent depths through C-V measurement at various frequencies at room and elevated temperatures. By comparison of these two C-V characteristics, the correlation between time constant of border traps and temperatures is obtained. Then the border trap density is determined by integration of capacitance difference against gate voltage at the regions where border traps dominate. The results reveal that border trap concentration a few nanometers away from the interface increases exponentially towards the interface, which is in good agreement with previous work. It has been proved that high temperature 1 MHz C-V method is effective for border trap evaluation.

  17. Validation of a temperature prediction model for heat deaths in undocumented border crossers.

    PubMed

    Ruttan, Tim; Stolz, Uwe; Jackson-Vance, Sara; Parks, Bruce; Keim, Samuel M

    2013-04-01

    Heat exposure is a leading cause of death in undocumented border crossers along the Arizona-Mexico border. We performed a validation study of a weather prediction model that predicts the probability of heat related deaths among undocumented border crossers. We analyzed a medical examiner registry cohort of undocumented border crosser heat- related deaths from January 1, 2002 to August 31, 2009 and used logistic regression to model the probability of one or more heat deaths on a given day using daily high temperature (DHT) as the predictor. At a critical threshold DHT of 40 °C, the probability of at least one heat death was 50 %. The probability of a heat death along the Arizona-Mexico border for suspected undocumented border crossers is strongly associated with ambient temperature. These results can be used in prevention and response efforts to assess the daily risk of deaths among undocumented border crossers in the region.

  18. The Migrant Border Crossing Study: A methodological overview of research along the Sonora-Arizona border.

    PubMed

    Martínez, Daniel E; Slack, Jeremy; Beyerlein, Kraig; Vandervoet, Prescott; Klingman, Kristin; Molina, Paola; Manning, Shiras; Burham, Melissa; Walzak, Kylie; Valencia, Kristen; Gamboa, Lorenzo

    2017-07-01

    Increased border enforcement efforts have redistributed unauthorized Mexican migration to the United States (US) away from traditional points of crossing, such as San Diego and El Paso, and into more remote areas along the US-Mexico border, including southern Arizona. Yet relatively little quantitative scholarly work exists examining Mexican migrants' crossing, apprehension, and repatriation experiences in southern Arizona. We contend that if scholars truly want to understand the experiences of unauthorized migrants in transit, such migrants should be interviewed either at the border after being removed from the US, or during their trajectories across the border, or both. This paper provides a methodological overview of the Migrant Border Crossing Study (MBCS), a unique data source on Mexican migrants who attempted an unauthorized crossing along the Sonora-Arizona border, were apprehended, and repatriated to Nogales, Sonora in 2007-09. We also discuss substantive and theoretical contributions of the MBCS.

  19. An innovative cascade system for simultaneous separation of multiple cell types.

    PubMed

    Pierzchalski, Arkadiusz; Mittag, Anja; Bocsi, Jozsef; Tarnok, Attila

    2013-01-01

    Isolation of different cell types from one sample by fluorescence activated cell sorting is standard but expensive and time consuming. Magnetic separation is more cost effective and faster by but requires substantial effort. An innovative pluriBead-cascade cell isolation system (pluriSelect GmbH, Leipzig, Germany) simultaneously separates two or more different cell types. It is based on antibody-mediated binding of cells to beads of different size and their isolation with sieves of different mesh-size. For the first time, we validated the pluriSelect system for simultaneous separation of CD4+- and CD8+-cells from human EDTA-blood samples. Results were compared with those obtained by magnetic activated cell sorting (MACS; two steps -first isolation of CD4+, then restaining of the residual cell suspension with anti-human CD8+ MACS antibody followed by the second isolation). pluriSelect separation was done in whole blood, MACS separation on density gradient isolated mononuclear cells. Isolated and residual cells were immunophenotyped by 7-color 9-marker panel (CD3; CD16/56; CD4; CD8; CD14; CD19; CD45; HLA-DR) using flow cytometry. Cell count, purity, yield and viability (7-AAD exclusion) were determined. There were no significant differences between both systems regarding purity (MACS (median[range]: 92.4% [91.5-94.9] vs. pluriSelect 95% [94.9-96.8])) of CD4+ cells, however CD8+ isolation showed lower purity by MACS (74.8% [67.6-77.9], pluriSelect 89.9% [89.0-95.7]). Yield was not significantly different for CD4 (MACS 58.5% [54.1-67.5], pluriSelect 67.9% [56.8-69.8]) and for CD8 (MACS 57.2% [41.3-72.0], pluriSelect 67.2% [60.0-78.5]). Viability was slightly higher with MACS for CD4+ (98.4% [97.8-99.0], pluriSelect 94.1% [92.1-95.2]) and for CD8+-cells (98.8% [98.3-99.1], pluriSelect 86.7% [84.2-89.9]). pluriSelect separation was substantially faster than MACS (1h vs. 2.5h) and no pre-enrichment steps were necessary. In conclusion, pluriSelect is a fast, simple and gentle system for efficient simultaneous separation of two and more cell subpopulation directly from whole blood and provides a simple alternative to magnetic separation.

  20. Causation by Diesel Exhaust Particles of Endothelial Dysfunctions in Cytotoxicity, Pro-inflammation, Permeability, and Apoptosis Induced by ROS Generation.

    PubMed

    Tseng, Chia-Yi; Wang, Jhih-Syuan; Chao, Ming-Wei

    2017-10-01

    Epidemiological studies suggest that an increase of diesel exhaust particles (DEP) in ambient air corresponds to an increase in hospital-recorded myocardial infarctions within 48 h after exposure. Among the many theories to explain this data are endothelial dysfunction and translocation of DEP into vasculature. The mechanisms for such DEP-induced vascular permeability remain unknown. One of the major mechanisms underlying the effects of DEP is suggested to be oxidative stress. Experiments have shown that DEP induce the generation of reactive oxygen species (ROS), such as superoxide anion and H 2 O 2 in the HUVEC tube cells. Transcription factor Nrf2 is translocated to the cell nucleus, where it activates transcription of the antioxidative enzyme HO-1 and sequentially induces the release of vascular permeability factor VEGF-A. Furthermore, a recent study shows that DEP-induced intracellular ROS may cause the release of pro-inflammatory TNF-α and IL-6, which may induce endothelial permeability as well by promoting VEGF-A secretion independently of HO-1 activation. These results demonstrated that the adherens junction molecule, VE-cadherin, becomes redistributed from the membrane at cell-cell borders to the cytoplasm in response to DEP, separating the plasma membranes of adjacent cells. DEP were occasionally found in endothelial cell cytoplasm and in tube lumen. In addition, the induced ROS is cytotoxic to the endothelial tube-like HUVEC. Acute DEP exposure stimulates ATP depletion, followed by depolarization of their actin cytoskeleton, which sequentially inhibits PI3K/Akt activity and induces endothelial apoptosis. Nevertheless, high-dose DEP augments tube cell apoptosis up to 70 % but disrupts the p53 negative regulator Mdm2. In summary, exposure to DEP affects parameters influencing vasculature permeability and viability, i.e., oxidative stress and its upregulated antioxidative and pro-inflammatory responses, which sequentially induce vascular permeability factor, VEGF-A release and disrupt cell-cell junction integrity. While exposure to a low dose of DEP actin triggers cytoskeleton depolarization, reduces PI3K/Akt activity, and induces a p53/Mdm2 feedback loop, a high dose causes apoptosis by depleting Mdm2. Addition of ROS scavenger N-acetyl cysteine suppresses DEP-induced oxidative stress efficiently and reduces subsequent damages by increasing endogenous glutathione.

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